Next Article in Journal
Molecular Mechanisms of Anti-Inflammatory Phytochemicals 2.0
Next Article in Special Issue
The miRNAs let-7b and miR-141 Coordinately Regulate Vitellogenesis by Modulating Methyl Farnesoate Degradation in the Swimming Crab Portunus trituberculatus
Previous Article in Journal
Maackiain Mimics Caloric Restriction through aak-2-Mediated Lipid Reduction in Caenorhabditis elegans
Previous Article in Special Issue
The Potential Role of the Piwi Gene in the Development and Reproduction of Plutella xylostella
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
IJMS
Volume 24
Issue 24
10.3390/ijms242417433
ijms-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles thumb_up
...
Endorse textsms
...
Comment
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Review

Monosex Populations of the Giant Freshwater Prawn Macrobrachium rosenbergii—From a Pre-Molecular Start to the Next Generation Era

by
Melody Wahl
1,
Tom Levy
2,3,
Tomer Ventura
4,5 and
Amir Sagi
1,6,*
1
Department of Life Sciences, Ben-Gurion University of the Negev, P.O. Box 653, Beer-Sheva 8410501, Israel
2
Institute for Stem Cell Biology and Regenerative Medicine, School of Medicine, Stanford University, Stanford, CA 94305, USA
3
Hopkins Marine Station, Stanford University, Pacific Grove, CA 93950, USA
4
Centre for Bioinnovation, University of the Sunshine Coast, Sippy Downs, QLD 4556, Australia
5
School of Science and Engineering, University of the Sunshine Coast, Sippy Downs, QLD 4556, Australia
6
National Institute for Biotechnology in the Negev, Ben-Gurion University of the Negev, P.O. Box 653, Beer-Sheva 8410501, Israel
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2023, 24(24), 17433; https://doi.org/10.3390/ijms242417433
Submission received: 22 April 2023 / Revised: 27 October 2023 / Accepted: 21 November 2023 / Published: 13 December 2023
(This article belongs to the Special Issue Neuropeptides and Endocrine Regulations in Insects and Crustaceans)
Browse Figures
Review Reports Versions Notes

Abstract

:
Sexual manipulation in the giant freshwater prawn Macrobrachium rosenbergii has proven successful in generating monosex (both all-male and all-female) populations for aquaculture using a crustacean-specific endocrine gland, the androgenic gland (AG), which serves as a key masculinizing factor by producing and secreting an insulin-like AG hormone (IAG). Here, we provide a summary of the advancements from the discovery of the AG and IAG in decapods through to the development of monosex populations in M. rosenbergii. We discuss the broader sexual development pathway, which is highly divergent across decapods, and provide our future perspective on the utility of novel genetic and genomic tools in promoting refined approaches towards monosex biotechnology. Finally, the future potential benefits of deploying monosex prawn populations for environmental management are discussed.

1. The AG and IAG-Switch in Crustaceans

A brief history of the discoveries of the AG and the IAG in crustaceans, with an emphasis on decapods.
In vertebrates, sexual development cascades are well characterized, including the pivotal role of sex steroids. Studies have investigated the presence of sex steroids and their potential role in sexual development in crustaceans. Sex steroids, including androgen, estrogen, and progestogen, have been identified in decapod tissues, such as the gonads and eyestalk [1,2,3,4]. Although some studies have explored the effects of hormone treatments on sexual development and reproductive processes [5,6,7,8], the literature regarding the underlying mechanisms and factors influencing the outcomes remains limited and lacks clear evidence.
Instead, an endocrine gland unique to male crustaceans from the class Malacostraca was shown to establish masculinization. The first mention of the gland was in 1947 in the male reproductive system of the blue swimming crab Callinectes sapidus Rathbun [9], followed by its discovery in many other decapod crustaceans (Figure 1A,B), while its function was deduced by the early work of [10] in the amphipod Orchestia gammarella, demonstrating for the first time an androgenic function of the unique gland, thus named the androgenic gland (AG) [10]. Bilateral removal of the AGs from O. gammarella males ceased the differentiation of secondary characteristics and caused a decrease in spermatogenesis. In the reciprocal study, AG implantation into O. gammarella females caused masculinization of the primary and secondary sexual characteristics. However, gonads implanted into the opposite sex had no effect on the host. These studies led to the conclusion that the AG is the exclusive source of a hormone responsible for the development of primary and secondary male characteristics [10,11,12]. In decapods, similar results validated the conserved key role of the AG in masculinization; immature and mature giant freshwater prawn (Macrobrachium rosenbergii) females implanted with AG developed appendices masculinae (male-specific protrusions that develop bilaterally on the second pair of swimming legs), while females implanted with vas deferens or testicular tissue developed normally [13]. In the red swamp crayfish, Procambarus clarkii, AG implantation into immature females inhibited vitellogenesis [14], while ovarian regression ensued in AG implanted females of the mud crab Scylla paramamosain [15]. In the Chinese mitten crab, Eriocheir sinensis, AG extracts from S. paramamosain and E. sinensis injected into females caused the development of external male characteristics [16].
The attempts to isolate the AG hormone raised a debate with regard to the chemical nature of the active substance, with evidence suggesting either a protein or a lipidic nature. In isopods, Katakura (1975) partially purified the active protein from the AG of Armadillidium vulgare and showed that a single injection of the extract into females induced masculinization of the secondary sexual characteristics [17]. The lipidic and steroidal extracts of the AG did not induce masculinization [18]. A series of studies in the isopod, A. vulgare, purified and elucidated the structure of an androgenic hormone [19,20]. Isolation and characterization of a cDNA encoding a precursor of the hormone showed a resemblance to the insulin superfamily structure, comprised of a signal peptide, B chain, C peptide, and A chain [21]. Later, similar androgenic hormones with highly conserved protein sequences were identified in two additional isopod species, Porcellio scaber and Porcellio dilatatus [22]. In decapods, biochemical studies [23] and histological observations [24] of lipidic substances in the AG suggested that steroids act as androgenic hormones. Efforts to find genes orthologous to the isopods’ androgenic hormones in the group of decapods have failed. It was not until 2007 when the first AG-specific gene, expressed exclusively in males, was discovered in the crayfish Cherax quadricarinatus, termed insulin-like AG factor (IAG) [25] (Figure 1C), followed by its discovery in M. rosenbergii [26], demonstrating structural similarity to the insulin superfamily (Supplementary Data S1 and S2). The identification of the first decapod IAG-encoding gene signifies a longstanding gap in our understanding of the AG and its function in decapods, following decades since the discovery of an androgenic hormone in isopods. The finding was enabled through a subtractive cDNA library prior to the prevalent use of next-generation sequencing. Since the discovery of IAG in these two species, IAG-encoding sequences have been identified in a wide range of decapod species including gonochoristic, hermaphrodites [27,28], and parthenogenic [29], of commercial and ecological significance, greatly facilitated by the next-generation sequencing, which is commonly used nowadays.

2. Manipulating the IAG-Switch

From early surgical attempts at the AG level up to state-of-the-art molecular manipulations to achieve full and functional sex reversal
Early studies of AG ablation and implantation established the crustacean AG as a key sexual differentiating organ. The IAG hormone secreted from the AG serves as a master switch in decapod crustaceans, in which IAG expression induces masculinization and its absence results in feminization, thus termed the ”IAG-switch” [30].
Moreover, unlike vertebrates, in male crustaceans, the endocrine and gametogenic functions are separated into two distinct organs, the AG and the testis, respectively [31,32]; thus, manipulation of the AG can be performed without affecting the gonad.
With respect to sexual plasticity, one of the most studied crustaceans is M. rosenbergii, which displays a fascinating social hierarchy [33], with distinctive morphotypic differentiation of the males and clear differences between males and females in growth patterns (Figure 2A,B). Since males grow faster than females, an interest in developing all-male populations was explored based on a small-scale cage experiment showing that all-male populations might provide double the yield compared with mixed and all-female populations [34] (Figure 2C). The above triggered experiments of bilateral AG ablation at an early juvenile developmental stage, which resulted in complete sex reversal in functional neo-females. When mated with normal males, these neo-females produced all-male progeny [35]. The timing of AG removal was shown to be critical for the manipulation to succeed. M. rosenbergii males that were andrectomized in the youngest developmental stage exhibited complete feminization, including the development of oviducts and female gonopores and the initiation of oogenesis, in contrast to later developmental stages in which males were either partially feminized or not feminized at all [36]. The first mass production of all-male populations was established by microsurgical AG ablation at early differentiation stages; however, the production process had drawbacks such as low microsurgery success rates, long duration between microsurgery, and identification of neo-females and intensive labor demand [37]. It became evident, though, that a biotechnology based on manipulation of this endocrine gland could provide the long-sought solution for monosex population aquaculture.
The needed transformation from surgical manipulations to molecular methods was completed through the use of RNA interference (RNAi) gene knockdown [38]. This method has contributed to clarifying the functionality of many crustacean genes of importance to metabolism, development, growth, and reproduction [39]. The RNAi-based biotechnology was a game-changer in the production of M. rosenbergii monosex populations. Initial use of this method to silence the IAG in mature M. rosenbergii males showed reduced spermatogenesis and caused AG hypertrophy [26]. Given the temporal effect and non-GMO nature of the intervention [40], manipulating the expression of the IAG-switch was suggested as a replacement for the microsurgical removal of the AG by using IAG silencing prior to sexual differentiation.
M. rosenbergii, like many Malacostraca species, bears the WZ/ZZ system of sexual heritability, in which females are heterogametic (WZ) and males are homogametic (ZZ) [41,42,43,44,45]. A key component in detecting successful IAG-switch manipulations was the use of genetic sex markers. These were not available for M. rosenbergii until 2010 [46], when the use of the cumbersome amplified fragment length polymorphism (AFLP) enabled the identification of the first genetic sex markers for this species. With the advent of restriction-assisted DNA sequencing (RAD-Seq) and RNA-Seq, genetic sex marker identification is streamlined and has indeed contributed to the discovery of these markers in multiple decapod species, either within the WZ/ZZ or XX/XY systems [30,47]. Using genetic sex markers, successful sex reversal of genetic ZZ males into functional ZZ neo-females was first achieved through gene silencing in 2012 [48]. This technology now enables large-scale production of all-male populations using RNAi, the first application of this method in the entire field of aquaculture.
M. rosenbergii presents clear differences between sexes. Males display morphotypic differentiation while females grow more uniformly, making monosex a clear advantage. The AG is known to be key for this change in behavior, as was exemplified in C. quadricarinatus, where AG implanted into females induces male-like behaviors [49]. While sex-specific behaviors remain largely unexplored across farmed decapods, evidence suggests monosex would be advantageous across decapods, as, for example, in the case of the commercially most important marine shrimp, Litopenaeus vannamei, where subtle sex-biased behaviors were recorded [50].
Although in M. rosenbergii, some of the males grow larger than females and reach the highest sizes in a mixed population, thus providing economic benefits [51,52,53], all-female culture was suggested to be more favorable under intensified conditions [54] (Figure 2D). Production of all-male populations requires stocking in low densities due to aggressiveness and territoriality. Males display a high variation in size distribution with strong dominance in hierarchical social structure [33,55]. While females do not reach the size of the dominant males, they grow more uniformly, as initially proposed in 1992 [43]. Manipulation of the AG using surgical implantation of AG tissue from adult males into early developmental stage females resulted in a fully functional reversal of sex to neo-males. However, the survival and success rates of the functional neo-males were only ∼10% [43]. Malecha (2012) claimed that the efficiency of producing all-female populations depends on increasing the success rate of producing WZ neo-males via the use of exogenous AG materials [54]. In 2016, through molecular manipulation of the IAG-switch, Levy et al. (2016) demonstrated a fully functional sex reversal of WZ females into WZ neo-males by a single injection of suspended hypertrophied AG cells [42]. Verification of the successful generation of neo-males was enabled using genetic sex markers [46]. Above 80% of cell-injected females had developed both appendix masculina and male gonopores. The injected sex-reversed females functioned as males and were reproductive, thus considered neo-males [42]. This successful all-female biotechnology has one drawback of being dependent on the relatively small amount of AG cells found in a donor body; thus, lentiviral-transduced ectopic expression of IAG in non-AG primary cell culture was attempted to increase the volume of relevant cells for the process and avoid the male donor dependency in this biotechnology [56]. This is a preliminary step to be further tested for applicability.

