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Molecular Advance on Reproduction and Fertility of Aquatic Animals 2.0

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A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Molecular Biology".

Deadline for manuscript submissions: closed (20 January 2025) | Viewed by 17341

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Prof. Dr. Hongtuo Fu

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Guest Editor

1. Wuxi Fisheries College, Nanjing Agricultural University, Wuxi 214081, China
2. Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture and Rural Affairs, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi 214081, China
Interests: biological breeding; reproduction of the oriental river prawn (Macrobrachium nipponense); aquatic animal
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Dr. Hui Qiao

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Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture and Rural Affairs, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi 214081, China
Interests: aquaculture; biotechnology; germplasm improvement; genetic breeding; gene function; genomics; gene editing; population genetics
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Reproduction and fertility are complex mechanisms in aquatic animals. Many aquatic animals have sexual dimorphism between male and female individuals, showing significant differences in growth. Thus, monosex culture may have dramatic economic benefits. Many economic aquatic animals are widely cultured in many countries and regions. However, both rapid and slow gonad development have negative effects on sustainable development. Slow gonad development will extend the breeding cycle, while rapid gonad development will result in inbreeding between new-born animals, leading to short life span, small size and low disease resistance. Therefore, analyses of their reproductive mechanisms are equally important, including but not limited to molecular functional studies of sex differentiation and gonadal development. In addition, several biological or chemical reagents have been shown to have significant effects on aquatic animal reproduction and fertility, such as steroid hormones, sex-determining genes, etc. Thus, the mechanisms of sex-determination and reproduction needed to be fully understood in order to establish techniques to produce monosex cultures and regulate gonad development in aquatic animals. Significant progress has been made over the past few decades in areas such as biological reproduction and developmental processes. However, relatively little has been conducted in fish, crustaceans and shellfish. The effects of steroid hormones on the process of sex-determination and reproduction in aquatic animals will continue to be unraveled, as well as identifying the complex signaling networks and genes regulated by the steroid hormone. Identification of the roles of nutritional and environmental factors on hormone secretion, hormone balance and corresponding sex-determination and reproduction outcomes altered by changed hormone levels are areas of focus.

Reviews or research articles in this fascinating area, which discuss the reproduction and fertility of aquatic animals and their applications or which aim to discern the bottlenecks in the field, are welcome.

Prof. Dr. Hongtuo Fu
Dr. Hui Qiao
Guest Editors

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Keywords

reproduction
aquatic animals
sex determination
gonadal development
steroid hormones
sex-related genes
monosex culture

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Research

11 pages, 3118 KiB

Open AccessArticle

Infectious Spleen and Kidney Necrosis Virus ORF093R and ORF102R Regulate Glutamate Metabolic Reprogramming to Support Virus Proliferation by Interacting with c-Myc

by Yinjie Niu, Caimei Ye, Qiang Lin, Hongru Liang, Xia Luo, Baofu Ma, Ningqiu Li and Xiaozhe Fu

Int. J. Mol. Sci. 2025, 26(2), 718; https://doi.org/10.3390/ijms26020718 - 16 Jan 2025

Viewed by 794

Abstract

Glutamine metabolism is essential for infectious spleen and kidney necrosis virus (ISKNV) replication. Glutaminase 1 (GLS1), the key enzyme of the glutamine metabolism, and c-Myc positively regulate ISKNV infection, while c-Myc is closely correlated with GLS1. However, the regulatory mechanism among ISKNV, c-Myc and glutamine metabolism remains unclear. Here, we indicated that c-Myc increased glutamine uptake by increasing the GLS1, glutamate dehydrogenase (GDH) and isocitrate dehydrogenase (IDH2) expression of glutamine metabolism. ISKNV ORF102R, ORF093R and ORF118L co-located with c-Myc in CPB cells. Co-IP results showed that ISKNV ORF102R and ORF093R interacted with c-Myc, while ORF118L did not interact with c-Myc. The expression levels of c-Myc, GLS1 and IDH2 were increased in ISKNV ORF093R expression cells, and the mRNA and protein levels of GLS1 were upregulated in ISKNV 102R-expressing cells. These results indicated that ISKNV reconstructed glutamine metabolism to satisfy the energy and macromolecule requirements for virus proliferation by ORF093R and ORF102R interacting with c-Myc, which provides the foundation for innovative antiviral strategies. Full article

(This article belongs to the Special Issue Molecular Advance on Reproduction and Fertility of Aquatic Animals 2.0)

