Trudel Eric, Bourque Charles W
Centre for Research in Neuroscience, Montreal General Hospital and McGill University, 1650 Cedar Avenue, Montreal, QC, Canada H3G 1A4.
J Neurosci Methods. 2003 Sep 30;128(1-2):67-77. doi: 10.1016/s0165-0270(03)00149-3.
The organum vasculosum lamina terminalis (OVLT), the suprachiasmatic nucleus (SCN) and the supraoptic nucleus (SON) are three hypothalamic structures involved in the osmotic and circadian control of neurohypophysial secretion. Recent experiments have suggested that interactions between osmotic and circadian factors may be important for homeostasis. The existence of an in vitro slice preparation retaining these nuclei and their interconnections would therefore be useful for the analysis of synaptic interactions. In the rat, the OVLT, SCN and SON are found at increasingly ventral and lateral positions along the rostro-caudal axis, such that conventional 400 microm slices taken in the pure coronal or horizontal planes do not retain all three nuclei. Here we show that horizontal slices cut at angles of 38-42 degrees relative to the dorsal surface of the cortex retain large fractions of the three nuclei. Intracellular recordings revealed membrane properties consistent with those previously published for OVLT, SCN and SON neurons. Moreover, antidromic and synaptic responses evoked by electrical stimulation revealed that extensive axonal projections are retained between these nuclei. Finally, chemical and osmotic stimulation of the OVLT exerted powerful influences on the rate of spontaneous synaptic events in SON neurons. We therefore conclude that angled horizontal hypothalamic slices represent a useful preparation for the analysis of physiological interactions between the OVLT, SCN and SON.
终板血管器(OVLT)、视交叉上核(SCN)和视上核(SON)是下丘脑的三个结构,参与神经垂体分泌的渗透压和昼夜节律控制。最近的实验表明,渗透压和昼夜节律因素之间的相互作用可能对体内平衡很重要。因此,保留这些核及其相互连接的体外切片制备方法对于分析突触相互作用将是有用的。在大鼠中,OVLT、SCN和SON沿头尾轴在越来越腹侧和外侧的位置被发现,以至于在纯冠状面或水平面获取的传统400微米切片不能保留所有三个核。在这里我们表明,相对于皮质背表面以38 - 42度角切割的水平切片保留了大部分的这三个核。细胞内记录显示的膜特性与先前发表的OVLT、SCN和SON神经元的膜特性一致。此外,电刺激诱发的逆向和突触反应表明,这些核之间保留了广泛的轴突投射。最后,对OVLT的化学和渗透压刺激对SON神经元中自发突触事件的发生率产生了强大影响。因此,我们得出结论,倾斜的水平下丘脑切片是分析OVLT、SCN和SON之间生理相互作用的有用制备方法。