3. Controlling Elements of the IAG-Switch

Since the discovery of the first decapod IAG [25], it has been found in a variety of crustacean species, including prawns, shrimp, crayfish, lobsters, and crabs [30], providing a wide context within crustaceans and a starting point for the challenge of discovering the bridging cascade upstream of the IAG-switch. To date, IAG has enabled full sex change only in M. rosenbergii, suggesting this molecular switch needs to be further explored to fulfill its potential in other species. The path towards such discoveries of mechanisms upstream of the IAG-switch is now open and could be studied using functional genomics tools such as RNAi and CRISPR. Another interesting species with the above respects is the crayfish C. quadricarinatus, in which sexual plasticity is reflected in naturally occurring WZ-genotyped intersexual animals with an active male reproductive system and male secondary sexual characteristics, along with an inactive ovary (i.e., naturally born WZ neo-males). Intersexuality was manipulated by IAG silencing, which resulted in a sexual shift. This IAG-switch manipulation resulted in male feature feminization, vitellogenin expression, and oocytes with yolk accumulation [57]. Moreover, findings in hermaphrodites, particularly in protandric species, suggest the IAG-switch plays a critical role in the natural sex reversal path. Unlike gonochoristic species, the sexual differentiation process in hermaphrodites does not relate to the early developmental stages but happens at the mature life stage with a sexual transformation from maleness to femaleness. A study of the northern spot shrimp, Pandalus platyceros, describes the IAG-switch using four stages: juveniles, adult males, transitionals, and in adult females [28]. The IAG temporal expression pattern in the AG was the highest in juveniles, declined in adult males, and was found to be negligible during the transitional phase and adult females. This suggests that the IAG has a crucial role in the early male differentiation and maturation stage. Moreover, IAG loss of function through RNAi in mature P. platyceros males induced the masculine to feminine sexual transformation that naturally occurs in this protandric species, supporting the pivotal role of the IAG-switch in regulating the transformation between adult stages in hermaphrodite species [28]. In M. rosenbergii adult stages, IAG was found to be correlated with the reproductive readiness of male morphotypes. The expression levels of Mr-IAG in the reproductively less active orange-claw males were significantly lower than in the blue-clawed males and small males, suggesting a key role for IAG in regulating morphotypic differentiation [58].
The research on the upstream controlling elements of sexual differentiation through the IAG-switch is in progress, with functional studies suggesting diverse candidates for this role (Table 1). In mature specimens of gonochoristic species, it seems that the IAG is controlled as part of the eyestalk-AG-testis endocrine axis [59]. Within this axis, IAG is thought to be controlled by upstream neuropeptides that are produced in the X-organ (XO) and accumulated in the sinus gland (SG; both the XO and SG reside in the eyestalk, forming a neuroendocrine complex known as the XO-SG), from where they are secreted. The fact that eyestalk ablation in males causes hypertrophy of the AG [59,60] and IAG over-expression [61] indicates that eyestalk neuropeptides serve as upstream controlling elements of AG activity in adults [59,62]. Recent studies in E. sinensis [63] and S. paramamosain [64] suggest regulatory feedback between the Crustacean Female Sex Hormone (CFSH) in the eyestalk and IAG. In EsCFSH-1-silenced E. sinensis mature males, the expression of EsIAG in the AG was significantly increased, while EsIAG knockdown significantly increased the expression of EsCFSH-1 in the male eyestalk [63]. In S. paramamosain, in vitro treatment with recombinant SpCFSH protein in AG significantly decreased the mRNA levels of the signal transducers and activators of transcription (STAT)-binding site in the IAG promoter. Based on these results, it was suggested that CFSH acts as an inhibiting factor by suppressing the expression of SpSTAT, which then regulates SpIAG expression [64]. In the protandric hermaphrodite peppermint shrimp, Lysmata vittata, injection of recombinant CFSH1b suppressed IAG expression, and Lvit-CFSH knockdown stimulated the male phenotype development [65]. Other eyestalk-borne neuropeptides suggested to play a role in the eyestalk-AG-testis axis are members of the crustacean hyperglycemic hormone (CHH) superfamily of neuropeptides. In the Pacific white leg shrimp L. vannamei, RNAi of two CHH genes, LvCHH1 or LvCHH2 induced elevated LvIAG expression, while injection of their recombinant protein reduced LvIAG expression, suggesting inhibitory regulation of CHHs over IAG [66].
Sexual development is a succession of three processes, starting with sex determination occurring at the establishment of the zygote, which leads to the sexual differentiation process, followed by sexual maturation (Figure 3). While the AG is regarded as a sexual differentiating organ, the sex-determining mechanism is considered the primary process that regulates the development and function of the AG [67,68]. Several candidates were suggested as the transmitters between sex determination and sexual differentiation in decapods. In C. quadricarinatus, Sxl (sex-lethal) alternatively-spliced variants were suggested to be involved in the mechanism of male sex determination/differentiation, with a gradually increased expression pattern in embryonic stages, higher transcript levels at early-stage testis development, and significantly reduced CqIAG expression levels following CqSxl3 silencing [69]. In the mud crab, S. paramamosain, the transcription factor Doublesex (Dsx) promotes Sp-IAG expression, while the transcription factor forkhead L2 (foxl-2) inhibits it, as demonstrated in both in vitro cell experiments and in vivo RNAi studies [70]. Knockdown of the invertebrate double-sex and mab-3 related transcription factor 2 (idmrt-2) in S. paramamosain decreased the expression of Dmrt-like and foxl-2 genes in the testis, and IAG in the AG [71]. In the black tiger shrimp, Penaeus monodon, RNAi of Dsx significantly decreased the expression of PmIAG in the testis and was suggested as a positive regulator of IAG by specific binding upstream of the IAG promoter [72]. Similarly, Dsx was suggested to regulate IAG in the shrimp Fenneropenaeus chinensis, as knockdown of FcDsx resulted in FcIAG transcript down-regulation [73]. In E. sinensis, RNAi of EsDsx1, EsiDMY, and EsiDmrt1a reduced IAG transcription levels [74]. Direct interaction of the EsDsx-like protein with the EsIAG promoter suggests its role as an upstream regulator of IAG [75]. Contrary to this, knockdown of C. quadricarinatus Dsx induced CqIAG transcript levels [76]. In M. rosenbergii, MrDsx expression was significantly induced in the testis and AG following eyestalk ablation, and gene knockdown of MrDsx resulted in a significant decrease in MrIAG transcript levels in the AG, suggesting that eyestalk elements negatively control MrDsx, which regulates the activation of MrIAG [77]. As part of the extensive research aimed at elucidating the IAG signaling cascade in decapod species, IAG loss-of-function studies have played a significant role in the understanding of the IAG downstream factors [78,79,80,81,82,83]. Still, in most of the studies on candidates upstream of the IAG-switch, it seems that the intervention (usually RNAi) is too late to effectively influence sexual differentiation. That can be potentially resolved by an oocyte-specific delivery (OSDel) tool for gene silencing at the oocyte stage prior to egg laying [84]. Yet, the barrier of sex identification at an early stage might be the main obstacle to understanding the sexual differentiation mechanism in other decapod species that might be even more variable.
Table 1. Genes suggested as sexual development regulation upstream of the IAG-Switch.
Table 1. Genes suggested as sexual development regulation upstream of the IAG-Switch.
GeneSpeciesTested Tissue or StageSex Heritability MechanismEffect on Sexual Differentiation ProcessesReference
Dmrt11EMacrobrachium rosenbergiiPost larvaeWZ/ZZMroDmrt11E knockdown induced a functional sex reversal[85]
Terminal ampullaeWZ/ZZMroDmrt11E knockdown reduced MrIAG expression[86]
Macrobrachium nipponenseOvary and hepatopancreas (females)
Abdominal ganglia (males)		WZ/ZZMniDMRT11E knockdown reduced MnVG expression in females and increased MnIAG expression in males[87]
DsxFenneropenaeus chinensisMale cephalothoraxWZ/ZZTwo isoforms of FcIAG gene were down-regulated following FcDsx knockdown[73]
Macrobrachium rosenbergiiAGWZ/ZZMrDsx knockdown reduced MrIAG expression[77]
Cherax quadricarinatusCephalothoraxes of undifferentiated crayfishWZ/ZZCqDsx knockdown decreased CqIAG expression[76]
SOXE Portunus
trituberculatus		AG and testisXY/XXPtSoxE siRNA reduced PtIAG and PtIR expression[88]
SOXB2-1Eriocheir sinensisTestisWZ/ZZEsSoxB2-1 knockdown led to abnormal nucleus transformation during spermiogenesis[89]
SOX9Scylla paramamosainCell cultureWZ/ZZSox9 binding site mutation in SpVIH promoter reduced activity[90]
CHH1 Litopenaeus vannameiAGWZ/ZZKnockdown of either LvCHH1 or LvCHH2 resulted in increased LvIAG expression; injection of their recombinant protein led to decreased LvIAG expression.[66]
CHH2
GC receptor Litopenaeus vannameiAGWZ/ZZLvGC knockdown increased LvIAG expression[66]
MIH Macrobrachium nipponenseAGWZ/ZZGene knockdown increased IAG expression[91]
GIH (VIH)
MascMacrobrachium rosenbergiiPost larvaeWZ/ZZMrMasc knockdown obtained full sex reversal; Insulin-like signal pathway has been identified in MrMasc knockdown prawns.[92]
SxlCherax quadricarinatusUndifferentiated juvenile prawnsWZ/ZZSxl3 knockdown decreased CqIAG expression[69]
Foxl2Scylla paramamosainOvariesWZ/ZZSpfoxl2 knockdown increased SpVG expression[93]
CFSHPortunus
trituberculatus		Eyestalk, AG and testisXY/XXPtCFSH knockdown increased PtIAG expression; injection of its recombinant protein led to decreased PtIAG expression.[94]
Scylla paramamosainCultured AGWZ/ZZRecombinant SpCFSH reduced SpIAG expression[95]
Cultured AGWZ/ZZRecombinant SpCFSH reduced SpSTAT expression[64]
STAT Scylla paramamosainCultured AGWZ/ZZSpSTAT knockdown reduced SpIAG expression[64]
Tra2Macrobrachium nipponenseGonadsWZ/ZZMnTra2 Inhibited MnSxl expression[96]
GEM Macrobrachium nipponenseAG and testisWZ/ZZMnGEM knockdown increased MnIAG expression and testosterone content[97]
BMP receptor Scylla paramamosainPre/early/late vitellogenic femalesWZ/ZZSpBMPR knockdown decreased vitellogenin receptor expression[98]
Abbreviations: AG, androgenic gland; BMP, bone morphogenetic protein; CFSH, crustacean female sex hormone; CHH, crustacean hyperglycemic hormone; Dmrt11E, doublesex and mab-3 related transcription factor 11E; Dsx, doublesex; Foxl2, forkhead box protein L2; GC, Guanylate cyclase; GEM, gem-associated protein 2-like isoform X1; GIH, gonad inhibiting hormone; IAG, insulin-like androgenic gland hormone; IR, insulin-like receptor; Masc, masculinizer; MIH, molt-inhibiting hormone; SOX9, SRY-Box Transcription Factor 9; SOXB2-1, SRY-related HMG box group B2; SOXE SRY-related HMG box group E; STAT, signal transducers and activators of transcription; Sxl, sex-lethal; Tra2, transformer-2; VG, vitellogenin; VIH, vitellogenin inhibiting hormone.