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13 pages, 3689 KiB

Open AccessArticle

Integrated Analysis of Transcriptome and Metabolome in the Brain After Cold Stress of Red Tilapia During Overwintering

by Chenxi Zhu, Haoran Yang, Wenbin Zhu, Qichen Jiang, Zaijie Dong and Lanmei Wang

Int. J. Mol. Sci. 2024, 25(24), 13372; https://doi.org/10.3390/ijms252413372 - 13 Dec 2024

Cited by 1 | Viewed by 844

Abstract

Cold stress during overwintering is considered a bottleneck problem limiting the development of the red tilapia (Oreochromis spp.) industry, and the regulation mechanism is currently not well understood. In this study, the fish (initial weight: 72.71 ± 1.32 g) were divided into the cold stress group (cold) and the control (normal) group. In the control group, the water temperature was maintained at 20 °C, which is basically consistent with the overwintering water temperature in greenhouses of local areas. In the cold group, the water temperature decreased from 20 °C to 8 °C by 2 °C per day during the experiment. At the end of the experiment, the levels of fish serum urea nitrogen, glucose, norepinephrine, alkaline phosphatase, total bilirubin, and total cholesterol in the cold group changed significantly compared with that in the control group (P < 0.05). Then transcriptome sequencing and LC–MS metabolome of brain tissue were further employed to obtain the mRNA and metabolite datasets. We found that the FoxO signaling pathway and ABC transporters played an important role by transcriptome–metabolome association analysis. In the FoxO signaling pathway, the differentially expressed genes were related to cell cycle regulation, apoptosis and immune-regulation, and oxidative stress resistance and DNA repair. In the ABC transporters pathway, the ATP-binding cassette (ABC) subfamily abca, abcb, and abcc gene expression levels, and the deoxycytidine, L-lysine, L-glutamic acid, L-threonine, ornithine, and uridine metabolite contents changed. Our results suggested that the cold stress may promote apoptosis through regulation of the FoxO signaling pathway. The ABC transporters may respond to cold stress by regulating amino acid metabolism. The results provided a comprehensive understanding of fish cold stress during overwintering, which will facilitate the breeding of new cold-resistant varieties of red tilapia in the future. Full article

(This article belongs to the Special Issue Molecular Advance on Reproduction and Fertility of Aquatic Animals 2.0)

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18 pages, 3544 KiB

Open AccessArticle

A Repetitive Acipenser gueldenstaedtii Genomic Region Aligning with the Acipenser baerii IGLV Gene Cluster Suggests a Role as a Transcription Termination Element Across Several Sturgeon Species

by Alexander V. Chouljenko, Brent A. Stanfield, Tetiana O. Melnyk, Ojasvi Dutta and Vladimir N. Chouljenko

Int. J. Mol. Sci. 2024, 25(23), 12685; https://doi.org/10.3390/ijms252312685 - 26 Nov 2024

Viewed by 712

Abstract

This study focuses on the common presence of repetitive sequences within the sturgeon genome that may contribute to enhanced immune responses against infectious diseases. A repetitive 675 bp VAC-2M sequence in Russian sturgeon DNA that aligns with the Siberian sturgeon IGLV gene cluster was identified. A specific 218 bp long portion of the sequence was found to be identical between Acipenser gueldenstaedtii, A. baerii and A. stellatus species, and NCBI blast analysis confirmed the presence of this DNA segment in the A. ruthenus genome. Multiple mutated copies of the same genomic region were detected by PCR analysis, indicating that different versions of this highly repetitive sequence exist simultaneously within the same organism. The selection toward specific genetic differences appears to be highly conserved based on the sequence variations within DNA originating from fish grown at distant geographical regions and individual caviar grains from the same fish. The corresponding A. baerii genomic region encompassing the 357 bp DNA sequence was cloned either ahead or after the human cytomegalovirus immediate early promoter (HCMV-IE) into a pBV-Luc reporter vector expressing the luciferase gene. The DNA segment significantly reduced luciferase expression in transient transfection/expression experiments. The results indicate that this genomic region functions as a transcription termination element that may affect antibody production in sturgeons. Full article

(This article belongs to the Special Issue Molecular Advance on Reproduction and Fertility of Aquatic Animals 2.0)

