Department of Physiology, School of Medicine, University of Occupational and Environmental Health, Kitakyushu 807-8555, Japan.
J Neurosci. 2010 Jan 20;30(3):876-84. doi: 10.1523/JNEUROSCI.2986-09.2010.
The release of arginine vasopressin (AVP) from the magnocellular neurosecretory cells (MNCs) in the supraoptic nucleus (SON) is crucial for body fluid homeostasis. The MNC activity is modulated by synaptic inputs and humoral factors. A recent study demonstrated that an N-terminal splice variant of the transient receptor potential vanilloid type 1 (TRPV1) is essential for osmosensory transduction in the SON. In the present study, we examined the effects of mannitol and angiotensin II on miniature EPSCs (mEPSCs) in the supraoptic MNCs using whole-cell patch-clamp recording in in vitro slice preparation. Mannitol (60 mm) and angiotensin II (0.1 microm) increased the frequency of mEPSCs without affecting the amplitude. These effects were attenuated by pre-exposure to a nonspecific TRPV channel blocker, ruthenium red (10 microm) and enhanced by pre-exposure to cannabinoid type1 receptor antagonist, AM251 (2 microm). Mannitol-induced potentiation of mEPSCs was not attenuated by angiotensin II receptor antagonist, losartan (10 microm), indicating independent pathways of mannitol and angiotensin II to the TRPV channels. The potentiation of mEPSCs by mannitol was not mimicked by a TRPV1 agonist, capsaicin, and also not attenuated by TRPV1 blockers, capsazepine (10 microm). PKC was involved in angiotensin II-induced potentiation of mEPSCs. The effects of mannitol and angiotensin II on the supraoptic MNCs in trpv1 knock-out mice were significantly attenuated compared with those in wild-type mice counterparts. The results suggest that hyperosmotic stimulation and angiotensin II independently modulate mEPSCs through capsaicin-insensitive TRPV1 channel in the presynaptic terminals of the SON.
精氨酸加压素(AVP)从视上核(SON)的大细胞神经分泌细胞(MNC)中释放对于体液平衡至关重要。MNC 的活动受到突触输入和体液因素的调节。最近的一项研究表明,瞬时受体电位香草酸类型 1(TRPV1)的 N 端剪接变体对于 SON 中的渗透压感应至关重要。在本研究中,我们使用体外切片制备中的全细胞膜片钳记录技术,检查甘露醇和血管紧张素 II 对视上核 MNC 中小型 EPSC(mEPSC)的影响。甘露醇(60mm)和血管紧张素 II(0.1μm)增加了 mEPSC 的频率,而不影响幅度。这些作用被非特异性 TRPV 通道阻滞剂钌红(10μm)预先暴露所减弱,并被大麻素 1 型受体拮抗剂 AM251(2μm)预先暴露所增强。血管紧张素 II 受体拮抗剂洛沙坦(10μm)并不能减弱甘露醇诱导的 mEPSC 增强,表明甘露醇和血管紧张素 II 作用于 TRPV 通道的途径是独立的。TRPV1 激动剂辣椒素不能模拟甘露醇对 mEPSC 的增强作用,也不能被 TRPV1 阻滞剂辣椒素(10μm)减弱。PKC 参与了血管紧张素 II 诱导的 mEPSC 增强。与野生型小鼠相比,trpv1 敲除小鼠的 SON 中 mEPSC 对甘露醇和血管紧张素 II 的作用明显减弱。结果表明,高渗刺激和血管紧张素 II 通过 SON 突触前末梢中辣椒素不敏感的 TRPV1 通道独立调节 mEPSC。