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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 #339198

Title: Effects of xanthosine in isoform switch and splice variants of expressed genes in mammary epithelial cells

Author
item CHOUDHARY, RATAN - Punjab Agricultural University
item Li, Wenli
item CHOUDHARY, SHANTI - Punjab Agricultural University
item RAMNEEK, VERMA - Punjab Agricultural University
item CAPUCO, ANTHONY - Retired ARS Employee

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 4/24/2017
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Although intramammary xanthosine (XS) treatment was reported to increase the mammary stem cell population and milk yield in bovine and caprine, underlying molecular mechanisms remain unclear. The goal of this study was to evaluate effects of XS treatment on the mammary transcriptome in early-lactation dairy goats. Primiparous Beetle goats were used. Five d after kidding, one gland was infused with XS (treatment [TRT]) twice daily for 3 d, and the other gland served as control (CON). Milk from TRT and CON glands was collected 10 d after the last treatment, followed by mammary epithelial cells (MSCs) isolation and RNA extraction. We characterized XS-induced isoform expression changes in MSCs of 2 goats, using RNA-seq and associated softwares. Relative expression level was analyzed at both net transcript and isoform levels. Overall, 56 genes showed differential expression at net transcript production level. Among them, 46% had two or more annotated isoforms. Using isoform-level quantification, a total of 340 genes showed significant isoform expression changes upon XS treatment. A predominant portion of these genes would not be identified if only net transcript production was considered. Some (14) genes showed major isoform switch in response to treatment. On average, each of these genes had 9 annotated isoforms. A potential pathway impacted by these genes includes cellular reponse to growth simulation. Of note, PRLR, a prolactin receptor important in proliferation and differentiation of secretory mammary epithelium, was among these identified with major isoform switch. A further analysis of splicing sites of expressed genes revealed 54 genes with an increase in observed spicing sites, and 4,162 genes with a decrease in observed splicing sites. This observation indicated that XS treatment has substantial impact on transcript spicing patterns that will likely affect the gloal transcriptome production profile upon treatment of XS. This study documented extensive transcriptome changes induced by XS infusion in milk-producing cells of goats. Further, our analysis indicated that differential expression of isoforms and associated splicing pattern changes are likely an integral part of the physiological changes in goat mammary gland cells in reponse to XS treatment.