DMRT1 Research Articles

The effects of 11-ketotestosterone implants on transcript levels of gonadotropin receptors, and foxl2 and dmrt1 genes in the Previtellogenic ovary of cultured beluga (Huso huso).

The in vivo effect of 11-ketotestosterone (11KT) on transcript levels of the gonadotropin receptors (fshr and lhr) and sex differentiation-related genes (dmrt1 and foxl2) was examined in the ovaries of immature female beluga. For this purpose, six fish were treated with implants containing 2.5 mg 11KT and a placebo group of six females of the same age and gametogenic stage were given a blank implant. The implants were intraperitoneally inserted into 4-year-old females at the previtellogenic stage (mean body weight 5580 ± 165 g) and maintained under culture conditions for 8 weeks. Ovary samples for gene expression analysis of lhr, fshr, dmrt1 and foxl2 were collected by biopsy at 3 and 8 weeks post implantation. Diameters of oocytes increased in response to 11KT treatment, both at 3 and at 8 weeks post implantation, but no obvious changes were evident in cytology. Three weeks of 11KT treatment did not affect target gene expression, but a tendency for a time-dependent decrease of lhr and dmrt1 mRNA levels was observed in both treatment and placebo groups. By 8 weeks of treatment, however, 11KT implants provoked the upregulation of fshr and foxl2 transcript levels. Furthermore, lhr and dmrt1 transcript abundances recovered by 8 weeks of exposure in both blank- and 11KT-implanted beluga. These results suggest that 11KT, either directly or indirectly, may affect gametogenesis and regulate some key components of the reproductive axis in female beluga.

Effects of the DMRT1 genotype on the body weight and gut microbiota in the broiler chicken

Intestinal microbiota is a critical determinant of growth and risk of metabolic diseases. Our previous studies showed that the locus rs16775833 within the DMRT1 gene is significantly associated with variation in the population structure of the gut microbiota, which is involved in determining the BW of the chicken. To assess the accuracy of correlation of rs16775833 located in the DMRT1 gene on microbial population and BW in birds, 2 genotypes GG and TT in the rs16775833 were identified in Chinese Yellow broiler breeders. We found that BW in the TT genotype group was significantly higher than in the GG genotype group at 7 and 13 wk of age in 777 female chickens. A full-length 16S rRNA sequencing approach was used to further evaluate the fecal bacterial composition of female broilers in 11 TT genotype chickens with high weight (HW-TT) and 11 GG genotype chickens with low weight (LW-GG) at 91 D of age. Partial least squares discriminant analysis revealed that the microbiota of the HW-TT and LW-GG females were clearly separated into 2 clusters. Furthermore, we identified 13 significantly different (P < 0.05) microbes at the genus level and 17 significantly different (P < 0.05) species between the HW-TT and LW-GG groups. Our data show that rs16775833 can modulate the microbial community structure and is associated with the BW of birds. To our knowledge, this is the first time that DMRT1 has been identified as a specific host factor, which is not only involved in sex determination but also has an effect on microbial function that might regulate animal growth.

Starvation causes female-to-male sex reversal through lipid metabolism in the teleost fish, medaka (Olyzias latipes).

ABSTRACTThe teleost fish, medaka (Oryzias latipes), employs the XX/XY genetic sex determination system. We show here that the phenotypic sex of medaka is affected by changes in lipid metabolism. Medaka larvae subjected to 5 days of starvation underwent female-to-male sex reversal. Metabolomic and RT-qPCR analyses indicated that pantothenate metabolism was suppressed by starvation. Consistently, inhibiting the pantothenate metabolic pathway caused sex reversal. The final metabolite in this pathway is coenzyme A, an essential factor for lipogenesis. Inhibiting fatty acid synthesis, the first step of lipogenesis, also caused sex reversal. The expression of dmrt1, a critical gene for male development, was suppressed by starvation, and a dmrt1 (Δ13) mutant did not show sex reversal under starvation. Collectively, these results indicate that fatty acid synthesis is involved in female-to-male sex reversal through ectopic expression of male gene dmrt1 under starvation.