4. Distinct Biotechnologies for Monosex Populations of Prawns

Description of the two biotechnologies and their respective uses at different husbandry options.
Through manipulations of the IAG-switch, a high degree of induced sexual plasticity in M. rosenbergii has been demonstrated. The manipulations afforded cases of functional WZ males, ZZ females, and, surprisingly, even WW males and females [42,48,99]. IAG-switch manipulations led to the pioneering development of two biotechnologies for M. rosenbergii: all-male [48] and all-female [42] aquaculture. The RNAi-based biotechnology for all-male aquaculture through dsIAG injection [100] facilitated a large-scale production of all-male M. rosenbergii with 86% success sex-reversal of ZZ male to ZZ neo-female, then mating the neo-females with normal males, yielding 100% of all ZZ male progeny exhibiting typical population structure of M. rosenbergii male morphotype differentiation [33,40]. Together with the population structure, the temporal effect of the dsRNA in the target tissue proved the safety of RNAi usage in all-male production [40], the first commercial use of RNAi in the entire field of aquaculture. Shpak et al. showed the application of the technology beyond one all-male generation and established three successive generations of W chromosome-free ZZ prawn lines [101]. Up to date, over 15 consecutive W-free generations have been reported in this population (Wahl, personal communication). The third generation of all-males was comparable to males from a normal mixed population. The typical morphology of the testicular lobes, spermatophore, and AG was demonstrated by a histology of the testis and terminal ampulla in males from both populations. Gene expression patterns of male-specific genes such as Mr-IAG and Mr-Mrr were similar in males from the all-male third generations compared to males from a normal mixed population [101].
Correspondingly, the first biotechnology for all-female aquaculture by a single injection of hypertrophied AG cell suspension enabled, for the first time, all-female mass production in three steps: (1) WZ females were sex reversed into WZ neo-males by injection of suspended hypertrophied AG cells. (2) crossing of WZ neo-males with normal WZ females yielding a progeny with 25% WW females; (3) crossing the WW females with normal ZZ males yielding a 100% WZ female population [42]. This technology enabled, for the first time, a comparative large-scale field study of all-female versus mixed populations under extensive and intensive stocking conditions without the need for manual segregation. This field study, in which all-female progenies were cultured without males from Day 0, showed that all-female culture had superior performance to that of mixed culture with respect to feed conversion ratios, survival rates, total yield, and uniform body size, with stocking densities up to 4 times those usually practiced with mixed sex and all-male populations of the species [102]. However, the road to a viable breeding stock for all-female M. rosenbergii production necessitated a more efficient way to produce WW females via further manipulations of the IAG-switch to achieve WW neo-males. Indeed, this final manipulation of the sexual switch yielded WW females that were sex-reversed by injection of suspended hypertrophied AG cells into fully functional WW neo-males, which were then crossed with WW females to produce a Z chromosome-free breeding stock population [99]. The case of all-female consecutive generations without the Z chromosome was further investigated for three generations, showing that the performances of the WW all-female population were comparable to the WZ all-female population in survival rate, size uniformity, body weight, and yield [103]. This Z-free population has been managed with no visible abnormalities for over 8 generations (Wahl, personal communication).

5. Molecular and Genomic Implications

The availability of NGS technologies yielded novel molecular and genomic tools that were applied to several crustacean species [104]. Ample transcriptomic libraries of decapod crustaceans were studied, with several referring to sexually biased transcripts [27,28,105,106,107,108], including the description of different stages in the life cycle of M. rosenbergii [109,110,111]. This is true also at the genomic level, with published genomes of decapod species, including the crayfish Procambarus virginalis [112] and C. quadricarinatus [113], the crabs E. sinensis [114,115], Birgus latro and Paralithodes camtschaticus [116], the ridgetail white prawn Exopalaemon carinicauda [117], the shrimp Neocaridina denticulata [118], L. vannamei [119], Marsupenaeus japonicus and Penaeus monodon [120], and the lobsters Panulirus ornatus [116] and Homarus americanus [121]. A high-quality M. rosenbergii genome, exhibiting distinguishable paternal and maternal sequences and enabling the identification of W/Z-specific sequences [99], was also studied for future mapping at the chromosome level with resolved chromatids and SNP mapping.
A major reliance of molecular functional research is based on the RNAi procedure, which is readily available and frequently used as an instrumental tool for the study of the transcriptional regulation mechanism upstream of the IAG-switch at early post-larval stages [30]. However, when studying the upstream sex-determining factors that affect the IAG-switch, the post-larval stage might prove to be too late for the extensive functional genomic efforts needed for such a challenging study in which some earlier interventions seem to be needed. Thus, a genomic editing platform based on CRISPR is required for manipulations at early embryonic stages. In recent years, the CRISPR-Cas genome-editing tool was developed [122] and revolutionized life science research, enabling the induction of targeted mutagenesis to determine the role of studied genes. In crustaceans, a few cases were described using CRISPR to determine the role of genes in different processes such as eye development [123,124], limb specification [125], chitinase activity [126], and more. Recently, Molcho et al. reported the first genomic editing platform in M. rosenbergii, presenting a CRISPR protocol through direct injection into one- to four-cell embryos, which results in entire organism genome editing [127]. In the prawn E. carinicauda, knocking out EcIAG and obtaining homozygous mutants with biallelic mutations led to sex reversal from males to neo-females, suggesting CRISPR/Cas9 genome editing as an effective tool for sex manipulation in crustaceans, further supporting monosex aquaculture [128]. However, the transformation of applied technologies from RNAi interventions that do not affect the genome into genetically modifying methods is debatable. Indeed, the first such modified organism has been approved for aquaculture [129], while regulators in several countries are debating whether minor CRISPR modifications will be more easily approved [130]. Given that the IAG-encoding gene was found to be residing on an autosome, gene editing can potentially be harnessed to manipulate the IAG-switch even in WW superfemales [42], which lack the male sex chromosome. Further research is therefore required to better understand the upstream regulation of IAG expression (comparing promoter methylation and acetylation patterns in males and females, and so forth).

6. Future Environmental Applications of the IAG Switch

Crustaceans’ hardiness and adaptability place them among the worst-known invasive species, subsequently threatening biodiversity [131] and ecosystem stability [132]. One of the leading invasive species on earth is the red swamp crayfish, P. clarkii, which is known to cause considerable environmental and economic damage [133]. Production of P. clarkii neo-females through IAG-switch manipulation, using RNAi-based biotechnology for all-male aquaculture similar to prawns [100], was suggested as a sustainable solution with the potential to greatly impact the aquaculture industry [134]. All-male crayfish aquaculture is expected to have higher incomes and is safer than a mixed population by preventing escapees from becoming invasive in regions where P. clarkii is non-native [134]. Savaya et al. (2020) also suggested a demographic model that explores the potential of stocking neo-females to control the invasive population in the wild by skewing the sex ratio of the population [134]. Utilizing additional control tools would be important for ensuring the efficacy of sex-skewing strategy in invasive population management [135]. To fulfill these potential solutions, further study is required on IAG-switch manipulations in P. clarkii to achieve fully functional sex reversal and monosex population production or the use of tools such as gene drive [136,137,138]. With respect to managing invasive species, the availability of M. rosenbergii monosex populations is not only advantageous for aquaculture mass production but also for better safeguarding the environment from aquaculture escapees in areas where M. rosenbergii could become an invasive species.
Another interesting environmental management strategy that was unlocked by the availability of monosex populations is the use of prawns as biological control agents. M. rosenbergii are voracious predators of pest freshwater snails. The possible use of non-breeding monosex prawn populations poses them as excellent biocontrol agents against such snails without the hazard of prawns establishing invasive populations that could harm the environment and its natural biodiversity. Among the pest snails most noted are freshwater snails that host the flatworm Schistosoma spp., which are responsible for the parasitic disease schistosomiasis (bilharzia). Schistosomiasis is a serious human health concern, mainly in Africa, with hundreds of millions of infected people [139]. Applying monosex biotechnology in areas with abundant snail hosts can break the disease cycle [140]. Apart from human health, snails are involved in the transmission of fish diseases in the aquaculture industry. Extensive damage is caused to freshwater aquaculture by parasitic diseases, such as those transmitted by the snails of the Thiaridae family, which hosts the disease-causing parasitic Centrocestus species [141]. The genus Macrobrachium is not vulnerable to Trematode infection [142,143], and studies demonstrated the high ability of the prawn to abolish snail hatchlings [140,144,145]. Savaya et al. (2020) demonstrated the first application of M. rosenbergii monosex populations as biocontrol agents in commercial aquaculture ponds. A field experiment in Tilapia aquaculture ponds in Israel included prawns as biocontrol agents, which significantly reduced the total biomass of Melanoides tuberculate and Thiara scabra snails of all size classes, and the number of parasites per fish was lower in ponds with prawns than in the control (no-prawns) ponds [146]. Another invasive snail with a high impact on global agriculture is the invasive freshwater apple snail (Pomacea spp.), one of the major unresolved problems worldwide with a significant impact on global rice production, affecting not only economics but also natural ecosystems and potentially also causing health issues [147]. Utilizing monosex prawn populations has been suggested as a sustainable solution against the invasive apple snails and presents an effective biocontrol method under different conditions. Future field validation experiments are required for environmental risk assessment [144,148]. With respect to the treatment of pest snails in agriculture, aquaculture, and even human disease, prawns as biocontrol agents could become a crop by themselves, contributing additional quality protein sources or extra income, thus suggesting a win-win solution and providing a sustainable solution that has the cultural, economic, and environmental incentives to be propagated in perpetuity.

Supplementary Materials

The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/ijms242417433/s1.

Funding

This research was partially funded by the Gruss Lipper Postdoctoral Fellowship awarded to TL. This research was partially funded by the Israel Science Foundation (ISF) within the ISF-NSFC (Grant No. 2368/18) and by a grant from the Ministry of Science & Technology, under The Taiwan-Israel Collaboration program (grant no 001994).

