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Characterization of Testicular Angiotensin-Converting Enzyme before and after Semen Cryopreservation and in the Acrosome Reaction of Spermatozoids of Nelore Bulls

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Agricultural Sciences 2019年第4期10卷 518-529页

作者： Ricardo G. Almeida Fabio J. C. Faria Fernando H. G. Furtado Elisvania F. Santos Deiler S. Costa lab of assisted reproduction-famez Federal University of Mato Grosso do Sul Campo Grande Brazil Institute of Biology-INBIO Federal University of Mato Grosso do Sul Campo Grande Brazil

The aim of this study was to characterize the testicular isoform of angio-tensin-converting enzyme (tACE) before and after semen cryopreservation, and in the acrosome reaction of sperm from Nelore bulls in vitro. Ejaculates of 10 sexually mature Nelore bulls were used. After semen was collected, 1.0 mL of the ejaculate was used for the analysis and the rest was subjected to cryopreservation. Fresh semen before freezing, and frozen/thawed semen were centrifuged twice and the pellet was resuspended intyrode’s albumin lactate pyruvate (TALP). Thereafter, 100 μL aliquots containing 100 × 106 spermatozoa were prepared. Aliquots of samples were used for western blot analysis, subjected to capacitation, and thereafter, acrosome reaction assays were performed in vitro. With the help of an anti-ACE monoclonal antibody, a 100 kDa protein band was identified in the spermatozoa of Nelore bulls. Cryopreservation reduced the intensity of the protein bands obtained by western blot assay to less than half of that observed prior to freezing (P P < 0.05), indicating the involvement of ACE in these processes. It is concluded that tACE can be found in the spermatozoa of Nelore bulls, and cryopreservation process decreases the intensity of bands of this enzyme;and that the inactivation of tACE reduces the capacity of spermatozoa to undergo the acrosome reaction.

关键词： Bovine Semen Proteomic

Hormone-like activities of Kuntai capsule in the uteri of ovariectomized rats and immature rabbits

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Journal of Traditional Chinese Medicine 2018年第6期38卷 853-861页

作者： Zhong Ruihua Xie Shuwu Li Guoting Yang Wenjie Guo Xiangjie Li Zhao Zhou Jieyun Ma Aying Zhu Yan laboratory of Reproductive Pharmacology National Health Commision Key Lab of Reproduction RegulationShanghai Institute of Planned Parenthood Research Fudan University

OBJECTIVE: To investigate the hormone-like activities of Kuntai capsule(KTC) in the uteri of ovariectomized rats and immature ***: Following bilateral ovariectomy, rats were randomly divided into six groups including sham group, control group, estradiol valerate group, KTC 0.24, 0.6, and 1.5 g/kg groups. The rats were treated with 0.5% CMC-Na, estradiol valerate and KTC(0.24, 0.6, and 1.5 g/kg), respectively for 28 consecutive days. Then the estrous cycle, uterine changes and pathological changes were *** levels of estradiol(E2), and progesterone(P4)were measured by enzyme-linked immunosorbent assay(ELISA). Protein levels of estradiol receptor(ER), progesterone receptor(PR), vascular endothelial growth factor(VEGF), proliferating cell nuclear antigen(PCNA) and nuclear-associated antigen-67(Ki-67) in uterine tissues were detected by western blot. Immature rabbits were estrogen-primed prior to intragastric administration with KTC for 6 d consecutively. Then, the uteri underwent hematoxylin-eosin staining to observe endometrial ***: Compared with the control group, 0.6and 1.5 g/kg KTC markedly decreased the uterine organ coefficient and endometrial thickness(P <0.05). The serum level of P4 was increased in the KTC 0.6 g/kg group(P < 0.05). There were no significant variations in the serum level of E2 in the KTC groups compared with the control group. ERβ, but not ERα, was markedly upregulated after KTC administration(P < 0.05). Furthermore, 1.5 g/kg KTC significantly decreased the protein level of PRA(P <0.05) and 0.6 g/kg KTC increased the protein level of PRB in the uteri(P < 0.05). VEGF was highly expressed after treatment with 0.24 and 0.6 g/kg KTC,and Ki-67 was markedly reduced in ovariectomized rats treated with 1.5 g/kg KTC. No difference was found in the expression of PCNA. KTC 0.24 and0.6 g/kg promoted endometrial transformation in immature rabbit ***: KTC does not demonstrate obvious estrogen-like effect

