Accumulation of piRNAs in the chromatoid bodies purified by a novel isolation protocol

• Published in: Experimental cell research Vol. 316; no. 9; pp. 1567 - 1575
• Main Authors: Meikar, Oliver, Da Ros, Matteo, Liljenbäck, Heidi, Toppari, Jorma, Kotaja, Noora
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 15.05.2010; Elsevier BV
• Subjects: Animals; Blotting, Northern; Cell Nucleus - chemistry; Cell Nucleus - metabolism; Cellular biology; Chromatin; Chromatoid body; Cytoplasm; Cytoplasm - metabolism; Cytoplasmic Granules - metabolism; Fluorescent Antibody Technique; Germ Cells - physiology; Immunoblotting; Immunoprecipitation; In Situ Hybridization; Male; Mice; Mice, Inbred C57BL; mRNA regulation; piRNA; Ribonucleic acid; RNA; RNA, Small Interfering - physiology; Round spermatid; Small RNA; Spermatogenesis; Testis - cytology; Testis - metabolism; mRNA regulation; Small RNA; Chromatoid body; Spermatogenesis; Round spermatid; piRNA; MVH; PIWI; CB
• ISSN: 0014-4827; 1090-2422
• DOI: 10.1016/j.yexcr.2010.02.023

Haploid male germ cells are featured by an intriguing cytoplasmic cloud-like structure that has been named as chromatoid body (CB) on the basis of its staining properties and appearance under a microscope. Notwithstanding its early discovery in the late 19th century, the function of the CB is still largely obscure. Emerging evidence suggests a role for the CB and other similar RNA-containing granules, such as germ plasm in lower organism and processing bodies in somatic cells, in the control and organization of RNA processing and/or storage. Despite the increasing scientific demand, the lack of CB purification protocols has still been the main obstacle in the functional characterization of this structure. We have successfully isolated CBs from mouse testis by a novel immunoaffinity purification procedure and validated by several different methods that pure CB fractions are obtained. Analysis of the CB RNA content reveals enrichment of PIWI-interacting RNAs (piRNAs), further emphasizing the role of CB as the RNA processing body.