Effects of vasopressin on isolated rat adrenal chromaffin cells.

It has been demonstrated that arginine vasopressin (AVP) is synthesized not only in specific hypothalamic nuclei, but also in the adrenal medulla where it is thought to regulate adrenal functions by autocrine and paracrine mechanisms. In order to further characterise the effects of AVP on rat adrenal chromaffin cells, we examined: (a) the mRNA expression for V(1a) and V(1b) AVP receptors in these cells; (b) the effects of AVP on the membrane potential and membrane currents measured with the whole-cell patch-clamp technique; and (c) effect of AVP on catecholamine release from single adrenal chromaffin cells measured with carbon fibre microelectrodes. Reverse transcription-polymerase chain reaction (RT-PCR) on tissue punch samples obtained from the adrenal medulla demonstrated message for both the V(1a) and V(1b) receptors, while material obtained from the adrenal cortex showed expression of the V(1a) receptor only. Single-cell RT-PCR conducted on acutely isolated chromaffin cells showed message for the V(1a) receptor in 84% of cells, while 38% of cells also contained message for the V(1b) receptor (n=45). Under current-clamp recording, responses to AVP application (4-40 microM) were variable; 22/34 (65%) tested cells were depolarised, 29% hyperpolarised, and the remaining cells showed a biphasic response. Changes in membrane potential of either direction were dose-dependent and accompanied by a decrease in cell membrane resistance. Under voltage-clamp (V(hold)=-60 mV), AVP evoked inward current in 27/52 (52%) and outward current in 16/52 (31%) chromaffin cells. Both types of AVP-evoked responses were blocked by co-application of a nonselective V(1a)/V(1b) antagonist. Application of AVP evoked prolonged bursts of amperometric currents (indicative of catecholamine release) in 4/9 tested cells, but reduced the currents evoked by ACh application in all tested cells (n=7). These findings demonstrate a complex action of AVP on adrenal chromaffin cells, with individual adrenal chromaffin cells responding with either excitation or inhibition. This response pattern may be related to the expression of V(1) receptor subtypes.

Receptor subtype-specific modulation by dopamine of glutamatergic responses in striatal medium spiny neurons.

The output of GABAergic medium-sized spiny neurons in the dorsal striatum is controlled in part by glutamatergic input from the neocortex and the thalamus, and dopaminergic input from ventral midbrain. We acutely isolated these neurons from juvenile (P14-24) rats to study the consequences of the interaction between glutamate and dopamine for neuronal excitability. Single-cell RT-PCR analysis was used to identify the expression patterns of dopamine receptors. D1 and D2 dopamine receptor mRNA was detected in 11/22 and 3/22 of isolated neurons, respectively. Receptor mRNA co-expression was detected in 1/22 cells tested. Whole-cell voltage clamp recording (V(h)=-70 mV) was combined with local or bath application of dopaminergic and glutamatergic agonists to explore dopamine receptor modulation of glutamatergic excitation. Glutamate-evoked inward currents (5 microM, Mg(2+)-free, 1 microM glycine) were attenuated by dopamine (5 microM) to 83.2+/-3.6% (n=31). NMDA-evoked (20 microM), APV-sensitive currents were attenuated by dopamine to 80.9+/-4.5% (n=24). NMDA-induced responses were also attenuated by the D1 receptor agonist SKF 38393 (1 microM; n=28), while the D2/3 receptor agonist quinpirole (10 microM) had no effect. The currents evoked by application of AMPA (5 microM) displayed a steady rundown. Application of dopamine abolished or significantly reduced the rundown in the cells tested (n=17). A similar effect was observed after the application of SKF 38393 (1 microM), while quinpirole (10 microM) had no significant effect. Our results provide direct evidence for modulation by dopamine of glutamatergic responses of striatal medium spiny neurons, and demonstrate that the effects of this neuromodulator are receptor subtype specific. Disruption of this modulatory effect is likely to contribute to movement disorders associated with Parkinson's disease.