Accumulation of piRNAs in the chromatoid bodies purified by a novel isolation protocol.
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.
Meikar
Oliver
O
Department of Physiology, Institute of Biomedicine, University of Turku, Turku, Finland. oliver.meikar@utu.fi
Da Ros
Matteo
M
Liljenbäck
Heidi
H
Toppari
Jorma
J
Kotaja
Noora
N
D000818:Animals; D015152:Blotting, Northern; D002467:Cell Nucleus / Q000737:chemistry* / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D003594:Cytoplasmic Granules / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D005854:Germ Cells / Q000502:physiology*; D015151:Immunoblotting; D047468:Immunoprecipitation; D017403:In Situ Hybridization; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D034741:RNA, Small Interfering / Q000502:physiology*; D013091:Spermatogenesis; D013737:Testis / Q000166:cytology / Q000378:metabolism*
D034741:RNA, Small Interfering