Случай интерстициальной делеции короткого плеча хромосомы 9, ассоциированной с нарушением формирования пола

Делеции короткого плеча хромосомы 9 представляют собой клинически и генетически гетерогенную группу. Большинство описанных случаев представляют собой терминальные делеции или несбалансированные транслокации с различными с точками разрыва на коротком плече хромосомы 9. Интерстициальные делеции короткого плеча хромосомы 9 - крайне редко встречающаяся хромосомная патология. Мы сообщаем о пациенте с задержкой психомоторного развития, гипоплазией мозжечка и гипоспадией у которого при проведении хромосомного микроматричного анализа диагностирована интерстициальная делеция 9p24.3-p23, затрагивающая ген DMRT1. Deletions of the short arm of chromosome 9 are a clinically and genetically heterogeneous group. Most of the cases described are terminal deletions or unbalanced translocations with different break points on the short arm of chromosome 9. Interstitial deletions of the short arm of chromosome 9 are an extremely rare chromosome pathology. We report a patient with developmental and psychomotor delay, cerebellar hypoplasia and hypospadias, who was diagnosed with interstitial 9p24.3-p23 deletion affecting the DMRT1 gene during chromosome microarray analysis (СMA).

Dmrt genes participate in the development of Cajal-Retzius cells derived from the cortical hem in the telencephalon.

During development, Cajal-Retzius (CR) cells are the first generated and essential pioneering neurons that control neuronal migration and arealization in the mammalian cortex. CR cells are derived from specific regions within the telencephalon, that is, the pallial septum in the rostromedial cortex, the pallial-subpallial boundary, and the cortical hem (CH) in the caudomedial cortex. However, the molecular mechanism underlying the generation of CR cell subtypes in distinct regions of origin is poorly understood. We found that double-sex and mab-3 related transcription factor (Dmrt) genes, that is, Dmrta1 and Dmrt3, were expressed in the progenitor domains that produce CR cells. The number of CH-derived CR cells was severely decreased in Dmrt3 mutants, especially in Dmrta1 and Dmrt3 double mutants. The reduced production of the CR cells was consistent with the developmental impairment of the CH structures in the medial telencephalon from which the CR cells are produced. Dmrta1 and Dmrt3 cooperatively regulate patterning of the CH structure and production of the CR cells from the CH during cortical development.

Molecular characterization, computational analysis and expression profiling of Dmrt1 gene in Indian major carp, Labeo rohita (Hamilton 1822)

Sexual dimorphism of fish morphology, physiology and behavior is diverse and complex in nature. Doublesex and mab-3 related transcription factor (Dmrt) is a large protein family whose function is sexual development and differentiation in vertebrates. Here, we report a full-length cDNA sequence of Labeo rohita (rohu) Dmrt1 of 907 bp length having 798 bp of open reading frame encoding 265 amino acids. The molecular weight of rohu DMRT1 protein was found to be 28.74 KDa and isoelectric point was 7.53. DMRT1 protein contains 23 positively and 24 negatively charged amino acids with a GRAVY score of -0.618. A characteristic DM domain was found in DMRT1 protein, which is a novel DNA-binding domain. Phylogenetic analysis showed maximum similarity with Cyprinus carpio when compared with DMRT1 of other vertebrates. Molecular docking study identified active sites to be targeted for drug designing. Rohu DMRT1 was observed to interact with other proteins such as FOXL2, CYP19a1a, AMH and SOX9a. Differential expression study revealed higher expression in testis tissue implying its role in male sex differentiation and testicular development. The information generated in the present work could facilitate further research to resolve the issues related to gonadal maturation and reproduction of commercially important aquaculture species.

