Huabin Zhu

Northwest A&F University

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Xueming Zhao

Zhejiang Chinese Medical University

Shanjiang Zhao

Chinese Academy of Agricultural Sciences

Weihua Du

Institute of Animal Sciences

Yunwei Pang

Chinese Academy of Agricultural Sciences

Haisheng Hao

Chinese Academy of Agricultural Sciences

Huiying Zou

Chinese Academy of Agricultural Sciences

Hai‐Sheng Hao

Chinese Academy of Agricultural Sciences

Nuo Heng

Chinese Academy of Agricultural Sciences

Zhihui Hu

Chinese Academy of Medical Sciences & Peking Union Medical College

Bahlibi Weldegebriall Sahlu

Chinese Academy of Agricultural Sciences

Cooperative Institutions

Chinese Academy of Agricultural Sciences

Ministry of Agriculture and Rural Affairs

Institute of Animal Sciences

China Agricultural University

Chinese Academy of Sciences

Northwest A&F University

Nanjing Agricultural University

Feed Research Institute

Chinese Academy of Medical Sciences & Peking Union Medical College

Peking University

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Long Non-Coding RNA and mRNA Profiling in Early-Stage Bovine Embryos Treated with Glutathione

Antioxidants (2020)

Qinqin Guo Lixin Cui Weijun Sun Li Feng Haisheng Hao

We measured differential expression profiles of genes and long non-coding RNA (lncRNA) using RNA sequencing in bovine embryos with or without glutathione (GSH) treatment. Bovine embryos fertilized in vitro were treated with GSH to blastocyst. Embryos at the 8-16-cell and morula stages were collected, with embryos without GSH treatment as the control. RNA was isolated, amplified, and sequenced. Differentially expressed genes (DEGs) and lncRNAs (DElncRNAs) were identified and bioinformatic analyses carried out. Transcript levels were confirmed using quantitative RT-PCR. A total of 4100 DEGs were identified, of which 3952 were in GSH-treated morulae and 884 in untreated morulae. More gene ontology (GO) terms were associated with GSH treatment than with control conditions. KEGG analysis showed that glutathione metabolism, citrate cycle, and metabolic pathways involving glycine, serine, and threonine were observed only in GSH-treated embryos. Among 4273 DElncRNAs identified, 59 were potentially important in GSH-treated embryo development, including 14 involved in glutathione metabolism. The 59 DElncRNAs co-expressed with protein-coding mRNAs involved similar GO terms and pathways as the DEGs. This appears to be the first comprehensive profiling of DEGs and DElncRNAs in bovine embryos fertilized in vitro with or without GSH, and the first systematic screen of potential lncRNAs in bovine embryos.

KEGG

10.3390/antiox9050402

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Citations (2)

Profiling of miRNAs in neonatal cloned bovines with collapsed lungs and respiratory distress

Reproduction in Domestic Animals (2018)

Zhang Fy Na Yang YF Rao W‐H Du H‐S Hao

Neonatal respiratory distress is a major mortality factor in cloned animals; however, the pathogenesis of this disease has rarely been investigated. Previous studies have shown that miRNAs regulate critical genes related to lung development, cell differentiation, surfactant synthesis, secretion and lung disease. This study aimed to examine differentially expressed miRNAs in collapsed lungs of cloned bovine neonates and normal lungs in order to identify key pathways and functions that might be related to the pathogenesis of neonatal respiratory distress. In this study, miRNA transcriptomes of collapsed lungs of neonatal cloned bovines and normal lungs were analysed by next-generation sequencing and the results were validated using quantitative real-time PCR (qRT-PCR). A total of 177 differentially expressed miRNAs were identified in the two groups (fold change > 2, RPM ≥ 5), some of which were associated with type II cell differentiation, for example, mmu-miR-29a-5p_L-2R+1, hsa-miR-200c-5p_L-1R+1 and mmu-miR-18a-3p_R+1. The differentially expressed miRNAs were predicted to 6,031 target genes. By Gene Ontology (GO) and Kyoto Encyclopeida of Genes and Genomes (KEGG) DATA base, 133 significant GO terms (p < .05) and 13 significant KEGG pathways (p < .05) were obtained. Many of them were associated with lung development and surfactant homoeostasis, such as lipid biosynthetic processes, protein transport, endocytosis, lysosome, endosome, Golgi apparatus and membrane. Our results of miRNAs express profiles may partially explain the respiratory distress and lung collapse in neonatal bovine clones and could provide novel insights into roles of miRNAs in regulation of lung collapse and neonatal respiratory distress in cloned farm animals.

