Axonal tubulin and axonal microtubules: biochemical evidence for cold stability

Nerve extracts containing tubulin labeled by axonal transport were analyzed by electrophoresis and differential extraction. We found that a substantial fraction of the tubulin in the axons of the retinal ganglion cell of guinea pigs is not solubilized by conventional methods for preparation of microtubules from whole brain. In two-dimensional polyacrylamide gel electrophoresis this cold-insoluble tubulin was biochemically distinct from tubulin obtained from whole brain microtubules prepared by cold cycling. Cleveland peptide maps also indicated some differences between the cold-extractable and cold- insoluble tubulins. The demonstration of cold-insoluble tubulin that is specifically axonal in origin permits consideration of the physiological role of cold-insoluble tubulin in a specific cellular structure. It appears likely that much of this material is in the form of cold-stable microtubules. We propose that the physiological role of cold-insoluble tubulin in the axon may be associated with the regulation of the axonal microtubule complexes in neurons.

[1]  M. Black,et al.  Solubility properties of neuronal tubulin: Evidence for labile and stable microtubules , 1984, Brain Research.

[2]  R. Lasek,et al.  Axonal transport of a subclass of tau proteins: evidence for the regional differentiation of microtubules in neurons. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[3]  T. Tashiro,et al.  Subunit composition specific to axonally transported tubulin , 1983, Neuroscience.

[4]  I. Gozes.  Tubulin in the nervous system , 1982, Neurochemistry International.

[5]  L. Greene,et al.  Changes in the colchicine susceptibility of microtubules associated with neurite outgrowth: studies with nerve growth factor-responsive PC12 pheochromocytoma cells , 1982, The Journal of cell biology.

[6]  R. Margolis,et al.  Recycling of cold-stable microtubules: evidence that cold stability is due to substoichiometric polymer blocks. , 1982, Biochemistry.

[7]  J. R. Morris,et al.  Stable polymers of the axonal cytoskeleton: the axoplasmic ghost , 1982, The Journal of cell biology.

[8]  S. Heidemann,et al.  Polarity orientation of axonal microtubules , 1981, The Journal of cell biology.

[9]  D. Bray,et al.  Serial analysis of microtubules in cultured rat sensory axons , 1981, Journal of neurocytology.

[10]  R. Margolis,et al.  Rapid disassembly of cold-stable microtubules by calmodulin. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[11]  H. Ishikawa,et al.  THE CYTOSKELETON IN MYELINATED AXONS: SERIAL SECTION STUDY , 1981 .

[12]  R. Margolis,et al.  Characterization of rat brain crude extract microtubule assembly: correlation of cold stability with the phosphorylation state of a microtubule-associated 64K protein. , 1981, Biochemistry.

[13]  R. Lasek,et al.  Axonal transport of calmodulin: a physiologic approach to identification of long-term associations between proteins , 1981, The Journal of cell biology.

[14]  R. Lasek,et al.  Nerve-specific enolase and creatine phosphokinase in axonal transport: soluble proteins and the axoplasmic matrix , 1981, Cell.

[15]  Lasek Rj,et al.  The dynamic ordering of neuronal cytoskeletons. , 1981 .

[16]  Shelanski Ml,et al.  Evidence for interactions between neurofilaments and microtubules. , 1981 .

[17]  R. Lasek,et al.  Slow components of axonal transport: two cytoskeletal networks , 1980, The Journal of cell biology.

[18]  J. Barron,et al.  Cold stable microtubules in brain studied in fractions and slices , 1980, Journal of neurocytology.

[19]  M. Kirschner.  Implications of treadmilling for the stability and polarity of actin and tubulin polymers in vivo , 1980, The Journal of cell biology.

[20]  R. Goldman,et al.  In vitro reassembly of squid brain intermediate filaments (neurofilaments): purification by assembly-disassembly. , 1980, Science.

[21]  E. Wieben,et al.  Increased levels of mRNAs for tubulin and other flagellar proteins after amputation or shortening of Chlamydomonas Flagella , 1980, Cell.

[22]  L. Wilson,et al.  Cold-stable microtubules from brain. , 1980, Biochemistry.

[23]  R. Lasek,et al.  Axonal transport of actin: Slow component b is the principal source of actin for the axon , 1979, Brain Research.

[24]  R. Raff,et al.  Mutation in a structural gene for a beta-tubulin specific to testis in Drosophila melanogaster. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[25]  S. Berkowitz,et al.  Separation and characterization of microtubule proteins from calf brain. , 1977, Biochemistry.

[26]  M. Kirschner,et al.  Physical and chemical properties of purified tau factor and the role of tau in microtubule assembly. , 1977, Journal of molecular biology.

[27]  D. Raybin,et al.  Modification of tubulin by tyrosylation in cells and extracts and its effect on assembly in vitro , 1977, The Journal of cell biology.

[28]  M. Kirschner,et al.  Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. , 1977, The Journal of biological chemistry.

[29]  R. Lasek,et al.  The slow component of axonal transport. Identification of major structural polypeptides of the axon and their generality among mammalian neurons , 1975, The Journal of cell biology.

[30]  B. Brinkley,et al.  COLD‐LABILE AND COLD‐STABLE MICROTUBULES IN THE MITOTIC SPINDLE OF MAMMALIAN CELLS * , 1975, Annals of the New York Academy of Sciences.

[31]  P. O’Farrell.  High resolution two-dimensional electrophoresis of proteins. , 1975, The Journal of biological chemistry.

[32]  W. Bonner,et al.  A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. , 1974, European journal of biochemistry.

[33]  G. Borisy,et al.  Characterization of microtubule assembly in porcine brain extracts by viscometry. , 1973, Biochemistry.

[34]  D. Rodbard,et al.  INSTABILITY OF pH GRADIENTS FORMED BY ISOELECTRIC FOCUSING IN POLYACRYLAMIDE GEL , 1973, Annals of the New York Academy of Sciences.

[35]  C. Cantor,et al.  Microtubule assembly in the absence of added nucleotides. , 1973, Proceedings of the National Academy of Sciences of the United States of America.

[36]  S. Barondes,et al.  COLCHICINE‐BINDING ACTIVITY IN PARTICULATE FRACTIONS OF MOUSE BRAIN 1 , 1970, Journal of neurochemistry.

[37]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[38]  R. Friede,et al.  Axon caliber related to neurofilaments and microtubules in sciatic nerve fibers of rats and mice , 1970, The Anatomical record.

[39]  V. Tennyson.  The Fine Structure of the Nervous System. , 1970 .

[40]  I. Mcquarrie.  ROLE OF THE AXONAL CYTOSKELETON IN THE REGENERATING NERVOUS SYSTEM , 1983 .

[41]  R. Lasek,et al.  Axonal transport: a cell-biological method for studying proteins that associate with the cytoskeleton. , 1982, Methods in cell biology.

[42]  R. Lasek.  The dynamic ordering of neuronal cytoskeletons. , 1981, Neurosciences Research Program bulletin.

[43]  R. Liem,et al.  Evidence for interactions between neurofilaments and microtubules. , 1981, Neurosciences Research Program bulletin.

[44]  E. Gray.  The Fine Structure of the Nervous System , 1971 .