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Wang, T.; He, K.; Blaney, L.; Chung, J.S. Testosterone and steroidogenic genes in the male blue crab Callinectes sapidus and their relationship with insulin-like androgenic gland factor (IAG) and crustacean female sex hormone (CFSH). Aquaculture 2023, 568, 739297. [Google Scholar] [CrossRef]
  2. Wang, T.; He, K.; Blaney, L.; Chung, J.S. 17β-Estradiol (E2) may be involved in the mode of crustacean female sex hormone (CFSH) action in the blue crab, Callinectes sapidus. Front. Endocrinol. 2022, 13, 962576. [Google Scholar] [CrossRef] [PubMed]
  3. Okumura, T.; Sakiyama, K. Hemolymph levels of vertebrate-type steroid hormones in female kuruma prawn Marsupenaeus japonicus (Crustacea: Decapoda: Penaeidae) during natural reproductive cycle and induced ovarian development by eyestalk ablation. Fish. Sci. 2004, 70, 372–380. [Google Scholar] [CrossRef]
  4. Lin, J.; Shi, X.; Fang, S.; Zhang, Y.; You, C.; Ma, H.; Lin, F. Comparative transcriptome analysis combining SMRT and NGS sequencing provides novel insights into sex differentiation and development in mud crab (Scylla paramamosain). Aquaculture 2019, 513, 734447. [Google Scholar] [CrossRef]
  5. Sugestya, I.N.G.; Widodo, M.S.; Soeprijanto, A. Effect of 17β-estradiol on feminization, growth rate and survival rate of pacific white shrimp (Litopenaeus vannamei, Boone 1931) postlarvae. J. Exp. Life Sci. 2018, 8, 37–42. [Google Scholar] [CrossRef]
  6. Yano, I. Effect of 17α-hydroxy-progesterone on vitellogenin secretion in kuruma prawn, Penaeus japonicus. Aquaculture 1987, 61, 49–57. [Google Scholar] [CrossRef]
  7. Yano, I. Induced ovarian maturation and spawning in greasyback shrimp, Metapenaeus ensis, by progesterone. Aquaculture 1985, 47, 223–229. [Google Scholar] [CrossRef]
  8. Liu, M.; Pan, J.; Liu, Z.; Cheng, Y.; Gong, J.; Wu, X. Effect of estradiol on vitellogenesis and oocyte development of female swimming crab, Portunus trituberculatus. Aquaculture 2018, 486, 240–245. [Google Scholar] [CrossRef]
  9. Cronin, L.E. Anatomy and histology of the male reproductive system of Callinectes sapidus Rathbun. J. Morphol. 1947, 81, 209–239. [Google Scholar] [CrossRef]
  10. Charniaux-Cotton, H. Découverte chez un Crustacé Amphipode (Orchestia gammarella) d’une glande endocrine responsable de la différenciation des caractères sexuels primaires et secondaires mâles. Comptes Rendus Hebd. Des Seances De L’academie Des Sci. Paris 1954, 239, 780–782. [Google Scholar]
  11. Charniaux-Cotton, H. Le determinisme hormonal des caracteres sexuels d’Orchestia gammarella (Crustace Amphipode). CR Acad. Sci. Paris 1955, 240, 1487–1489. [Google Scholar]
  12. Charniaux-Cotton, H. Croissance, régénération et déterminisme endocrinien des caractères sexuels d’Orchestia gammarella Pallas (Crustacé Amphipode). Ann. Sci. Nat. Zool. Biol. Anim. 1957, 19, 411–559. [Google Scholar]
  13. Nagamine, C.; Knight, A.W.; Maggenti, A.; Paxman, G. Masculinization of female Macrobrachium rosenbergii (de Man) (Decapoda, Palaemonidae) by androgenic gland implantation. Gen. Comp. Endocrinol. 1980, 41, 442–457. [Google Scholar] [CrossRef]
  14. Taketomi, Y.; Nishikawa, S. Implantation of androgenic glands into immature female crayfish, Procambarus clarkii, with masculinization of sexual characteristics. J. Crustac. Biol. 1996, 16, 232–239. [Google Scholar] [CrossRef]
  15. Cui, Z.; Liu, H.; Lo, T.S.; Chu, K.H. Inhibitory effects of the androgenic gland on ovarian development in the mud crab Scylla paramamosain. Comp. Biochem. Physiol. Part A Mol. Integr. Physiol. 2005, 140, 343–348. [Google Scholar] [CrossRef]
  16. Liu, H.; Cai, S.-L.; Zhang, C.-F.; Chu, K.-H. Masculinization of female Eriocheir sinensis by injecting the extract of androgenic gland of E. sinensis and Scylla paramamosain. J. Fish. China 2006, 30, 577–585. [Google Scholar]
  17. Katakura, Y. Partial purification and characterization of androgenic gland hormone from the isopod crustacean, Armadillidium vulgare. Annot. Zool. 1975, 48, 203–209. [Google Scholar]
  18. Katakura, Y.; Hasegawa, Y. Masculinization of females of the isopod crustacean, Armadillidium vulgare, following injections of an active extract of the androgenic gland. Gen. Comp. Endocrinol. 1983, 49, 57–62. [Google Scholar] [CrossRef] [PubMed]
  19. Hasegawa, Y.; Haino-Fukushima, K.; Katakura, Y. Isolation and properties of androgenic gland hormone from the terrestrial isopod, Armadillidium vulgare. Gen. Comp. Endocrinol. 1987, 67, 101–110. [Google Scholar] [CrossRef] [PubMed]
  20. Okuno, A.; Hasegawa, Y.; Nagasawa, H. Purification and properties of androgenic gland hormone from the terrestrial isopod Armadillidium vulgare. Zool. Sci. 1997, 14, 837–842. [Google Scholar] [CrossRef]
  21. Okuno, A.; Hasegawa, Y.; Ohira, T.; Katakura, Y.; Nagasawa, H. Characterization and cDNA cloning of androgenic gland hormone of the terrestrial isopod Armadillidium vulgare. Biochem. Biophys. Res. Commun. 1999, 264, 419–423. [Google Scholar] [CrossRef] [PubMed]
  22. Ohira, T.; Hasegawa, Y.; Tominaga, S.; Okuno, A.; Nagasawa, H. Molecular cloning and expression analysis of cDNAs encoding androgenic gland hormone precursors from two porcellionidae species, Porcellio scaber and P. dilatatus. Zool. Sci. 2003, 20, 75–81. [Google Scholar] [CrossRef]
  23. Godbillon, G.; Balesdent, M. Specific substance of steroid nature in the male genital tract of the decapod crustacean, Carcinus maenas. Comptes Rendus Seances L’academie Sci. 1974, 278, 2823–2826. [Google Scholar]
  24. Veith, W.; Malecha, S. Histochemical-study of the distribution of lipids, 3-alpha-hydroxysteroid and 3-beta-hydroxysteroid dehydrogenase in the androgenic gland of the cultured prawn, Macrobachium-rosenbergii (de man)(Crustacea, Decapoda). S. Afr. J. Sci. 1983, 79, 84–85. [Google Scholar]
  25. Manor, R.; Weil, S.; Oren, S.; Glazer, L.; Aflalo, E.D.; Ventura, T.; Chalifa-Caspi, V.; Lapidot, M.; Sagi, A. Insulin and gender: An insulin-like gene expressed exclusively in the androgenic gland of the male crayfish. Gen. Comp. Endocrinol. 2007, 150, 326–336. [Google Scholar] [CrossRef] [PubMed]
  26. Ventura, T.; Manor, R.; Aflalo, E.D.; Weil, S.; Raviv, S.; Glazer, L.; Sagi, A. Temporal silencing of an androgenic gland-specific insulin-like gene affecting phenotypical gender differences and spermatogenesis. Endocrinology 2009, 150, 1278–1286. [Google Scholar] [CrossRef] [PubMed]
  27. Levy, T.; Zupo, V.; Mutalipassi, M.; Somma, E.; Ruocco, N.; Costantini, M.; Abehsera, S.; Manor, R.; Chalifa-Caspi, V.; Sagi, A.; et al. Protandric Transcriptomes to Uncover Parts of the Crustacean Sex-Differentiation Puzzle. Front. Mar. Sci. 2021, 8, 745540. [Google Scholar] [CrossRef]
  28. Levy, T.; Tamone, S.L.; Manor, R.; Aflalo, E.D.; Sklarz, M.Y.; Chalifa-Caspi, V.; Sagi, A. The IAG-switch and further transcriptomic insights into sexual differentiation of a protandric shrimp. Front. Mar. Sci. 2020, 7, 587454. [Google Scholar] [CrossRef]
  29. Levy, T.; Rosen, O.; Simons, O.; Savaya Alkalay, A.; Sagi, A. The gene encoding the insulin-like androgenic gland hormone in an all-female parthenogenetic crayfish. PLoS ONE 2017, 12, e0189982. [Google Scholar] [CrossRef]
  30. Levy, T.; Sagi, A. The “IAG-switch”—A key controlling element in decapod crustacean sex differentiation. Front. Endocrinol. 2020, 11, 651. [Google Scholar] [CrossRef]
  31. Charniaux-Cotton, H.; Payen, G. Reproduction in malacostraca crustacea. In Endocrinology of Selected Invertebrate Types; Liss: New York, NY, USA, 1988; pp. 177–186. [Google Scholar]
  32. Ginsburger-Vogel, T.; Charniaux-Cotton, H. Sex determination. In The Biology of Crustacea; Bliss, D.E., Ed.; Academic Press: New York, NY, USA, 1982; pp. 257–281. [Google Scholar]
  33. Kuris, A.M.; Ra’anan, Z.; Sagi, A.; Cohen, D. Morphotypic differentiation of male Malaysian giant prawns, Macrobrachium rosenbergii. J. Crustac. Biol. 1987, 7, 219–237. [Google Scholar] [CrossRef]
  34. Sagi, A.; Ra’anan, Z.; Cohen, D.; Wax, Y. Production of Macrobrachium rosenbergii in monosex populations: Yield characteristics under intensive monoculture conditions in cages. Aquaculture 1986, 51, 265–275. [Google Scholar] [CrossRef]
  35. Sagi, A.; Cohen, D. Growth, maturation and progeny of sex-reversed Macrobrachium rosenbergii males. World Aquac. Rep. 1990, 21, 87–90. [Google Scholar]
  36. Nagamine, C.; Knight, A.W.; Maggenti, A.; Paxman, G. Effects of androgenic gland ablation on male primary and secondary sexual characteristics in the Malaysian prawn, Macrobrachium rosenbergii (de Man) (Decapoda, Palaemonidae), with first evidence of induced feminization in a nonhermaphroditic decapod. Gen. Comp. Endocrinol. 1980, 41, 423–441. [Google Scholar] [CrossRef] [PubMed]
  37. Aflalo, E.D.; Hoang, T.T.T.; Nguyen, V.; Lam, Q.; Nguyen, D.; Trinh, Q.; Raviv, S.; Sagi, A. A novel two-step procedure for mass production of all-male populations of the giant freshwater prawn Macrobrachium rosenbergii. Aquaculture 2006, 256, 468–478. [Google Scholar] [CrossRef]
