RESUMO
Gamete production is a fundamental process for reproduction; however, exposure to stress, such as increased environmental temperature, can decrease or even interrupt this process, affecting fertility. Thus, the survival of spermatogonial stem cells (SSCs) is crucial for the recovery of spermatogenesis upon stressful situations. Here, we show that the Notch pathway is implicated in such survival, by protecting the SSCs against thermal stress. First, we corroborated the impairment of spermatogenesis under heat stress in medaka, observing an arrest in metaphase I at 10 days of heat treatment, an increase in the number of spermatocytes, and downregulation of ndrg1b and sycp3. In addition, at 30 days of treatment, an interruption of spermatogenesis was observed with a strong loss of spermatocytes and spermatids. Then, the exposure of adult males to thermal stress condition induced apoptosis mainly in spermatogenic and supporting somatic cells, with the exception of the germinal region, where SSCs are located. Concomitantly, the Notch pathway-related genes were upregulated, including the ligands (dll4, jag1-2) and receptors (notch1a-3). Moreover, during thermal stress presenilin enhancer-2 (pen-2), the catalytic subunit of γ-secretase complex of the Notch pathway was restricted to the germinal region of the medaka testis, observed in somatic cells surrounding type A spermatogonia (SGa). The importance of Notch pathway was further supported by an ex vivo approach, in which the inhibition of this pathway activity induced a loss of SSCs. Overall, this study supports the importance of Notch pathways for the protection of SSCs under chronic thermal stress.
Assuntos
Oryzias , Masculino , Animais , Diferenciação Celular , Testículo/metabolismo , Espermatogênese/genética , Espermatogônias/fisiologia , Células-Tronco , Resposta ao Choque TérmicoRESUMO
Atrazine (ATZ) is a frequent contaminant in freshwater ecosystems within agricultural regions. The capacity of this herbicide to interfere with the vertebrate endocrine system is broadly recognized, but the mechanisms and responses usually differ among species. In this study, ATZ effects on hypothalamus-pituitary-gonadal (HPG) axis key genes expression and early gonadal development were evaluated in Odontesthes bonariensis larvae waterborne exposed during the gonadal differentiation period. Fish were treated to 0, 0.7, 7.0, and 70 µg ATZ/L at 25 °C from the 2nd to 6th week after hatching (wah), and a group was kept in clean water until the 12th wah. Parallelly, a group was submitted to 0.05 µg/L of ethinylestradiol (EE2) as a positive estrogenic control. From each treatment, eight larvae were sampled at 6 wah for gene expression analysis and twelve larvae at 12 wah for phenotypic sex histological determination. The expression of gnrh1, lhb, fshb, and cyp19a1b was assessed in the head, and the ones of amha, 11ßhsd2, and cyp19a1a in the trunk. Fish growth was significantly higher in fish exposed to 7 and 70 µg ATZ/L in the 6 wah, but the effect vanished at the 12 wah. The expression of lhb was upregulated in both sex larvae exposed from 7 µg ATZ/L. However, a dimorphic effect was induced on cyp19a1a expression at 70 µg ATZ/L, up or downregulating mRNA transcription in males and females, respectively. Delayed ovarian development and increased number of testicular germ cells were histologically observed from 7 to 70 µg ATZ/L, respectively, and a sex inversion (genotypic male to phenotypic female) was found in one larva at 70 µg ATZ/L. The lhb expression was also upregulated by EE2, but the cyp19a1a expression was not affected, and a complete male-to-female reversal was induced. Further, EE2 upregulated gnrh1 in females and cyp19a1b in both sexes, but it did not alter any assessed gene in the trunk. In conclusion, ATZ disrupted HPG axis physiology and normal gonadal development in O. bonariensis larvae at environmentally relevant concentrations. The responses to ATZ only partially overlapped and were less active when compared to the model estrogenic compound EE2.
Assuntos
Atrazina , Poluentes Químicos da Água , Animais , Masculino , Feminino , Atrazina/toxicidade , Ecossistema , Poluentes Químicos da Água/toxicidade , Peixes/fisiologia , Desenvolvimento Sexual , Larva , Transcrição Gênica , América do SulRESUMO
The exposure of adult fish to warm or high temperatures is known to impair reproduction, yet the long-term reproductive impacts for treatments at early life are not well clarified. This study aimed to evaluate the effects of warm temperature (WT) during juvenile stage on gonad maturation, gamete quality, and offspring thermotolerance in rainbow trout. While the comparison of basic reproductive parameters in WT females did not reveal any kind of impairment, many WT males showed an atrophied, undeveloped gonad, or a smaller testis with lower milt volume; sperm quality parameters in WT males and deformity rates in the respective progeny were also highly affected. However, despite of such negative effects, many of the remaining progeny presented better rates of survival and growth when exposed to the same conditions as those of parental fish (WT), suggesting that thermal stress in parr stage males elicited intergenerational thermotolerance after a single generation. The present results support that prolonged warming stress during early life stages can adversely affect key reproductive aspects, but contrastingly increase offspring performance at upper thermal ranges. These findings have implications on the capacity of fish to adapt and to cope with global warming.
