Measurement of dendritic mRNA transport using ribosomal markers.
mRNA is transported to the dendritic regions by forming RNA granules, an aggregate of mRNA, ribosomal proteins, rRNA, and RNA-binding proteins such as Staufen. In this study, the dendritic transport of RNA granules was measured using the individual antibodies to ribosome-specific markers such as ribosomal L4 or S6 protein, and Y10B, a monoclonal antibody specific to rRNA. All the markers showed significant immunoreactivity in the dendritic regions of the hippocampal neurons. In addition, a GFP-tagged Staufen, a marker protein of the RNA granules, was colocalized with the Y10B and S6 signals in the dendrites. The S6 signals were also colocalized with the Y10B signals in the dendrites. Consistent with previous studies, the depolarization induced by KCl stimulation increased the ribosomal level, revealed by the S6 or Y10B immunostaining in the distal dendrites. These results demonstrate the utility of ribosomal markers for detecting the RNA granules or mRNA transport in dendrites.
Kim
Hyong Kyu
HK
Department of Medicine and Microbiology, College of Medicine and Medical Research Institute, Chungbuk National University, Cheongju 361-768, Republic of Korea. hkkim69@chungbuk.ac.kr
Kim
Yun-Bae
YB
Kim
Eung-Gook
EG
Schuman
Erin
E
D000818:Animals; D000831:Animals, Newborn; D001693:Biological Transport, Active / Q000502:physiology; D015415:Biomarkers / Q000378:metabolism; D002478:Cells, Cultured; D003594:Cytoplasmic Granules / Q000378:metabolism; D003712:Dendrites / Q000378:metabolism* / Q000648:ultrastructure*; D016231:Fluorescence Polarization Immunoassay / Q000379:methods*; D006624:Hippocampus / Q000378:metabolism; D009474:Neurons / Q000378:metabolism / Q000648:ultrastructure; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D012269:Ribosomal Proteins / Q000032:analysis / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism*
D015415:Biomarkers; D012333:RNA, Messenger; D012269:Ribosomal Proteins