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1.
Gene ; 741: 144541, 2020 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-32165303

RESUMO

Melanocortin-4 receptor (MC4R) is a G protein-coupled receptor with multiple functions in mammals. However, the functions of MC4R in fish have not been investigated extensively. The purpose of this study was to determine potential regulation of reproduction by the MC4R. We cloned the black rockfish MC4R and analyzed its tissue distribution and function. The results showed that black rockfish mc4r cDNA consisted of 981 nucleotides encoding a protein of 326 amino acids. The quantitative PCR data showed that mc4r mRNA was primarily expressed in the brain, gonad, stomach and intestine. In the brain, mc4r was found to be primarily located in the hypothalamus. Both α-MSH and ß-MSH increased gnih expression and decreased sgnrh and cgnrh expression (P < 0.05). α-MSH and ß-MSH had opposite effects on kisspeptin expression. In contrast, α-MSH and ß-MSH increased the expression of cyp11, cyp19, 3ß-hsd and star. In summary, our study shows that MC4R in black rockfish might regulate reproductive function and that the effects of α-MSH and ß-MSH might differ.


Assuntos
Peixes/genética , Perciformes/genética , Receptor Tipo 4 de Melanocortina/genética , Reprodução/genética , Sequência de Aminoácidos/genética , Animais , Clonagem Molecular , Peixes/crescimento & desenvolvimento , Regulação da Expressão Gênica no Desenvolvimento/genética , Gônadas/crescimento & desenvolvimento , Hipotálamo/crescimento & desenvolvimento , Perciformes/crescimento & desenvolvimento , Filogenia , RNA Mensageiro/genética , alfa-MSH/genética , beta-MSH/genética
2.
Artigo em Inglês | MEDLINE | ID: mdl-32017989

RESUMO

Steroid hormones have been proven as a key drive of sex change in sequentially hermaphroditic organisms. However, the upstream mechanism of sex steroid hormones regulation that affect sex change remain unknown. The main glucocorticoid in teleost fish is cortisol, which both regulates steroidogenesis and has antistress action. Thus, cortisol might be one of the prime factors in sex change. In this study, the glucocorticoid-induced leucine zipper (GILZ) gene, was proven to have a dramatic effect in orange-spotted groupers (Epinephelus coioides) during sex change at the early stage of gonadal transition. The specific action of the GILZ protein is at the pouch-shaped proliferative spermatogonia instead of the degenerative oocyte at the onset of sex change. Immunohistochemical (IHC) evidence revealed that GILZ performs intensively at undifferentiated spermatogonia in the early testis stage. These results imply that cortisol provokes a rise of GILZ through regulation caused by steroid hormones leading to sex change.


Assuntos
Bass/metabolismo , Proteínas de Peixes/metabolismo , Zíper de Leucina/fisiologia , Fatores de Transcrição/metabolismo , Sequência de Aminoácidos , Animais , Bass/genética , Bass/crescimento & desenvolvimento , Feminino , Proteínas de Peixes/biossíntese , Proteínas de Peixes/genética , Gônadas/crescimento & desenvolvimento , Gônadas/metabolismo , Organismos Hermafroditas , Masculino , Filogenia , Homologia de Sequência de Aminoácidos , Diferenciação Sexual/fisiologia , Fatores de Transcrição/biossíntese , Fatores de Transcrição/genética
3.
Aquat Toxicol ; 221: 105441, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32045789

RESUMO

Previous studies have shown that BDE-47, one of the most abundant polybrominated diphenyl ethers (PBDEs) congeners, has a weak estrogenic activity, but it has remained unclear whether BDE-47 disrupts gonadal development and causes male-to-female sex reversal in lower vertebrates, with limited and controversial data. The present study aimed to determine the effects of BDE-47 on gonadal development in Xenopus laevis, a model amphibian species for studying adverse effects of estrogenic chemicals on reproductive development. X. laevis at stage 45/46 were exposed to BDE-47 (0.5, 5, 50 nM) in semi-static system, with 1 nM 17ß-estradiol (E2) as the positive control. When reaching stage 53, tadpoles were examined for gonadal morphology, histology and sex-dimorphic gene expression. The phenotypic sex (gonadal morphology and histology) of each BDE-47-treated tadpole matched its genetic sex, showing no sex-reversal, whereas one half of genetic males treated with E2 displayed ovarian-like features. However, some genetic males (26%) in the 50 nM BDE-47 treatment group were found to contain more germ cells clumping together in the medulla, along with an increasing tendency of the gonad length/kidney length ratio in males, resembling feminizing outcomes of E2. These observations seem to suggest that BDE-47 exerted weak feminizing effects. However, BDE-47 induced increases in expression of both female-biased genes and male-biased genes in two sexes, which disagrees with feminizing outcomes, suggesting complicated effects of BDE-47 on gonadal development. Taken together, all results demonstrate that nanomolar BDE-47 disrupted gonadal development and exerted weak feminizing effects, but not resulted in male-to-female sex reversal in X. laevis.


