Combined analysis of mRNA–miRNA from testis tissue in Tibetan sheep with different FecB genotypes

Abstract Testis size is important for identifying breeding animals with adequate sperm production. The aim of this study was to survey the expression profile of mRNA and miRNA in testis tissue from rams carrying different FecB genotypes, including the wild-type and heterozygous genotypes in Tibetan sheep. Comparative transcriptome profiles for ovine testes were established for wild-type and heterozygote Tibetan sheep by next-generation sequencing. RNA-seq results identified 3,910 (2,034 up- and 1,876 downregulated) differentially expressed (DE) genes and 243 (158 up- and 85 downregulated) DE microRNAs (miRNAs) in wild-type vs heterozygote sheep, respectively. Combined analysis of mRNA-seq and miRNA-seq revealed that 20 miRNAs interacted with 48 true DE target genes in wild-type testes compared to heterozygous genotype testes. These results provide evidence for a functional series of genes operating in Tibetan sheep testis. In addition, quantitative real-time PCR analysis showed that the expression trends of randomly selected DE genes in testis tissues from different genotypes were consistent with high-throughput sequencing results.

[1]  M. Ryten,et al.  ggtranscript: an R package for the visualization and interpretation of transcript isoforms using ggplot2 , 2022, bioRxiv.

[2]  Fadi Li,et al.  Identification of Key Genes Related to Postnatal Testicular Development Based on Transcriptomic Data of Testis in Hu Sheep , 2022, Frontiers in Genetics.

[3]  M. Chu,et al.  The expression and mutation of BMPR1B and its association with litter size in small-tail Han sheep (Ovis aries) , 2021, Archives animal breeding.

[4]  yongqing guo,et al.  Polymorphisms in BMPR-IB gene and their association with litter size trait in Chinese Hu sheep , 2020, Animal biotechnology.

[5]  W. Bai,et al.  Correlation analysis of candidate gene SNP for high-yield in Liaoning cashmere goats with litter size and cashmere performance , 2019, Animal biotechnology.

[6]  Junyi Luo,et al.  MiR-125b-2 Knockout in Testis Is Associated with Targeting to the PAP Gene, Mitochondrial Copy Number, and Impaired Sperm Quality , 2019, International journal of molecular sciences.

[7]  Ruiyang Zhao,et al.  MicroRNA-299-3p regulates proliferation, migration and invasion of human ovarian cancer cells by modulating the expression of OCT4. , 2018, Archives of biochemistry and biophysics.

[8]  Wangsheng Zhao,et al.  Differentially expressed microRNAs between cattleyak and yak testis , 2018, Scientific Reports.

[9]  K. Hagiya,et al.  Genetic correlations between yield traits or days open measured in cows and semen production traits measured in bulls. , 2017, Animal : an international journal of animal bioscience.

[10]  S. Armstrong,et al.  miR-99 regulates normal and malignant hematopoietic stem cell self-renewal , 2017, The Journal of experimental medicine.

[11]  Eun-Young Kim,et al.  The miR-125 family is an important regulator of the expression and maintenance of maternal effect genes during preimplantational embryo development , 2016, Open Biology.

[12]  Changchun Li,et al.  Genome-wide differential expression of genes and small RNAs in testis of two different porcine breeds and at two different ages , 2016, Scientific Reports.

[13]  I. Igbokwe,et al.  Testicular and Related Size Evaluations in Nigerian Sahel Goats with Optimal Cauda Epididymal Sperm Reserve , 2015, Veterinary medicine international.

[14]  A. Sosnowska,et al.  Phenotypic Correlations of Testes Size with Semen Traits and the Productive Traits of Young Boars. , 2015, Reproduction in domestic animals = Zuchthygiene.

[15]  Keyi Tang,et al.  Selection of endogenous reference microRNA genes for quantitative reverse transcription polymerase chain reaction studies of boar spermatozoa cryopreservation. , 2015, Theriogenology.

[16]  J. Hua,et al.  CD49f‐Positive Testicular Cells in Saanen Dairy Goat Were Identified as Spermatogonia‐Like Cells by miRNA Profiling Analysis , 2014, Journal of cellular biochemistry.

[17]  Pradeep G. Kumar,et al.  Dramatic Changes in 67 miRNAs During Initiation of First Wave of Spermatogenesis in Mus musculus Testis: Global Regulatory Insights Generated by miRNA-mRNA Network Analysis1 , 2014, Biology of reproduction.

[18]  Paul Theodor Pyl,et al.  HTSeq – A Python framework to work with high-throughput sequencing data , 2014, bioRxiv.

[19]  H. Okano,et al.  Small RNA profiling and characterization of piRNA clusters in the adult testes of the common marmoset, a model primate , 2014, RNA.