  38. Agrawal, N.; Dasaradhi, P.; Mohmmed, A.; Malhotra, P.; Bhatnagar, R.K.; Mukherjee, S.K. RNA interference: Biology, mechanism, and applications. Microbiol. Mol. Biol. Rev. 2003, 67, 657–685. [Google Scholar] [CrossRef] [PubMed]
  39. Sagi, A.; Manor, R.; Ventura, T. Gene silencing in crustaceans: From basic research to biotechnologies. Genes 2013, 4, 620–645. [Google Scholar] [CrossRef]
  40. Lezer, Y.; Aflalo, E.D.; Manor, R.; Sharabi, O.; Abilevich, L.K.; Sagi, A. On the safety of RNAi usage in aquaculture: The case of all-male prawn stocks generated through manipulation of the insulin-like androgenic gland hormone. Aquaculture 2015, 435, 157–166. [Google Scholar] [CrossRef]
  41. Katakura, Y. Endocrine and genetic control of sex differentiation in the malacostracan Crustacea. Invertebr. Reprod. Dev. 1989, 16, 177–181. [Google Scholar] [CrossRef]
  42. Levy, T.; Rosen, O.; Eilam, B.; Azulay, D.; Aflalo, E.D.; Manor, R.; Shechter, A.; Sagi, A. A single injection of hypertrophied androgenic gland cells produces all-female aquaculture. Mar. Biotechnol. 2016, 18, 554–563. [Google Scholar] [CrossRef]
  43. Malecha, S.R.; Nevin, P.A.; Ha, P.; Barck, L.E.; Lamadrid-Rose, Y.; Masuno, S.; Hedgecock, D. Sex-ratios and sex-determination in progeny from crosses of surgically sex-reversed freshwater prawns, Macrobrachium rosenbergii. Aquaculture 1992, 105, 201–218. [Google Scholar] [CrossRef]
  44. Parnes, S.; Khalaila, I.; Hulata, G.; Sagi, A. Sex determination in crayfish: Are intersex Cherax quadricarinatus (Decapoda, Parastacidae) genetically females? Genet. Res. 2003, 82, 107–116. [Google Scholar] [CrossRef]
  45. Waiho, K.; Shi, X.; Fazhan, H.; Li, S.; Zhang, Y.; Zheng, H.; Liu, W.; Fang, S.; Ikhwanuddin, M.; Ma, H. High-density genetic linkage maps provide novel insights into ZW/ZZ sex determination system and growth performance in mud crab (Scylla paramamosain). Front. Genet. 2019, 10, 298. [Google Scholar] [CrossRef] [PubMed]
  46. Ventura, T.; Aflalo, E.; Weil, S.; Kashkush, K.; Sagi, A. Isolation and characterization of a female-specific DNA marker in the giant freshwater prawn Macrobrachium rosenbergii. Heredity 2011, 107, 456–461. [Google Scholar] [CrossRef] [PubMed]
  47. Chandler, J.C.; Elizur, A.; Ventura, T. The decapod researcher’s guide to the galaxy of sex determination. Hydrobiologia 2018, 825, 61–80. [Google Scholar] [CrossRef]
  48. Ventura, T.; Manor, R.; Aflalo, E.D.; Weil, S.; Rosen, O.; Sagi, A. Timing sexual differentiation: Full functional sex reversal achieved through silencing of a single insulin-like gene in the prawn, Macrobrachium rosenbergii. Biol. Reprod. 2012, 86, 90. [Google Scholar] [CrossRef] [PubMed]
  49. Barki, A.; Karplus, I.; Khalaila, I.; Manor, R.; Sagi, A. Male-like behavioral patterns and physiological alterations induced by androgenic gland implantation in female crayfish. J. Exp. Biol. 2003, 206, 1791–1797. [Google Scholar] [CrossRef]
  50. Bardera, G.; Owen, M.A.; Façanha, F.N.; Sloman, K.A.; Alexander, M.E. The influence of sex on feeding behaviour in Pacific white shrimp (Litopenaeus vannamei). Appl. Anim. Behav. Sci. 2020, 224, 104946. [Google Scholar] [CrossRef]
  51. Nair, C.; Salin, K.; Raju, M.; Sebastian, M. Economic analysis of monosex culture of giant freshwater prawn (Macrobrachium rosenbergii De Man): A case study. Aquac. Res. 2006, 37, 949–954. [Google Scholar] [CrossRef]
  52. Phansawat, P.; Keetanon, A.; Rairat, T.; Pichitkul, P.; Poldetch, P.; Chuchird, N. Benefits of Male Monosex Culture of Giant Freshwater Prawn (Macrobrachium rosenbergii): Improving Growth Performance, Production Yield, and Profitability. J. Fish. Environ. 2022, 46, 157–168. [Google Scholar]
  53. Islam, S.S.; Golder, M.R.; Bir, J.; Mistry, S.K.; Siddique, M.N.A.; Ali, M.R.; Sabbir, W.; Azad, M.A.K.; Huq, K.A. Growth comparison of mono and mixed-sex giant freshwater prawn (Macrobrachium rosenbergii, De man) and stocking optimization of male monosex farming. Khulna Univ. Stud. 2022, 19, 43–54. [Google Scholar] [CrossRef]
  54. Malecha, S. The case for all-female freshwater prawn, Macrobrachium rosenbergii (De Man), culture. Aquac. Res. 2012, 43, 1038–1048. [Google Scholar] [CrossRef]
  55. Karplus, I.; Sagi, A. The biology and management of size variation. In Freshwater Prawns Biology and Farming; Wiley-Blackwell: Hoboken, NJ, USA, 2010; pp. 316–347. [Google Scholar]
  56. Rotem-Dai, N.; Weil, S.; Greenshpan, Y.; Abehsera, S.; Manor, R.; Ofir, R.; Gazit, R.; Rosental, B.; Aflalo, E.D.; Sagi, A. Lentiviral-Transduced Ectopic Expression of Androgenic Hormone in a Crustacean Hematopoietic Primary Cell Culture. Front. Mar. Sci. 2021, 8, 677679. [Google Scholar] [CrossRef]
  57. Rosen, O.; Manor, R.; Weil, S.; Gafni, O.; Linial, A.; Aflalo, E.D.; Ventura, T.; Sagi, A. A sexual shift induced by silencing of a single insulin-like gene in crayfish: Ovarian upregulation and testicular degeneration. PLoS ONE 2010, 5, e15281. [Google Scholar] [CrossRef] [PubMed]
  58. Ventura, T.; Manor, R.; Aflalo, E.D.; Weil, S.; Khalaila, I.; Rosen, O.; Sagi, A. Expression of an androgenic gland-specific insulin-like peptide during the course of prawn sexual and morphotypic differentiation. Int. Sch. Res. Not. Endocrinol. 2011, 2011, 476283. [Google Scholar] [CrossRef] [PubMed]
  59. Khalaila, I.; Manor, R.; Weil, S.; Granot, Y.; Keller, R.; Sagi, A. The eyestalk–androgenic gland–testis endocrine axis in the crayfish Cherax quadricarinatus. Gen. Comp. Endocrinol. 2002, 127, 147–156. [Google Scholar] [CrossRef]
  60. Sroyraya, M.; Chotwiwatthanakun, C.; Stewart, M.J.; Soonklang, N.; Kornthong, N.; Phoungpetchara, I.; Hanna, P.J.; Sobhon, P. Bilateral eyestalk ablation of the blue swimmer crab, Portunus pelagicus, produces hypertrophy of the androgenic gland and an increase of cells producing insulin-like androgenic gland hormone. Tissue Cell 2010, 42, 293–300. [Google Scholar] [CrossRef]
  61. Chung, J.S.; Manor, R.; Sagi, A. Cloning of an insulin-like androgenic gland factor (IAG) from the blue crab, Callinectes sapidus: Implications for eyestalk regulation of IAG expression. Gen. Comp. Endocrinol. 2011, 173, 4–10. [Google Scholar] [CrossRef]
  62. Keller, R. Crustacean neuropeptides: Structures, functions and comparative aspects. Experientia 1992, 48, 439–448. [Google Scholar] [CrossRef]
  63. Zhu, D.; Feng, T.; Mo, N.; Han, R.; Lu, W.; Shao, S.; Cui, Z. New insights for the regulatory feedback loop between type 1 crustacean female sex hormone (CFSH-1) and insulin-like androgenic gland hormone (IAG) in the Chinese mitten crab (Eriocheir sinensis). Front. Physiol. 2022, 13, 1054773. [Google Scholar] [CrossRef]
  64. Jiang, Q.; Lu, B.; Wang, G.; Ye, H. Transcriptional inhibition of Sp-IAG by crustacean female sex hormone in the mud crab, Scylla paramamosain. Int. J. Mol. Sci. 2020, 21, 5300. [Google Scholar] [CrossRef] [PubMed]
  65. Liu, F.; Liu, A.; Zhu, Z.; Wang, Y.; Ye, H. Crustacean female sex hormone: More than a female phenotypes-related hormone in a protandric simultaneous hermaphroditism shrimp. Int. J. Biol. Macromol. 2023, 238, 124181. [Google Scholar] [CrossRef] [PubMed]
  66. Guo, Q.; Li, S.; Lv, X.; Xiang, J.; Manor, R.; Sagi, A.; Li, F. Sex-biased CHHs and their putative receptor regulate the expression of IAG gene in the shrimp Litopenaeus vannamei. Front. Physiol. 2019, 10, 1525. [Google Scholar] [CrossRef]
  67. Toyota, K.; Miyakawa, H.; Hiruta, C.; Sato, T.; Katayama, H.; Ohira, T.; Iguchi, T. Sex determination and differentiation in decapod and cladoceran crustaceans: An overview of endocrine regulation. Genes 2021, 12, 305. [Google Scholar] [CrossRef] [PubMed]
  68. Li, J. A review of sexual determination and differentiation in crustacean. J. Biosci. Med. 2022, 10, 19–37. [Google Scholar] [CrossRef]
  69. Zheng, J.; Cheng, S.; Jia, Y.; Gu, Z.; Li, F.; Chi, M.; Liu, S.; Jiang, W. Molecular identification and expression profiles of four splice variants of Sex-lethal gene in Cherax quadricarinatus. Comp. Biochem. Physiol. Part B Biochem. Mol. Biol. 2019, 234, 26–33. [Google Scholar] [CrossRef] [PubMed]
  70. Liao, J.; Wan, H.; Zhang, Z.; Sheng, Y.; Jia, X.; Wang, Y. Transcriptional regulation of IAG by dsx and foxl-2 in mud crab (Scylla paramamosain). Gen. Comp. Endocrinol. 2024, 345, 114396. [Google Scholar] [CrossRef] [PubMed]
  71. Peng, B.; Wan, H.; Zhang, Z.; Jia, X.; Liu, C.; Wang, Y. A novel Dmrt gene of crustacean: Functional analysis of idmrt-2 gene in the male reproductive system from Scylla paramamosain. Gene 2023, 850, 146922. [Google Scholar] [CrossRef]
  72. Wei, W.-Y.; Huang, J.-H.; Zhou, F.-L.; Yang, Q.-B.; Li, Y.-D.; Jiang, S.; Jiang, S.-G.; Yang, L.-S. Identification and Expression Analysis of Dsx and Its Positive Transcriptional Regulation of IAG in Black Tiger Shrimp (Penaeus monodon). Int. J. Mol. Sci. 2022, 23, 12701. [Google Scholar] [CrossRef]
  73. Li, S.; Li, F.; Yu, K.; Xiang, J. Identification and characterization of a doublesex gene which regulates the expression of insulin-like androgenic gland hormone in Fenneropenaeus chinensis. Gene 2018, 649, 1–7. [Google Scholar] [CrossRef]
