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RNA ; 20(4): 483-95, 2014 Apr.
Article in English | MEDLINE | ID: mdl-24554440

ABSTRACT

The genome of male germ cells is actively transcribed during spermatogenesis to produce phase-specific protein-coding mRNAs and a considerable amount of different noncoding RNAs. Ribonucleoprotein (RNP) granule-mediated RNA regulation provides a powerful means to secure the quality and correct expression of the requisite transcripts. Haploid spermatids are characterized by a unique, unusually large cytoplasmic granule, the chromatoid body (CB), which emerges during the switch between the meiotic and post-meiotic phases of spermatogenesis. To better understand the role of the CB in male germ cell differentiation, we isolated CBs from mouse testes and revealed its full RNA and protein composition. We showed that the CB is mainly composed of RNA-binding proteins and other proteins involved RNA regulation. The CB was loaded with RNA, including pachytene piRNAs, a diverse set of mRNAs, and a number of uncharacterized long noncoding transcripts. The CB was demonstrated to accumulate nascent RNA during all the steps of round spermatid differentiation. Our results revealed the CB as a large germ cell-specific RNP platform that is involved in the control of the highly complex transcriptome of haploid male germ cells.


Subject(s)
Cytoplasmic Granules/physiology , Germ Cells/metabolism , RNA, Messenger/metabolism , RNA-Binding Proteins/metabolism , Spermatids/metabolism , Spermatogenesis/physiology , Animals , Biomarkers/metabolism , Cytoplasmic Granules/ultrastructure , Fluorescent Antibody Technique , Gene Expression Profiling , Germ Cells/ultrastructure , Male , Mice , Oligonucleotide Array Sequence Analysis , RNA, Messenger/genetics , Real-Time Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain Reaction , Spermatids/ultrastructure
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