Timing of gonadal development and dimorphic expression of sex-related genes in gonads during early sex differentiation in the Yellow River carp

The common carp, a gonochoristic teleost fish with an XX/XY sex-determining system, provides an excellent model for studying cyprinidae gonadal sex differentiation. The present study aims at investigating the detailed process of early gonadal sex differentiation of common carp by combining the expression patterns of sex related genes (cyp19a1a, foxl2, amh, dmrt1, sox9b, sycp3, dmc1) with gonadal histological changes using mono-sex female and male Yellow River carp. H&E staining, immunohistochemistry and immunofluorescence of gonads indicated that the ovaries and testis of common carp were both directly differentiated from undifferentiated gonads. Oogonia and spermatogonia were first observed at 40 dph (days post hatching) and 70 dph in female and male gonads, respectively. Female related genes cyp19a1a and foxl2 and male related genes amh, dmrt1 and sox9b increased significantly in 30–40 dph female, and male gonads, respectively. These suggest that the critical timing of sex determination of common carp takes place between 30 and 40 dph or possibly earlier. Meiotic marker gene dmc1 and sycp3 increased expression significantly in 55 dph female gonads while sycp3 increased expression significantly in 100 dph male gonads. Oocytes in meiosis and spermatocytes were first observed in 55 dph female and 100 dph male gonads, respectively. These indicate that the initiation time of gonadal sex differentiation is 50–55 dph in female common carp and 90–100 dph in male common carp. To our knowledge, this is the first study investigating the detailed process of early gonadal sex differentiation of common carp by combining the expression patterns of sex related genes with gonadal histological changes. We have identified the critical window of sex determination and the initiation times of oogenesis and spermatogenesis in the Yellow River carp. These results provide researchers with valuable information to possibly reveal the mechanisms of sex determination and differentiation of common carp.

Phylogeography, more than elevation, accounts for sex chromosome differentiation in Swiss populations of the common frog (Rana temporaria).

Sex chromosomes in vertebrates range from highly heteromorphic (as in most birds and mammals) to strictly homomorphic (as in many fishes, amphibians, and nonavian reptiles). Reasons for these contrasted evolutionary trajectories remain unclear, but species such as common frogs with polymorphism in the extent of sex chromosome differentiation may potentially deliver important clues. By investigating 92 common frog populations from a wide range of elevations throughout Switzerland, we show that sex chromosome differentiation strongly correlates with alleles at the candidate sex-determining gene Dmrt1. Y-specific Dmrt1 haplotypes cluster into two main haplogroups, YA and YB , with a phylogeographic signal that parallels mtDNA haplotypes: YA populations, with mostly well-differentiated sex chromosomes, occur primarily south of the main alpine ridge that bisects Switzerland, whereas YB populations, with mostly undifferentiated (proto-)sex chromosomes, occur north of this ridge. Elevation has only a marginal effect, opposing previous suggestions of a major role for climate on sex chromosome differentiation. The Y-haplotype effect might result from differences in the penetrance of alleles at the sex-determining locus (such that sex reversal and ensuing X-Y recombination are more frequent in YB populations), and/or fixation of an inversion on YA (as supported by the empirical observation that YA haplotypes might not recombine in XYA females).

Parallel Evolution of Two dmrt1-Derived Genes, dmy and dm-W, for Vertebrate Sex Determination.

SummaryAnimal sex-determining genes, which bifurcate for female and male development, are diversified even among closely related species. Most of these genes emerged independently from various sex-related genes during species diversity as neofunctionalization-type genes. However, the common mechanisms of this divergent evolution remain poorly understood. Here, we compared the molecular evolution of two sex-determining genes, the medaka dmy and the clawed frog dm-W, which independently evolved from the duplication of the transcription factor-encoding masculinization gene dmrt1. Interestingly, we detected parallel amino acid substitutions, from serine (S) to threonine (T), on the DNA-binding domains of both ancestral DMY and DM-W, resulting from positive selection. Two types of DNA-protein binding experiments and a luciferase reporter assay demonstrated that these S-T substitutions could strengthen the DNA-binding abilities and enhance the transcriptional regulation function. These findings suggest that the parallel S-T substitutions may have contributed to the establishment of dmy and dm-W as sex-determining genes.

