J. Cuevas et al., Two distinct classes of functional alpha 7-containing nicotinic receptor on rat superior cervical ganglion neurons, J PHYSL LON, 525(3), 2000, pp. 735-746
1. Nicotinic acetylcholine receptors (nAChRs) that bind alpha-bungarotoxin
(alpha Bgt) were studied on isolated rat superior cervical ganglion (SCG) n
eurons using whole-cell patch clamp recording techniques.
2. Rapid application of ACh onto the soma of voltage clamped neurons evoked
a slowly desensitizing current that was reversibly blocked by alpha Bgt (5
0 nM). The toxin-sensitive current constituted on average about half of the
peak whole-cell response evoked by ACh.
3. Nanomolar concentrations of methyllycaconitine blocked the alpha Bgt-sen
sitive component of the ACh-evoked current as did intracellular dialysis wi
th an anti-alpha 7 monoclonal antibody. The results indicate that the slowl
y reversible toxin-sensitive response elicited by ACh arises from activatio
n of an unusual class of alpha 7-containing receptor (alpha 7-nAChR) simila
r to that reported previously fur rat intracardiac ganglion neurons.
4. A second class of functional alpha 7-nAChR was identified on some SCG ne
urons by using rapid application of choline to elicit responses. In these c
ases a biphasic response was obtained, which included a rapidly desensitizi
ng component that was blocked by alpha Bgt in a pseudo-irreversible manner.
The pharmacology and kinetics of the responses resembled those previously
attributed to alpha 7-nAChRs in a number of other neuronal cell types.
5. Experiments measuring the dissociation rate of I-125-labelled alpha Bgt
from SCG neurons revealed two classes of toxin-binding site. The times for
toxin dissociation were consistent with those required to reverse blockade
of the two kinds of alpha Bgt-sensitive response.
6. These results indicate that rat SCG neurons express two types of functio
nal alpha 7-nAChR, differing in pharmacology, desensitization and reversibi
lity of alpha Bgt blockade.