Differential Expression of Newly Identified Long Intergenic Non‐Coding RNAs in Buffalo Oocytes Indicating Their Possible Role in Maturation and Embryonic Development

Abstract

Female germ cell and its intricate milieu regulate key processes of folliculogenesis and early embryonic development. However, the composition and dynamics of the oocyte transcriptome defines its future fertilizing ability which in turn depends on a number of oocyte specific genes whose identities are still unknown. In this context, the construction of buffalo oocyte specific subtracted cDNA library has raised fresh challenges of defining the importance of a battery of oocyte expressed transcripts in oocyte maturation. The present study tried to characterize these hitherto unknown transcripts and further to assess their expression dynamics in buffalo oocytes of different quality. For this purpose, three ESTs were selected from the library and subjected to 5' and 3' RACE for generating their full length sequences. These constructed full length sequences were validated by amplifying them in oocytes. Further these sequences were extensively analyzed for their coding potential and possible role using coding potential calculator and miRNA database. Besides, their expression was monitored during in vitro maturation in good (BCB+) and poor quality (BCB-) oocytes which was interestingly found to be differing significantly. All the three sequences under study were interpreted as long intergenic non-coding RNAs with the possibility of two of them acting as a miRNA precursors. Also, their differential expression trends in competitively diverse oocytes hints at their possible involvement in oocyte maturation and future embryonic development which needs to be explored further. J. Cell. Biochem. 118: 1712-1721, 2017. © 2016 Wiley Periodicals, Inc.

Long Noncoding RNA lncARSR Promotes Doxorubicin Resistance in Hepatocellular Carcinoma via Modulating PTEN‐PI3K/Akt Pathway

“…With more than 200 nucleotides in length, lncRNAs have limited protein coding potential [Novak Kujundzic et al, 2008;Ponting et al, 2009;Huarte, 2015]. However, accumulating evidences reveal that lncRNAs have critical functions in various pathophysiological processes and their dysregulation contributes to many disease states, particular for cancers [Dholpuria et al, 2017;Lin et al, 2016;Liu et al, 2016;Schmitt and Chang, 2016;Zhang et al, 2016;Chen et al, 2017]. In HCC, several lncRNAs have been reported to play oncogenic or tumor suppressive roles through diverse mechanisms.…”

Single‐cell RNA‐seq reveals mRNAs and lncRNAs important for oocytes in vitro matured in pigs

“…In addition, our group has reported a novel lncRNA regulating meiosis of pig oocytes through epigenetic modification and cytoskeleton organization (Yang, Wang, et al., 2020). Accumulating evidences of lncRNAs functioning in mouse oocytes (Dholpuria et al., 2017; Ganesh et al., 2020) also imply biological importance of lncRNAs in mammalian oocytes. Our findings provide the resource of lncRNAs specific for maturing pig oocytes for additional future functional study.…”

Analysis of amplification and association polymorphisms in the bovine beta-defensin 129 (BBD129) gene revealed its function in bull fertility

“…The bovine BBD129 gene is an ortholog of the primate DEFB126 gene 23 . RLM-RACE has been utilized to amplify matured capped and tailed mRNAs 80 , 81 , correctly detecting the 5' capped and 3' polyadenylation ends of mRNA 43 , 44 . The Bos taurus BBD129 gene consists of two exons and 50 nucleotide UTRs at both ends of the mRNA 82 .…”

“…RLM-RACE of BBD129. The 5' and 3' ends of the BBD129 mRNA were amplified by the RLM-RACE kit (#AM1700M, Invitrogen, Waltham, Massachusetts, USA) as per company suggested protocol 43 . The RNA processing steps for the 5' RLM-RACE were following:A) Calf Intestine Alkaline Phosphate (CIP) treatment-Dephosphorylation of incomplete and non-mRNA, B) Termination of CIP enzyme, C) Tobacco Acid Pyrophosphatase (TAP) associated enzymatic decapping of mRNA molecules, and D) 5' RLM-RACE Adapter ligation 43,44 .…”