2,4-Dichlorophenol induces feminization of zebrafish (Danio rerio) via DNA methylation

2,4-Dichlorophenol (2,4-DCP) is a ubiquitous contaminant of aquatic environments with an estrogenic effect on fish. However, the molecular mechanism underlying this effect remains elusive. To this end, the present study aimed to explore the effect of 2,4-DCP on sex differentiation and its relevant mechanism in zebrafish (Danio rerio). The results showed that a female-biased sex ratio was induced after exposing larval zebrafish to 2,4-DCP (0–160 μg/L) from 20 to 50 days post fertilization (dpf). The feminization of zebrafish was accompanied by decreased expression of male-related genes (sox9a, amh and dmrt1) under 2,4-DCP from 20 to 50 dpf. However, the expression of female-related genes (cyp19a1a, foxl2 and esr1) was also suppressed. Nevertheless, it is noteworthy that the methylation level of sox9a promoter was significantly increased, which may result in the significantly decreased expression of sox9a and ultimately the feminization effect of 2,4-DCP on zebrafish. In addition, 5-aza-2′-deoxycytidine (5-AZA), a methyltransferase inhibitor, significantly reduced the methylation level, increased the expression of sox9a, and partly impaired the feminization effect caused by 2,4-DCP, which further confirmed the importance of DNA methylation of sox9a in 2,4-DCP-induced feminization. These findings provide novel insights into the epigenetic mechanisms of DCP-induced estrogenic effect in fish.

Evolution of the Proto Sex-Chromosome in Solea senegalensis.

Solea senegalensis is a flatfish belonging to the Soleidae family within the Pleuronectiformes order. It has a karyotype of 2n = 42 (FN = 60; 6M + 4 SM + 8 St + 24 T) and a XX/XY system. The first pair of metacentric chromosomes has been proposed as a proto sex-chromosome originated by a Robertsonian fusion between acrocentric chromosomes. In order to elucidate a possible evolutionary origin of this chromosome 1, studies of genomic synteny were carried out with eight fish species. A total of 88 genes annotated within of 14 BACs located in the chromosome 1 of S. senegalensis were used to elaborate syntenic maps. Six BACs (BAC5K5, BAC52C17, BAC53B20, BAC84K7, BAC56H24, and BAC48P7) were distributed in, at least, 5 chromosomes in the species studied, and a group of four genes from BAC53B20 (grsf1, rufy3, slc4a4 and npffr2) and genes from BAC48K7 (dmrt2, dmrt3, dmrt1, c9orf117, kank1 and fbp1) formed a conserved cluster in all species. The analysis of repetitive sequences showed that the number of retroelements and simple repeat per BAC showed its highest value in the subcentromeric region where 53B20, 16E16 and 48K7 BACs were localized. This region contains all the dmrt genes, which are associated with sex determination in some species. In addition, the presence of a satellite “chromosome Y” (motif length: 860 bp) was detected in this region. These findings allowed to trace an evolutionary trend for the large metacentric chromosome of S. senegalensis, throughout different rearrangements, which could be at an initial phase of differentiation as sex chromosome.

FOXL2 antagonises the male developmental pathway in embryonic chicken gonads.

