Subtype-selective reconstitution of synaptic transmission in sympathetic ganglion neurons by expression of exogenous calcium channels.
Fast cholinergic neurotransmission between superior cervical ganglion neurons (SCGNs) in cell culture is initiated by N-type Ca(2+) currents through Ca(v)2.2 channels. To test the ability of different Ca(2+)-channel subtypes to initiate synaptic transmission in these cells, SCGNs were injected with cDNAs encoding Ca(v)1.2 channels, which conduct L-type currents, Ca(v)2.1 channels, which conduct P/Q-type Ca(2+) currents, and Ca(v)2.3 channels, which conduct R-type Ca(2+) currents. Exogenously expressed Ca(v)2.1 channels were localized in nerve terminals, as assessed by immunocytochemistry with subtype-specific antibodies, and these channels effectively initiated synaptic transmission. Injection with cDNA encoding Ca(v)2.3 channels yielded a lower level of presynaptic labeling and synaptic transmission, whereas injection with cDNA encoding Ca(v)1.2 channels resulted in no presynaptic labeling and no synaptic transmission. Our results show that exogenously expressed Ca(2+) channels can mediate synaptic transmission in SCGNs and that the specificity of reconstitution of neurotransmission (Ca(v)2.1 > Ca(v)2.3 >> Ca(v)1.2) follows the same order as in neurons in vivo. The specificity of reconstitution of neurotransmission parallels the specificity of trafficking of these Ca(v) channels to nerve terminals.
Mochida
Sumiko
S
Department of Physiology, Tokyo Medical University, Tokyo 160-8402, Japan.
Westenbroek
Ruth E
RE
Yokoyama
Charles T
CT
Itoh
Kanako
K
Catterall
William A
WA
D060848:Agatoxins; D000818:Animals; D001921:Brain / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D015220:Calcium Channels / Q000378:metabolism*; D020864:Calcium Channels, N-Type / Q000378:metabolism; D020908:Calcium Channels, R-Type; D027682:Cation Transport Proteins; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000378:metabolism; D005728:Ganglia, Sympathetic / Q000378:metabolism*; D005982:Glutathione Transferase / Q000378:metabolism; D007150:Immunohistochemistry; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D018408:Patch-Clamp Techniques; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013111:Spider Venoms / Q000378:metabolism; D013569:Synapses / Q000502:physiology*; D013997:Time Factors
D060848:Agatoxins; D002118:Calcium; D015220:Calcium Channels; D020864:Calcium Channels, N-Type; D020908:Calcium Channels, R-Type; D027682:Cation Transport Proteins; D018076:DNA, Complementary; D005982:Glutathione Transferase; D011993:Recombinant Fusion Proteins; D013111:Spider Venoms; C498917:Cacna1e protein, rat; C060465:omega-agatoxin I