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Cao Z.,Anhui Agricultural University | Wu R.,Anhui Agricultural University | Song D.,Anhui Agricultural University | Han F.,Anhui Agricultural University | And 6 more authors.
Journal of Animal and Plant Sciences | Year: 2014

The present study was designed to examine whether the frozen storage of porcine zygote medium (PZM) with 3 mg/ml of BSA was feasible for culturing porcine embryos. In experiment 1, the effect of PZM3 that was frozen and stored within one week on the in vitro developmental competence of porcine parthenotes was evaluated. PZM3 that was stored at 4°C served as control. The results show that the cleavage (83.9±1.2% vs. 84.5±1.8%, P> 0.05) and blastocyst (65.2±2.1% vs. 63.1±3.8%, P>0.05) rates are similar between frozen-warmed PZM3 and the control. In addition, the total cell numbers per blastocyst (50±7 cells vs. 47±5 cells, P>0.05) were similar between the groups. In experiment 2, we tested whether PZM3 in frozen storage for 5 months was able to support the in vitro development of parthenotes similar to that supported by freshly made PZM3. The results show that no statistical differences were observed although the cleavage (97.8±2.7% vs. 90.7 ±3.1%, P>0.05) and blastocyst (75.4 ±1.6% vs. 65.1±2.3,P>0.05) rates in the control group were slightly higher than that in the test group. Also, we found no significant differences in the total cell numbers per blastocyst (48±7 cells vs. 46±6 cells, P>0.05) between the groups. In experiment 3, the effect of PZM3 frozen storage on the pre-implantation development of porcine-cloned embryos was investigated. Our results indicate no significant differences in the rates of cleavage (71.5±5.1% vs. 78.1±1.9%, P>0.05) and blastocyst formation (34.6±7.6% vs. 38.2±3.5%, P>0.05) as well as the total cell numbers per blastocyst (40±11 vs. 48±9, P>0.05) between the test and control groups. Taken together, our results imply that the frozen storage of PZM3 is feasible and of practical value for culturing parthenote and cloned porcine embryos. © 2014, Pakistan Agricultural Scientists Forum. All rights reserved. Source


Ling Y.-H.,Anhui Agricultural University | Ling Y.-H.,Anhui Provincial Laboratory of Local Animal Genetic Resources Conservation and Biobreeding | Ren C.-H.,Anhui Agricultural University | Ren C.-H.,Anhui Provincial Laboratory of Local Animal Genetic Resources Conservation and Biobreeding | And 13 more authors.
BMC Genomics | Year: 2014

Background: Superior kidding rate is an important economic trait in production of meat goat, and ovulation rate is the precondition of kidding rate. MicroRNAs (miRNAs) play critical roles in almost all ovarian biological processes, including folliculogenesis, follicle development, follicle atresia, luteal development and regression. To find out the different ovarian activity and follicle recruitment with miRNA-mediated posttranscriptional regulation, the small RNAs expressed pattern in the ovarian tissues of multiple and uniparous Anhui White goats during follicular phase was analyzed using Solexa sequencing data.Results: 1008 miRNAs co-expressed, 309 and 433 miRNAs specifically expressed in the ovaries of multiple and uniparous goats during follicular phase were identified. The 10 most highly expressed miRNAs in the multiple library were also the highest expressed in the uniparous library, and there were no significantly different between each other. The highest specific expressed miRNA in the multiple library was miR-29c, and the one in the uniparous library was miR-6406. 35 novel miRNAs were predicted in total. GO annotation and KEGG Pathway analyses were implemented on target genes of all miRNA in two libraries. RT-PCR was applied to detect the expression level of 5 randomly selected miRNAs in multiple and uniparous hircine ovaries, and the results were consistent with the Solexa sequencing data.Conclusions: In the present study, the different expression of miRNAs in the ovaries of multiple and uniparous goats during follicular phase were characterized and investigated using deep sequencing technology. The result will help to further understand the role of miRNAs in kidding rate regulation and also may help to identify miRNAs which could be potentially used to increase hircine ovulation rate and kidding rate in the future. © 2014 Ling et al.; licensee BioMed Central Ltd. Source