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12 pages, 2735 KiB

Open AccessArticle

Identification of Genes and Long Non-Coding RNAs Putatively Related to Portunus trituberculatus Sex Determination and Differentiation Using Oxford Nanopore Technology Full-Length Transcriptome Sequencing

by Shaoting Jia, Guang Li, Yuchao Huang, Yashi Hou, Baoquan Gao and Jianjian Lv

Int. J. Mol. Sci. 2024, 25(21), 11845; https://doi.org/10.3390/ijms252111845 - 4 Nov 2024

Cited by 1 | Viewed by 942

Abstract

The swimming crab (Portunus trituberculatus) is an economically important species in China, and its growth traits show obvious sexual dimorphism. Thus, it is important to study the mechanism of sex determination and differentiation in this species. Herein, we identified 2138 differentially expressed genes and 132 differentially expressed long non-coding RNAs (lncRNAs) using Oxford Nanopore Technology full-length transcriptome sequencing. We predicted 561 target genes of the differentially expressed lncRNAs according to their location and base pair complimentary principles. Furthermore, pathways related to sex determination, differentiation, and reproduction were enriched for lncRNAs according to gene ontology and Kyoto Encyclopedia of Genes and Genomes analyses. This indicated that lncRNAs might play regulatory roles in these pathways. Our results could form the basis for future studies of sex determination and differentiation in P. trituberculatus. Full article

(This article belongs to the Special Issue Molecular Advance on Reproduction and Fertility of Aquatic Animals 2.0)

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18 pages, 8094 KiB

Open AccessArticle

Unveiling the Molecular Mechanisms Regulating Muscle Elasticity in the Large Yellow Croaker: Insights from Transcriptomics and Metabolomics

by Mengyang Liu, Guangde Qiao, Yabing Wang, Shengyu Liu, Xiaoshan Wang, Yanfeng Yue and Shiming Peng

Int. J. Mol. Sci. 2024, 25(20), 10924; https://doi.org/10.3390/ijms252010924 - 11 Oct 2024

Viewed by 1180

Abstract

The large yellow croaker (Larimichthys crocea) is an important economic fish in China. However, intensive farming practices, such as high stocking densities, suboptimal water quality, and imbalanced nutrition, have led to a decline in muscle quality. Muscle elasticity is a key texture property influencing muscle quality. Herein, transcriptomic and metabolomic analyses were performed on four groups: male high muscle elasticity (MEHM), female high muscle elasticity (MEHF), male low muscle elasticity (MELM), and female low muscle elasticity (MELF), to explore the molecular regulation underlying muscle elasticity in the large yellow croaker. Transcriptomics identified 2594 differentially expressed genes (DEGs) across the four groups, while metabolomics revealed 969 differentially expressed metabolites (DEMs). Association analysis indicated that the valine, leucine, and isoleucine biosynthesis pathways were significantly enriched between the MELF and MEHF groups; 2-Oxoisovalerate and L-Valine were DEMs; and the gene encoding L-threonine ammonia-lyase was a DEG. In the MELM and MEHM groups, pathways such as arginine biosynthesis; arginine and proline metabolism; and valine, leucine, and isoleucine degradation were significantly enriched. 4-guanidinobutanoate, L-aspartate, N-acetylornithine, and L-leucine were among the DEMs, while the DEGs included glul, gls, srm, hmgcs, and aacs. These findings provide insights into the molecular mechanisms controlling muscle elasticity, representing a theoretical foundation to breed high-quality large yellow croakers. Full article

(This article belongs to the Special Issue Molecular Advance on Reproduction and Fertility of Aquatic Animals 2.0)

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20 pages, 7174 KiB

Open AccessArticle

Genome-Wide Association Studies (GWAS) and Transcriptome Analysis Reveal Male Heterogametic Sex-Determining Regions and Candidate Genes in Northern Snakeheads (Channa argus)

by Haiyang Liu, Jin Zhang, Tongxin Cui, Weiwei Xia, Qing Luo, Shuzhan Fei, Xinping Zhu, Kunci Chen, Jian Zhao and Mi Ou

Int. J. Mol. Sci. 2024, 25(20), 10889; https://doi.org/10.3390/ijms252010889 - 10 Oct 2024