Assuntos
Gônadas/efeitos dos fármacos , Éteres Difenil Halogenados/toxicidade , Diferenciação Sexual/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Animais , Estradiol/toxicidade , Feminino , Gônadas/crescimento & desenvolvimento , Humanos , Larva/efeitos dos fármacos , Masculino , Reprodução/efeitos dos fármacos , Diferenciação Sexual/genética , Xenopus laevis
4.
Environ Pollut ; 260: 113980, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-31991354

RESUMO

Developmental exposures to estrogenic chemicals possibly cause structural and functional abnormalities of reproductive organs in vertebrates. Bisphenol AF (BPAF), a bisphenol A (BPA) analogue, has been shown to have higher estrogenic activity than BPA, but little is known about the effects of BPAF on gonadal development, particularly gonadal differentiation. We aimed to determine whether low concentrations of BPAF could disrupt gonadal differentiation and subsequent development using Xenopus laevis, a model species for studying feminizing effects of estrogenic chemicals. X. laevis tadpoles were exposed to BPAF (1, 10, 100 nM) or 17ß-estradiol (E2, positive control) from stages 45/46 to 53 and 66 in a semi-static exposure system, with a prolonged treatment with the highest concentration to the eighth week post-metamorphosis (WPM8). Gonadal morphology and histology as well as sexually dimorphic gene expression were examined to evaluate the effects of BPAF. All concentrations of BPAF caused changes in testicular morphology at different developmental stages compared with controls. Specifically, at stage 53, BPAF like E2 resulted in decreases in both the size and the number of gonadal metameres (gonomeres) in testes, looking like ovaries. Some of BPAF-treated testes remained segmented and even became discontinuous and fragmented at subsequent stages. Histological abnormalities were also observed in BPAF-treated testes, such as ovarian cavity at stages 53 and 66 and poorly developed seminiferous tubules on WPM8. At the molecular level, BPAF inhibited expression of male highly expressed genes in testes at stage 53. Correspondingly, BPAF, like E2, inhibited cell proliferation in testes at stage 50. All results show that low concentrations of BPAF inhibited testicular differentiation and subsequent development in X. laevis, along with feminizing effects to some degree. Our finding implies a risk of BPAF to the male reproductive system of vertebrates including humans.


Assuntos
Compostos Benzidrílicos/toxicidade , Poluentes Ambientais/toxicidade , Gônadas/crescimento & desenvolvimento , Animais , Diferenciação Celular , Feminino , Feminização , Gônadas/efeitos dos fármacos , Humanos , Masculino , Fenóis , Testículo , Xenopus laevis
5.
PLoS One ; 15(1): e0227411, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31910233

RESUMO

Disorders/differences of sex development (DSD) cause profound psychological and reproductive consequences for the affected individuals, however, most are still unexplained at the molecular level. Here, we present a novel gene, 3-hydroxy-3-methylglutaryl coenzyme A synthase 2 (HMGCS2), encoding a metabolic enzyme in the liver important for energy production from fatty acids, that shows an unusual expression pattern in developing fetal mouse gonads. Shortly after gonadal sex determination it is up-regulated in the developing testes following a very similar spatial and temporal pattern as the male-determining gene Sry in Sertoli cells before switching to ovarian enriched expression. To test if Hmgcs2 is important for gonad development in mammals, we pursued two lines of investigations. Firstly, we generated Hmgcs2-null mice using CRISPR/Cas9 and found that these mice had gonads that developed normally even on a sensitized background. Secondly, we screened 46,XY DSD patients with gonadal dysgenesis and identified two unrelated patients with a deletion and a deleterious missense variant in HMGCS2 respectively. However, both variants were heterozygous, suggesting that HMGCS2 might not be the causative gene. Analysis of a larger number of patients in the future might shed more light into the possible association of HMGCS2 with human gonadal development.


Assuntos
Transtornos do Desenvolvimento Sexual/genética , Disgenesia Gonadal/genética , Gônadas/crescimento & desenvolvimento , Hidroximetilglutaril-CoA Sintase/genética , Adolescente , Animais , Transtornos do Desenvolvimento Sexual/patologia , Feminino , Regulação da Expressão Gênica no Desenvolvimento/genética , Disgenesia Gonadal/patologia , Gônadas/patologia , Heterozigoto , Humanos , Masculino , Camundongos , Mutação de Sentido Incorreto/genética , Ovário/crescimento & desenvolvimento , Ovário/patologia , Células de Sertoli/metabolismo , Proteína da Região Y Determinante do Sexo/genética , Testículo/crescimento & desenvolvimento , Testículo/patologia
6.
Gen Comp Endocrinol ; 285: 113270, 2020 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-31525374

RESUMO

Metazoans have evolved a complexity of sexual system and gonad development, however, sexual reproduction of scleractinian corals is not well understood. This study aimed to address the sexual system and gametogenesis in Porites lichen, a common species in the Indo-West Pacific. This study represents the first description of sexual system, which were determined by histological analysis of the samples collected in northern Taiwan. In addition, female and hermaphroditic colonies were separately cultured in aquarium to further monitor the release of eggs/larvae and thereby confirm the breeding system. The results demonstrate that P. lichen is a polygamodioecious brooder and displays seasonal gametogenesis and embryogenesis that ends in late summer. In hermaphroditic colonies, male polyps are predominant and hermaphroditic polyps make up a very small percent (1%-19.3%). In addition, two new gametogenic features were observed from the histological analysis: 1) oocytes developed within the spermaries in hermaphroditic polyps during the early stage of gametogenesis and 2) melanin granular cells were clustered in spermaries in both male and hermaphroditic colonies. This study demonstrated the plasticity of gametogenesis and melanin related cells appeared in corals, which provides an important information to explore hormones and molecular mechanism involving in gonadal arrangement and production of melanin for further studies.


