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Shahid Y Khan, Sean F Hackett, Mei-Chong Wendy Lee, C. Conover Talbot, Nader Pourmand, S Amer Riazuddin; Transcriptome Profiling of Developing Murine Lens through RNA Sequencing. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):4005.
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© ARVO (1962-2015); The Authors (2016-present)
Transcriptome is the entire repertoire of all transcripts present in a cell at any particular time. We undertook next-generation whole transcriptome sequencing approach to gain insight of the transcriptional landscape of developing mouse lens.
We ascertained mice lenses at six developmental time points including two embryonic (E15 and E18) and four postnatal stages (P0, P3, P6, and P9). The ocular tissue at each time point was maintained as two distinct pools serving as biological replicates for each developmental stage. The mRNA and small RNA libraries were paired-end sequenced on Illumina HiSeq 2000 and subsequently analyzed using different bioinformatics platforms.
The mapping of mRNA and small RNA libraries generated 187.56 and 154.22 million paired-end reads, respectively. We detected a total of 14,465 genes in the mouse ocular lens. Of these, 46 genes exhibited 40-fold differential expression compared to transcriptional levels at E15. Likewise, small RNA profiling identified 379 microRNAs (miRNAs) expressed in mouse lens. Of these, 49 miRNAs manifested an 8-fold or higher differential expression when compared, as above to the microRNA expression at E15.
In conclusion, we report the first comprehensive profile of developing murine lens transcriptome including both mRNA and miRNA through next-generation RNA sequencing. These data will add to our understanding of lens development and elucidate processes essential for maintenance of lens transparency.
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