Ling Y.H.,Anhui Agricultural University | Ling Y.H.,Anhui Provincial Laboratory of Local Animal Genetic Resources Conservation and Biobreeding | Guo X.F.,Anhui Agricultural University | Guo X.F.,Innovation Farm | And 11 more authors.
Animal Reproduction Science | Year: 2016

Ovarian activity, which is mainly controlled by follicle-stimulating hormone and luteinizing hormone, is vital to successful reproduction and maintaining reproductive efficiency in livestock. To determine if the regulation of follicular-luteal transition occurs at the post-transcriptional level in hircine ovaries, the expression patterns of small RNAs in the ovarian tissues of Anhui white goats in the follicular and luteal phases were analyzed using Solexa sequencing. In total, 1039 miRNAs were co-expressed in the two libraries, and 278 and 469 miRNAs were specifically expressed in the hircine ovaries during the follicular and luteal phases, respectively. A total of 43 potential novel miRNAs were predicted in the two libraries. GO annotation and KEGG pathway analysis were applied to analyze the target genes of all miRNAs predicted in the two libraries. The highly and differentially expressed miRNAs included miR-26-5p, miR-145-5p, miR-145, miR-145a-5p, miR-125a-5p, miR-320d, and miR-320c, which may participate in follicular-luteal transition. Five co-expressed miRNAs, of which 2 were differentially expressed between the two libraries, were randomly selected to validate the expression pattern using RT-PCR, and the results were consistent with the Solexa sequencing data. Our present results help to clarify the roles of miRNAs in the regulation of follicular-luteal transition in goat ovaries, which may further enhance the reproductive efficiency of commercially important animals in the future. © 2016 Elsevier B.V. Source


Zhang X.-D.,Anhui Provincial Laboratory of Local Animal Genetic Resources Conservation and Biobreeding | Zhang X.-D.,Anhui Agricultural University | Zhang Y.-H.,Anhui Provincial Laboratory of Local Animal Genetic Resources Conservation and Biobreeding | Zhang Y.-H.,Anhui Agricultural University | And 12 more authors.
BMC Genomics | Year: 2013

Background: Ovarian follicular development and hormone secretion are complex and coordinated biological processes which will usually be altered during pregnancy. Ovarian function is tightly regulated by a multitude of genes, and also by some specific miRNAs. It is necessary to identify the differentially expressed miRNAs in the ovaries of pregnant and non-pregnant mammals, in order to further understand the role of miRNA-mediated post-transcriptional regulation in mammalian reproduction. Here, we performed a comprehensive search for hircine miRNAs using two small RNA sequencing libraries prepared from the ovaries of pregnant and non-pregnant goats.Results: 617 conserved and 7 putative novel miRNAs were identified in the hircine ovaries. A total of 471 conserved miRNAs (76.34%) were co-expressed in both pregnant and non-pregnant libraries, and 90 pregnancy-specific and 56 non-pregnancy-specific conserved miRNAs were identified. Additionally, 407 unique miRNAs (65.96%) were significantly differentially expressed in the pregnant and non-pregnant libraries, of which 294 were upregulated and 113 were downregulated in the pregnant library compared to the non-pregnant library. Further analysis showed that miR-143 was predicted to bind to the target sequences of Frizzled-6 and -3 receptor genes in the Wnt/beta-catenin signaling pathway, and let-7b may target the Activin receptor I and Smad 2/3 genes in the TGF-beta signaling pathway. The expression level of 5 randomly selected miRNAs were analyzed by quantitative real-time PCR (q-PCR), and the results demonstrated that the expression patterns were consistent with the Solexa sequencing results. Conclusions: The identification and characterization of differentially expressed miRNAs in the ovaries of pregnant and non-pregnant goats provides important information on the role of miRNA in the regulation of the ovarian development and function. This data will be helpful to facilitate studies on the regulation of miRNAs during mammalian reproduction. © 2013 Zhang et al; licensee BioMed Central Ltd. Source

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