  74. Zhang, P.; Yang, Y.; Xu, Y.; Cui, Z. Analyses of the Dmrt family in a decapod crab, Eriocheir sinensis uncover new facets on the evolution of DM domain genes. Front. Physiol. 2023, 14, 1201846. [Google Scholar] [CrossRef]
  75. Wang, L.; Liu, Z.-Q.; Ma, K.-Y.; Tao, J.-R.; Fang, X.; Qiu, G.-F. A testis-specific gene doublesex is involved in spermatogenesis and early sex differentiation of the Chinese mitten crab Eriocheir sinensis. Aquaculture 2023, 569, 739401. [Google Scholar] [CrossRef]
  76. Zheng, J.; Cai, L.; Jia, Y.; Chi, M.; Cheng, S.; Liu, S.; Li, F.; Gu, Z. Identification and functional analysis of the doublesex gene in the redclaw crayfish, Cherax quadricarinatus. Gene Expr. Patterns 2020, 37, 119129. [Google Scholar] [CrossRef] [PubMed]
  77. Zhong, P.; Zhou, T.; Zhang, Y.; Chen, Y.; Yi, J.; Lin, W.; Guo, Z.; Xu, A.; Yang, S.; Chan, S. Potential involvement of a DMRT family member (Mr-Dsx) in the regulation of sexual differentiation and moulting in the giant river prawn Macrobrachium rosenbergii. Aquac. Res. 2019, 50, 3037–3049. [Google Scholar] [CrossRef]
  78. Lv, Q.; Li, S.; Miao, M.; Jin, S.; Li, F. IAG Regulates the Expression of Cytoskeletal Protein-Encoding Genes in Shrimp Testis. Genes 2023, 14, 564. [Google Scholar] [CrossRef]
  79. Sun, R.; Yi, S.; Shi, L.; Tan, K.; Shi, R.; Yang, S.; Li, Y. Analysis of mRNA and MicroRNA Expression Profiles of Nervous Tissues and Reproductive Tissues in Male Procambarus clarkii After Silencing IAG. Mar. Biotechnol. 2023, 1–14. [Google Scholar] [CrossRef] [PubMed]
  80. Smith, G.; Glendinning, S.; Ventura, T. Transcriptomic Changes Following Induced De-Masculinisation of Australian Red Claw Crayfish Cherax quadricarinatus. Int. J. Mol. Sci. 2023, 24, 3292. [Google Scholar] [CrossRef]
  81. Lin, D.; Wang, M.; Yu, F.; Shi, W.; Luo, F.; Wu, C.; Yang, J.; Ma, W. Identification and expression pattern of three sex-related genes in the shrimp Neocaridina denticulata sinensis (Decapoda, Caridea). Crustaceana 2022, 95, 723–746. [Google Scholar] [CrossRef]
  82. Chen, Y.-L.; Wang, Y.-M.; Xu, H.-J.; Li, J.-W.; Luo, J.-Y.; Wang, M.-R.; Ma, W.-M. The characterization and knockdown of a male gonad-specific insulin-like receptor gene in the white shrimp Penaeus vannamei. Aquac. Rep. 2022, 27, 101345. [Google Scholar] [CrossRef]
  83. Qian, H.; Ma, K.; Feng, J.; Guo, Z.; Gong, J.; Chen, H.; Bai, H.; Qiu, G. Transcriptome analysis of the post-larvae of giant freshwater prawn (Macrobrachium rosenbergii) after IAG gene knockdown with microRNA interference. Gen. Comp. Endocrinol. 2022, 325, 114054. [Google Scholar] [CrossRef]
  84. Cohen, S.; Hasan, M.; Frishman, N.; Khalaila, I. A crustacean vitellogenin-derived peptide as an oocyte-specific delivery vehicle for gene silencing. Front. Mar. Sci. 2023, 10, 1128524. [Google Scholar] [CrossRef]
  85. Xu, H.-J.; Chen, Y.-L.; Wang, Y.-M.; Luo, J.-Y.; Li, J.-W.; Shen, S.-Q.; Yang, J.-S.; Ma, W.-M. Full functional sex reversal achieved through silencing of MroDmrt11E gene in Macrobrachium rosenbergii: Production of all-male monosex freshwater prawn. Front. Endocrinol. 2022, 12, 772498. [Google Scholar] [CrossRef] [PubMed]
  86. Yu, Y.-Q.; Ma, W.-M.; Zeng, Q.-G.; Qian, Y.-Q.; Yang, J.-S.; Yang, W.-J. Molecular cloning and sexually dimorphic expression of two Dmrt genes in the giant freshwater prawn, Macrobrachium rosenbergii. Agric. Res. 2014, 3, 181–191. [Google Scholar] [CrossRef]
  87. Wang, Y.; Jin, S.; Fu, H.; Qiao, H.; Sun, S.; Zhang, W.; Jiang, S.; Gong, Y.; Xiong, Y.; Wu, Y. Identification and characterization of the DMRT11E gene in the oriental river prawn Macrobrachium nipponense. Int. J. Mol. Sci. 2019, 20, 1734. [Google Scholar] [CrossRef] [PubMed]
  88. Jiang, Q.; Xu, D.; Wang, M.; Xie, X.; Zhu, D. Molecular Characterization of a Male-specific SoxE Gene in the Swimming Crab, Portunus trituberculatus, and Transcriptional Interaction with Insulin-like Androgenic Gland Hormone. Fishes 2023, 8, 351. [Google Scholar] [CrossRef]
  89. Liu, Z.-Q.; Jiang, X.-H.; Qi, H.-Y.; Xiong, L.-W.; Qiu, G.-F. A novel SoxB2 gene is required for maturation of sperm nucleus during spermiogenesis in the Chinese mitten crab, Eriocheir sinensis. Sci. Rep. 2016, 6, 32139. [Google Scholar] [CrossRef] [PubMed]
  90. Liu, C.; Jia, X.; Zou, Z.; Wang, X.; Wang, Y.; Zhang, Z. VIH from the mud crab is specifically expressed in the eyestalk and potentially regulated by transactivator of Sox9/Oct4/Oct1. Gen. Comp. Endocrinol. 2018, 255, 1–11. [Google Scholar] [CrossRef]
  91. Li, F.; Bai, H.; Zhang, W.; Fu, H.; Jiang, F.; Liang, G.; Jin, S.; Sun, S.; Qiao, H. Cloning of genomic sequences of three crustacean hyperglycemic hormone superfamily genes and elucidation of their roles of regulating insulin-like androgenic gland hormone gene. Gene 2015, 561, 68–75. [Google Scholar] [CrossRef]
  92. Luo, J.-Y.; Shen, S.-Q.; Xu, H.-J.; Yang, J.-S.; Ma, W.-M. The transcription factor masculinizer in sexual differentiation and achieved full functional sex reversal in prawn. iScience 2023, 26, 106968. [Google Scholar] [CrossRef]
  93. Wan, H.; Zhong, J.; Zhang, Z.; Xie, Y.; Wang, Y. Characterization of the foxl2 gene involved in the vtg expression in mud crab (Scylla paramamosain). Gene 2021, 798, 145807. [Google Scholar] [CrossRef]
  94. Wang, M.; Xu, R.; Tu, S.; Yu, Q.; Xie, X.; Zhu, D. Putative Role of CFSH in the Eyestalk-AG-Testicular Endocrine Axis of the Swimming Crab Portunus trituberculatus. Animals 2023, 13, 690. [Google Scholar] [CrossRef]
  95. Liu, A.; Liu, J.; Liu, F.; Huang, Y.; Wang, G.; Ye, H. Crustacean female sex hormone from the mud crab Scylla paramamosain is highly expressed in prepubertal males and inhibits the development of androgenic gland. Front. Physiol. 2018, 9, 924. [Google Scholar] [CrossRef] [PubMed]
  96. Chen, S.; Fu, H.; Qiao, H.; Sun, S.; Zhang, W.; Jin, S.; Gong, Y.; Jiang, S.; Xiong, Y.; Wu, Y. RNA interference studies on the sex-linked genes transformer-2 and sex-lethal in the oriental river prawn Macrobrachium nipponense. Isr. J. Aquac. Bamidgeh 2017, 69. [Google Scholar] [CrossRef]
  97. Jin, S.; Hu, Y.; Fu, H.; Jiang, S.; Xiong, Y.; Qiao, H.; Zhang, W.; Gong, Y.; Wu, Y. Potential functions of Gem-associated protein 2-like isoform X1 in the oriental river prawn Macrobrachium nipponense: Cloning, qPCR, in situ hybridization, and RNAi analysis. Int. J. Mol. Sci. 2019, 20, 3995. [Google Scholar] [CrossRef] [PubMed]
  98. Shu, L.; Yang, Y.; Huang, H.; Ye, H. A bone morphogenetic protein ligand and receptors in mud crab: A potential role in the ovarian development. Mol. Cell. Endocrinol. 2016, 434, 99–107. [Google Scholar] [CrossRef]
  99. Levy, T.; Rosen, O.; Manor, R.; Dotan, S.; Azulay, D.; Abramov, A.; Sklarz, M.Y.; Chalifa-Caspi, V.; Baruch, K.; Shechter, A. Production of WW males lacking the masculine Z chromosome and mining the Macrobrachium rosenbergii genome for sex-chromosomes. Sci. Rep. 2019, 9, 12408. [Google Scholar] [CrossRef]
  100. Ventura, T.; Sagi, A. The insulin-like androgenic gland hormone in crustaceans: From a single gene silencing to a wide array of sexual manipulation-based biotechnologies. Biotechnol. Adv. 2012, 30, 1543–1550. [Google Scholar] [CrossRef]
  101. Shpak, N.; Manor, R.; Aflalo, E.D.; Sagi, A. Three generations of cultured prawn without W chromosome. Aquaculture 2017, 467, 41–48. [Google Scholar] [CrossRef]
  102. Levy, T.; Rosen, O.; Eilam, B.; Azulay, D.; Zohar, I.; Aflalo, E.D.; Benet, A.; Naor, A.; Shechter, A.; Sagi, A. All-female monosex culture in the freshwater prawn Macrobrachium rosenbergii–A comparative large-scale field study. Aquaculture 2017, 479, 857–862. [Google Scholar] [CrossRef]
  103. Molcho, J.; Levy, T.; Benet, A.; Naor, A.; Savaya, A.; Manor, R.; Abramov, A.; Aflalo, E.D.; Shechter, A.; Sagi, A. Three generations of prawns without the Z chromosome: Viable WW Macrobrachium rosenbergii all-female populations in polyculture with Oreochromis niloticus. Aquaculture 2020, 515, 734531. [Google Scholar] [CrossRef]
  104. Yuan, J.; Yu, Y.; Zhang, X.; Li, S.; Xiang, J.; Li, F. Recent advances in crustacean genomics and their potential application in aquaculture. Rev. Aquac. 2023, 15, 1501–1521. [Google Scholar] [CrossRef]
  105. González-Castellano, I.; Manfrin, C.; Pallavicini, A.; Martínez-Lage, A. De novo gonad transcriptome analysis of the common littoral shrimp Palaemon serratus: Novel insights into sex-related genes. BMC Genom. 2019, 20, 757. [Google Scholar] [CrossRef] [PubMed]
  106. Zhang, W.; Lv, J.; Lan, W.; Gao, B.; Liu, P. Discovery of Sex-Determining Genes in Portunus trituberculatus: A Comparison of Male and Female Transcriptomes During Early Developmental Stages. Front. Mar. Sci. 2022, 8, 811052. [Google Scholar] [CrossRef]
  107. Wang, Y.; Yu, Y.; Li, S.; Zhang, X.; Xiang, J.; Li, F. Sex-Specific transcriptome sequencing of zoea I larvae and identification of sex-linked genes using bulked segregant analysis in Pacific white shrimp Litopenaeus vannamei. Mar. Biotechnol. 2020, 22, 423–432. [Google Scholar] [CrossRef] [PubMed]
