Expression profiles of high voltage-activated calcium channels in sympathetic and parasympathetic pelvic ganglion neurons innervating the urogenital system.
Among the autonomic ganglia, major pelvic ganglia (MPG) innervating the urogenital system are unique because both sympathetic and parasympathetic neurons are colocalized within one ganglion capsule. Sympathetic MPG neurons are discriminated from parasympathetic ones by expression of low voltage-activated Ca2+ channels that primarily arise from T-type alpha1H isoform and contribute to the generation of low-threshold spikes. Until now, however, expression profiles of high voltage-activated (HVA) Ca2+ channels in these two populations of MPG neurons remain unknown. Thus, in the present study, we dissected out HVA Ca2+ channels using pharmacological and molecular biological tools. Reverse transcription-polymerase chain reaction analysis showed that MPG neurons contained transcripts encoding all of the known HVA Ca2+ channel isoforms (alpha1B, alpha1C, alpha1D and alpha1E), with the exception of alpha1A. Western blot analysis and pharmacology with omega-agatoxin IVA (1 microM) confirmed that MPG neurons lack the alpha1A Ca2+ channels. Unexpectedly, the expression profile of HVA Ca2+ channel isoforms was identical in the sympathetic and parasympathetic neurons of the MPG. Of the total Ca2+ currents, omega-conotoxin GVIA-sensitive N-type (alpha1B) currents constituted 57 +/- 5% (n = 9) and 60 +/- 3% (n = 6), respectively; nimodipine-sensitive L-type (alpha1C and alpha1D) currents made up 17 +/- 4% and 14 +/- 2%, respectively; and nimodipine-resistant and omega-conotoxin GVIA-resistant R-type currents were 25 +/- 3% and 22 +/- 2%, respectively. The R-type Ca2+ currents were sensitive to NiCl2 (IC50 = 22 +/- 0.1 microM) but not to SNX-482, which was able to potently (IC50 = 76 +/- 0.4 nM) block the recombinant alpha1E/beta2a/alpha2delta Ca2+ currents expressed in human embryonic kidney 293 cells. Taken together, our data suggest that sympathetic and parasympathetic MPG neurons share a similar but unique profile of HVA Ca2+ channel isoforms.
Won
Yu-Jin
YJ
Department of Physiology, Yonsei University Wonju College of Medicine, Wonju, Kangwon-Do 220-701, Republic of Korea.
Whang
Kum
K
Kong
In Deok
ID
Park
Kyu-Sang
KS
Lee
Joong-Woo
JW
Jeong
Seong-Woo
SW
D000818:Animals; D015153:Blotting, Western; D015220:Calcium Channels / Q000096:biosynthesis*; D002460:Cell Line; D005726:Ganglia, Parasympathetic / Q000378:metabolism*; D005728:Ganglia, Sympathetic / Q000378:metabolism*; D006801:Humans; D008297:Male; D009474:Neurons / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D010388:Pelvis / Q000294:innervation*; D020033:Protein Isoforms; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014566:Urogenital System / Q000294:innervation*
D015220:Calcium Channels; D020033:Protein Isoforms