RESUMO
Estrogens regulate key aspects of sexual determination and differentiation, and exposure to exogenous estrogens can alter ovarian development. Alligators inhabiting Lake Apopka, FL, are historically exposed to estrogenic endocrine disrupting contaminants and are characterized by a suite of reproductive abnormalities, including altered ovarian gene expression and abated transcriptional responses to follicle stimulating hormone. Here, we test the hypothesis that disrupting estrogen signaling during gonadal differentiation results in persistent alterations to ovarian gene expression that mirror alterations observed in alligators from Lake Apopka. Alligator embryos collected from a reference site lacking environmental contamination were exposed to estradiol-17 beta or a nonaromatizable androgen in ovo and raised to the juvenile stage. Changes in basal and gonadotropin-challenged ovarian gene expression were then compared to Apopka juveniles raised under identical conditions. Assessing basal transcription in untreated reference and Apopka animals revealed a consistent pattern of differential expression of key ovarian genes. For each gene where basal expression differed across sites, in ovo estradiol treatment in reference individuals recapitulated patterns observed in Apopka alligators. Among those genes affected by site and estradiol treatment were three aryl hydrocarbon receptor (AHR) isoforms, suggesting that developmental estrogen signaling might program sensitivity to AHR ligands later in life. Treatment with gonadotropins stimulated strong ovarian transcriptional responses; however, the magnitude of responses was not strongly affected by steroid hormone treatment. Collectively, these findings demonstrate that precocious estrogen signaling in the developing ovary likely underlies altered transcriptional profiles observed in a natural population exposed to endocrine disrupting contaminants.
Assuntos
Jacarés e Crocodilos , Embrião não Mamífero/efeitos dos fármacos , Disruptores Endócrinos/toxicidade , Estrogênios/toxicidade , Exposição Materna/efeitos adversos , Ovário/efeitos dos fármacos , Jacarés e Crocodilos/embriologia , Jacarés e Crocodilos/genética , Animais , Reprogramação Celular/efeitos dos fármacos , Reprogramação Celular/genética , Embrião não Mamífero/metabolismo , Desenvolvimento Embrionário/efeitos dos fármacos , Desenvolvimento Embrionário/genética , Exposição Ambiental/efeitos adversos , Feminino , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Lagos , Modelos Animais , Ovário/metabolismo , Oviparidade/efeitos dos fármacos , Oviparidade/genética , Transcriptoma/efeitos dos fármacos , Transcriptoma/genética , Poluentes Químicos da Água/toxicidadeRESUMO
The laying quail is a worldwide breed which exhibits high economic value. In our current study, the vasoactive intestinal peptide receptor-1 (VIPR-1) was selected as the candidate gene for identifying traits of egg production. A single nucleotide polymorphism (SNP) detection was performed in 443 individual quails, including 196 quails from the H line, 202 quails from the L line, and 45 wild quails. The SNPs were genotyped using a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Two mutations (G373T, A313G) were detected in all the tested quail populations. The associated analysis showed that the SNP genotypes of the VIPR-1 gene were significantly linked with the egg weight of G373T and A313G in 398 quails. The quails with the genotype GG always exhibited the largest egg weight for the two mutations in the H and L lines. Linkage disequilibrium (LD) analysis indicated that G373T and A313G loci showed the weakest LD. Seven main diplotypes from the four main reconstructed haplotypes were observed, indicating a significant association of diplotypes with egg weight. Quails with the h1h2 (GGGT) diplotype always exhibited the smallest egg weight and largest egg number at 20 weeks of age. The overall results suggest that the alterations in quails may be linked with potential major loci or genes affecting reproductive traits.
Assuntos
Haplótipos , Oviparidade/genética , Polimorfismo de Nucleotídeo Único , Receptores Tipo I de Polipeptídeo Intestinal Vasoativo/genética , Animais , Genótipo , Desequilíbrio de Ligação , CodornizRESUMO
The Sichuan White goose is a Chinese breed well known for the quality of its meat. However, reproductive performance in this goose is not ideal, and little information is available regarding the abundance of transcripts. To better understand the molecular mechanism(s) underpinning prelaying and laying periods in the Sichuan white goose, high-throughput RNA sequencing was performed to analyze the transcriptome in the hypothalamus. After sequencing and annotation, 26,921 unigenes were obtained, with 48 transcripts up-regulated in the prelaying period and 180 transcripts up-regulated during the laying period. These transcripts were primarily related to diseases, cancers, signaling molecules and interactions, the nervous system and the immune system. Eight transcripts were selected for further analyses with quantitative reverse-transcription polymerase chain reaction (qRT-PCR). The qRT-PCR results were mostly consistent with those from the high-throughput RNA sequencing. Among these transcripts, serine/threonine-protein kinase (AMPK), heat shock protein 70 (HSP70) and NADH dehydrogenase 1 (ND1) were differentially expressed during the prelaying and laying periods. The results of this study provide a useful resource for future studies examining the hypothalamus in geese.
Assuntos
Gansos/genética , Gansos/fisiologia , Perfilação da Expressão Gênica , Hipotálamo/fisiologia , Oviparidade/genética , Transcriptoma/genética , Animais , Complexo I de Transporte de Elétrons/genética , Complexo I de Transporte de Elétrons/metabolismo , Feminino , Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento/genética , Proteínas de Choque Térmico HSP70/genética , Proteínas de Choque Térmico HSP70/metabolismo , Sequenciamento de Nucleotídeos em Larga Escala , Proteínas Serina-Treonina Quinases/genética , Proteínas Serina-Treonina Quinases/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
The brine shrimp, Artemia, reproduces either oviparously, producing encysted embryos (diapause cysts), or ovoviviparously, producing free-swimming nauplii. Environmental factors, such as photoperiod, have been applied to control the reproduction mode of Artemia, but when the determination of a reproductive mode occurs remains unknown. We analysed the differential gene expression between oocytes from oviparous and ovoviviparous Artemia reared under different photoperiods. A total of 692 qualified cDNA clones were obtained by subtractive hybridization, 327 of which matched GenBank® Nucleotide Sequence Database entries. Gene expressions of 44 cDNAs (representing 56 clones) were analysed in oocytes using real-time PCR. Among these genes, 11 (21 clones) were significantly (P<0.05) up-regulated and 7 (9 clones) down-regulated in Artemia oocytes that subsequently enter diapause. Remarkably, known diapause-related proteins such as ArHsp22 (Artemia heat-shock protein 22) and chitin-binding proteins are found to be already differentially expressed. Furthermore, RNAi (RNA interference) knockdown of a differentially expressed gene, polo-like kinase 1, in oocyte of ovoviviparous Artemia led to the production of white embryos rather than free-swimming nauplii. In summary, our results provide evidence at the molecular level that the reproductive mode of Artemia is already determined at the oocyte stage of their life cycle.