  108. Ventura, T.; Chandler, J.C.; Nguyen, T.V.; Hyde, C.J.; Elizur, A.; Fitzgibbon, Q.P.; Smith, G.G. Multi-Tissue transcriptome analysis identifies key sexual development-related genes of the ornate spiny lobster (Panulirus ornatus). Genes 2020, 11, 1150. [Google Scholar] [CrossRef]
  109. Grinshpan, N.; Abayed, F.A.A.; Wahl, M.; Ner-Gaon, H.; Manor, R.; Sagi, A.; Shay, T. The transcriptional landscape of the giant freshwater prawn: Embryonic development and early sexual differentiation mechanisms. Front. Endocrinol. 2022, 13, 1059936. [Google Scholar] [CrossRef]
  110. Abayed, F.A.A.; Manor, R.; Aflalo, E.D.; Sagi, A. Screening for Dmrt genes from embryo to mature Macrobrachium rosenbergii prawns. Gen. Comp. Endocrinol. 2019, 282, 113205. [Google Scholar] [CrossRef]
  111. Sharabi, O.; Manor, R.; Weil, S.; Aflalo, E.; Lezer, Y.; Levy, T.; Aizen, J.; Ventura, T.; Mather, P.; Khalaila, I. Identification and characterization of an insulin-like receptor involved in crustacean reproduction. Endocrinology 2016, 157, 928–941. [Google Scholar] [CrossRef]
  112. Gutekunst, J.; Andriantsoa, R.; Falckenhayn, C.; Hanna, K.; Stein, W.; Rasamy, J.; Lyko, F. Clonal genome evolution and rapid invasive spread of the marbled crayfish. Nat. Ecol. Evol. 2018, 2, 567–573. [Google Scholar] [CrossRef]
  113. Tan, M.H.; Gan, H.M.; Lee, Y.P.; Grandjean, F.; Croft, L.J.; Austin, C.M. A giant genome for a giant crayfish (Cherax quadricarinatus) with insights into cox1 pseudogenes in decapod genomes. Front. Genet. 2020, 11, 201. [Google Scholar] [CrossRef]
  114. Liu, B.-H.; Zhang, Y.-Q.; Ma, K.-Y.; Wu, X.-G.; Qiu, G.-F. Identification of sex-specific markers and ZW-chromosome DNA clones from the genomic BAC library of the Chinese mitten crab Eriocheir sinensis. Aquaculture 2022, 560, 738576. [Google Scholar] [CrossRef]
  115. Song, L.; Bian, C.; Luo, Y.; Wang, L.; You, X.; Li, J.; Qiu, Y.; Ma, X.; Zhu, Z.; Ma, L. Draft genome of the Chinese mitten crab, Eriocheir sinensis. GigaScience 2016, 5, s13742-13016-10112-y. [Google Scholar] [CrossRef] [PubMed]
  116. Veldsman, W.P.; Ma, K.Y.; Hui, J.H.L.; Chan, T.F.; Baeza, J.A.; Qin, J.; Chu, K.H. Comparative genomics of the coconut crab and other decapod crustaceans: Exploring the molecular basis of terrestrial adaptation. BMC Genom. 2021, 22, 313. [Google Scholar] [CrossRef] [PubMed]
  117. Li, J.; Lv, J.; Liu, P.; Chen, P.; Wang, J.; Li, J. Genome survey and high-resolution backcross genetic linkage map construction of the ridgetail white prawn Exopalaemon carinicauda applications to QTL mapping of growth traits. BMC Genom. 2019, 20, 598. [Google Scholar] [CrossRef] [PubMed]
  118. Kenny, N.J.; Sin, Y.W.; Shen, X.; Zhe, Q.; Wang, W.; Chan, T.F.; Tobe, S.S.; Shimeld, S.M.; Chu, K.H.; Hui, J.H. Genomic sequence and experimental tractability of a new decapod shrimp model, Neocaridina denticulata. Mar. Drugs 2014, 12, 1419–1437. [Google Scholar] [CrossRef] [PubMed]
  119. Zhang, X.; Yuan, J.; Sun, Y.; Li, S.; Gao, Y.; Yu, Y.; Liu, C.; Wang, Q.; Lv, X.; Zhang, X. Penaeid shrimp genome provides insights into benthic adaptation and frequent molting. Nat. Commun. 2019, 10, 356. [Google Scholar] [CrossRef] [PubMed]
  120. Yuan, J.; Zhang, X.; Liu, C.; Yu, Y.; Wei, J.; Li, F.; Xiang, J. Genomic resources and comparative analyses of two economical penaeid shrimp species, Marsupenaeus japonicus and Penaeus monodon. Mar. Genom. 2018, 39, 22–25. [Google Scholar] [CrossRef]
  121. Polinski, J.M.; Zimin, A.V.; Clark, K.F.; Kohn, A.B.; Sadowski, N.; Timp, W.; Ptitsyn, A.; Khanna, P.; Romanova, D.Y.; Williams, P. The American lobster genome reveals insights on longevity, neural, and immune adaptations. Sci. Adv. 2021, 7, eabe8290. [Google Scholar] [CrossRef]
  122. Anzalone, A.V.; Koblan, L.W.; Liu, D.R. Genome editing with CRISPR–Cas nucleases, base editors, transposases and prime editors. Nat. Biotechnol. 2020, 38, 824–844. [Google Scholar] [CrossRef]
  123. Gao, Y.; Zhang, X.; Zhang, X.; Yuan, J.; Xiang, J.; Li, F. CRISPR/Cas9-mediated mutation reveals Pax6 is essential for development of the compound eye in Decapoda Exopalaemon carinicauda. Dev. Biol. 2020, 465, 157–167. [Google Scholar] [CrossRef]
  124. Nakanishi, T.; Kato, Y.; Matsuura, T.; Watanabe, H. CRISPR/Cas-mediated targeted mutagenesis in Daphnia magna. PLoS ONE 2014, 9, e98363. [Google Scholar] [CrossRef] [PubMed]
  125. Martin, A.; Serano, J.M.; Jarvis, E.; Bruce, H.S.; Wang, J.; Ray, S.; Barker, C.A.; O’Connell, L.C.; Patel, N.H. CRISPR/Cas9 mutagenesis reveals versatile roles of Hox genes in crustacean limb specification and evolution. Curr. Biol. 2016, 26, 14–26. [Google Scholar] [CrossRef] [PubMed]
  126. Gui, T.; Zhang, J.; Song, F.; Sun, Y.; Xie, S.; Yu, K.; Xiang, J. CRISPR/Cas9-mediated genome editing and mutagenesis of EcChi4 in Exopalaemon carinicauda. G3 Genes Genomes Genet. 2016, 6, 3757–3764. [Google Scholar] [CrossRef]
  127. Molcho, J.; Manor, R.; Shamsian, M.; Sudarshan, G.; Ofir, R.; Parker, D.; Weil, S.; Wattad, H.; Hayun, E.; Levy, T. On genome editing in embryos and cells of the freshwater prawn Macrobrachium rosenbergii. Aquaculture 2022, 558, 738391. [Google Scholar] [CrossRef]
  128. Miao, M.; Li, S.; Yuan, J.; Liu, P.; Fang, X.; Zhang, C.; Zhang, X.; Li, F. CRISPR/Cas9-mediated gene mutation of EcIAG leads to sex reversal in the male ridgetail white prawn Exopalaemon carinicauda. Front. Endocrinol. 2023, 14, 1266641. [Google Scholar] [CrossRef]
  129. Stotish, R.L. Navigating the FDA approval process for genetically engineered animals. Transgenic Res 2014, 23, 198–199. [Google Scholar]
  130. Bratlie, S.; Halvorsen, K.; Myskja, B.K.; Mellegård, H.; Bjorvatn, C.; Frost, P.; Heiene, G.; Hofmann, B.; Holst-Jensen, A.; Holst-Larsen, T. A novel governance framework for GMO: A tiered, more flexible regulation for GMO s would help to stimulate innovation and public debate. EMBO Rep. 2019, 20, e47812. [Google Scholar] [CrossRef]
  131. Gherardi, F.; Aquiloni, L.; Diéguez-Uribeondo, J.; Tricarico, E. Managing invasive crayfish: Is there a hope? Aquat. Sci. 2011, 73, 185–200. [Google Scholar] [CrossRef]
  132. Kolar, C.S.; Lodge, D.M. Progress in invasion biology: Predicting invaders. Trends Ecol. Evol. 2001, 16, 199–204. [Google Scholar] [CrossRef]
  133. Gherardi, F. Crayfish invading Europe: The case study of Procambarus clarkii. Mar. Freshw. Behav. Physiol. 2006, 39, 175–191. [Google Scholar] [CrossRef]
  134. Savaya, A.; De Leo, G.; Aalto, E.; Levy, T.; Rosen, O.; Manor, R.; Aflalo, E.D.; Tricarico, E.; Sagi, A. The IAG gene in the invasive crayfish Procambarus clarkii–towards sex manipulations for biocontrol and aquaculture. Manag. Biol. Invasions 2020, 11, 237. [Google Scholar] [CrossRef]
  135. Erickson, R.A.; Thompson, H.M.; Kageyama, S.A.; Andriacchi, G.M.; Cupp, A.R.; Patiño, R.; Amberg, J.J. Assessing the suitability of YY males and ZZ females as an invasive species population control method across life histories. Biol. Invasions 2023, 25, 3737–3751. [Google Scholar] [CrossRef]
  136. Dearden, P.K.; Gemmell, N.J.; Mercier, O.R.; Lester, P.J.; Scott, M.J.; Newcomb, R.D.; Buckley, T.R.; Jacobs, J.M.; Goldson, S.G.; Penman, D.R. The potential for the use of gene drives for pest control in New Zealand: A perspective. J. R. Soc. N. Z. 2018, 48, 225–244. [Google Scholar] [CrossRef]
  137. Gierus, L.; Birand, A.; Bunting, M.D.; Godahewa, G.I.; Piltz, S.G.; Oh, K.P.; Piaggio, A.J.; Threadgill, D.W.; Godwin, J.; Edwards, O. Leveraging a natural murine meiotic drive to suppress invasive populations. Proc. Natl. Acad. Sci. USA 2022, 119, e2213308119. [Google Scholar] [CrossRef] [PubMed]
  138. Hammond, A.; Galizi, R.; Kyrou, K.; Simoni, A.; Siniscalchi, C.; Katsanos, D.; Gribble, M.; Baker, D.; Marois, E.; Russell, S. A CRISPR-Cas9 gene drive system targeting female reproduction in the malaria mosquito vector Anopheles gambiae. Nat. Biotechnol. 2016, 34, 78–83. [Google Scholar] [CrossRef]
  139. Sokolow, S.H.; Jones, I.J.; Jocque, M.; La, D.; Cords, O.; Knight, A.; Lund, A.; Wood, C.L.; Lafferty, K.D.; Hoover, C.M. Nearly 400 million people are at higher risk of schistosomiasis because dams block the migration of snail-eating river prawns. Philos. Trans. R. Soc. B Biol. Sci. 2017, 372, 20160127. [Google Scholar] [CrossRef]
  140. Savaya-Alkalay, A.; Ndao, P.D.; Jouanard, N.; Diane, N.; Aflalo, E.D.; Barki, A.; Sagi, A. Exploitation of reproductive barriers between Macrobrachium species for responsible aquaculture and biocontrol of schistosomiasis in West Africa. Aquac. Environ. Interact. 2018, 10, 487–499. [Google Scholar] [CrossRef]
  141. Acosta-Pérez, V.J.; Ángeles-Hernández, J.C.; Vega-Sánchez, V.; Zepeda-Velázquez, A.P.; Añorve-Morga, J.; Ponce-Noguez, J.B.; Reyes-Rodríguez, N.E.; De-La-Rosa-Arana, J.L.; Ramírez-Paredes, J.G.; Gómez-De-Anda, F.R. Prevalence of Parasitic Infections with Zoonotic Potential in Tilapia: A Systematic Review and Meta-Analysis. Animals 2022, 12, 2800. [Google Scholar] [CrossRef]
  142. Lee, C.; Fielder, D. Maintenance and reproductive behaviour in the freshwatre prawn Macrobrachium australiense Holthuis (Crustacea: Decapoda: Palaemonidae). Mar. Freshw. Res. 1982, 33, 629–646. [Google Scholar] [CrossRef]
