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The Journal of Neuroscience, December 1, 2002, 22(23):10201-10208

Glial Cell Line-Derived Neurotrophic Factor and Target-Dependent Regulation of Large-Conductance K_Ca Channels in Developing Chick Lumbar Motoneurons

Miguel Martin-Caraballo and Stuart E. Dryer

Department of Biology and Biochemistry, University of Houston, Houston, Texas 77204-5513

The functional expression of large-conductance Ca²⁺-activated K⁺ (K_Ca) channels in lumbar motoneurons (LMNs) of the developing chick embryo is regulated in part by interactions with striated muscle target tissues. Here we show that the functional expression of K_Ca channels in LMNs developing in vitro can be stimulated by application of a skeletal muscle extract (MEX) or by coculture with hindlimb myotubes. A similar stimulation of K_Ca channels in vitro can be produced by the trophic factors glial cell line-derived neurotrophic factor (GDNF) and brain-derived neurotrophic factor but not by neurotrophin (NT)-3 or NT-4. The actions of MEX and hindlimb myotubes are blocked by a GDNF-neutralizing antiserum. Moreover, injection of this same antiserum into the embryonic hindlimb reduced the functional expression of K_Ca channels in vivo to levels seen in LMNs deprived of interactions with the hindlimb. The effects of GDNF on K_Ca channel expression in LMNs require 24 hr of continuous exposure to reach maximum and are blocked by the translation inhibitor anisomycin, indicating the need for synthesis of new proteins. GDNF actions are also blocked by the farnesyl transferase inhibitor manumycin, suggesting a role for Ras in the actions of GDNF. Finally, the actions of GDNF are inhibited by PP2, an inhibitor of Src family tyrosine kinases, and by LY29003, an inhibitor of phosphatidylinositol 3 kinases, but not by PD98059, an inhibitor of the Erk signaling cascade. None of these treatments alter expression of voltage-activated Ca²⁺ channels. Thus, the actions of GDNF on LMN K_Ca channel expression appear to use a transduction pathway similar to that used for regulation of apoptosis.

Key words: motoneuron; development; Ca²⁺-activated K⁺ channels; trophic factors; GDNF; kinase

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Copyright © 2002 Society for Neuroscience  0270-6474/02/222310201-08$05.00/0

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