FOXL2 is a conserved transcription factor with a central role in ovarian development and function. Studies in humans and mice indicate that the main role of FOXL2 is in the postnatal ovary, namely folliculogenesis. To shed light on the function and evolution of FOXL2 in the female gonad, we examined its role in embryonic avian gonads, using in ovo over-expression and knockdown. FOXL2 mRNA and protein are expressed female-specifically in the embryonic chicken gonad, just prior to the onset of sexual differentiation. FOXL2 is expressed in the medullary cord cells, in the same cell type as aromatase (CYP19A1). In addition, later in development, expression also becomes localised in a subset of cortical cells, distinct from those expressing estrogen receptor alpha. Mis-expression of FOXL2 in the male chicken embryonic gonad suppresses the testis developmental pathway, abolishing local expression of the male pathway genes, SOX9, DMRT1 and AMH, and repressing Sertoli cell development. Conversely, knockdown of FOXL2 expression allows ectopic activation of SOX9 in female gonads. However, mis-expression of FOXL2 alone was insufficient to activate aromatase expression in male gonads, while FOXL2 knockdown did not affect aromatase expression in females. These results indicate that FOXL2 plays an important role in embryonic differentiation of the avian ovary via antagonism of SOX9, but may be dispensable for aromatase activation at embryonic stages. The data suggest that FOXL2 has different roles in different species, more central for embryonic ovarian differentiation in egg-laying vertebrates.

Independent evolution for sex determination and differentiation in the DMRT family in animals.

ABSTRACTSome DMRT family genes including arthropod dsx, nematode mab-3, and vertebrate dmrt1 are involved in sex determination and/or differentiation in bilaterian animals. Although there have been some reports about evolutionary analyses of the family by using its phylogenetic trees, it is still undecided as to whether these three sex determination-related genes share orthologous relationships or not. To clarify this question, we analyzed evolutional relationships among the family members in various bilaterians by using not only phylogenetic tree analysis, but also synteny analysis. We found that only four genes, dmrt2a/2b, dmrt3, dmrt4/5 and dmrt93B were commonly present in invertebrate bilateria. The syntenies of dmrt2a/2b-dmrt3 and dmrt4/5-dmrt93B are conserved before and after two rounds of whole genome duplication in the ancestral vertebrate. Importantly, this indicates that dmrt1 must have appeared in the common vertebrate ancestor. In addition, dmrt1, dsx, or mab-3 formed each different cluster at a distance in our phylogenetic tree. From these findings, we concluded that the three sex determination-related genes, dmrt1, dsx, and mab-3 have no orthologous relationships, and suggested independent evolution for sex determination and differentiation in the DMRT gene family. Our results may supply clues about why sex-determining systems have diverged during animal evolution.

Dmrt factors determine the positional information of cerebral cortical progenitors via differential suppression of homeobox genes.

The spatiotemporal identity of neural progenitors and the regional control of neurogenesis are essential for the development of cerebral cortical architecture. Here, we report that mammalian DM domain factors (Dmrt) determine the identity of cerebral cortical progenitors. Among the Dmrt family genes expressed in the developing dorsal telencephalon, Dmrt3 and Dmrta2 show a medialhigh/laterallow expression gradient. Their simultaneous loss confers a ventral identity to dorsal progenitors, resulting in the ectopic expression of Gsx2 and massive production of GABAergic olfactory bulb interneurons in the dorsal telencephalon. Furthermore, double-mutant progenitors in the medial region exhibit upregulated Pax6 and more lateral characteristics. These ventral and lateral shifts in progenitor identity depend on Dmrt gene dosage. We also found that Dmrt factors bind to Gsx2 and Pax6 enhancers to suppress their expression. Our findings thus reveal that the graded expression of Dmrt factors provide positional information for progenitors by differentially repressing downstream genes in the developing cerebral cortex.

Potential involvement of a DMRT family member ( Mr‐Dsx ) in the regulation of sexual differentiation and moulting in the giant river prawn Macrobrachium rosenbergii

