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ARS Home » Midwest Area » Madison, Wisconsin » U.S. Dairy Forage Research Center » Cell Wall Biology and Utilization Research » Research » Publications at this Location » Publication #345118
Title: Examination of the xanthosine response on gene expression of mammary epithelial cells using RNA-seq technology

Author
item CHOUDHARY, SHANTI - Guru Angad Dev Veterinary & Animal Sciences University
item Li, Wenli
item Bickhart, Derek
item RAMNEEK, VERM - Guru Angad Dev Veterinary & Animal Sciences University
item SETHI, R - Guru Angad Dev Veterinary & Animal Sciences University
item MUKHOPADHYAY, C - Guru Angad Dev Veterinary & Animal Sciences University
item CHOUDHARY, RATAN - Guru Angad Dev Veterinary & Animal Sciences University

Submitted to: Journal of Animal Science and Technology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/9/2018
Publication Date: 7/13/2018
Citation: Choudhary, S., Li, W., Bickhart, D.M., Ramneek, V., Sethi, R.S., Mukhopadhyay, C.S., Choudhary, R.K. 2018. Examination of the xanthosine response on gene expression of mammary epithelial cells using RNA-seq technology. Journal of Animal Science and Technology. 60. Article 18. https://doi.org/10.1186/s40781-018-0177-5.
DOI: https://doi.org/10.1186/s40781-018-0177-5

Interpretive Summary: Xanthosine treatment was previously reported to increase mammary stem cell population and milk production in both cattle and goats. However, the underlying molecular mechanisms behind this increase in production remain unclear. Using RNA-sequencing technology, we set out to study the genome-wide transcriptome changes in goat mammary glands using milk fat globules upon xanthosine treatment. This study provides valuable insights regarding whole-transcriptome changes in milk-producing cells in response to xanthosine treatment in goats. Further functional follow-up on differentially expressed genes is highly likely to help delineate probable molecular mechanisms of enhanced cell proliferation in goat milk-producing cells through administration of xanthosine.

Technical Abstract: Xanthosine treatment was previously reported to increase mammary stem cell population and milk production in both bovine and caprine. However, the underlying molecular mechanisms behind this increase in production remain unclear. Using whole-transcriptome RNA sequencing technology, the goal of this study was to examine the effects of xanthosine on the transcriptome changes in mammary epithelial cells (MECs) during early lactation in dairy goats. Primiparous Beetal goats were assigned to the study. Five days post-partum, one gland was infused xanthosine (TRT) twice daily (2×) for 3 d consecutively, and the other gland served as a control (CON). Milk samples from the TRT and CON glands were collected on the 10th day after the last xanthosine infusion, and RNA was isolated from milk fat globules (MFGs). Total RNA in MFGs was found to be mainly derived from the MECs. We characterized xanthosine-induced transcriptome changes in MECs between the TRT and CON samples on two goats. After quality control, more processed reads were aligned to goat genome using STAR. Transcript raw read counts were obtained using HTseq. Differential transcript analysis was performed using both cuffdiff and DESeq2. Significantly differentially expressed genes (DEGs) were subjected to Gene Ontology (GO) terms using PANTHER, and gene network was generated using STRING db. Preliminary analysis indicated that individual goats respond to xanthosine treatment differently reflected by varied number of DEGs within the same animal’s mammary gland. Several pathways were impacted by these DEGs, including cell communication, cell cycle and cell proliferation. A predominant portion of the DEGs showed down-regulation upon xanthosine treatment. Functional prediction of these genes include: (i) regulation of inflammation signalling mediated by chemokine and cytokines (CXCR1, CXCR2, PTAFR, RGS14, C5AR1, SAMD9 and JUNB), and (ii) anti-proliferative signals by dual-specificity phosphatases (DUSP1 and DUSP2) and NOTCH1. The rest of the DEGs showed up-regulation of anti-bacterial and anti-tumorigenic genes (LTF and B2M) upon xanthosine treatment. Notably, down-regulation of adhesion molecules (PECAM1, SSH2, SYNE1, CYTIP, SELL) highlighted its potential role in promoting cell motility in response to endogenous stimulus. This study provides valuable insights into whole-transcriptome changes in milk-producing cells in response to xanthosine treatment in goats. Further functional follow-up on the DEGs is highly likely to help delineate probable molecular mechanisms of enhanced cell proliferation in goat milk-producing cells through administration of xanthosine.