Assuntos
Antozoários/crescimento & desenvolvimento , Antozoários/fisiologia , Gônadas/crescimento & desenvolvimento , Animais , Feminino , Geografia , Células Germinativas/metabolismo , Masculino , Melaninas/metabolismo , Oogênese , Reprodução/fisiologia , Espermatogênese , Taiwan
7.
Int J Mol Sci ; 20(21)2019 Nov 04.
Artigo em Inglês | MEDLINE | ID: mdl-31690065

RESUMO

Sex development is a complex process involving many genes and hormones. Defects in this process lead to Differences of Sex Development (DSD), a group of heterogeneous conditions not as rare as previously thought. Part of the obstacles in proper management of these patients is due to an incomplete understanding of the genetics programs and molecular pathways involved in sex development and DSD. Several challenges delay progress and the lack of a proper model system for the single patient severely hinders advances in understanding these diseases. The revolutionary techniques of cellular reprogramming and guided in vitro differentiation allow us now to exploit the versatility of induced pluripotent stem cells to create alternatives models for DSD, ideally on a patient-specific personalized basis.


Assuntos
Técnicas de Reprogramação Celular/métodos , Transtornos do Desenvolvimento Sexual/metabolismo , Células-Tronco Pluripotentes Induzidas/metabolismo , Animais , Transtornos do Desenvolvimento Sexual/patologia , Transtornos do Desenvolvimento Sexual/terapia , Gônadas/citologia , Gônadas/crescimento & desenvolvimento , Gônadas/metabolismo , Humanos , Células-Tronco Pluripotentes Induzidas/citologia , Modelagem Computacional Específica para o Paciente , Cultura Primária de Células/métodos
8.
Zhongguo Zhong Yao Za Zhi ; 44(15): 3239-3245, 2019 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-31602878

RESUMO

The present study was conducted to explores the effects of short-term addition of 17ß-E2 on the growth,gonad development and internal quality of overwintering Whitmania pigra. Before overwintering,0. 0,1. 0,10. 0,25. 0,50. 0,100. 0 µg·L~(-1) of 17ß-E2 were added to the aquaculture water for 6 weeks and then hibernated for 60 days. The changes of growth performance,gonad index,morphological structure of spermary( ovary),endogenous steroid hormones level and internal quality were measured. The results showed that the body weight,weight gain rate,specific growth rate,female gonad index,oocyte development and endogenous estrogen level of the leech increased first and then decreased with the increase of the concentration of exogenous 17ß-E2,which were higher than those of the control group. The body weight,weight gain rate and specific growth rate of the leech at the concentration of 25 µg·L~(-1)17ß-E2 were significantly higher than those of the other groups( P<0. 05),oocyte development and endogenous estrogen levels were significantly higher than those of other groups at the concentration of 50 µg·L~(-1)( P<0. 05). When the concentration of exogenous 17ß-E2 was higher than 50 µg·L~(-1),the levels of male gonad index,spermatocyte development,endogenous androgen and progesterone were significantly inhibited( P< 0. 05). There was no significant difference in endogenous corticosteroid levels among the groups. In conclusion,short-term addition of exogenous 17ß-E2 of 10-25 µg·L~(-1) could promote the growth of overwintering leeches,oocyte development and antithrombin activity without inhibiting the development of male gonads.


Assuntos
Estradiol/farmacologia , Gônadas/crescimento & desenvolvimento , Sanguessugas/efeitos dos fármacos , Sanguessugas/crescimento & desenvolvimento , Androgênios/análise , Animais , Estrogênios/análise , Feminino , Gônadas/efeitos dos fármacos , Hibernação , Masculino , Progesterona/análise
9.
Genes Genomics ; 41(12): 1397-1415, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31485990

RESUMO

BACKGROUND: Sea urchin gonads of both sexes, commonly termed "roe", are highly valued seafood delicacies, and Strongylocentrotus intermedius is considered one of the tastiest sea urchins. In order to produce high-quality gonads for consumption and clarify the mechanism of gonad growth and development of the sea urchin, more genetic information, especially at the transcriptome level, is needed. OBJECTIVE: A more thorough understanding of sea urchin gonad growth and development in both sexes could enable regulation of these processes at several stages with the aim of suppressing gametogenesis in order to produce high-quality gonads for consumption. METHODS: The adult sea urchins S. intermedius were cultured for 3 months, and were sampled for the gonadal transcriptome analysis which has been performed on the RNAs of three male and female adults of S. intermedius in each gonad development stage. RESULTS: Illumina sequencing raw sequence data was deposited in the NCBI Sequence Read Archive (SRA) database (PRJNA532998). It generated 560,196,356 raw reads and 548,956,944 clean reads were acquired, which were assembled into 107,850 transcripts with 44,124 genes. Comparative analysis showed the differentially expressed genes (DEGs) from 114 to 2566. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were used to determine the functional significance of these DEGs. We have selected 9 genes related to growth and 12 genes related to fatty acid biosynthesis and metabolism in sea urchin gonads. CONCLUSION: These data for sea urchins were intended to provide markers for gonad growth and development that can be accumulated for use in aquaculture applications.