关键词： Kuntai capsule Ovariectomiy Endometritis Receptors,progesterone Receptors,estradiol Vascular endothelial growth factors Allen-Doisy bioassay McPhail score

Prohibitin(PHB)interacts with AKT in mitochondria to coordinately modulate sperm motility

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Asian Journal of Andrology 2020年第6期22卷 583-589页

作者： Xiao-Hui Li Ran-Ran Chai Guo-Wu Chen Ling-Fei Zhang Wen-Jing Tan-Tai Hui-Juan Shi Patricia A Martin-DeLeon Wai-Sum O Hong Chen Department of Anatomy Histology and EmbryologyKey Laboratory of Medical Imaging Computing and Computer Assisted Intervention of ShanghaiSchool of Basic Medical SciencesShanghai Medical CollegeFudan UniversityShanghai 200032China Shanghai Ji'Ai Genetics and IVF Institute Obstetrics and Gynecology HospitalInstitute of Reproduction and DevelopmentFudan UniversityShanghai 200011China Key lab of reproduction Regulation of NPFPC-Shanghai Institute of Planned Parenthood Research Fudan University Reproduction and Development InstitutionShanghai 200032China Department of Biological Sciences University of DelawareNewarkDE 19716-2590USA School of Biomedical Sciences The University of Hong KongHong Kong SARChina

Prohibitin(PHB),an evoluti on arily con served mitochondrial inner membra ne protein,is highly expressed in cells that require strong mitoch on drial ***,we dem on strated that the deleti on of Phb in spermatocytes results in impaired mitochondrial *** addition,PHB expression in the mitochondrial sheath of human sperm has a significantly negative correlation with mitochondrial reactive oxygen species levels,but a positive one with mitochondrial membrane potential and sperm *** results suggest that mitochondrial PHB expression plays a role in sperm ***,the mechanism of PHB-mediated regulation of sperm motility remai ns unk ***,we dem on strate for the first time that PHB interacts with protei n kinase B(AKT)and exists in a complex with phospho-PHB(pT258)and phospho-AKT in the mitochondrial sheath of murine sperm,as determined using colocalization and coimmunoprecipitation *** blocking AKT activity using wortmannin(a phosphatidylinositol 3-kinase[PI3K]inhibitor),murine sperm have significantly(P<0.05)decreased levels of phospho-PHB(pT258)and the total and progressive ***,significantly(P<0.05)lower levels of phospho-PI3K P85 subunit a+γ(pY199 and pY46)and phospho-AKT(pS473;pT308)are found in sperm from infertile asthenospermic and oligoasthenospermic men compared with no rmospermic subjects,which suggest a reduced activity of the PI3K/AKT pathway in these in fertile ***,these sperm from infertile subjects also have a significantly(P<0.05)lower level of phospho-PHB(pT258).Collectively,our findings suggest that the interaction of PHB with AKT in the mitochondrial sheath is critical for sperm motility,where PHB phosphorylation(pT258)level and PI3K/AKT activity are key regulatory factors.

关键词： male in fertility prohibitin(PHB) protein kinase B(AKT) sperm motility

An efficient and rapid method to detect and verify natural antisense transcripts of animal genes

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Journal of Integrative Agriculture 2016年第9期15卷 2070-2076页

作者： ZHANG Li ZHAO Rui XIAO Mei LIN Shu-dai LI Bi-xiao QIU Feng-fang MA Jing-e ZHANG De-xiang NIE Qing-hua AN Li-long ZHANG Xi-quan Agricultural College Guangdong Ocean University Guangdong Provincial Key lab of Agro-Animal Genomics and Molecular Breeding/Key lab of Chicken Genetics Breeding and Reproduction Ministry of Agriculture/South China Agricultural University

High-throughput sequencing has identified a large number of sense-antisense transcriptional pairs, which indicates that these genes were transcribed from both directions. Recent reports have demonstrated that many antisense RNAs, especially lnc RNA（long non-coding RNA）, can interact with the sense RNA by forming an RNA duplex. Many methods, such as RNA-sequencing, Northern blotting, RNase protection assays and strand-specific PCR, can be used to detect the antisense transcript and gene transcriptional orientation. However, the applications of these methods have been constrained, to some extent, because of the high cost, difficult operation or inaccuracy, especially regarding the analysis of substantial amounts of data. Thus, we developed an easy method to detect and validate these complicated RNAs. We primarily took advantage of the strand specificity of RT-PCR and the single-strand specificity of S1 endonuclease to analyze sense and antisense transcripts. Four known genes, including mouse β-actin and Tsix（Xist antisense RNA）, chicken LXN（latexin） and GFM1（Gelongation factor, mitochondrial 1）, were used to establish the method. These four genes were well studied and transcribed from positive strand, negative strand or both strands of DNA, respectively, which represented all possible cases. The results indicated that the method can easily distinguish sense, antisense and sense-antisense transcriptional pairs. In addition, it can be used to verify the results of high-throughput sequencing, as well as to analyze the regulatory mechanisms between RNAs. This method can improve the accuracy of detection and can be mainly used in analyzing single gene and was low cost.

