title: Chromatin and extracellular vesicle associated sperm RNAs.
abstract: A diverse pool of RNAs remain encapsulated within the transcriptionally silent spermatozoon despite the dramatic reduction in cellular and nuclear volume following cytoplasm/nucleoplasm expulsion. The impact of this pronounced restructuring on the distribution of transcripts inside the sperm essentially remains unknown. To define their compartmentalization, total RNA >100 nt was extracted from sonicated (SS) mouse spermatozoa and detergent demembranated sucrose gradient fractionated (Cs/Tx) sperm heads. Sperm RNAs predominately localized toward the periphery. The corresponding distribution of transcripts and thus localization and complexity were then inferred by RNA-seq. Interestingly, the number of annotated RNAs in the CsTx sperm heads exhibiting reduced peripheral enrichment was restricted. However this included Cabyr, the calcium-binding tyrosine phosphorylation-regulated protein encoded transcript. It is present in murine zygotes prior to the maternal to the zygotic transition yet absent in oocytes, consistent with the delivery of internally positioned sperm-borne RNAs to the embryo. In comparison, transcripts enriched in sonicated sperm contributed to the mitochondria and exosomes along with several nuclear transcripts including the metastasis associated lung adenocarcinoma transcript 1 (Malat1) and several small nucleolar RNAs. Their preferential peripheral localization suggests that chromatin remodeling during spermiogenesis is not limited to nucleoproteins as part of the nucleoprotein exchange.
authors.affiliation: Center for Molecular Medicine and Genetics, Wayne State University School of Medicine, Detroit, MI 48201, USA.
authors.forename: Graham D
authors.initials: GD
authors.lastname: Johnson
authors.affiliation: CReATe Fertility Centre, Toronto, ON, M5G 1N8, Canada.
authors.forename: Paula
authors.initials: P
authors.lastname: Mackie
authors.affiliation: Center for Molecular Medicine and Genetics, Wayne State University School of Medicine, Detroit, MI 48201, USA Department of Obstetrics and Gynaecology, University of Toronto, ON, M5G 1E2, Canada.
authors.forename: Meritxell
authors.initials: M
authors.lastname: Jodar
authors.affiliation: CReATe Fertility Centre, Toronto, ON, M5G 1N8, Canada Department of Obstetrics and Gynecology, Wayne State University School of Medicine, Detroit, MI 48201, USA.
authors.forename: Sergey
authors.initials: S
authors.lastname: Moskovtsev
authors.affiliation: Center for Molecular Medicine and Genetics, Wayne State University School of Medicine, Detroit, MI 48201, USA Department of Obstetrics and Gynaecology, University of Toronto, ON, M5G 1E2, Canada steve@compbio.med.wayne.edu.
authors.forename: Stephen A
authors.initials: SA
authors.lastname: Krawetz
mesh_terms: D000818:Animals; D002451:Cell Compartmentation; D002843:Chromatin / Q000737:chemistry*; D055354:Exosomes / Q000737:chemistry*; D006801:Humans; D008297:Male; D008822:Mice, Transgenic; D012313:RNA / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D000077278:RNA, Mitochondrial; D012335:RNA, Ribosomal / Q000032:analysis; D012091:Repetitive Sequences, Nucleic Acid; D013094:Spermatozoa / Q000737:chemistry* / Q000648:ultrastructure
chemical_list: D002843:Chromatin; D000077278:RNA, Mitochondrial; D012335:RNA, Ribosomal; D012313:RNA