Assuntos
Ácidos Graxos/metabolismo , Strongylocentrotus/genética , Animais , Ácidos Graxos/biossíntese , Feminino , Perfilação da Expressão Gênica , Gônadas/anatomia & histologia , Gônadas/crescimento & desenvolvimento , Gônadas/metabolismo , Masculino , Repetições de Microssatélites , Polimorfismo de Nucleotídeo Único , Strongylocentrotus/crescimento & desenvolvimento , Strongylocentrotus/metabolismo , Transcriptoma
10.
Anim Reprod Sci ; 208: 106131, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31405476

RESUMO

Gonadal development usually involves alternative splicing of sex-related genes. Vasa, a highly conserved ATP-dependent RNA helicase present mainly in germ cells, has an important function in gonadal development. As an important sex-related gene, recent evidence indicates that different splice variants of vasa exist in many species. In this study, there was identification of two types of vasa splice variants in the Chinese mitten crab Eriocheir sinensis, termed Esvasa-l and Esvasa-s, respectively. Furthermore, splice variants of Esvasa-s were sub-divided into Esvasa-s1, Esvasa-s2, Esvasa-s3, Esvasa-s4, and Esvasa-s5, based on differing numbers of TGG repeats. Results from genomic structure analyses indicated that these forms are alternatively spliced transcripts from a single vasa gene. Results from tissue distribution assessments indicate the vasa splice variants were exclusively expressed in the gonads of male and female adult crabs. In situ hybridization results indicate Esvasa mRNA was mainly present in the cytoplasm of previtellogenic oocytes. As oocyte size increased, relative abundance of Esvasa mRNA decreased and became distributed near the cellular membrane. The Esvasa mRNA was not detectable in mature oocytes. In testis, Esvasa mRNA was detected in spermatids and spermatozoa, but not in spermatogonia and spermatocytes. Notably, results from qPCR analysis of Esvasa-l and Esvasa-s indicate there are different relative proportions during gametogenesis, implying that splice variants of the Esvasa gene may have different biological functions during crab gonadal development.


Assuntos
Processamento Alternativo , Braquiúros/genética , RNA Helicases DEAD-box/metabolismo , Gônadas/crescimento & desenvolvimento , RNA Mensageiro/metabolismo , Sequência de Aminoácidos , Animais , Sequência de Bases , RNA Helicases DEAD-box/genética , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Regulação Enzimológica da Expressão Gênica/fisiologia , RNA Mensageiro/genética , Maturidade Sexual
11.
Insect Biochem Mol Biol ; 113: 103211, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31425852

RESUMO

Wolbachia are intracellular bacteria that manipulate host reproduction by several mechanisms including cytoplasmic incompatibility (CI). However, the underlying mechanisms of Wolbachia-induced CI are not entirely clear. Here, we monitored the Wolbachia distribution in the male gonads of the small brown planthopper (Laodelphax striatellus, SBPH) at different development stages, and investigated the influence of Wolbachia on male gonads by a quantitative proteomic analysis. A total of 276 differentially expressed proteins were identified, with the majority of them participating in metabolism, modification, and reproduction. Knocking down the expression of outer dense fiber protein (ODFP) and venom allergen 5-like (VA5L) showed decreased egg reproduction, and these two genes might be responsible for Wolbachia improved fecundity in infected L. striatellus; whereas knocking down the expression of cytosol amino-peptidase-like (CAL) significantly decreased the egg hatch rate in Wolbachia-uninfected L. striatellus, but not in the Wolbachia-infected one. Considering that the mRNA/protein level of CAL was downregulated by Wolbachia infection and dsCAL treatment closely mimicked Wolbachia-induced CI, we presumed that CAL might be one of the factors determining the CI phenotype.


Assuntos
Hemípteros/fisiologia , Proteínas de Insetos/genética , Wolbachia/fisiologia , Animais , Gônadas/crescimento & desenvolvimento , Gônadas/microbiologia , Hemípteros/genética , Hemípteros/crescimento & desenvolvimento , Proteínas de Insetos/metabolismo , Masculino , Ninfa/genética , Ninfa/crescimento & desenvolvimento , Proteoma , Proteômica , Reprodução
12.
Mar Biotechnol (NY) ; 21(5): 697-706, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31372794

