OMEGA-CONOTOXIN AND PREJUNCTIONAL MODULATION OF THE BIPHASIC RESPONSE OF THE RAT ISOLATED URINARY-BLADDER TO SINGLE PULSE ELECTRICAL-FIELD STIMULATION

1 Single pulse electrical field stimulation (EFS) produces a biphasic response of muscle strips of the rat isolated urinary bladder consisting of an early and a late contraction which were atropine-resistant and atropine-sensitive, respectively. Repeated application of desensitizing doses of the P2 purinoceptor agonist, alpha,beta-methylene ATP (mATP) inhibited the early response while leaving unaffected the late component. 2 Omega conotoxin (CTX, 0.1-mu-M) inhibited both the early and the late response either in control conditions or after enhancement by physostigmine (0.1-mu-M). The effect of CTX was, in both cases, more pronounced on the late than the early response to EFS. CTX (0.1-mu-M) failed to affect contraction produced by ATP or acetylcholine at concentrations (0.3 mM and 0.5-mu-M) which produced a response similar to that to EFS. 3 The effect of physostigmine was more intense for the late than the early response and was abolished by atropine. In the presence of CTX, physostigmine enhanced both the early and the late components of the mechanical response to EFS. 4 Nifedipine (0.1-mu-M) reduced to a similar extent both the early and late responses. Bay K 8644 (1-mu-M) produced a marked enhancement of the response to EFS, which, however, did not have a distinct late peak. In the presence of Bay K 8644, either atropine (3-mu-M) or tetrodotoxin (1-mu-M) had minor inhibitory effects indicating the myogenic origin of the response. 5 Neurokinin A (0.1-1 nM) enhanced both the early and late responses to EFS without affecting the contraction produced by exogenous acetylcholine or ATP. A consistent potentiation was evident also in the presence of CTX and for the early response, in the presence of atropine. Clonidine (3-mu-M) inhibited the response to EFS either in the absence or the presence of physostigmine. The inhibitory effect of clonidine, shown previously to depend upon activation of prejunctional alpha-2-adrenoceptors, was still observed in presence of CTX or atropine. 6 It is concluded that CTX-sensitive voltage dependent calcium channels play a more important role in determining the cholinergic rather than the non-cholinergic, putatively purinergic, component of the biphasic response of the rat bladder to single pulse EFS. The action of CTX is likely to be exerted on N-type rather than L-type (dihydropyridine-sensitive) calcium channels. Prejunctional modulation (enhancement by neurokinin A, inhibition by clonidine) occurs even in the presence of CTX-sensitive channels blockade.