Abstract #125

Section: Gene Expression
Session: Gene Expression
Format: Poster
Location: Rio Exhibit Hall B
# 125
J. Duan*1, N. K. Jue2, Z. Jiang1, R. O’Neill2, E. Wolf3, L. A. Blomberg4, H. Dong5, X. Zheng5, J. Chen5, X. Tian1, 1Center for Regenerative Biology, Department of Animal Science, University of Connecticut, Storrs, CT, USA;, 2Department of Molecular and Cell Biology, University of Connecticut, Storrs, CT, USA;, 3Laboratory for Functional Genome Analysis, Ludwig-Maximilians-Universität München, Munich, Germany;, 4Animal Biosciences and Biotechnology Laboratory, USDA, Agricultural Research Service, Beltsville, MD, USA;, 5Institute of Animal Science, Xinjiang Academy of Animal Science, Urumqi, Xinjiang, P.R. China.

The maintenance of a proper gene dosage is essential in cellular networks. To resolve the dosage imbalance between eutherian females (XX) and male (XY), X chromosome inactivation (XCI) occurs in females, while X-chromosome dosage compensation up-regulates the active X to balance its expression with that of autosome pairs [Ohno’s hypothesis; Ohno 1967 Sex Chromosomes and Sex-linked Genes (Springer-Verlag), p. 99]. These phenomena have been well studied in humans and mice, despite many controversies over the existence of such up-regulation. Using RNA sequencing data, we determined X chromosome dosage compensation in the bovine by analysing the global expression profiles of germ cells, embryos, and somatic tissues. Eight bovine RNA-seq data sets were obtained in from the Gene Expression Omnibus, covering bovine immature/mature oocytes (GSE59186 and GSE52415), pre-implantation conceptuses (GSE59186, GSE52415, and GSE56513), extra-embryonic tissues (PRJNA229443), and male/female somatic tissues (GSE74076, GSE63509, PRJEB6377, and GSE65125). The RNAseq data were trimmed and non-uniquely (paralogs included) mapped to the bovine reference genome assembly UMD3.1.1 using Hisat2 (version 2.0.5) aligner. The mRNA level of each gene, estimated by transformed transcripts per kilobase million was quantified by IsoEM (version 1.1.5). These RNA-seq data sets represented 4 chromosome scenarios in cells: XXXX:AAAA (diploid immature oocyte with DNA duplication), XX:AA (haploid mature oocyte with DNA duplication), XX:AA and X:AA (gradual changed X status in bovine pre-implantation conceptuses), and X:AA (extra-embryonic tissues and somatic cells in female with one active X or XY male) were analysed for dosage compensation. A total of 959 X-linked genes and 20,316 autosome genes were used to calculate the relative X to autosomal gene (A) expression (RXE): log2 (X expression) – log2 (A expression). The following dosage determinations were made: RXE values ≥ 0: complete dosage compensation (or X: A ratio ≥ 1); RXE values < 0: in-complete dosage compensation; RXE value = −1: no dosage compensation (or X: A ratio = 0.5). Our analyses showed a decreased RXE after fertilization, from −0.33 in matured oocytes to −0.50 at the 2-cell stage, indicating that the sperm that undergo meiotic sex chromosome inactivation (MSCI) bring in inactive X chromosomes to the matured oocytes. Subsequently, the activation of the bovine embryonic genome at the 4- to 8-cell stage increased RXE from −0.54 to −0.05. This was followed by a sharp RXE decline from −0.02 at the 16-cell stage, 0.1 at the 32-cell stage to −0.29 at the compact morula stage, which is known as paternal X inactivation stage in the bovine. Finally, RXE was stabilised from blastocysts −0.19 through the Day 19 conceptuses −0.25 to somatic tissue average −0.21 with a pattern of incomplete X compensation. These findings support X expression up-regulation as proposed by Ohno. No significant RXE differences were observed between bovine female and male somatic tissues, further supporting Ohno’s hypothesis, which predicts a balance in the expression of X-linked genes to that of autosomes. This study confirms Ohno’s hypothesis of X dosage compensation in bovine germ cells, early embryos, and somatic tissues.