RESUMO

The availability of sexually mature fish often dictates the success of its captive breeding. In this study, we induced reproductive development in juvenile protogynous tiger grouper through oral administration of a plasmid (p) containing an engineered follicle-stimulating hormone (FSH). An expression construct (pcDNA3.1) was designed to express a single-chain FSH consisting of giant grouper FSH ß-subunit and glycoprotein subunit-α (CGα), linked by the carboxy-terminal peptide (CTP) sequence from the human chorionic gonadotropin (hCG). Single oral delivery of pFSH encapsulated in liposome and chitosan to tiger grouper yielded a significant increase in plasma FSH protein level after 4 days. Weekly pFSH feeding of juvenile tiger groupers for 8 weeks stimulated ovarian development as indicated by a significant increase in oocyte diameter and progression of oocytes to cortical alveolar stage. As the pFSH treatment progressed from 20 to 38 weeks, female to male sex change was initiated, characterized by oocyte regression, proliferation of spermatogonial cells, and occurrence of spermatogenic cysts. It was also associated with significantly lower mRNA expression of steroidogenic genes (cyp11b, cyp19a1a, and foxl2) and basal plasma levels of sex steroid hormones 17ß-estradiol (E2), testosterone (T), and 11-ketotestosterone (11KT). Results suggest that pFSH stimulates ovarian development up to cortical alveolar stage and then initiates sex change in tiger grouper. These findings significantly contribute to our knowledge on the role of FSH in the development of protogynous hermaphroditic fish. This study is the first to demonstrate induction of reproductive development in fish through oral delivery of plasmid gonadotropin.


Assuntos
Gonadotropina Coriônica/genética , Hormônio Foliculoestimulante/genética , Gônadas/efeitos dos fármacos , Organismos Hermafroditas/efeitos dos fármacos , Perciformes/genética , Processos de Determinação Sexual/efeitos dos fármacos , Diferenciação Sexual/efeitos dos fármacos , Administração Oral , Animais , Quitosana/química , Gonadotropina Coriônica/administração & dosagem , Gonadotropina Coriônica/biossíntese , Composição de Medicamentos , Feminino , Proteínas de Peixes/biossíntese , Proteínas de Peixes/genética , Hormônio Foliculoestimulante/administração & dosagem , Hormônio Foliculoestimulante/biossíntese , Hormônios Esteroides Gonadais/biossíntese , Hormônios Esteroides Gonadais/genética , Gônadas/crescimento & desenvolvimento , Gônadas/metabolismo , Organismos Hermafroditas/genética , Humanos , Lipossomos/administração & dosagem , Lipossomos/química , Masculino , Oogênese/efeitos dos fármacos , Oogênese/genética , Perciformes/crescimento & desenvolvimento , Perciformes/metabolismo , Plasmídeos/química , Plasmídeos/metabolismo , Proteínas Recombinantes de Fusão/administração & dosagem , Proteínas Recombinantes de Fusão/biossíntese , Proteínas Recombinantes de Fusão/genética , Pré-Seleção do Sexo/métodos , Espermatogênese/efeitos dos fármacos , Espermatogênese/genética
13.
Mar Biotechnol (NY) ; 21(5): 718-730, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31392593

RESUMO

The major causal factors for the irreversible decline in physical vitality during organismal aging are postulated to be a chronic state of cellular redox imbalance, metabolic toxicity, and impaired energy homeostasis. We assessed whether the relevant enzyme activity, oxidative stress, and intracellular ATP might be causally involved in the aging of short-lived Chlamys farreri (life span 4~5 years). A total of eight related biochemical and cellular indicators were chosen for the subsequent analysis. All the indicators were measured in seven different tissues from scallops aged one to four years, and our data support that the aging of C. farreri is associated with attenuated tissue enzyme activity as well as a decreased metabolic rate. Through principal component analysis, we developed an integrated vigor index for each tissue for comprehensive age-related fitness evaluation. Remarkably, all tissue-integrated vigor indexes significantly declined with age, and the kidney was observed to be the most representative tissue. Further transcriptional profiling of the enzymatic genes provided additional detail on the molecular responses that may underlie the corresponding biochemical results. Moreover, these critical molecular responses may be attributed to the conserved hierarchical regulators, e.g., FOXO, AMPKs, mTOR, and IGF1R, which were identified as potentially novel markers for chronic fitness decline with age in bivalves. The present study provides a systematic approach that could potentially benefit the global assessment of the aging process in C. farreri and provide detailed evaluation of the biochemical, cellular, and genetic indicators that might be involved. This information may assist in a better understanding of bivalve adaptability and life span.


Assuntos
Envelhecimento/genética , Bivalves/genética , Regulação da Expressão Gênica no Desenvolvimento , Transcriptoma , Proteínas Quinases Ativadas por AMP/genética , Proteínas Quinases Ativadas por AMP/metabolismo , Trifosfato de Adenosina/metabolismo , Animais , Bivalves/crescimento & desenvolvimento , Bivalves/metabolismo , Metabolismo Energético/genética , Proteína Forkhead Box O1/genética , Proteína Forkhead Box O1/metabolismo , Perfilação da Expressão Gênica , Brânquias/crescimento & desenvolvimento , Brânquias/metabolismo , Gônadas/crescimento & desenvolvimento , Gônadas/metabolismo , Hepatopâncreas/crescimento & desenvolvimento , Hepatopâncreas/metabolismo , Homeostase/genética , Rim/crescimento & desenvolvimento , Rim/metabolismo , Especificidade de Órgãos , Oxirredução , Estresse Oxidativo , Análise de Componente Principal , Receptor IGF Tipo 1/genética , Receptor IGF Tipo 1/metabolismo , Serina-Treonina Quinases TOR/genética , Serina-Treonina Quinases TOR/metabolismo
14.
Genetics ; 213(2): 529-553, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31399485

