Abstract
BackgroundTranscriptionally quiescent spermatozoa have been established to be a repository of mRNA coding for several functionally essential cellular proteins. This entourage of mRNA is envisaged to be involved in post-fertilization and early embryogenesis. Minisatellites tagged with mRNA transcripts have been implicated with gene organization, regulation and function. However, the organization and expression of the minisatellite tagged transcript diversity, particularly in spermatozoa, remains unclear.ResultsIn the present study, we identified and characterized 12 mRNA transcripts from the spermatozoa of water buffalo Bubalus bubalis employing minisatellite associated sequence amplification (MASA) and a consensus sequence of 33.15 repeat loci. Of these 33.15 tagged transcripts, only one was found to be homologous to Bovine steroid 21-hydroxylase (P-450-c21) gene. Other ten transcripts showed significant similarity with various mRNAs or chromosomal contigs across the species. The remaining one construed to be novel since this was unreported in the database (NCBI GenBank). All these uncharacterized and known transcripts showed highest expression in testis and spermatozoa compared to that in somatic tissues and ovary. Of these 12 mRNA transcripts, 4 showed differential expression in the forebrain and hindbrain of buffalo. Moreover, genes corresponding to all the 33.15 tagged spermatozoal transcripts were found to be conserved across 13 other species analyzed.ConclusionOur results show MASA as an important tool to capture mRNA transcript diversity tagged with minisatellites in the spermatozoa. Comprehensive characterization of these transcripts is envisaged to augment our understanding on the genes involved in testicular functions and sustenance of a viable paternal genome during pre- and post- fertilization events and early stages of development. Prospects of this approach in genome analysis in general and comparative genomics in particular are highlighted.
Highlights
Quiescent spermatozoa have been established to be a repository of mRNA coding for several functionally essential cellular proteins
Differential distribution of the consensus sequence of 33.15 repeat loci within the spermatozoal and somatic transcriptomes In previous study, we identified 6 different mRNA transcripts from various tissues of water buffalo using oligos based on the consensus of 33.15 repeat loci [20]
Comparative analysis of these spermatozoal mRNA transcripts showed that none was common to those identified from different somatic tissues, testis and ovary in earlier study [20]
Summary
Quiescent spermatozoa have been established to be a repository of mRNA coding for several functionally essential cellular proteins. Despite the transcriptionally inactive state, spermatozoa retain an entourage of mRNA transcripts encoding transcription factors and proteins involved in signal transduction, cell proliferation, chromatin condensation, regulation of sperm motility, capacitation and acrosome reaction [2,3,4,5,6,7]. Delivery of such spermatozoal transcripts to ooplasm is envisaged to have their potential significance during fertilization, embryogenesis and morphogenesis. These remain to be characterized for their organization, expression and association with different regulatory elements and repetitive sequences
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