Department of Physiology, Oregon Health and Science University, Portland, OR, United States.
Neurosci Lett. 2012 Oct 31;529(1):55-9. doi: 10.1016/j.neulet.2012.08.089. Epub 2012 Sep 19.
Heart rate is controlled by stimulatory sympathetic and inhibitory parasympathetic nerves innervating the sino-atrial node and cardiac conduction system. Sympathetic release of norepinephrine (NE) and parasympathetic release of acetylcholine (ACh) are controlled by the central nervous system, and by pre-synaptic inhibition of transmitter release within the atria. An increase in cardiac sympathetic transmission relative to parasympathetic transmission is pathological as it can lead to disturbances in heart rhythm, catecholaminergic toxicity and development of arrhythmias or fibrillation. Mice lacking the p75 neurotrophin receptor (p75(-/-)) have elevated atrial NE but a low heart rate suggesting autonomic dysregulation. Similarly, mice whose sympathetic neurons lack the gp130 cytokine receptor (gp130 KO) have a normal heart rate but enhanced bradycardia after vagal nerve stimulation. What is unclear is whether cardiac autonomic disturbances in these animals reflect systemic alterations in nerve activity or whether localized defects in neurotransmitter stores or release are involved. To examine local stimulus-evoked release of neurotransmitters, we have developed a novel method for simultaneous quantification of both NE and ACh after ex vivo atrial field stimulation. Using HPLC with electrochemical detection for NE, and HPLC with mass spectrometry for ACh, we found that following field stimulation NE release was impaired in p75(-/-) atria while ACh content and release was elevated in gp130 KO atria. Thus, alterations in localized transmitter release from atrial explants are consistent with in vivo deficits in heart rate control, suggesting peripheral alterations in autonomic transmission in these mice.
心率由支配窦房结和心脏传导系统的兴奋性交感神经和抑制性副交感神经控制。交感神经释放去甲肾上腺素(NE)和副交感神经释放乙酰胆碱(ACh)受中枢神经系统控制,并通过在心房内抑制递质释放的突触前抑制来控制。与副交感神经释放相比,心脏交感神经传递的增加是病理性的,因为它可能导致心律紊乱、儿茶酚胺毒性和心律失常或颤动的发展。缺乏 p75 神经生长因子受体(p75(-/-))的小鼠心房内 NE 升高,但心率较低,提示自主神经失调。同样,缺乏 gp130 细胞因子受体(gp130 KO)的交感神经元的小鼠心率正常,但迷走神经刺激后心动过缓增强。尚不清楚这些动物的心脏自主神经紊乱是反映全身神经活动的改变,还是涉及局部神经递质储存或释放的缺陷。为了检查神经递质的局部刺激诱发释放,我们开发了一种新的方法,用于在离体心房场刺激后同时定量测定 NE 和 ACh。我们使用高效液相色谱法(HPLC)电化学检测 NE,和高效液相色谱法(HPLC)质谱检测 ACh,发现 p75(-/-)心房中的 NE 释放在刺激后受损,而 gp130 KO 心房中的 ACh 含量和释放增加。因此,心房外植体中局部递质释放的改变与心率控制的体内缺陷一致,提示这些小鼠的自主神经传递存在外周改变。