RESUMO

Fetal mammalian testes secrete Anti-Müllerian hormone (Amh), which inhibits female reproductive tract (Müllerian duct) development. Amh also derives from mature mammalian ovarian follicles, which marks oocyte reserve and characterizes polycystic ovarian syndrome. Zebrafish (Danio rerio) lacks Müllerian ducts and the Amh receptor gene amhr2 but, curiously, retains amh To discover the roles of Amh in the absence of Müllerian ducts and the ancestral receptor gene, we made amh null alleles in zebrafish. Results showed that normal amh prevents female-biased sex ratios. Adult male amh mutants had enormous testes, half of which contained immature oocytes, demonstrating that Amh regulates male germ cell accumulation and inhibits oocyte development or survival. Mutant males formed sperm ducts and some produced a few offspring. Young female mutants laid a few fertile eggs, so they also had functional sex ducts. Older amh mutants accumulated nonvitellogenic follicles in exceedingly large but sterile ovaries, showing that Amh helps control ovarian follicle maturation and proliferation. RNA-sequencing data partitioned juveniles at 21 days postfertilization (dpf) into two groups that each contained mutant and wild-type fish. Group21-1 upregulated ovary genes compared to Group21-2, which were likely developing as males. By 35 dpf, transcriptomes distinguished males from females and, within each sex, mutants from wild types. In adult mutants, ovaries greatly underexpressed granulosa and theca genes, and testes underexpressed Leydig cell genes. These results show that ancestral Amh functions included development of the gonadal soma in ovaries and testes and regulation of gamete proliferation and maturation. A major gap in our understanding is the identity of the gene encoding a zebrafish Amh receptor; we show here that the loss of amhr2 is associated with the breakpoint of a chromosome rearrangement shared among cyprinid fishes.


Assuntos
Hormônio Antimülleriano/genética , Genitália Feminina/crescimento & desenvolvimento , Processos de Determinação Sexual , Peixe-Zebra/genética , Animais , Feminino , Gônadas/crescimento & desenvolvimento , Ductos Paramesonéfricos/crescimento & desenvolvimento , Folículo Ovariano/crescimento & desenvolvimento , Ovário/crescimento & desenvolvimento , RNA-Seq , Receptores de Peptídeos/genética , Receptores de Fatores de Crescimento Transformadores beta/genética , Peixe-Zebra/crescimento & desenvolvimento
15.
Stem Cell Reports ; 13(2): 419-433, 2019 08 13.
Artigo em Inglês | MEDLINE | ID: mdl-31412286

RESUMO

In vertebrates, estrogen receptors are essential for estrogen-associated early gonadal sex development. Our previous studies revealed sexual dimorphic expression of estrogen receptor ß2 (ERß2) during embryogenesis of medaka, and here we investigated the functional importance of ERß2 in female gonad development and maintenance using a transgenerational ERß2-knockdown (ERß2-KD) line and ERß2-null mutants. We found that ERß2 reduction favored male-biased gene transcription, suppressed female-responsive gene expression, and affected SDF1a and CXCR4b co-assisted chemotactic primordial germ cell (PGC) migration. Co-overexpression of SDF1a and CXXR4b restored the ERß2-KD/KO associated PGC mismigration. Further analysis confirmed that curtailment of ERß2 increased intracellular Ca2+ concentration, disrupted intra- and extracellular calcium homeostasis, and instigated autophagic germ cell degradation and germ cell loss, which in some cases ultimately affected the XX female sexual development. This study is expected improve our understanding of germ cell maintenance and sex spectrum, and hence open new avenues for reproductive disorder management.


Assuntos
Receptor beta de Estrogênio/metabolismo , Proteínas de Peixes/metabolismo , Gônadas/crescimento & desenvolvimento , Diferenciação Sexual , Animais , Cálcio/metabolismo , Proliferação de Células , Quimiocina CXCL12/genética , Quimiocina CXCL12/metabolismo , Embrião não Mamífero/metabolismo , Receptor beta de Estrogênio/antagonistas & inibidores , Receptor beta de Estrogênio/genética , Feminino , Proteínas de Peixes/antagonistas & inibidores , Proteínas de Peixes/genética , Regulação da Expressão Gênica no Desenvolvimento , Células Germinativas/citologia , Células Germinativas/metabolismo , Gônadas/metabolismo , Masculino , Oryzias/crescimento & desenvolvimento , Oryzias/metabolismo , RNA Antissenso/genética , RNA Antissenso/metabolismo , Receptores CXCR4/genética , Receptores CXCR4/metabolismo
16.
Dokl Biol Sci ; 485(1): 37-39, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31197591

RESUMO

During the growth of hydrothermal vestimentiferan Oasisia alvinae the trunk part of body was found to be elongated (from 51 to 83.4% of the overall body length), while the relative dimensions of all other body regions decreased. This was related to the enhanced trophosome and gonad development in the trunk part. We suppose that predominant trunk growth is a common feature of all vestimentiferans.


