Czech J. Anim. Sci., 2016, 61(3):99-107 | DOI: 10.17221/8782-CJAS

Combination of let-7d-5p, miR-26a-5p, and miR-15a-5p is suitable normalizer for studying microRNA expression in skin tissue of Liaoning cashmere goat during hair follicle cycleOriginal Paper

W.L. Bai¹, Y.L. Dang¹, R.H. Yin¹, R.L. Yin², W.Q. Jiang¹, Z.Y. Wang¹, Y.B. Zhu¹, J.J. Wang¹, Z.H. Zhao³, G.B. Luo¹

¹ College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang, P.R. China

² Research Academy of Animal Husbandry and Veterinary Medicine Sciences of Jilin Province, Changchun, P.R. China

³ College of Animal Science and Veterinary Medicine, Jilin University, Changchun, P.R. China

The microRNAs are non-coding RNA molecules of approximately 20-22 nucleotides that are found to be implicated in a wide range of physiological processes. In this study, the suitability of 10 candidate reference RNAs was evaluated for microRNA expression data in the skin tissue of Liaoning cashmere goat including 1 small nuclear RNA (snRNA; RNU6B), 1 small nucleolar RNA (snoRNA; Z30), 1 rRNA (5S), 1 transfer RNA (tRNA; Met-tRNA), and 6 microRNAs (miR; let-7d-5p, miR-15a-5p, miR-26a-5p, miR-125a-5p, miR-214-3p, and miR-221-3p). Based on geNorm and NormFinder algorithms, we identified let-7d-5p, miR-26a-5p, and miR-15a-5p as the most stable reference RNAs. Also, three reference RNAs (let-7d-5p, miR-26a-5p, and miR-15a-5p) were sufficient for the normalization of microRNA expression data in the skin of this breed. We further assessed the suitability of let-7d-5p, miR-26a-5p, and miR-15a-5p in a combination as reference RNAs through detecting the relative expression of miR-24-3p, miR-29a-3p, miR-145a-5p, and miR-205-5p as putative genes of interest. Significant differences were revealed in the relative expression of miR-24-3p, miR-29a-3p, miR-145a-5p, and miR-205-5p at telogen stage of hair follicle cycle when a combination of let-7d-5p, miR-26a-5p, and miR-15a-5p vs a single let-7d-5p were used as reference RNA. Based on the results from this study, we suggested that the combination of let-7d-5p, miR-26a-5p, and miR-15a-5p as normalizers for microRNA expression data would be more reliable than that of single let-7d-5p, and the geometric mean of these three microRNAs (let-7d-5p, miR-26a-5p, and miR-15a-5p) can be used for the normalization of microRNAs expression data in the skin of Liaoning cashmere goat.

Keywords: cashmere goat skin; microRNA expression reference; expression stability

Published: March 31, 2016  Show citation