KEGG

Pathogenesis

10.1111/rda.13144

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Mitofusins: from mitochondria to fertility

Cellular and Molecular Life Sciences (2022)

Shanjiang Zhao Nuo Heng Huan Wang Haoyu Wang Haobo Zhang

Germ cell formation and embryonic development require ATP synthesized by mitochondria. The dynamic system of the mitochondria, and in particular, the fusion of mitochondria, are essential for the generation of energy. Mitofusin1 and mitofusin2, the homologues of Fuzzy onions in yeast and Drosophila, are critical regulators of mitochondrial fusion in mammalian cells. Since their discovery mitofusins (Mfns) have been the source of significant interest as key influencers of mitochondrial dynamics, including membrane fusion, mitochondrial distribution, and the interaction with other organelles. Emerging evidence has revealed significant insight into the role of Mfns in germ cell formation and embryonic development, as well as the high incidence of reproductive diseases such as asthenospermia, polycystic ovary syndrome, and gestational diabetes mellitus. Here, we describe the key mechanisms of Mfns in mitochondrial dynamics, focusing particularly on the role of Mfns in the regulation of mammalian fertility, including spermatogenesis, oocyte maturation, and embryonic development. We also highlight the role of Mfns in certain diseases associated with the reproductive system and their potential as therapeutic targets.

10.1007/s00018-022-04386-z

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Long noncoding RNAs: new insights in modulating mammalian spermatogenesis

Journal of Animal Science and Biotechnology/Journal of animal science and biotechnology (2020)

Bahlibi Weldegebriall Sahlu Shanjiang Zhao Xiuge Wang Saqib Umer Huiying Zou

Abstract Spermatogenesis is a complex differentiating developmental process in which undifferentiated spermatogonial germ cells differentiate into spermatocytes, spermatids, and finally, to mature spermatozoa. This multistage developmental process of spermatogenesis involves the expression of many male germ cell-specific long noncoding RNAs (lncRNAs) and highly regulated and specific gene expression. LncRNAs are a recently discovered large class of noncoding cellular transcripts that are still relatively unexplored. Only a few of them have post-meiotic; however, lncRNAs are involved in many cellular biological processes. The expression of lncRNAs is biologically relevant in the highly dynamic and complex program of spermatogenesis and has become a research focus in recent genome studies. This review considers the important roles and novel regulatory functions whereby lncRNAs modulate mammalian spermatogenesis.

10.1186/s40104-019-0424-8

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Long Noncoding RNAs: Recent Insights into Their Role in Male Infertility and Their Potential as Biomarkers and Therapeutic Targets

International Journal of Molecular Sciences (2021)

Shanjiang Zhao Nuo Heng Bahlibi Weldegebriall Sahlu Huan Wang Huabin Zhu

Long noncoding RNAs (lncRNAs) are composed of nucleotides located in the nucleus and cytoplasm; these are transcribed by RNA polymerase II and are greater than 200 nt in length. LncRNAs fulfill important functions in a variety of biological processes, including genome imprinting, cell differentiation, apoptosis, stem cell pluripotency, X chromosome inactivation and nuclear transport. As high throughput sequencing technology develops, a substantial number of lncRNAs have been found to be related to a variety of biological processes, such as development of the testes, maintaining the self-renewal and differentiation of spermatogonial stem cells, and regulating spermatocyte meiosis. These indicate that lncRNAs can be used as biomarkers and potential therapeutic targets for male infertility. However, only a few comprehensive reviews have described the role of lncRNAs in male reproduction. In this paper, we summarize recent findings relating to the role of lncRNAs in spermatogenesis, their potential as biomarkers for male infertility and the relationship between reproductive arrest and transgenerational effects. Finally, we suggest specific targets for the treatment of male infertility from the perspective of lncRNAs.

