Microtubules are involved in the localization of tau mRNA in primary neuronal cell cultures.
Subcellular localization of neuronal mRNAs contributes to the development of identifiable microdomains. In differentiated neurons, tau mRNA is localized in the cell body and the proximal portion of the axon, and MAP2 mRNA is localized in the cell body and dendrites, whereas tubulin mRNA is restricted to the cell body. To investigate the mechanism(s) leading to segregation of mictrotubule-associated protein mRNA, we examined the role of the cytoskeleton in this process. Detergent extraction of primary neuronal cells in culture followed by in situ hybridization analysis demonstrated that tau mRNA remains bound to cytoskeleton of the treated cells. In addition, biochemical fractionation showed that tau and MAP2 mRNAs are preferentially associated with the fraction of assembled microtubules. In contrast, mRNAs restricted to the neuronal cell body, such as those of tubulin, the 68 kDa neurofilament, and mouse GAPDH, are preferentially found in the supernatant. Using cytoskeletal inhibitors, we demonstrate that tau mRNA is associated with the microtubule system, and not with the actin filaments, thus supporting the hypothesis that the mechanism of mRNA localization is a multistep pathway in which the microtubules play a crucial role.
Litman
P
P
Department of Neurobiology, Weizmann Institute of Science, Rehovot, Israel.
Barg
J
J
Ginzburg
I
I
D008841:Actin Cytoskeleton / Q000378:metabolism; D000818:Animals; D001921:Brain / Q000166:cytology; D002451:Cell Compartmentation; D002478:Cells, Cultured; D003078:Colchicine / Q000494:pharmacology; D003571:Cytochalasin B / Q000494:pharmacology; D005786:Gene Expression Regulation; D017403:In Situ Hybridization; D066298:In Vitro Techniques; D008869:Microtubule-Associated Proteins / Q000235:genetics; D008870:Microtubules / Q000502:physiology*; D016900:Neurofilament Proteins / Q000235:genetics; D009474:Neurons / Q000378:metabolism / Q000648:ultrastructure*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D014404:Tubulin / Q000235:genetics; D016875:tau Proteins / Q000235:genetics*
D008869:Microtubule-Associated Proteins; D016900:Neurofilament Proteins; D012333:RNA, Messenger; D014404:Tubulin; D016875:tau Proteins; D003571:Cytochalasin B; D003078:Colchicine