TY - JOUR
T1 - Effect of subtype-specific Ca2+-antagonists and Ca2+-free media on the field stimulation-evoked release of ATP and [3H]acetylcholine from rat habenula slices
AU - Sperlágh, Beáta
AU - András, Ibolya
AU - Vizi, E. Sylvester
N1 - Funding Information:
This work was supported by grants from the Hungarian Research Fund (OTKA F013019) the Hungarian Medical Research Council (ETT-144/1993) and Cooperation in Science and Technology with Central and Eastern European Countries (ERB CIPA-CT 92-3014).
PY - 1997
Y1 - 1997
N2 - The involvement of different subtypes of voltage-sensitive Ca2+ channels in the initiation of field stimulation-induced endogenous adenosine triphosphate (ATP) and [3H]acetylcholine ([3H]ACh) release was investigated in the superfused rat habenula slices. ATP, measured by the luciferin- luciferase assay, and [3H]ACh were released simultaneously from the tissue in response to low frequency electrical stimulation (2 Hz, 2.5 msec, 360 shocks). The N-type Ca2+ channel blocker ω-conotoxin GVIA (ω-CgTX, 0.01- 1 μM) reduced the stimulation-evoked release of ATP and [3H]ACh in a dose- dependent manner. Similarly, the P-type Ca2+ channel antagonist ω-agatoxin IVA (ω-Aga IVA) (0.05 μM) and the inorganic Ca2+ channel blocker Cd2+ (0.2 mM) inhibited the outflow of both transmitters, while Ni2+ (0.1 mM) was without significant effect. A high correlation was observed between the percent inhibition of ATP release and percent inhibition of ACh release caused by the different Ca2+ antagonists. Long-term perfusion (i.e., 90 min) with Ca2+ free solution inhibited the evoked-release of ATP and [3H]ACh. In contrast, perfusion of slices with the same media for a shorter time (i.e., 20 min) did not reduce the release of [3H]ACh and ATP but even increased the evoked-release of ATP about fourfold. The breakdown of extracellular ATP was not blocked under low [Ca2+]0 condition, measured by the creatine phosphokinase assay and HPLC-UV technique. Application of extra- or intracellular Ca2+ chelators, and dipyridamole (2 μM), the nucleoside transporter inhibitor, did not reduce the excess release of ATP after short- term perfusion with Ca2+-free media. Tetrodotoxin (TTX, 1 μM), while inhibiting the majority of ATP release under normal conditions, was also unable to reduce release under low [Ca2+]0 conditions. In summary, we showed that both N- and P-type Ca2+ channels are involved in the initiation of electrical stimulation-evoked release of ATP and [3H]ACh in the rat habenula under normal extracellular calcium concentration. Under low [Ca2+]0 conditions an additional release of ATP occurs, which is not associated with action potential propagation.
AB - The involvement of different subtypes of voltage-sensitive Ca2+ channels in the initiation of field stimulation-induced endogenous adenosine triphosphate (ATP) and [3H]acetylcholine ([3H]ACh) release was investigated in the superfused rat habenula slices. ATP, measured by the luciferin- luciferase assay, and [3H]ACh were released simultaneously from the tissue in response to low frequency electrical stimulation (2 Hz, 2.5 msec, 360 shocks). The N-type Ca2+ channel blocker ω-conotoxin GVIA (ω-CgTX, 0.01- 1 μM) reduced the stimulation-evoked release of ATP and [3H]ACh in a dose- dependent manner. Similarly, the P-type Ca2+ channel antagonist ω-agatoxin IVA (ω-Aga IVA) (0.05 μM) and the inorganic Ca2+ channel blocker Cd2+ (0.2 mM) inhibited the outflow of both transmitters, while Ni2+ (0.1 mM) was without significant effect. A high correlation was observed between the percent inhibition of ATP release and percent inhibition of ACh release caused by the different Ca2+ antagonists. Long-term perfusion (i.e., 90 min) with Ca2+ free solution inhibited the evoked-release of ATP and [3H]ACh. In contrast, perfusion of slices with the same media for a shorter time (i.e., 20 min) did not reduce the release of [3H]ACh and ATP but even increased the evoked-release of ATP about fourfold. The breakdown of extracellular ATP was not blocked under low [Ca2+]0 condition, measured by the creatine phosphokinase assay and HPLC-UV technique. Application of extra- or intracellular Ca2+ chelators, and dipyridamole (2 μM), the nucleoside transporter inhibitor, did not reduce the excess release of ATP after short- term perfusion with Ca2+-free media. Tetrodotoxin (TTX, 1 μM), while inhibiting the majority of ATP release under normal conditions, was also unable to reduce release under low [Ca2+]0 conditions. In summary, we showed that both N- and P-type Ca2+ channels are involved in the initiation of electrical stimulation-evoked release of ATP and [3H]ACh in the rat habenula under normal extracellular calcium concentration. Under low [Ca2+]0 conditions an additional release of ATP occurs, which is not associated with action potential propagation.
KW - ATP
KW - Acetylcholine
KW - Ca channels
KW - Habenula
KW - Release
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U2 - 10.1023/A:1022470725132
DO - 10.1023/A:1022470725132
M3 - Article
C2 - 9239752
AN - SCOPUS:0030809254
VL - 22
SP - 967
EP - 975
JO - Neurochemical Research
JF - Neurochemical Research
SN - 0364-3190
IS - 8
ER -