Differential expression of microtubule associated protein MAP-2 in developing cochleovestibular neurons and its modulation by neurotrophin-3.
Microtubule associated proteins (MAPs) are essential cytoskeletal proteins in developing neurons. The present study was undertaken to analyze the expression of MAP2 and its isoforms (a,b,c) during the embryonal and early post-hatching development of chicken cochleovestibular ganglion (CVG) neurons. Moreover, we have investigated MAP2 expression in primary cultures of CVG neurons, and whether it is regulated by neurotrophin-3 (NT3). The expression of MAP2 immunoreactivity (IR) was studied using both Western blot and immunohistochemistry on tissue sections and primary cultures. In vivo MAP2c was expressed from incubation day 4 (E4) to E10, and MAP2b was found in all embryonal stages studied and at post-hatching day 10 (P10), whereas MAP2a was restricted to the post-hatching periods. The cellular localization of IR was in the neuronal perikarya and their peripheral processes (dendrites) but not in axons. Primary cultures matched the in vivo pattern of MAP2 expression, and IR was localized in neuronal cell bodies and the initial segment of the neuronal processes. Exogenous NT3 regulated the expression of MAP2 isoforms in a dose dependent manner. At the survival dose of 0.5 ng/ml NT3, the main MAP2 expression was MAP2c. Conversely, at the neuritogenic dose of 5 ng/ml NT3 increased MAP2b and MAP2a expression, but not MAP2c. The present results demonstrate that MAP2 isoforms are developmentally regulated, thus suggesting that each isoform is specifically involved in CVG neuron maturation. Furthermore, we provide evidence of MAP2 regulation in culture by the neurotrophic factor NT3.
San José
I
I
Departamento de Anatomía Humana, Instituto de Biologia y Genética Molecular, Universidad de Valladolid-Consejo Superior de Investigaciones Cientificas, Spain.
Vázquez
E
E
García-Atares
N
N
Huerta
J J
JJ
Vega
J A
JA
Represa
J
J
D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D002642:Chick Embryo; D003056:Cochlear Nerve / Q000187:drug effects / Q000196:embryology / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects*; D007150:Immunohistochemistry; D008869:Microtubule-Associated Proteins / Q000096:biosynthesis*; D009414:Nerve Growth Factors / Q000494:pharmacology*; D009475:Neurons, Afferent / Q000378:metabolism*; D020933:Neurotrophin 3; D014725:Vestibular Nerve / Q000187:drug effects / Q000196:embryology / Q000378:metabolism*
D008869:Microtubule-Associated Proteins; D009414:Nerve Growth Factors; D020933:Neurotrophin 3