关键词： natural antisense transcripts transcription orientation detection method RNA sequencing long non-coding RNA

Determination of Residual Feed Intake and Its Associations with Single Nucleotide Polymorphism in Chickens

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Journal of Integrative Agriculture 2014年第1期13卷 148-157页

作者： XU Zhen-qiang CHEN Jie ZHANG Yan JI Cong-liang ZHANG De-xiang ZHANG Xi-quan Guangdong Provincial Key lab of Agro-Animal Genomics and Molecular Breeding/Key lab of Chicken Genetics Breeding and ReproductionMinistry of Agriculture/South China Agricultural University Wens Nanfang Poultry Breeding Co.Ltd.

Marker assisted selection （MAS） for residual feed intake （RFI） is considered to be one of the powerful means to improve feed conversion efficiency, and therefore reduce production costs. To test the inner relationship among body compositions, growth traits and RFI, four models were proposed to assess the extensively explanatory variables accounting for partial variables in feed intake besides metabolic body weight and growth rate. As a result, the original model （Koch＇s model） had the lowest R2 （80.78%） and the highest Bayesian information criterion （1 323.3） value among the four models. Moreover, the effects on RFI caused by single nucleotide polymorphisms （SNPs） were assessed in this study. Twelve SNPs from 7 candidate genes were genotyped in 2 Chinese native strains, rs14743490 of RPLP2 gene showed suggestively significant association with initial body weight in both strains （P〈0.10）. rs15047274 of TAF15 was significantly associated with growth weight, final weight, and feed intake （P〈0.05） in N301 strain, in contrast, it was only suggestively significant associated with feed intake （P〈0.10） in N414 strain, rs15869967 was significantly associated with RFI in N414 strain but not in N301 strain. This study has identified potential genetic markers suitable for MAS in improving the above mentioned traits, but these associations need to be rectified in other larger populations in future.

关键词： RFI model SNPs growth traits association

The genetic regulation of skeletal muscle development:insights from chicken studies

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Frontiers of Agricultural Science and Engineering 2017年第3期4卷 295-304页

作者： Wen LUO Bahareldin A.ABDALLA Qinghua NIE Xiquan ZHANG Department of Animal Genetics Breeding and ReproductionCollege of Animal ScienceSouth China Agricultural University Guangdong Provincial Key lab of Agro-Animal Genomics and Molecular Breeding and Key Lab of Chicken GeneticsBreeding and ReproductionMinistry of AgricultureSouth China Agricultural University

Skeletal muscle development is a complex multi-process trait regulated by various genetic *** chicken embryo is an ideal model system for studying skeletal muscle development. However, only a small proportion of the genetic factors affecting skeletal muscle development have been identified in chicken. The aim of this review is to summarize recent knowledge about the genetic factors involved in the regulation of skeletal muscle development in the chicken, such as gene polymorphisms, epigenetic modification, noncoding RNAs and transcription factors, which can influence skeletal muscle development at the genome, epigenome,transcriptome and proteome levels. Research on the regulation of skeletal muscle development in chicken is not yet comprehensive and most of the candidate genes and single nucleotide polymorphisms related to chicken muscle growth remain to be verified in experimental studies. In addition, the data derived from transcriptome sequencing and genome-wide association studies still require further investigation and analysis and comprehensive studies on the regulation of chicken skeletal muscle development will continue as a major research focus.