Assuntos
Poliquetos/crescimento & desenvolvimento , Animais , Gônadas/crescimento & desenvolvimento , Fontes Hidrotermais , Poliquetos/anatomia & histologia
17.
Artigo em Inglês | MEDLINE | ID: mdl-31125834

RESUMO

Tudor domain-containing proteins (TDRDs) are highly conserved among organisms and have a function in gonads to regulate gametogenesis and genome stability through the piwi-interacting RNA (piRNA) pathway. With diverse sexual development patterns in teleost species, the evolution and function of Tdrd genes among teleosts remain unclear. Here, we identified and characterized 12 Tdrd genes (PoTdrds) in Japanese flounder (Paralichthys olivaceus) which represents dramatic sexual dimorphic metrics and sex reversal during sex differentiation. Phylogenetic and comparative synteny indicated the gain and loss of Tdrd genes after teleost-specific whole-genome duplication (3R-WGD). Tdrd1, Tdrd5, Tdrd6 and Ecat8 were abundantly expressed in their gonads. Four PoTdrds (Tdrd6, Tdrd7b, Tdrd9 and Ecat8) represented significant male-biased expression in gynogenetic and wild-type Japanese flounder gonads (p < .01). This finding indicated their important roles in spermatogenesis of P. olivaceus. Some PoTdrds were either highly up-regulated in gynogenetic testis (Tdrd3, Tdrd5, Tdrd7b and Ecat8) or down-regulated in gynogenetic ovary (Tdrkh, Tdrd3, Tdrd6l) compared with wild-type gonads (p < .05). Molecular evolution tests revealed that the selective pressure of Tdrd6/6l differed between ancestral aquatic and terrestrial organisms with 13 positively selected sites found in the ancestral lineages of teleost Tdrd6. Expression profile analysis suggested that PoTdrd6 differed significantly from PoTdrd6l, indicating sub-functionalization after 3R-WGD. All these results are important for the functional annotation of Tdrd genes and can benefit the further deciphering of Tdrd functions during gonadal development and gametogenesis of teleost fish.


Assuntos
Proteínas de Peixes/genética , Linguado/genética , Sequência de Aminoácidos , Animais , Evolução Molecular , Feminino , Proteínas de Peixes/química , Linguado/crescimento & desenvolvimento , Regulação da Expressão Gênica no Desenvolvimento , Gônadas/crescimento & desenvolvimento , Gônadas/metabolismo , Japão , Masculino , Filogenia , Alinhamento de Sequência , Transcriptoma , Domínio Tudor
18.
Hum Mol Genet ; 28(14): 2319-2329, 2019 07 15.
Artigo em Inglês | MEDLINE | ID: mdl-30985895

RESUMO

Disorders of sex development (DSDs) are defined as congenital conditions in which chromosomal, gonadal or anatomical sex is atypical. In many DSD cases, genetic causes remain to be elucidated. Here, we performed a case-control exome sequencing study comparing gene-based burdens of rare damaging variants between 26 DSD cases and 2625 controls. We found exome-wide significant enrichment of rare heterozygous truncating variants in the MYRF gene encoding myelin regulatory factor, a transcription factor essential for oligodendrocyte development. All three variants occurred de novo. We identified an additional 46,XY DSD case of a de novo damaging missense variant in an independent cohort. The clinical symptoms included hypoplasia of Müllerian derivatives and ovaries in 46,XX DSD patients, defective development of Sertoli and Leydig cells in 46,XY DSD patients and congenital diaphragmatic hernia in one 46,XY DSD patient. As all of these cells and tissues are or partly consist of coelomic epithelium (CE)-derived cells (CEDC) and CEDC developed from CE via proliferaiton and migration, MYRF might be related to these processes. Consistent with this hypothesis, single-cell RNA sequencing of foetal gonads revealed high expression of MYRF in CE and CEDC. Reanalysis of public chromatin immunoprecipitation sequencing data for rat Myrf showed that genes regulating proliferation and migration were enriched among putative target genes of Myrf. These results suggested that MYRF is a novel causative gene of 46,XY and 46,XX DSD and MYRF is a transcription factor regulating CD and/or CEDC proliferation and migration, which is essential for development of multiple organs.


Assuntos
Transtornos 46, XX do Desenvolvimento Sexual/genética , Transtorno 46,XY do Desenvolvimento Sexual/genética , Proteínas de Membrana/genética , Fatores de Transcrição/genética , Transtornos 46, XX do Desenvolvimento Sexual/patologia , Adolescente , Estudos de Casos e Controles , Movimento Celular , Proliferação de Células , Pré-Escolar , Estudos de Coortes , Biologia Computacional , Transtorno 46,XY do Desenvolvimento Sexual/patologia , Feminino , Ontologia Genética , Gônadas/crescimento & desenvolvimento , Haploinsuficiência , Humanos , Masculino , Proteínas de Membrana/metabolismo , Mutação , Mutação de Sentido Incorreto , Análise de Célula Única , Fatores de Transcrição/metabolismo , Sequenciamento Completo do Exoma , Adulto Jovem
19.
Aquat Toxicol ; 211: 124-136, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30965180

