Baclofen inhibits guinea pig magnocellular neurones via activation of an inwardly rectifying K+ conductance

The GABA B receptors GABA B -R1 and GABA B -R2 have been cloned in several mammalian species, and the functional receptor has been shown to exist as a heterodimeric complex. We have cloned guinea pig GABA B -R1 and GABA B -R2 receptor sequences and, using in situ hybridization and immunocytochemistry for vasopressin (AVP), we found that GABA B -R1 and -R2 receptors are expressed in vasopressin neurones of the supraoptic (SON) and paraventricular nuclei (PVN). Therefore, we used both sharp electrode and whole-cell patch recording techniques to examine the effects of the selective GABA B agonist baclofen on SON and PVN magnocellular neurones and to determine the coupling of the GABA B receptor to effector pathways. Recordings were made in coronal hypothalamic slices from both female (ovariectomized) and male guinea pigs. In the presence of tetrodotoxin (TTX), baclofen hyperpolarized (Δ V max = 5.6 mV, EC 50 = 2.3 μM) SON magnocellular neurones ( n = 27) under current clamp, or induced an outward current that reversed at E K (Δ I max = 24.2 pA) in PVN magnocellular neurones ( n = 33) under voltage clamp. Seventeen of the 24 biocytin-labelled SON magnocellular neurones were identified as AVP neurones, and ten of the 33 biocytin-labelled PVN neurones were identified as AVP or neurophysin-containing neurones, although all of the cells were clustered in the vasopressin-rich core. In the absence of TTX, baclofen activated an outward K + current that hyperpolarized SON and PVN neurones and significantly reduced their firing rate. The outward current showed inward rectification and was blocked by the K + channel blocker barium and the GABA B receptor antagonist CGP 35348. Therefore, GABA B receptors are coupled to inwardly rectifying K + channels in SON and PVN magnocellular neurones and may play a prominent role in modulating phasic bursting activity in guinea pig vasopressin neurones.