Spermatocyte

Genomic Imprinting

10.3390/ijms222413579

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Neonatal respiratory distress syndrome and underlying mechanisms in cloned cattle

Molecular Reproduction and Development (2018)

Yifan Rao Huabin Zhu Na Yang Fanyi Zhang Xiaojing Jiang

Neonatal respiratory distress is a major mortality factor in cloned animals, but the pathogenesis of this disease is rarely investigated. In this study, four neonatal cloned cattle, born after full‐term gestation, exhibited symptoms of neonatal respiratory distress syndrome (NRDS), which included symptoms of hyaline membrane disease as well as disordered surfactant homeostasis in their collapsed lungs. No differences in DNA methylation or histone modifications correlated with the suppressed SPB and SPC transcription observed in the cloned cattle group ( p > 0.05), whereas TTF‐1 occupancy at SPB and SPC promoter regions in cloned cattle was significantly reduced to 24% and 20% that of normal lungs, respectively ( SPB , p < 0.05; SPC , p < 0.01). Decreased TTF1 expression, dysregulation of SPB and SPC transcription by TTF‐1, and disordered proteolytic processing of Surfactant protein B precursor together potentially contribute to the disruption of surfactant homeostasis and NRDS in bovine clones. Elucidation of the associated mechanisms should facilitate the development of novel preventive or therapeutic strategies to reduce the mortality rate of cloned animals and to improve the efficiency of SCNT technology.

Homeostasis

Pathogenesis

10.1002/mrd.22956

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Effect of vitrification on promoter methylation and the expression of pluripotency and differentiation genes in mouse blastocysts

Molecular Reproduction and Development (2012)

Xueming Zhao Weihua Du Hai‐Sheng Hao Dong Wang Tong Qin

The present study was designed to determine the effects of vitrification on promoter methylation and the expression levels of pluripotency and differentiation genes in mouse blastocysts. Promoter region CpG methylation patterns and the expression levels of octamer-binding transcription factor (Oct4), Nanog homeobox (Nanog), caudal-type homeobox 2 (Cdx2), and heart and neural crest derivatives-expressed transcript 1 (Hand1) were analyzed in fresh and vitrified mouse blastocysts. Methylation was measured by bisulphate mutagenesis and sequencing; gene expression was determined by real-time reverse transcription-PCR. The results showed that vitrification significantly reduced the methylation levels of the Oct4 (85% vs. 62.5%), Nanog (77.5% vs. 55%), and Cdx2 promoters (4.6% vs. 1.4%; P < 0.05) in mouse blastocysts, which correlated with increased expression of Oct4 and Nanog in vitrified blastocysts. Hand1 promoter methylation was not significantly different in the fresh (17.9%) versus vitrification group (21.4%; P > 0.05). The expression levels of Cdx2 and Hand1 were not significantly different in fresh and vitrified blastocysts. In conclusion, vitrification significantly decreased Oct4, Nanog, and Cdx2 promoter methylation in mouse blastocysts, which correlated with increased expression of Oct4 and Nanog.

Homeobox protein NANOG

CDX2

Bisulfite sequencing

Rex1

Vitrification

DLX5

10.1002/mrd.22052

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Citations (41)

Detection of Insertion/Deletions (InDel) Within Five Clock Genes and Their Associations with Growth Traits in Four Chinese Sheep Breeds

Veterinary Sciences (2025)

Wang Ziteng Xiaohua Yi Mengzhe Yang Huabin Zhu Shuhui Wang

Organisms have the capacity to detect day-night fluctuations through oscillators regulated by circadian clock genes, which are crucial for regulating various biological processes. Numerous studies have demonstrated a marked association between these genes and various growth traits of sheep. This study identified polymorphisms at 23 potential loci within five clock genes in four Chinese sheep breeds. Only two polymorphic insertion/deletions (InDels) were detected in CLOCK and PER3 genes, respectively. The distribution of these two loci in four Chinese sheep breeds and their association with growth traits were further explored. A 12 bp deletion was found in the intron of the CLOCK gene (rs604230640), which was significantly associated with body height (p < 0.05), body oblique length (p < 0.05) and cannon girth (p < 0.05) in Hu sheep (HS). A 22 bp insertion in the intron of the PER3 gene (rs600537720) with a dominant genotype of insertion/insertion (II) was found to have a significant association with chest depth (p < 0.05) in Small-Tail Han sheep (STHS), tail width (p < 0.05) in Tong Sheep (TS), and in Lanzhou fat-tailed sheep (LFTS). In conclusion, this study has elucidated the polymorphisms of CLOCK and PER3 genes and has examined the influence of these two genes on the growth traits of sheep. Concurrently, the two molecular markers identified in CLOCK and PER3 could potentially serve in the marker-assisted selection of growing-related traits in local Chinese sheep breeds.