  143. Roberts, J.K.; Kuris, A. Predation and control of laboratory populations of the snail Biomphalaria glabrata by the freshwater prawn Macrobrachium rosenbergii. Ann. Trop. Med. Parasitol. 1990, 84, 401–412. [Google Scholar] [CrossRef]
  144. Savaya-Alkalay, A.; Ovadia, O.; Barki, A.; Sagi, A. Size-selective predation by all-male prawns: Implications for sustainable biocontrol of snail invasions. Biol. Invasions 2018, 20, 137–149. [Google Scholar] [CrossRef]
  145. Sokolow, S.H.; Lafferty, K.D.; Kuris, A.M. Regulation of laboratory populations of snails (Biomphalaria and Bulinus spp.) by river prawns, Macrobrachium spp.(Decapoda, Palaemonidae): Implications for control of schistosomiasis. Acta Trop. 2014, 132, 64–74. [Google Scholar] [CrossRef] [PubMed]
  146. Savaya, A.; Glassner, H.; Livne-Luzon, S.; Chishinski, R.; Molcho, J.; Aflalo, E.D.; Zilberg, D.; Sagi, A. Prawn monosex populations as biocontrol agents for snail vectors of fish parasites. Aquaculture 2020, 520, 735016. [Google Scholar] [CrossRef]
  147. Azmi, W.A.; Khoo, S.C.; Ng, L.C.; Baharuddin, N.; Abd Aziz, A.; Ma, N.L. The current trend in biological control approaches in the mitigation of golden apple snail Pomacea spp. Biol. Control 2022, 175, 105060. [Google Scholar] [CrossRef]
  148. Wang, Y.; Zhang, Y.; Lv, H.; Sheng, Q. Efficacy of Giant River Prawn Macrobrachium rosenbergii in Controlling the Invasive Snail Pomacea canaliculata: Implications for Ecological Farming. Diversity 2023, 15, 1001. [Google Scholar] [CrossRef]
Ijms 24 17433 g001
Figure 1. Monosex population aquaculture of the giant freshwater prawn Macrobrachium rosenbergii: ‘the tools of the trade’. (A) The androgenic gland (AG) was first discovered in a decapod in 1947, although it was not until 2007 that the first decapod AG-derived insulin-like peptide (IAG) encoding sequence (IAG) was identified. This photo shows the AG situated alongside the sperm duct in the Australian redclaw crayfish, Cherax quadricarinatus. (B) Proximal parts of the male reproductive systems of M. rosenbergii males; cross-section of the terminal ampullae (TA) with the sperm duct (SD) and AG area. The cells of this gland, following an endocrine manipulation to induce its hypertrophy, are dispersed and injected into females to induce sex change into neo-males, which generates all-female populations. Drawing—Dr. Shaul Raviv (C) Following decades of research prior to the advent of high-throughput sequencing, the IAG is now identified in multiple decapod species, where it is found to primarily express in the AG. The gene encodes a signal peptide, followed by a B chain, a C peptide, and an A chain. During translation, the signal peptide is cleaved off, followed by cleavage of the C peptide, leaving the B chain and A chain interlinked with two disulphide bonds and another disulphide bond within the A chain (linear model), rendering the mature IAG with an insulin-like three-dimensional structure signature. Silencing IAG in early stage of male development enables sex change into neo-females, which generates all-male populations.
Figure 1. Monosex population aquaculture of the giant freshwater prawn Macrobrachium rosenbergii: ‘the tools of the trade’. (A) The androgenic gland (AG) was first discovered in a decapod in 1947, although it was not until 2007 that the first decapod AG-derived insulin-like peptide (IAG) encoding sequence (IAG) was identified. This photo shows the AG situated alongside the sperm duct in the Australian redclaw crayfish, Cherax quadricarinatus. (B) Proximal parts of the male reproductive systems of M. rosenbergii males; cross-section of the terminal ampullae (TA) with the sperm duct (SD) and AG area. The cells of this gland, following an endocrine manipulation to induce its hypertrophy, are dispersed and injected into females to induce sex change into neo-males, which generates all-female populations. Drawing—Dr. Shaul Raviv (C) Following decades of research prior to the advent of high-throughput sequencing, the IAG is now identified in multiple decapod species, where it is found to primarily express in the AG. The gene encodes a signal peptide, followed by a B chain, a C peptide, and an A chain. During translation, the signal peptide is cleaved off, followed by cleavage of the C peptide, leaving the B chain and A chain interlinked with two disulphide bonds and another disulphide bond within the A chain (linear model), rendering the mature IAG with an insulin-like three-dimensional structure signature. Silencing IAG in early stage of male development enables sex change into neo-females, which generates all-male populations.
Ijms 24 17433 g001
Ijms 24 17433 g002
Figure 2. Monosex population aquaculture of the giant freshwater prawn Macrobrachium rosenbergii: both all-male and all-female populations might be advantageous. (A) Weight distribution of male morphotypes and females in a mixed population. Sample groups of females, small male (SM), orange-claw male (OC), blue-claw male (BC), and no-claw male (NC) under extensive stocking density. (B) A large dominant male harvested with four females from the same population. Note that all the females carry eggs and are of uniform size. Photograph—Tomer Ventura. (C) A small-scale pond study concluded that all-male giant freshwater prawn populations, when selectively harvested, produce double the cumulative yield when compared with mixed-sex and all-female populations. (D) Weight distribution of all-female and mixed population at the end of the grow-out season. Females grow uniformly, with better survival rates, even at high stocking densities.
Figure 2. Monosex population aquaculture of the giant freshwater prawn Macrobrachium rosenbergii: both all-male and all-female populations might be advantageous. (A) Weight distribution of male morphotypes and females in a mixed population. Sample groups of females, small male (SM), orange-claw male (OC), blue-claw male (BC), and no-claw male (NC) under extensive stocking density. (B) A large dominant male harvested with four females from the same population. Note that all the females carry eggs and are of uniform size. Photograph—Tomer Ventura. (C) A small-scale pond study concluded that all-male giant freshwater prawn populations, when selectively harvested, produce double the cumulative yield when compared with mixed-sex and all-female populations. (D) Weight distribution of all-female and mixed population at the end of the grow-out season. Females grow uniformly, with better survival rates, even at high stocking densities.
Ijms 24 17433 g002
Ijms 24 17433 g003
Figure 3. From fertilization to maturation in the giant freshwater prawn Macrobrachium rosenbergii through the IAG-switch. The process starts with the genotype determination, WZ or ZZ, followed by developmental stages of embryos, larvae, and early post-larvae. An individual with a ZZ set of chromosomes develops an androgenic gland (AG), which secretes the IAG, leading to the development of a functional, mature male phenotype. In an individual with the WZ set of chromosomes, the AG is absent, so there is no IAG secretion, and a functional, mature female phenotype is exhibited. This system could be manipulated as a sex-differentiating switch, thus termed the “IAG-switch”. However, the controlling elements upstream the IAG-switch are not yet elucidated.
Figure 3. From fertilization to maturation in the giant freshwater prawn Macrobrachium rosenbergii through the IAG-switch. The process starts with the genotype determination, WZ or ZZ, followed by developmental stages of embryos, larvae, and early post-larvae. An individual with a ZZ set of chromosomes develops an androgenic gland (AG), which secretes the IAG, leading to the development of a functional, mature male phenotype. In an individual with the WZ set of chromosomes, the AG is absent, so there is no IAG secretion, and a functional, mature female phenotype is exhibited. This system could be manipulated as a sex-differentiating switch, thus termed the “IAG-switch”. However, the controlling elements upstream the IAG-switch are not yet elucidated.
Ijms 24 17433 g003
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.

Share and Cite

MDPI and ACS Style

Wahl, M.; Levy, T.; Ventura, T.; Sagi, A. Monosex Populations of the Giant Freshwater Prawn Macrobrachium rosenbergii—From a Pre-Molecular Start to the Next Generation Era. Int. J. Mol. Sci. 2023, 24, 17433. https://doi.org/10.3390/ijms242417433

AMA Style

Wahl M, Levy T, Ventura T, Sagi A. Monosex Populations of the Giant Freshwater Prawn Macrobrachium rosenbergii—From a Pre-Molecular Start to the Next Generation Era. International Journal of Molecular Sciences. 2023; 24(24):17433. https://doi.org/10.3390/ijms242417433

Chicago/Turabian Style

Wahl, Melody, Tom Levy, Tomer Ventura, and Amir Sagi. 2023. "Monosex Populations of the Giant Freshwater Prawn Macrobrachium rosenbergii—From a Pre-Molecular Start to the Next Generation Era" International Journal of Molecular Sciences 24, no. 24: 17433. https://doi.org/10.3390/ijms242417433

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop