|LIN, SHUDAI - South China Agricultural University|
|FANG, LINGZHAO - University Of Edinburgh|
|KANG, XIAOLONG - Ningxia University|
|LIU, SHULI - China Agricultural University|
|LIU, MEI - Northwest Agricultural & Forestry University|
|Baldwin, Ransom - Randy|
|Liu, Ge - George|
Submitted to: Heliyon
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 5/28/2020
Publication Date: 6/1/2020
Citation: Lin, S., Fang, L., Kang, X., Liu, S., Liu, M., Connor, E.E., Baldwin, R.L., Liu, G., Li, C. 2020. Establishment and transcriptomic analyses of a cattle rumen epithelial primary cells (REPC) culture by bulk and single-cell RNA sequencing to elucidate interactions of butyrate and rumen development. Heliyon. 6(6):e041123. https://doi.org/10.1016/j.heliyon.2020.e04112.
Interpretive Summary: Rumen development after birth is important for the dairy cow. Butyrate is a nutrient for the cow. It is also involved in rumen development. However, the mechanism for this effect was not clear. We established a rumen cell culture from a two-week old calf to mimic the physiological states of cells to investigate the butyrate functions in rumen development. This report elucidate the putative roles and mechanisms of butyrate action in cattle and provides further evidence in support of the contention that butyrate plays a specific and central role in regulating genomic and epigenomic activities influencing rumen development.
Technical Abstract: Ruminant forestomachs are incompletely developed at birth and must fully develop both physically and metabolically prior to weaning. Butyrate, a short chain fatty acid produced as an end product of fermentation is an essential nutrient for cattle, plays a central role in regulating genomic and epigenomic elements which influence rumen development. To study the interaction of butyrate and rumen development, we established a rumen epithelial primary cell (REPC) culture from a two-week-old Holstein bull calf rumen epithelial tissue. Using transcriptomic analysis of REPC using bulk and single-cell RNA sequencing techniques, we tested the direct effects of butyrate addition to the culture media on gene expression and correlated networks identified to elucidate the putative roles and mechanisms of butyrate action in rumen epithelial development. The top networks perturbed by butyrate addition were four positively correlated gene networks (WNT, JNK, NF'B, and MAPK/ERK) predominantly associated with epithelial tissue development. Additionally, two key upstream regulators, E2F1 and TGFB1, were identified that have been previously shown to play critical roles in the differentiation, development, and growth of epithelial cells. Further, significant expression changes of CHGA, ONECUT2, OXT, PAX2, WNT1, WNT4, GRHL2, and ELF3 genes, presumably upregulated by E2F1 and TGFB1, provides further evidence in support of the contention that butyrate plays a specific and central role in regulating genomic and epigenomic activities influencing rumen development.