Macrobrachium rosenbergii is an important aquaculture species known as the king of freshwater prawn. Macrobrachium rosenbergii displays remarkable sexual dimorphism, yet our knowledge of the molecular mechanism underpinning sex determination and differentiation of M. rosenbergii is still fragmentary. In this study, through in silico prediction and experimental characterization, we identified a new Dmrt gene (Mr-Dsx) from M. rosenbergii. The cDNA of Mr-Dsx included a 115 bp 5′ untranslated region (UTR), a 2,568 bp open reading frame (ORF) and a 259 bp 3′ UTR. Mr-Dsx was expressed in a wide range of tissues, and showed dynamic expression changes at different moulting stages in the testis, androgenic gland (AG) and eyestalk. In response to unilateral eyestalk ablation (UEA), Mr-Dsx was significantly induced in the testis and AG. Gene knockdown of Mr-Dsx resulted in pronounced suppression of the insulin-like androgenic hormone (IAG) gene. The results suggest that Mr-Dsx is likely to be negatively regulated by the eyestalk neurohormones, and Mr-Dsx may participate in the transcriptional activation of IAG. The study characterized the potential roles of a newly identified Dmrt homologue from M. rosenbergii, and would contribute to a deeper understanding of the molecular regulatory network underlying sex differentiation and moulting of prawns.

Phylogenetic analysis and embryonic expression of panarthropod Dmrt genes

BackgroundOne set of the developmentally important Doublesex and Male-abnormal-3 Related Transcription factors (Dmrt) is subject of intense research, because of their role in sex-determination and sexual differentiation. This likely non-monophyletic group of Dmrt genes is represented by the Drosophila melanogaster gene Doublesex (Dsx), the Caenorhabditis elegans Male-abnormal-3 (Mab-3) gene, and vertebrate Dmrt1 genes. However, other members of the Dmrt family are much less well studied, and in arthropods, including the model organism Drosophila melanogaster, data on these genes are virtually absent with respect to their embryonic expression and function.ResultsHere we investigate the complete set of Dmrt genes in members of all main groups of Arthropoda and a member of Onychophora, extending our data to Panarthropoda as a whole. We confirm the presence of at least four families of Dmrt genes (including Dsx-like genes) in Panarthropoda and study their expression profiles during embryogenesis. Our work shows that the expression patterns of Dmrt11E, Dmrt93B, and Dmrt99B orthologs are highly conserved among panarthropods. Embryonic expression of Dsx-like genes, however, is more derived, likely as a result of neo-functionalization after duplication.ConclusionsOur data suggest deep homology of most of the panarthropod Dmrt genes with respect to their function that likely dates back to their last common ancestor. The function of Dsx and Dsx-like genes which are critical for sexual differentiation in animals, however, appears to be much less conserved.

Expression analysis and characterization of dmrt2 in Chinese tongue sole (Cynoglossus semilaevis)

Dmrt2 is a member of the dmrt gene family with a conserved zinc finger-like DNA-binding motif (DM domain). In the present study, CS-dmrt2 was cloned from the gonads of Chinese tongue sole (Cynoglossus semilaevis). The full-length cDNA of CS-dmrt2 is 2834 bp in length, with a 251 bp 5′-untranslated region (UTR), a 1086 bp 3′-UTR and a 1503 bp open reading frame (ORF) that encodes a 501-amino-acid peptide. qPCR revealed that CS-dmrt2 was mainly expressed in C. semilaevis testes. In situ hybridization (ISH) showed CS-dmrt2 expression throughout early gonadal development (36 days after hatching (dah) and 86 dah), but the expression was higher in male gonads than in female gonads. CS-dmrt2 mRNA was highly expressed in male germ cells. Comparison of methylation levels between females and males demonstrated hypo-methylated levels of the CS-dmrt2 promoter in the male gonads, which is consistent with the high mRNA expression. These results suggest that the CS-dmrt2 gene may play a functional role in gonadal differentiation/development and germ cell maturation in the testis.

