ABSTRACT. Although RNA-Seq is an effective method for identifying and exploring novel functional genes in mammals, it has rarely been applied to study fertility-related genes in the goat. In this study, RNA-Seq was used to screen the estrus ovaries of uniparous and multiparous Anhui white goats (AWGs). In total, 15,890 genes were identified and 2201 of these were found to be differentially expressed between the genetic libraries from uniparous and multiparous goats. Compared to the uniparous library, 1583 genes were upregulated and 618 genes were down-regulated in the multiparous library. The FER1L4 gene showed the level of highest up-regulation in the multiparous library, while SRD5A2 expression showed the greatest down-regulation. 18744©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 14 (4): 18743-18752 (2015) In order to determine the functions of FER1L4 and SRD5A2 in goats, the expression profiles of the two genes in different tissues from AWGs and Boer goats at diestrus were analyzed by quantitative PCR. FER1L4 and SRD5A2 showed tissue specific expression patterns and were highly expressed in ovaries from both AWGs and Boer goats. FER1L4 was more highly expressed in ovaries from multiparous than uniparous AWGs. In contrast, SRD5A2 was expressed at a lower level in multiparous AWGs. These results indicated that FER1L4 and SRD5A2 may be associated with the high fecundity of AWGs.

Background A multitude of genes tightly regulate ovarian follicular development and hormone secretion. These complex and coordinated biological processes are altered during pregnancy. In order to further understand the regulatory role of these genes during pregnancy, it is important to screen the differentially expressed genes (DEGs) in the ovaries of pregnant and nonpregnant mammals. To detect the genes associated with the development of pregnancy in goats, DEGs from the ovaries from pregnant and nonpregnant Anhui white goats (pAWGs and nAWGs, respectively) were analyzed using RNA sequencing technology (RNA-Seq). Results In this study, 13,676,394 and 13,549,560 clean reads were generated from pAWGs and nAWGs, respectively, and 1724 DEGs were identified between the two libraries. Compared with nAWGs, 1033 genes were upregulated and 691 genes were downregulated in pAWGs, including PGR , PRLR , STAR and CYP19A1 , which play important roles in goat reproduction. Gene Ontology analysis showed that the DEGs were enriched for 49 functional GO terms. Kyoto Encyclopedia of Genes and Genomes analysis revealed that 397 DEGs were significantly enriched in 13 pathways, including “cell cycle”, “cytokine–cytokine receptor interaction” and “steroid biosynthesis”, suggesting that the genes may be associated with cell cycle regulation, follicular development and hormone secretion to regulate the reproduction process. Additionally, quantitative real-time PCR was used to verify the reliability of the RNA-Seq data. Conclusions The data obtained in this work enrich the genetic resources of goat and provide a further understanding of the complex molecular regulatory mechanisms occurring during the development of pregnancy and reproduction in goats. The DEGs screened in this study may play an important role in follicular development and hormone secretion and they would provide scientific basis for related research in the future. Electronic supplementary material The online version of this article (10.1186/s40709-019-0095-9) contains supplementary material, which is available to authorized users.

Pregnancy is strictly regulated by neuronal and hormonal factors with an essential role being played by the pituitary gland. We screened for differentially expressed genes (DEGs) in the pituitary that function in goat gestational development. Pregnant (AWGp) and non-pregnant Anhui white goats (AWGn) were analysed by deep-sequencing technology. A total of 12 774 092 and 13 872 327 clear reads were obtained in the AWGp and AWGn libraries, respectively. A total of 2593 genes were labelled as significantly differentially expressed in AWGp compared to AWGn, including 2158 upregulated genes and 435 downregulated genes. These genes included follicle stimulating hormone beta (FSHB) and luteinizing hormone beta (LHB), which showed an involvement in reproductive regulation and downregulation (AWGp vs AWGn). Quantitative real-time PCR (qPCR) results validated the DEG data. Subsequent gene ontology analysis indicated that a large number of these DEGs function in cellular processes, cell structures, and cell binding. The DEGs were also found by Kyoto Gene and Genomic Encyclopaedia analysis to be significantly enriched in 54 pathways, including the GnRH and TGF-beta signalling pathways that affect cell proliferation and hormone secretion. These data also identify genes that may play a role in pregnancy and reproduction in the goat and thus provide avenues for future research.

The nucleotide excision repair (NER) system removes pyrimidine (6-4) pyrimidone photoproducts [(6-4)PPs] and cyclobutane pyrimidine dimers (CPDs) from DNA in the form of small (w30-nt-long), excised, damage-containing DNA oligonucleotides (sedDNAs). However, the detection of UV photoproduct-containing sedDNAs as a biochemical read-out for NER activity has thus far been limited to defined in vitro cell-free systems and to cultured cells. Using small punch biopsies of human skin, we show here that the sedDNA products of NER can be isolated, detected, and quantified in skin epidermis within minutes of UVB exposure and following low, sub-erythemal doses of UVB. Interestingly, a partially degraded form of the sedDNAs can be detected in human skin epidermis for several hours following UVB irradiation, which indicates that sedDNA degradation occurs slowly following their release from epidermal genomic DNA. Treatment of skin explants with spironolactone, which depletes the epidermis of the essential NER protein XPB (xeroderma pigmentosum group B), inhibits sedDNA generation following UVB exposure. Our results indicate that the detection of sedDNAs offers a sensitive and quantitative assay for nucleotide excision repair activity in human skin samples and may therefore be useful for determining how genetic, environmental, and pharmacological factors contribute to inter-individual differences in NER activity in human skin.

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