[20]  A. Calogero,et al.  Relationship between Testicular Volume and Conventional or Nonconventional Sperm Parameters , 2013, International journal of endocrinology.

[21]  J. Hua,et al.  Expression of miR‐34c in response to overexpression of Boule and Stra8 in dairy goat male germ line stem cells (mGSCs) , 2013, Cell biochemistry and function.

[22]  M. Zavolan,et al.  Analysis of CDS-located miRNA target sites suggests that they can effectively inhibit translation , 2013, Genome research.

[23]  Boyang Liu,et al.  A comparative profile of the microRNA transcriptome in immature and mature porcine testes using Solexa deep sequencing , 2012, The FEBS journal.

[24]  E. C. Mattos,et al.  Additive genetic relationships between scrotal circumference, heifer pregnancy, and stayability in Nellore cattle. , 2010, Journal of animal science.

[25]  K. Loveland,et al.  Activin A Balances Sertoli and Germ Cell Proliferation in the Fetal Mouse Testis1 , 2010, Biology of reproduction.

[26]  M. Gruber,et al.  EPAS1 Is Required for Spermatogenesis in the Postnatal Mouse Testis1 , 2010, Biology of reproduction.

[27]  Donald W. Harms,et al.  Ehd4 is required to attain normal prepubertal testis size but dispensable for fertility in male mice , 2010, Genesis.

[28]  M. Matzuk,et al.  Activin Bioactivity Affects Germ Cell Differentiation in the Postnatal Mouse Testis In Vivo1 , 2010, Biology of reproduction.

[29]  Matthew D. Young,et al.  Gene ontology analysis for RNA-seq: accounting for selection bias , 2010, Genome Biology.

[30]  K. Manova,et al.  Kit ligand cytoplasmic domain is essential for basolateral sorting in vivo and has roles in spermatogenesis and hematopoiesis. , 2010, Developmental biology.

[31]  Hakim Tafer,et al.  RNAplex: a fast tool for RNA-RNA interaction search , 2008, Bioinform..

[32]  H. Yao,et al.  Essential roles of inhibin beta A in mouse epididymal coiling , 2007, Proceedings of the National Academy of Sciences.

[33]  N. Lau,et al.  Characterization of the piRNA Complex from Rat Testes , 2006, Science.

[34]  M. Stoffel,et al.  MicroRNAs: a new class of regulatory genes affecting metabolism. , 2006, Cell metabolism.

[35]  Zuping He,et al.  Expression of Col1a1, Col1a2 and procollagen I in germ cells of immature and adult mouse testis. , 2005, Reproduction.

[36]  P. Shannon,et al.  Cytoscape: A Software Environment for Integrated Models of Biomolecular Interaction Networks , 2003 .

[37]  L. Du,et al.  [Studies of BMPR-IB and BMP15 as candidate genes for fecundity in little tailed han sheep]. , 2003, Yi chuan xue bao = Acta genetica Sinica.

[38]  R. Sharpe,et al.  Proliferation and functional maturation of Sertoli cells, and their relevance to disorders of testis function in adulthood. , 2003, Reproduction.

[39]  G. Spera,et al.  PDGF and the testis , 2002, Trends in Endocrinology & Metabolism.

[40]  Thomas D. Schmittgen,et al.  Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. , 2001, Methods.

[41]  Noi T. Tran,et al.  Characterization of EHD4, an EH Domain-containing Protein Expressed in the Extracellular Matrix* , 2001, The Journal of Biological Chemistry.

[42]  A. Yassen,et al.  Relationship between body weight and testicular size in buffalo bulls , 1972, The Journal of Agricultural Science.

[43]  Chon-Kit Kenneth Chan,et al.  Analysis of RNA-Seq Data Using TopHat and Cufflinks. , 2016, Methods in molecular biology.

[44]  J. Kastelic Understanding and evaluating bovine testes. , 2014, Theriogenology.

[45]  M. Gilbert,et al.  Akt1 protects against germ cell apoptosis in the postnatal mouse testis following lactational exposure to 6-N-propylthiouracil. , 2011, Reproductive toxicology.

[46]  M. Griswold,et al.  Fetal Testis Dysgenesis and Compromised Leydig Cell Function in Tgfbr3 (Betaglycan) Knockout Mice1 , 2010, Biology of reproduction.

[47]  Xuegong Zhang,et al.  DEGseq: an R package for identifying differentially expressed genes from RNA-seq data , 2010, Bioinform..

[48]  Di Ran,et al.  Establishment of molecular detection methods for high prolificacy major gene FecB in sheep and its application. , 2009 .