Effects of Phenylephrine on Spontaneous Activity and L-Type Ca2+ Current in Isolated Corpus Cavernosum Myocytes

Introduction. Norepinephrine is important in maintaining detumescent tone in the corpus cavernosum, although the mechanism is incompletely understood. As alpha-adrenoceptor-induced tone is antagonized by L-type Ca2+ channel blockers, it is usually assumed that direct modulation of this current is involved. However, the effects of alpha-adrenoceptor agonists have never been directly examined on L-type current in corpus cavernosum myocytes (CCSMC), leaving open other possibilities. In particular, CCSMC are now known to develop spontaneous tone via a pacemaker mechanism involving spontaneous Ca2+ waves that activate Cl- currents, causing depolarization and voltage-dependent activation of L-type channels. We hypothesized that phenylephrine modulates tone via this system, rather than by directly activating L-type channels. Aims. Examine in freshly isolated CCSMC the effect of phenylephrine on: (i) spontaneous Cl- currents and depolarizations; (ii) cytosolic Ca2+ waves; and (iii) L-type current. Methods. CCSMC were enzymatically dispersed from male New Zealand White rabbits for patch clamp recording and real time Ca2+ imaging. Main Outcome Measures. Spontaneous Cl- currents, spontaneous depolarizations, cytosolic Ca2+ and L-type current. Results. Phenylephrine enhanced the amplitude and frequency of spontaneous Cl- currents, increased the duration and frequency of spontaneous depolarizations and increased the frequency of spontaneous Ca2+ waves. These effects were blocked by 2-aminoethoxy diphenylborate (2-APB), suggesting that they were mediated by IP3-induced Ca2+ release from intracellular stores. In contrast, 2-APB had no effect on Ca2+ transients evoked by releasing stored Ca2+ with caffeine, suggesting that it had little effect on store Ca2+ content. Phenylephrine depressed L-type current by around 30%. This effect was removed by blocking with 2-APB. Notably, phenylephrine failed to enhance the current, even in the presence of 2-APB. Furthermore, the phorbol ester, phorbol 12-myristate 13-acetate, had no effect on L-type current. Conclusion. Phenylephrine effects on the corpus cavernosum are mediated by modulation of the spontaneous pacemaker mechanism, rather than by direct stimulation of L-type channels. Doyle C, Sergeant GP, Hollywood MA, McHale NG, and Thornbury KD. Effects of phenylephrine on spontaneous activity and L-type Ca2+ current in isolated corpus cavernosum myocytes. J Sex Med 2012;9:2795-2805.