RESUMO

Cu in surface waters has been demonstrated to affect aquatic animals at ecologically relevant concentrations. However, its effects on reproductive endocrine system and the underlying toxicological mechanisms are largely unknown. In this study, zebrafish (Danio rerio) were exposed to 0, 10, 20, 40 µg/L of Cu for 30 days. Growth, gonad histopathology, the hormone levels and the transcriptional profiles of genes in the hypothalamic-pituitary-gonadal (HPG) axis in both sexes were examined. The results indicated that body weight was significantly reduced, the gonadal development was affected, and the levels of E2, T and 11-KT were remarkably disturbed in Cu-exposed fish. Moreover, the expression profiles of steroidogenesis-related genes in gonad (3ßhsd, 17ßhsd, cyp11a1, cyp17, cyp19a, lhr, fshr, hmgra and star) and in brains (ar, cyp19b, erα, er2ß, lhß, fshß, gnrh2, gnrh3, gnrhr1, gnrh2 and gnrh4) displayed alterations after exposure to Cu. These results demonstrated that Cu could suppress the growth of zebrafish and significantly affect the reproductive biology in both sexes by damaging the structure of the gonads, altering the steroid hormone levels and the expressions of endocrine-related genes in HPG of zebrafish. This study suggests that Cu adversely affects the reproductive endocrine system in zebrafish and could pose a potential threat to fish populations inhabiting Cu-contaminated waters.


Assuntos
Cobre/toxicidade , Disruptores Endócrinos/toxicidade , Gônadas/efeitos dos fármacos , Reprodução/efeitos dos fármacos , Poluentes Químicos da Água/toxicidade , Peixe-Zebra/crescimento & desenvolvimento , Animais , Relação Dose-Resposta a Droga , Disruptores Endócrinos/metabolismo , Sistema Endócrino/efeitos dos fármacos , Sistema Endócrino/crescimento & desenvolvimento , Feminino , Gônadas/crescimento & desenvolvimento , Gônadas/metabolismo , Masculino , Peixe-Zebra/metabolismo , Proteínas de Peixe-Zebra/genética
20.
Genetics ; 212(2): 523-535, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30992386

RESUMO

Remodeling of the extracellular matrix supports tissue and organ development, by regulating cellular morphology and tissue integrity. However, proper extracellular matrix remodeling requires spatiotemporal regulation of extracellular metalloproteinase activity. Members of the ADAMTS (a disintegrin and metalloproteinase with thrombospondin motifs) family, including MIG-17 and GON-1, are evolutionarily conserved, secreted, zinc-requiring metalloproteinases. Although these proteases are required for extracellular matrix remodeling during gonadogenesis in Caenorhabditis elegans, their in vivo regulatory mechanisms remain to be delineated. Therefore, we focused on the C. elegans tissue inhibitors of metalloproteinases (TIMPs), TIMP-1 and CRI-2 Analysis of the transcription and translation products for GFP/Venus fusions, with TIMP-1 or CRI-2, indicated that these inhibitors were secreted and localized to the basement membrane of gonads and the plasma membrane of germ cells. A timp-1 deletion mutant exhibited gonadal growth defects and sterility, and the phenotypes of this mutant were fully rescued by a TIMP-1::Venus construct, but not by a TIMP-1(C21S)::Venus mutant construct, in which the inhibitor coding sequence had been mutated. Moreover, genetic data suggested that TIMP-1 negatively regulates proteolysis of the α1 chain of type IV collagen. We also found that the loss-of-function observed for the mutants timp-1 and cri-2 involves a partial suppression of gonadal defects found for the mutants mig-17/ADAMTS and gon-1/ADAMTS, and that this suppression was canceled upon overexpression of gon-1 or mig-17, respectively. Based on these results, we propose that both TIMP-1 and CRI-2 act as inhibitors of MIG-17 and GON-1 ADAMTSs to regulate gonad development in a noncell-autonomous manner.


Assuntos
Proteínas de Caenorhabditis elegans/metabolismo , Desintegrinas/metabolismo , Gônadas/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Metaloendopeptidases/metabolismo , Inibidor Tecidual de Metaloproteinase-1/fisiologia , Inibidores Teciduais de Metaloproteinases/metabolismo , Animais , Membrana Basal/metabolismo , Caenorhabditis elegans/genética , Caenorhabditis elegans/crescimento & desenvolvimento , Caenorhabditis elegans/metabolismo , Proteínas de Caenorhabditis elegans/genética , Membrana Celular/metabolismo , Colágeno Tipo IV/metabolismo , Desintegrinas/genética , Matriz Extracelular/metabolismo , Células Germinativas/metabolismo , Gônadas/crescimento & desenvolvimento , Peptídeos e Proteínas de Sinalização Intercelular/genética , Metaloendopeptidases/genética , Morfogênese/genética , Morfogênese/fisiologia , Inibidor Tecidual de Metaloproteinase-1/genética , Inibidor Tecidual de Metaloproteinase-1/metabolismo , Inibidores Teciduais de Metaloproteinases/genética
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