关键词： chicken epigenetic modification miRNAs skeletal muscle development SNP transcription factor

The SNPs C.513A>T in the MHC B-F gene and rs15001532 in the SPOCK1 gene are associated with Salmonella pullorum disease resistance in chickens

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Journal of Integrative Agriculture 2016年第8期15卷 1856-1862页

作者： ZHANG Ze-tang GAN Jian-kang ZHANG Wen-wu ZHANG De-xiang ZHANG Xi-quan LUO Qing-bin College of Animal Science South China Agricultural University Key lab of Chicken Genetics Breeding and Reproduction Ministry of Agriculture Xinguang Agriculture & Animal Husbandry Co. Ltd.

supported by the Earmarked Fund for the Modern Agroindustry Technology Research System, China （CARS-41）;the National High Technology Research and Development Program of China （2011AA100301）

关键词： MHC B-F gene SPOCK1 gene SNPs Salmonella pullorum partridge chicken

m^(6)A demethylase FTO regulate CTNNB1 to promote adipogenesis of chicken preadipocyte

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Journal of Animal Science and Biotechnology 2023年第2期14卷 578-592页

作者： Kan Li Weichen Huang Zhijun Wang Qinghua Nie Department of Animal Genetics Breeding and ReproductionCollege of Animal ScienceSouth China Agricultural UniversityGuangzhou 510642China National‑Local Joint Engineering Research Center for Livestock Breeding Guangdong Provincial Key Lab of Agro-Animal Genomics and Molecular Breedingand Key Laboratory of Chicken GeneticsBreeding and ReproductionMinistry of AgricultureGuangzhou 510642China

Background:N6-methyladenosine(m^(6)A)is an abundant post-transcriptional RNA modification that affects various biological *** fat mass and obesity-associated(FTO)protein,a demethylase encoded by the FTO gene,has been found to regulate adipocyte development in an m^(6)A-dependent manner in multiple ***,the effects of the m^(6)A methylation and FTO demethylation functions on chicken adipogenesis remain *** study aims to explore the association between m^(6)A modification and chicken adipogenesis and the underlying mechanism by which FTO affects chicken preadipocyte ***:The association between m^(6)A modification and chicken lipogenesis was assessed by treating chicken pread-ipocytes with different doses of methyl donor betaine and methylation inhibitor *** results showed that betaine significantly increased methylation levels and inhibited lipogenesis,and the inverse effect was found in preadipocytes after cycloleucine *** of FTO significantly inhibited m^(6)A levels and promoted proliferation and differentiation of chicken *** FTO showed opposite ***,FTO overexpression increased the expression of catenin beta 1(CTNNB1)by improving RNA stability in an m^(6)A-dependent manner,and we proved that FTO could directly target ***,CTNNB1 may be a positive regulator of adipogenesis in chicken ***:m^(6)A methylation of RNA was negatively associated with adipogenesis of chicken *** could regulate CTNNB1 expression in a demethylation manner to promote lipogenesis.

关键词： Adipogenesis Chicken CTNNB1 FTO m^(6)A

Transcriptional response of porcine skeletal muscle to feeding a linseed-enriched diet to growing pigs

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Journal of Animal Science and Biotechnology 2016年第2期7卷 238-247页

作者： Hongkui Wei Yuanfei Zhou Shuzhong Jiang Feiruo Huang Jian Peng Siwen Jiang Department of Animal Nutrition and Feed Science College of Animal Science and Technology Huazhong Agricultural University Key lab of Agricultural Animal Genetics Breeding and Reproduction of Ministry of Education & Key Lab of Swine Genetics and Breeding of Ministry of Agriculture Huazhong Agricultural University

Background： To investigate the effect of feeding a linseed-enriched diet to growing-finishing pigs on gene expression in skeletal muscle, pigs were fed with a linseed-enriched diet for 0, 30, 60 and 90 d. Transcriptional profiles of longissimus dorsi muscle were measured using Affymetrix ***： Results showed that 264 genes were identified as differentially expressed genes（DEGs）. The strongest transcriptional response was clearly observed at 30 d. DEGs were assigned to several main functional terms,including transcription, apoptosis, intracellular receptor-mediated signaling, muscle organ development, fatty acid metabolic process, cell motion, regulation of glucose metabolic process, spermatogenesis and regulation of myeloid cell differentiation. We also found that transcriptional changs of several transcription cofactors might contribute to n-3 PUFAs regulated gene expression. In addition, the increased expression of IGF-1, insulin signaling pathway and the metabolism of amino acids might involve in the muscle growth induced by feeding a linseedenriched diet. The results also provide the new evidence that the expression changes of PTPN1, HK2 and PGC-1αmight contribute to the regulation of insulin sensitivity by n-3 ***： Our finding provided correlative evidence that feeding the linseed enriched diet affact expression of genes involved in insulin signaling pathway and the metabolism of amino acids.

关键词： Linseed m RNA expression n-3 PUFAs Skeletal muscle