Cited by 1 | Viewed by 1567

Abstract

The Northern snakehead (Channa argus) is a significant economic aquaculture species in China. Exhibiting sexual dimorphism in the growth rate between females and males, mono-sex breeding holds substantial value for aquaculture. This study employed GWAS and transcriptome analysis were applied to identify sex determination genomic regions and develop sex-specific markers. A total of 270 single-nucleotide polymorphisms (SNPs) and 31 insertion-deletions (InDels) were identified as being sexually dimorphic through GWAS and fixation index (Fst) scanning. Based on GWAS results, two sex-specific InDel markers were developed, effectively distinguishing genetic sex for XX females, XY males, and YY super-males via (polymerase chain reaction) PCR amplification. A major genomic segment of approximately 115 kb on chromosome 3 (Chr 03) was identified as the sex-determination region. A comparative transcriptome analysis of gonads for three sexes identified 158 overlapping differentially expressed genes (DEGs). Additionally, three sex-related candidate genes were identified near the sex determination region, including id2, sox11, and rnf144a. Further studies are required to elucidate the functions of these genes. Overall, two sex-specific InDel markers support a male heterogametic XX/XY sex-determination system in Northern snakeheads and three candidate genes offer new insights into sex determination and the evolution of sex chromosomes in teleost fish. Full article

(This article belongs to the Special Issue Molecular Advance on Reproduction and Fertility of Aquatic Animals 2.0)

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11 pages, 1459 KiB

Open AccessCommunication

The Chronic Toxicity of Endocrine-Disrupting Chemical to Daphnia magna: A Transcriptome and Network Analysis of TNT Exposure

by Jun Lee, Hyun Woo Kim, Dong Yeop Shin, Jun Pyo Han, Yujin Jang, Ju Yeon Park, Seok-Gyu Yun, Eun-Min Cho and Young Rok Seo

Int. J. Mol. Sci. 2024, 25(18), 9895; https://doi.org/10.3390/ijms25189895 - 13 Sep 2024

Viewed by 1609

Abstract

Endocrine-disrupting chemicals (EDCs) impair growth and development. While EDCs can occur naturally in aquatic ecosystems, they are continuously introduced through anthropogenic activities such as industrial effluents, pharmaceutical production, wastewater, and mining. To elucidate the chronic toxicological effects of endocrine-disrupting chemicals (EDCs) on aquatic organisms, we collected experimental data from a standardized chronic exposure test using Daphnia magna (D. magna), individuals of which were exposed to a potential EDC, trinitrotoluene (TNT). The chronic toxicity effects of this compound were explored through differential gene expression, gene ontology, network construction, and putative adverse outcome pathway (AOP) proposition. Our findings suggest that TNT has detrimental effects on the upstream signaling of Tcf/Lef, potentially adversely impacting oocyte maturation and early development. This study employs diverse bioinformatics approaches to elucidate the gene-level toxicological effects of chronic TNT exposure on aquatic ecosystems. The results provide valuable insights into the molecular mechanisms of the adverse impacts of TNT through network construction and putative AOP proposition. Full article

(This article belongs to the Special Issue Molecular Advance on Reproduction and Fertility of Aquatic Animals 2.0)

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20 pages, 7963 KiB

Open AccessArticle

The Response of the Gut Physiological Function and Microbiome of a Wild Freshwater Fish (Megalobrama terminalis) to Alterations in Reproductive Behavior

by Yaqiu Liu, Chunni Kou, Jiayue Chen, Yuefei Li and Jie Li

Int. J. Mol. Sci. 2024, 25(13), 7425; https://doi.org/10.3390/ijms25137425 - 6 Jul 2024

Viewed by 1484

Abstract

The fish gut microbiome is well known for its role in degrading nutrients to improve the host’s digestion and absorption efficiency. In this study, we focused on the core physiological adaptability during the various reproductive stages of the black Amur bream (Megalobrama terminalis) to explore the interaction mechanisms among the fish host gut mucosal structure, gut enzyme activity, and gut microbial metabolism in the course of the host’s reproductive cycle. Our findings showed that M. terminalis exhibited locomotion metabolic type (aids in sporting) in the reproductive stage, and a change to visceral metabolic type (aids in digestion) during non-reproductive and post-reproductive stage phases. The impact of metabolic type selection and energy demand during various reproductive stages on fish nutrition strategy and digestive function was substantial. Our resulted showed that mitochondria in intestinal epithelial cells of reproductive M. terminalis appeared autophagy phenomenon, and the digestive enzyme activities in the intestines of reproductive M. terminalis were lower than those in the non-reproductive and post-reproductive individuals. Moreover, these differences in nutrition strategy have a prominent impact on the gut microbiome of reproductive M. terminalis, compared to non-reproductive and post-reproductive samples. Our findings showed that reproductive females had lower levels of alpha diversity compared to non-reproductive and post-reproductive females. Our results also showed a greater functional variety and an increase in functional genes related to carbohydrate, lipid, amino acid, cofactors, and vitamin metabolic pathways in the NRS and PRS group. It is noteworthy that an enrichment of genes encoding putative enzymes implicated in the metabolism of taurine and hypotaurine was observed in the RS samples. Our findings illustrated that the stability and resilience of the gut bacterial community could be shaped in the wild fish host–microbiome interactions during reproductive life history. Full article

