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Jiang S.,Anhui Agricultural University | Hong M.,Anhui Agricultural University | Su S.,Anhui Agricultural University | Song M.,Anhui Agricultural University | And 8 more authors.
Animal Science Journal | Year: 2015

This study was designed to explore the effect of active immunization against maltose binding protein-gonadotropin releasing hormone I hexamer (MBP-GnRH-I6) on the reproductive function in cats. Each immunized cat was administered twice intramuscularly in the neck at 16 and 20 weeks old. The concentrations of the testosterone and estradiol and the level of anti-GnRH-I antibody in the serum were measured by radioimmunoassay and ELISA, respectively. The results showed that the weight and size of testicles and ovaries, and the concentrations of serum testosterone and estradiol in the immunized animals were lower than those of the control cats (P<0.05), but that the levels of anti-GnRH-I antibody were significant higher compared to control animals (P<0.05). Testicular tissues from the immunized male cats showed that seminiferous tubules were depauperate with the lumen relatively empty and that the differentiation of spermatogonia was not obvious. Tissues from the immunized female cats showed that the ovaries had many primordial follicles and primary follicles, but no secondary follicle was observed. These results showed active immunization against MBP-GnRH-I6 could make the gonads atrophy and reduce the concentrations of gonadal hormones, which suggested that MBP-GnRH-I6 was a very effective immunogen in the cat. © 2015 Japanese Society of Animal Science.

Li D.,Anhui Agricultural University | Li D.,Fuyang Teachers College | Li D.,Anhui Provincial Laboratory for Local Livestock and Poultry Genetic Resource Conservation and Bio Breeding | Liu H.,Anhui Agricultural University | And 11 more authors.
Molecular Biology Reports | Year: 2014

Quantitative reverse transcription polymerase chain reaction (qRT-PCR) has become the preferred technique for studying low-abundance RNA expression. Proper normalization is a critical but often underappreciated aspect of quantitative gene expression analysis; popular endogenous control genes are usually selected with little knowledge of their real suitability. To date, there are very few reports regarding the general validation of endogenous control genes for microRNA (miRNA) expression analysis in bovine tissue. In the present study, eight candidate reference genes (U6, 18S rRNA, GAPDH, ACTB, miR-191, miR-15a, miR-18a, let-7f) were tested for use as normalizers of bovine miRNA in RT-qPCR assays. Their selection was based on publicly available data concerning normalization, hierarchical clustering and sequencing. Three of the genes (miR-191, U6-1 and let-7f) were found to be highly consistent in their expression across eight different bovine solid tissues. It is commonly accepted that gene expression studies should be normalized using more than one endogenous control gene. Based on our results, we propose using the combined results for miR-191, U6-1 and let-7f as the endogenous control for normalization of miRNA levels in qRT-PCR analysis of diverse bovine tissues. This result could act a guideline for future work on bovine miRNA expression. © 2014, Springer Science+Business Media Dordrecht.

Hua W.,Anhui Agricultural University | Luo L.,Anhui Agricultural University | Tian Y.,Anhui Agricultural University | Song M.,Anhui Agricultural University | And 7 more authors.
Animal Reproduction Science | Year: 2014

Kisspeptin and neurokinin B (NKB) have various functions. Their expression has been demonstrated in rat and human ovary, and estrogen affects their activity, suggesting a role for these molecules in the control of ovary function. However, whether these signaling systems are present in geese ovary, and the associated with serum estrogen remains largely unexplored. In this study we investigated the expression of kisspeptin and NKB in the ovary, and analysed their changes in the serum during the reproductive cycle and their association with serum estradiol in the geese. The results showed both kisspeptin and NKB immunoreactivity was found in the ovary, with marked expression in the granular layer and theca of the follicle, where intense coexpression of kisspeptin and NKB was also detected. The serum concentrations of kisspeptin and NKB in geese were significantly higher (P< 0.05) in broody period than in laying period and laying cessation stage. However, the level of estradiol was markedly higher (P< 0.05) in laying period than in broody and laying cessation stage. Serum kisspeptin was positively correlated with NKB (r= 0.866, P< 0.001), serum estradiol was negatively correlated with kisspeptin (r= -0.977, P< 0.05) and NKB (r= -0.887, P< 0.05). Overall, the existence of kisspeptin and NKB in geese ovary, and the difference of their serum concentrations during reproductive cycle and the inverse correlation with serum estradiol are highly suggestive of a role for kisspeptin and NKB in the regulation of geese reproductive function. © 2014 Elsevier B.V.

Yang C.,Anhui Agricultural University | Yang C.,Anhui Provincial Laboratory for Local Livestock and Poultry Genetic Resource Conservation and Bio Breeding | Ye J.,Anhui Agricultural University | Ye J.,Anhui Provincial Laboratory for Local Livestock and Poultry Genetic Resource Conservation and Bio Breeding | And 24 more authors.
PLoS ONE | Year: 2016

Female pubertal development is tightly controlled by complex mechanisms, including neuroendocrine and epigenetic regulatory pathways. Specific gene expression patterns can be influenced by DNA methylation changes in the hypothalamus, which can in turn regulate timing of puberty onset. In order to understand the relationship between DNA methylation changes and gene expression patterns in the hypothalamus of pubertal goats, wholegenome bisulfite sequencing and RNA-sequencing analyses were carried out. There was a decline in DNA methylation levels in the hypothalamus during puberty and 268 differentially methylated regions (DMR) in the genome, with differential patterns in different gene regions. There were 1049 genes identified with distinct expression patterns. High levels of DNA methylation were detected in promoters, introns and 3′-untranslated regions (UTRs). Levels of methylation decreased gradually from promoters to 5′-UTRs and increased from 5′-UTRs to introns. Methylation density analysis demonstrated that methylation level variation was consistent with the density in the promoter, exon, intron, 5′-UTRs and 3′-UTRs. Analyses of CpG island (CGI) sites showed that the enriched gene contents were gene bodies, intergenic regions and introns, and these CGI sites were hypermethylated. Our study demonstrated that DNA methylation changes may influence gene expression profiles in the hypothalamus of goats during the onset of puberty, which may provide new insights into the mechanisms involved in pubertal onset. © Copyright 2016 Yang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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