title: Solubility properties of neuronal tubulin: evidence for labile and stable microtubules.
abstract: The solubility properties of tubulin and microtubules in pure cultures of sympathetic neurons were examined by electron microscopic and biochemical techniques. For morphological analyses, neurons were extracted with Triton X-100 in the presence or absence of 1 mM CaCl2, and the resulting detergent-extracted residues were examined for microtubules. In parallel experiments, the solubility of tubulin was determined under various solution conditions. Detergent-extracted residues of neurons prepared without Ca2+ contained many microtubules. Neurite residues prepared in the presence of Ca2+ also contained microtubules, but at substantially lower numbers than in residues prepared without Ca2+. The biochemical data parallel the morphological observations. Following detergent-extraction under microtubule stabilizing conditions, 30% of the tubulin was detergent-soluble (i.e. unpolymerized), while 70% was detergent-insoluble (i.e. polymerized). A more detailed examination of the solubility properties of tubulin indicated that 62% was detergent-insoluble but soluble in buffers containing mM CaCl2, while 5-8% was detergent and Ca2+-insoluble. A variety of control experiments indicated that non-specific adsorption of tubulin onto detergent-insoluble components of the cultures, assembly of tubulin onto pre-existing microtubules, and incomplete extraction of tubulin from cells contributed minimally to the levels of Ca2+-soluble and insoluble tubulin obtained with the extraction conditions used. These results indicate that (a) the majority of neuronal tubulin is assembled into microtubules which disassemble upon treatment with Ca2+ and (b) a portion of the neuronal tubulin is assembled into microtubules which show the unusual property of Ca2+-stability.
mesh_terms: D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002478:Cells, Cultured; D008870:Microtubules / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D009474:Neurons / Q000032:analysis* / Q000166:cytology / Q000648:ultrastructure; D051381:Rats; D012995:Solubility; D014404:Tubulin / Q000032:analysis*
chemical_list: D014404:Tubulin; D002118:Calcium