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The Journal of Neuroscience, October 1, 1998, 18(19):7638-7649

Tyrosine Hydroxylase Expression in Primary Cultures of Olfactory Bulb: Role of L-Type Calcium Channels

Elena Cigola, Bruce T. Volpe, Jong Wha Lee, Linda Franzen, and Harriet Baker

Cornell University Medical College at The Burke Medical Research Institute, White Plains, New York, 10605

Sensory activity mediates regulation of tyrosine hydroxylase (TH), the first enzyme in the dopamine biosynthetic pathway, in the rodent olfactory bulb. The current studies established for the first time primary cultures of neonatal mouse olfactory bulb expressing TH and tested whether L-type calcium channels mediate the activity-dependent regulation of the dopamine phenotype. After 1 d in vitro (DIV), a small population of TH-immunostained neurons that lacked extensive processes could be demonstrated. After an additional 2 DIV in serum-free medium, the number of TH neurons had doubled, and they exhibited long interdigitating processes. Membrane depolarization for 48 hr with 50 mM KCl produced a further 2.4-fold increase in the number of TH-immunoreactive neurons compared with control cultures. Increased TH neuron number required at least 36 hr of exposure to KCl. Forskolin, which increases intracellular cAMP levels, induced a 1.5- to 1.6-fold increase in the number of TH-immunostained neurons. Combined treatment with KCl and forskolin was not additive. Nifedipine, an L-type calcium channel blocker, completely prevented the depolarization-mediated increase in TH expression but did not block the response to forskolin. Treatment with Bay K8644, an L-type calcium channel agonist, also significantly increased the number of TH-expressing neurons. Depolarization also induced alterations in neuritic outgrowth, resulting in a stellate versus an elongate morphology that, in contrast, was not prevented by nifedipine. These results are the first demonstration that in vitro, as in vivo, depolarization increases TH expression in olfactory bulb and that L-type calcium channels mediate this activity-dependent regulation of the dopamine phenotype.

Key words: tyrosine hydroxylase; olfactory bulb; calcium; depolarization; primary cultures; dopamine; cAMP; L-type calcium channel; Bay K8644

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Copyright © 1998 Society for Neuroscience  0270-6474/98/18197638-12$05.00/0

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