Heterogeneity of cold-stable and cold-labile tubulin in axon- and soma-enriched portions of the adult mouse brain

doi:10.1016/0304-3940(87)90580-5

Neuroscience Letters

Volume 77, Issue 2, 15 June 1987, Pages 166-170

https://doi.org/10.1016/0304-3940(87)90580-5 Get rights and content

Abstract

Microtubules from the optic nerve (axonal tubulin) and lateral geniculate nucleus (cell tubulin) were separated by cold treatment and the cold-soluble fraction was purified in the presence of Taxol. Isoforms of cold-stable and cold-soluble tubulin were resolved by the use of high-resolution isoelectric focusing. The cold-soluble fraction of axonal tubulin has only 14 of the 20 isotypes seen in the same fraction of cell tubulin. The two cold-stable fractions have 20 isotypes but axonal tubulin has a specific pattern of isotypes 1, 2 and 5. Cold-stable fractions of both axonal and cell tubulin display the existence of an intensely stained α-isotype, isotype 7, which seems associated with the property of cold stability. Our results highly favor the hypothesis of a physiological role of the heterogeneity of tubulin in neuronal microtubules.

References (19)

M.M. Black et al.
Solubility properties of neuronal tubulin: evidence for labile and stable microtubules
Brain Res.
(1984)
S.T. Brady et al.
Axonal tubulin and axonal microtubules: biochemical evidence for cold stability
J. Cell Biol.
(1984)
D.W. Cleveland et al.
Molecular biology and genetics of tubulin
Annu. Rev. Biochem.
(1985)
M.P. Daniels
Fine structural changes in neurons and nerve fibers associated with colchicine inhibition of nerve formation in vitro
J. Cell Biol.
(1973)
P. Dustin
Microtubules
H. Feit et al.
Colchicine binding activity in particulate fractions of mouse brain
J. Neurochem.
(1970)
D.J. Field et al.
Heterogeneity of vertebrate brain tubulins
H.J. George et al.
Polymorphism of brain tubulin
Biochemistry
(1981)
I. Gozes
Tubulin in the nervous system
Neurochem. Int.
(1982)

There are more references available in the full text version of this article.

Cited by (9)