(This article belongs to the Special Issue Molecular Advance on Reproduction and Fertility of Aquatic Animals 2.0)

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11 pages, 246 KiB

Open AccessArticle

Studies on the Relationships between Growth and Gonad Development during First Sexual Maturation of Macrobrachium nipponense and Associated SNPs Screening

by Sufei Jiang, Yinxiang Xie, Zijian Gao, Yunpeng Niu, Cheng Ma, Wenyi Zhang, Yiwei Xiong, Hui Qiao and Hongtuo Fu

Int. J. Mol. Sci. 2024, 25(13), 7071; https://doi.org/10.3390/ijms25137071 - 27 Jun 2024

Cited by 1 | Viewed by 1679

Abstract

In this study, we used full-sib families to investigate the association between growth and gonad development during first sexual maturation of M. nipponense. We found that male GSI was significantly negatively correlated with growth traits (p < 0.01) and there were no significant correlations between female GSI (Gonadosomatic index) and growth traits (p > 0.05). HSI (Hepatopancreas index) in both males and females showed no significant correlations with growth traits (p > 0.05). We furthermore investigated the association between the specific allele of Mn-CTS L1 polymorphism and gonad development and growth traits. In total, 35 mutation loci were screened and 16 high-quality single-nucleotide polymorphisms (SNPs) loci were obtained after validation. Four and two SNPs proved to be strongly associated with all growth traits in female and male M. nipponense separately, among which A+118T might be a candidate SNP positively associated with large growth traits. Two and one SNPs were screened, respectively, in males and females to associate with GSI, while three SNPs were detected to associate with female HSI, among which A+1379C may be applied as a potential molecular marker for gene-assisted selection to improve both reproduction speed and growth traits in M. nipponense. Full article

(This article belongs to the Special Issue Molecular Advance on Reproduction and Fertility of Aquatic Animals 2.0)

18 pages, 17319 KiB

Open AccessArticle

Genome-Wide Identification, Expression and Response to Estrogen of Vitellogenin Gene Family in Sichuan Bream (Sinibrama taeniatus)

by Zhe Zhao, Li Peng, Qiang Zhao and Zhijian Wang

Int. J. Mol. Sci. 2024, 25(12), 6739; https://doi.org/10.3390/ijms25126739 - 19 Jun 2024

Cited by 1 | Viewed by 1356

Abstract

To enhance our understanding of teleost reproductive physiology, we identified six Sichuan bream (Sinibrama taeniatus) vitellogenin genes (vtg1-6) and characterized their sequence structures. We categorized them into type Ⅰ (vtg1,4,5 and 6), type Ⅱ (vtg2) and type Ⅲ (vtg3) based on differences in their subdomain structure. The promoter sequence of vtgs has multiple estrogen response elements, and their abundance appears to correlate with the responsiveness of vtg gene expression to estrogen. Gene expression analyses revealed that the vitellogenesis of Sichuan bream involves both heterosynthesis and autosynthesis pathways, with the dominant pathway originating from the liver. The drug treatment experiments revealed that 17β-estradiol (E₂) tightly regulated the level of vtg mRNA in the liver. Feeding fish with a diet containing 100 μg/g E₂ for three weeks significantly induced vtg gene expression and ovarian development, leading to an earlier onset of vitellogenesis. Additionally, it was observed that the initiation of vtg transcription required E₂ binding to its receptor, a process primarily mediated by estrogen receptor alpha in Sichuan bream. The findings of this study provide novel insights into the molecular information of the vitellogenin gene family in teleosts, thereby contributing to the regulation of gonadal development in farmed fish. Full article

(This article belongs to the Special Issue Molecular Advance on Reproduction and Fertility of Aquatic Animals 2.0)