Assuntos
Reprodução , Termotolerância , Truta/fisiologia , Animais , Feminino , Infertilidade Masculina/etiologia , Masculino , Testículo/crescimento & desenvolvimento , Truta/crescimento & desenvolvimentoRESUMO
Morphological background adaptation is both an endocrine and a nervous response, involving changes in the amount of chromatophores and pigment concentration. However, whether this adaptation takes place at early developmental stages is largely unknown. Somatolactin (Sl) is a pituitary hormone present in fish, which has been associated to skin pigmentation. Moreover, growth hormone receptor type 1 (Ghr1) has been suggested to be the Sl receptor and was associated with background adaptation in adults. In this context, the aim of this work was to evaluate the ontogeny of morphological adaptation to background and the participation of ghr1 in this process. We found in larval stages of the cichlid Cichlasoma dimerus that the number of head melanophores and pituitary cells immunoreactive to Sl was increased in individuals reared with black backgrounds compared with that in fish grown in white tanks. In larval stages of the medaka Oryzias latipes, a similar response was observed, which was altered by ghr1 biallelic mutations using CRISPR/Cas9. Interestingly, melanophore and leucophore numbers were highly associated. Furthermore, we found that somatic growth was reduced in ghr1 biallelic mutant O. latipes, establishing the dual function of this growth hormone receptor. Taken together, these results show that morphological background adaptation is present at early stages during development and that is dependent upon ghr1 during this period.
Assuntos
Proteínas de Peixes , Receptores da Somatotropina , Aclimatação , Animais , Cor , Proteínas de Peixes/genética , Hormônio do Crescimento , Hormônios Hipofisários/genética , Receptores da Somatotropina/genéticaRESUMO
It has been traditionally accepted that the gonadotropins (GtHs), follicle stimulating hormone (FSH) and luteinizing hormone (LH), are synthesized and secreted only by the pituitary. However, the presence of theses hormones in extrapituitary tissues has been demonstrated in mammals, and more recently also in fish. In this study, we cloned the cDNAs and characterized the expression of FSH-ß, LH-ß, and glycoprotein hormone α (GPH-α) subunits from brain and gonads of male and female pejerrey Odontesthes bonariensis at different stages of gonadal maturation. In situ hybridization revealed that, in addition to their classical location in pituitary cells, the three GtH transcripts were also located in the gonads. FSH-ß and GPH-α subunits were found in the cytoplasm of oogonia, previtellogenic and vitellogenic oocytes in ovaries. LH-ß expression was detected in previtellogenic and vitellogenic oocytes but not in oogonia. In males, the three subunits were expressed in spermatogonia and to a lesser extent in spermatocytes. Exposure of fish to high water temperatures that impair pejerrey reproduction also induced a decrease of extrahypophyseal expression of GtH subunits.
Assuntos
Subunidade beta do Hormônio Folículoestimulante/metabolismo , Subunidade alfa de Hormônios Glicoproteicos/metabolismo , Hormônio Luteinizante Subunidade beta/metabolismo , Smegmamorpha/metabolismo , Estresse Fisiológico , Temperatura , Animais , Encéfalo/metabolismo , Feminino , Masculino , Smegmamorpha/fisiologiaRESUMO
Pejerrey is a teleost fish presenting a strong temperature-dependent sex determination. This study was conducted to clone pejerrey amh cDNA, analyze its expression profile during thermal and endocrine manipulation of gonadal differentiation, and compare its expression with that of gonadal aromatase (cyp19a1). Amh displayed higher expression at masculinizing than at feminizing temperatures during the gonadal differentiation period. Its expression at an intermediate temperature (females 1:1 males), was high in half of the larvae and low in the other half. Cyp19a1 showed a reciprocal expression pattern to that of amh both individually- and temperature-wise. Increased cyp19a1 and amh expression was observed before morphological gonadal differentiation. Amh expression in larvae feminized by administration of estradiol or masculinized by the administration of an aromatase inhibitor was down- and up-regulated, respectively. These results show that amh plays a critical role in testicular differentiation and it is apparently modulated by estrogens in this species.