Indel

INDEL Mutation

10.3390/vetsci12010039

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CREB1 Is Involved in miR-134-5p-Mediated Endometrial Stromal Cell Proliferation, Apoptosis, and Autophagy

Cells (2023)

Xiaodan Li Xiaolei Yao Kang Li Jiahe Guo Kaiping Deng

The successful establishment of endometrial receptivity is a key factor in ensuring the fertility of ewes and their economic benefits. Hu sheep have attracted attention due to their high fecundity and year-round estrus. In this study, we found that in the luteal phase, the uterine gland density, uterine coefficient, and number of uterine caruncles of high-fertility Hu sheep were higher than those of low-fertility Hu sheep. Thousands of differentially expressed genes were identified in the endometrium of Hu sheep with different fertility potential using RNA sequencing (RNA-Seq). Several genes involved in endometrial receptivity were screened using bioinformatics analysis. The qRT-PCR analysis further revealed the differential expression of cAMP reactive element binding protein-1 (CREB1) in the Hu sheep endometrium during the estrous cycle. Functionally, our results suggested that CREB1 significantly affected the expression level of endometrial receptivity marker genes, promoted cell proliferation by facilitating the transition from the G1 phase to the S phase, and inhibited cell apoptosis and autophagy. Moreover, we observed a negative linear correlation between miR-134-5p and CREB1 in the endometrium. In addition, CREB1 overexpression prevented the negative effect of miR-134-5p on endometrial stromal cell (ESC) growth. Taken together, these data indicated that CREB1 was regulated by miR-134-5p and may promote the establishment of uterine receptivity by regulating the function of ESCs. Moreover, this study provides new theoretical references for identifying candidate genes associated with fertility.

CREB1

10.3390/cells12212554

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Citations (2)

Protective effect of vitamin C and lycopene on the in vitro fertilization capacity of sex‐sorted bull sperm by inhibiting the oxidative stress

Reproduction in Domestic Animals (2020)

Peipei Zhang Jingjing Wang Chong‐Yang Li Hai‐Sheng Hao Haoyu Wang

Abstract The fertilization capacity of sex‐sorted sperms is seriously decreased, which inhibits its wide application. However, little information is still available about the effect of vitamin C (VC) and lycopene (Lyc) on the fertilization capacity of sex‐sorted bull sperm. In this study, the washing medium and fertilization medium of sex‐sorted sperm from three bull individuals were supplemented with different concentrations of VC (0, 1 × 10 –3 , 1 × 10 –4 , 1 × 10 –5 , 1 × 10 –6 M) or Lyc (0, 1 × 10 –4 , 1 × 10 –5 , 1 × 10 –6 , 1 × 10 –7 ). After washing twice and incubation for 1.5 hr, the malondialdehyde (MDA) level, phosphatidylserine (PS) translocation, membrane potential (Δψm) and IVF (in vitro fertilization) ability of sex‐sorted sperm were investigated. For the sex‐sorted sperm of bulls A, B and C, 1 × 10 –3 M VC or 1 × 10 –4 M Lyc treatment significantly decreased their MDA levels and PS translocation and increased their Δψm levels and cleavage rates after IVF. When blastocysts were concerned, 1 × 10 –4 M Lyc significantly improved the blastocyst rates and their IFN‐tau expression of bulls A and C. In conclusion, supplementation of 1 × 10 –3 M VC or 1 × 10 –4 M Lyc in washing and fertilization medium contributed greatly to improving the fertilization capacity of sex‐sorted bull sperm during IVF procedure.

Malondialdehyde

10.1111/rda.13748

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Citations (7)