Potent and voltage-dependent block by philanthotoxin-343 of neuronal nicotinic receptor/channels in PC12 cells.
1. Block by philanthotoxin-343 (PhTX-343), a neurotoxin from wasps, of ionic currents mediated through neuronal nicotinic acetylcholine (ACh) receptor/channels was characterized in rat phaeochromocytoma PC12 cells, by use of whole cell voltage-clamp techniques. 2. In the cells held at -60 mV, PhTX-343 at 0.1 and 1 microM inhibited an inward current activated by 100 microM ACh. The current inhibition was relieved by depolarizing steps, and augmented at negative potentials, suggesting that PhTX-343 blocks the channel in a voltage-dependent manner. Joro spider toxin-3 (JSTX-3) also exerted voltage-dependent inhibition of ACh-activated currents in a similar concentration range, but argiotoxin636 did not affect the currents. 3. Analysis of the current decay during hyperpolarizing steps indicated that the current inhibition by 100 nM PhTX-343 develops in an order of several hundres of milliseconds. On the other hand, the recovery from the current inhibition during depolarizing steps developed in an order of about 100 ms. 4. The results suggest that PhTX-343 blocks neuronal nicotinic receptor channels in PC12 cells at concentrations lower than those required for channel block in non-mammalian cells, and the block exhibits clear voltage-dependence. Estimated from the voltage-dependence, the binding site of PhTX-343 may be located near the outer mouth of the channel.
Liu
M
M
Division of Pharmacology, National Institute of Health Sciences, Tokyo, Japan.
Nakazawa
K
K
Inoue
K
K
Ohno
Y
Y
D000109:Acetylcholine / Q000494:pharmacology; D000818:Animals; D001180:Arthropod Venoms / Q000494:pharmacology*; D008564:Membrane Potentials; D018733:Nicotinic Antagonists / Q000494:pharmacology*; D016716:PC12 Cells; D018408:Patch-Clamp Techniques; D010636:Phenols / Q000494:pharmacology*; D011073:Polyamines / Q000494:pharmacology*; D051381:Rats; D011978:Receptors, Nicotinic / Q000187:drug effects* / Q000502:physiology
D001180:Arthropod Venoms; D018733:Nicotinic Antagonists; D010636:Phenols; D011073:Polyamines; D011978:Receptors, Nicotinic; C074003:philanthotoxin 343; D000109:Acetylcholine