T-Type calcium channel alpha1G and alpha1H subunits in human retinoblastoma cells and their loss after differentiation.
Human retinoblastoma cells are multipotent retinal precursor cells capable of differentiating into photoreceptors, neurons, and glia. The current-voltage relation of the undifferentiated cells is dominated by a transient inward current that disappears shortly after differentiation. In 20 mM Ba(2+)-containing bath solutions, the current has an activation midpoint near -25 mV and appears to be fully inactivated at -20 mV. Sr(2+) and Ca(2+) are preferred charge carriers relative to Ba(2+), and the current vanishes in the absence of these divalent cations. Cd(2+) blocks the current with an IC(50) of 160 microM, and Ni(2+) blocks in a biphasic manner with IC(50)s of 22 and 352 microM. The current is unaffected when sodium is replaced with other monovalent cations, and it is insensitive to nifedipine, omega-conotoxin GVIA, omega-agatoxin IVA, and omega-conotoxin MVIIC. RT-PCR revealed the presence of alpha 1G and alpha 1H mRNA in undifferentiated cells, but following differentiation, a striking reduction of both alpha 1G and alpha 1H mRNA was found, and this was paralleled by the loss of T-type Ca channel currents. alpha 1I subunit mRNA levels were low in undifferentiated and differentiated cells. These results suggest that T-type Ca channels could play a role in undifferentiated retinoblastoma cell physiology since alpha 1G and alpha 1H Ca channel subunit expression is reduced in cells that have differentiated and exited the cell cycle.
Hirooka
Kazuyuki
K
Department of Physiology and Biophysics, Dalhousie University, Halifax, Nova Scotia B3H 4H7.
Bertolesi
Gabriel E
GE
Kelly
Melanie E M
ME
Denovan-Wright
Eileen M
EM
Sun
Xiaolu
X
Hamid
Jawed
J
Zamponi
Gerald W
GW
Juhasz
Alexander E
AE
Haynes
Lawrence W
LW
Barnes
Steven
S
D002121:Calcium Channel Blockers / Q000494:pharmacology; D020747:Calcium Channels, T-Type / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D002413:Cations, Divalent / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology; D004553:Electric Conductivity; D004558:Electric Stimulation; D006801:Humans; D020748:Mibefradil / Q000494:pharmacology; D020033:Protein Isoforms / Q000378:metabolism / Q000502:physiology; D012175:Retinoblastoma / Q000378:metabolism* / Q000473:pathology; D014407:Tumor Cells, Cultured
D002121:Calcium Channel Blockers; D020747:Calcium Channels, T-Type; D002413:Cations, Divalent; D020033:Protein Isoforms; D020748:Mibefradil