Screening for Dmrt genes from embryo to mature Macrobrachium rosenbergii prawns

The doublesex and mab-3 related transcription factor (Dmrt) gene family is known to be related to the sexual regulators doublesex of arthropods and mab-3 of annelids and to hold highly conserved functions in sexual determination and differentiation across phyla. Here, we report a study of the Dmrt gene family in the freshwater prawn Macrobrachium rosenbergii, a crustacean whose sexual differentiation has been widely researched. A wide transcriptomic screen, from the embryo to the adult M. rosenbergii, identified five novel Dmrt genes (MroDmrts) and confirmed two known MroDmrts. The seven MroDmrts encode proteins of 275–855 amino acids; each protein contained at least one conserved DNA-binding DM domain, which is typical of Dmrt proteins, and five proteins contained 1–4 transactivation domains (TADs). Importantly, in the embryonic, larval and post-larval stages, MroDmrt genes exhibited time-dependent expression patterns rather than sex-specific expression. In-silico screening of the expression of the MroDmrt genes in adult males revealed the enrichment of MroiDmrt1b and MroiDmrt1c in the androgenic gland (AG) as compared to the eyestalks. In vivo silencing of the androgenic gland insulin-like (IAG) encoding gene significantly decreased the expression of the above two Dmrt genes, while not affecting the expression of control genes, thereby suggesting the possible role of these two genes in the IAG-switch and in sex-differentiation processes.

Discovery of sex-related genes from embryonic development stage based on transcriptome analysis in Eriocheir sinensis

Investigation of sex determination system in Eriocheir sinensis is important because of sex-dimorphism in its growth traits. However, little information about the sex-related genes in embryonic development stages were exposed. To obtain more information of sex determination in Chinese mitten crab, we performed the transcriptome analysis in embryonic development stage (fertilized egg stage, cleavage stage, blastula stage, gastrula stage and heartbeat stage) of Chinese mitten crab using nextgeneration sequencing technology. Thirty-one of 32,088 annotated unigenes were identified as sex-related genes including 16 genes involved in primary sex determination in model organisms and 8 genes of SOX family and 7 genes of DMRT gene family. Heatmap based on the RPKM value in five embryonic development stages indicated that these genes were clustered into two branches. Analysis of the differentially expressed 12 genes, including 3 genes of SOX family, 3 genes of DMRT gene family and 6 genes involved in primary sex determination in model organisms, showed significantly difference between the first three periods (fertilized egg stage–cleavage stage–blastula stage) and the last two periods (gastrula stage–heartbeat stage) and all 12 genes were up-regulated after blastula stage. In conclusion, we inferred that sex determination might be initiated after blastula stage in E. sinensis. Transcriptome analysis from embryonic development stage could provide a background information for further investigation in sex determination of Eriocheir sinensis.

Genomic organization and sexually dimorphic expression of the Dmrt1 gene in largemouth bass (Micropterus salmoides).

Doublesex and Mab-3 related transcription factor (Dmrt) genes play important roles in the process of sex determination and differentiation. In this study, a Dmrt1 gene open reading frame sequence was obtained from the gonadal transcriptome data of largemouth bass (Micropterus salmoides), and identified by cloning and sequencing. The ORF of Dmrt1 is 900 bp long, encodes 298 amino acids, and contains the DM region which is characteristic of Dmrt1. Full gDNA sequence of Dmrt1 was composed of five exons and four introns. RT-PCR and Q-PCR analysis of Dmrt1 were conducted in eight tissues and three developmental stages of mature male and female individuals. In situ hybridization was used to locate the expression of Dmrt1 in the testis and ovary of largemouth bass. The results showed that Dmrt1 was highly expressed in the testis of mature fish, but only weakly expressed in other tissues such as heart, liver, and brain, and exhibited gender dimorphism in the gonads of male and female fish at different stages. Furthermore, the expression level in female fish was very low and decreased gradually with ovary maturation. In situ hybridization indicated positive signals were found in early oocytes, but not in mature oocytes, while strong positive signals were found in all types of mature testis cells. The study showed that the sequence and structure of Dmrt1 were highly conserved and exhibited significant gender dimorphism in largemouth bass, as in other fish species. It is suggested that Dmrt1 plays an important role in sex determination and differentiation in largemouth bass.