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13 pages, 4297 KiB

Open AccessArticle

NPC Intracellular Cholesterol Transporter 1 Regulates Ovarian Maturation and Molting in Female Macrobrachium nipponense

by Sufei Jiang, Wenyi Zhang, Yiwei Xiong, Mengying Zhang, Huwei Yuan, Yunpeng Niu, Hui Qiao and Hongtuo Fu

Int. J. Mol. Sci. 2024, 25(11), 6049; https://doi.org/10.3390/ijms25116049 - 31 May 2024

Cited by 3 | Viewed by 1113

Abstract

NPC intracellular cholesterol transporter 1 (NPC1) plays an important role in sterol metabolism and transport processes and has been studied in many vertebrates and some insects, but rarely in crustaceans. In this study, we characterized NPC1 from Macrobrachium nipponense (Mn-NPC1) and evaluated its functions. Its total cDNA length was 4283 bp, encoding for 1344 amino acids. It contained three conserved domains typical of the NPC family (NPC1_N, SSD, and PTC). In contrast to its role in insects, Mn-NPC1 was mainly expressed in the adult female hepatopancreas, with moderate expression in the ovary and heart. No expression was found in the embryo (stages CS–ZS) and only weak expression in the larval stages from hatching to the post-larval stage (L1–PL15). Mn-NPC1 expression was positively correlated with ovarian maturation. In situ hybridization showed that it was mainly located in the cytoplasmic membrane and nucleus of oocytes. A 25-day RNA interference experiment was employed to illustrate the Mn-NPC1 function in ovary maturation. Experimental knockdown of Mn-NPC1 using dsRNA resulted in a marked reduction in the gonadosomatic index and ecdysone content of M. nipponense females. The experimental group showed a significant delay in ovarian maturation and a reduction in the frequency of molting. These results expand our understanding of NPC1 in crustaceans and of the regulatory mechanism of ovarian maturation in M. nipponense. Full article

(This article belongs to the Special Issue Molecular Advance on Reproduction and Fertility of Aquatic Animals 2.0)

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16 pages, 5093 KiB

Open AccessArticle

A Testis-Specific DMRT1 (Double Sex and Mab-3-Related Transcription Factor 1) Plays a Role in Spermatogenesis and Gonadal Development in the Hermaphrodite Boring Giant Clam Tridacna crocea

by Zohaib Noor, Zhen Zhao, Shuming Guo, Zonglu Wei, Borui Cai, Yanping Qin, Haitao Ma, Ziniu Yu, Jun Li and Yuehuan Zhang

Int. J. Mol. Sci. 2024, 25(11), 5574; https://doi.org/10.3390/ijms25115574 - 21 May 2024

Viewed by 2933

Abstract

The testis-specific double sex and mab-3-related transcription factor 1 (DMRT1) has long been recognized as a crucial player in sex determination across vertebrates, and its essential role in gonadal development and the regulation of spermatogenesis is well established. Here, we report the cloning of the key spermatogenesis-related DMRT1 cDNA, named Tc-DMRT1, from the gonads of Tridacna crocea (T. crocea), with a molecular weight of 41.93 kDa and an isoelectric point of 7.83 (pI). Our hypothesis is that DMRT1 machinery governs spermatogenesis and regulates gonadogenesis. RNAi-mediated Tc-DMRT1 knockdown revealed its critical role in hindering spermatogenesis and reducing expression levels in boring giant clams. A histological analysis showed structural changes, with normal sperm cell counts in the control group (ds-EGFP) but significantly lower concentrations of sperm cells in the experimental group (ds-DMRT1). DMRT1 transcripts during embryogenesis exhibited a significantly high expression pattern (p < 0.05) during the early zygote stage, and whole-embryo in-situ hybridization confirmed its expression pattern throughout embryogenesis. A qRT-PCR analysis of various reproductive stages revealed an abundant expression of Tc-DMRT1 in the gonads during the male reproductive stage. In-situ hybridization showed tissue-specific expression of DMRT1, with a positive signal detected in male-stage gonadal tissues comprising sperm cells, while no signal was detected in other stages. Our study findings provide an initial understanding of the DMRT1 molecular machinery controlling spermatogenesis and its specificity in male-stage gonads of the key bivalve species, Tridacna crocea, and suggest that DMRT1 predominantly functions as a key regulator of spermatogenesis in giant clams. Full article

(This article belongs to the Special Issue Molecular Advance on Reproduction and Fertility of Aquatic Animals 2.0)

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