Cold-Stable and Cold-Adapted Microtubules
1995, International Review of Cytology
Most mammalian microtubules disassemble at low temperatures, but some are cold stable. This probably has little to do with a need for cold-stable microtubules, but reflects that certain populations of microtubules must be stabilized for specific functions. There are several routes by which to achieve cold stability. Factors that interact with microtubules, such as microtubule-associated proteins, STOPs (stable tubule only polypeptides), histones, and possibly capping factors, are involved. Specific tubulin isotypes and posttranslational modifications might also be of importance. More permanent stable microtubules can be achieved by bundling factors, associations to membranes, as well as by assembly of microtubule doublets and triplets. This is, however, not the explanation for cold adaptation of microtubules from poikilothermic animals, that is, animals that must have all their microtubules adapted to low temperatures. All evidence so far suggests that cold adaptation is intrinsic to the tubulins, but it is unknown whether it depends on different amino acid sequences or posttranslational modifications.
Modifications of tubulin heterogeneity during embryonic and postnatal stages in a specific region of mouse brain
1991, Brain Research
An auditory nucleus (the inferior colliculus of the mouse) was used to study the modifications of the heterogeneity of tubulin which occur at various stages of development and maturation. Both the cold-stable (CS) and cold-labile (CL) fractions of tubulin were analyzed by high-resolution isoelectric focusing. Our results suggest that tubulin heterogeneity is modified at critical stages of development and maturation, with specific variations of the two fractions. Stage E₁₀ corresponding to the appearance of the first young neurons and axonal profiles is marked by the modification of the CS fraction with the emergence of isotypes α₅ to α₈ and β₁₂ and β₁₇. Stage E₁₂ is characterized by the modifications of the CL fraction, particularly the β-group; at this stage the first dendrites become visible. At birth, all isotypes increase in both the CL and CS fractions. At stages P₇–P₁₀ transient modifications of a group of both CS and CL fractions and of the β group of CS fraction occur. These are associated with the emergence of isotypes β₁₇ to β₂₀ in the CL fraction. This period precludes the initial period of functional maturation of the auditory system which occurs from P₁₀ to P₂₀. During this period, the CL fraction (α and β group) remains unmodified, whereas all isotypes of the CS fraction, except 3–4, display complex variations with an initial decrease until P₁₂, an increase until P₁₇, and a final decrease until P₂₀. Our results emphasize the importance of the CS fraction of the microtubules in the axonal domain and correlate the variations in tubulin isotypes patterns with some of the critical events occurring during the development of the nervous system.
Characteristics of Microtubules at the Different Stages of Neuronal Differentiation and Maturation
1989, International Review of Cytology
This chapter discusses the characteristics of microtubules (MT) in the developing nervous system by considering the morphological and biochemical organization of the MT and their components at the different stages of neurogenesis. The different stages and the morphogenetic events which occur during these stages are reviewed. The developing nervous system proves to be a powerful tool to analyze the involvement of MT in basic biological processes such as cell proliferation, cell migration, cell shaping, and transport. The extensive heterogeneity of microtubule-associated proteins (MAP) and tubulin in the central nervous system (CNS) confers on the neurons a wide range of capabilities of assembly of these proteins and suggests that the neuron has a unique potential of a relation between MT composition and cell function. The dynamics of MT and the morphological aspects of MT in the different cell types which are involved in neurogenesis and in the two major differentiating domains of the neurons, that is, the axonal and the dendritic-perikaryal domains are described in the chapter.
Compartmentation of α-tubulin in neurons: Identification of a somatodendritic-specific variant of α-tubulin
1989, Neuroscience
The composition of α-tubulin in axons was compared with that in cell bodies and dendrites in the rat. Pure axons and highly enriched preparations of cell bodies and dendrites were obtained from expiant cultures of sympathetic neurons, dissolved directly into sodium dodecyl sulfate, and analysed by isoelectric focusing gel electrophoresis. Immunoblots using two different monoclonal antibodies against α-tubulin and fluorographs of [³⁵S]methionine- and [³H]tyrosine-labeled α-tubulin all show several variants common to both compartments, and an additional variant, more basic than the others, prominent in the somatodendritic compartment but absent from the axonal compartment. This variant, termed alpha_s, partitions with both cytoskeletal and soluble tubulin pools, indicating that microtubules in the somatodendritic compartment differ from microtubules in the axonal compartment with respect to α-tubulin composition. Pulse-chase and immunoprecipitation analyses suggest that extensive post-translational modification of α-tubulin occurs in the neuron. Newly synthesized α-tubulin (10-min pulse-label) is relatively simple in profile, consisting of a major band with the mobility of alpha_s, and a minor band slightly lower in isoelectric point. The profile is somewhat more complex after 5 h chase time, but is much more complex after 20 h.
The generation of additional bands appears to be at the expense of the band co-migrating ith alpha_s, suggesting that alpha_s is the unmodified form of a primary translation product that undergoes extensive post-translational modification. The unmodified form of the primary translation product is restricted to the somatodendritic compartment, while many of its derivatives are present throughout the neuron.
Modifications of tubulin heterogeneity during axonal growth in the embryonic nervous system
1988, Developmental Brain Research
Tectal plates of mouse embryos were used to analyze the heterogeneity of tubulin with high resolution isoelectric focusing, at time of appearance of the young neurons and axons. In the cold-labile pool of tubulin, isotypes 6 and 7 appear, and isotypes 1, 3, 11, 13 increase their relative quantities. The increase of the prominence of the α-group with regard to the β-group in the cold-stable pool of tubulin raises the question as to the exact role of tubulin in the cold stability. A role of other specific microtubule components also appears probable.
Accumulation of delta 2-tubulin, a major tubulin variant that cannot be tyrosinated, in neuronal tissues and in stable microtubule assemblies
1994, Journal of Cell Science

View all citing articles on Scopus

View full text

Heterogeneity of cold-stable and cold-labile tubulin in axon- and soma-enriched portions of the adult mouse brain

Abstract

Solubility properties of neuronal tubulin: evidence for labile and stable microtubules

Brain Res.

Axonal tubulin and axonal microtubules: biochemical evidence for cold stability

J. Cell Biol.

Molecular biology and genetics of tubulin

Annu. Rev. Biochem.

Fine structural changes in neurons and nerve fibers associated with colchicine inhibition of nerve formation in vitro

J. Cell Biol.

Microtubules

Colchicine binding activity in particulate fractions of mouse brain

J. Neurochem.

Heterogeneity of vertebrate brain tubulins

Polymorphism of brain tubulin

Biochemistry

Tubulin in the nervous system

Neurochem. Int.