Neumann I D
Department of Behavioral Neuroendocrinology, Max Planck Institute of Psychiatry, Kraepelinstrasse 2, Munich, D-80804, Germany.
Methods. 2000 Nov;22(3):227-37. doi: 10.1006/meth.2000.1074.
The possibility of sequence-dependent, transient, and local inhibition of neuropeptide or neuropeptide receptor expression within the brain makes antisense targeting an attractive approach for those interested in the involvement of brain neuropeptide systems in behavioral and neuroendocrine regulation. Here, I describe our attempts to manipulate the synthetic activity of peptidergic systems of the hypothalamic-neurohypophysial system, i.e. , oxytocin and vasopressin, and the hypothalamic-pituitary-adrenal (HPA) axis by antisense oligodeoxynucleotides. Detailed experimental protocols including different approaches for intracerebral antisense application in anesthetized or conscious rats are provided. As a consequence of local oxytocin or vasopressin antisense treatment within the hypothalamic supraoptic nucleus, various aspects of the neuronal activity are already altered after a few hours. Thus, we monitored electrophysiological parameters of oxytocinergic and vasopressinergic neurons, stimulus-induced expression of the Fos protein in oxytocin neurons, and stimulated release of oxytocin or vasopressin into blood as well as within the hypothalamus by dendrites and cell bodies as measured by simultaneous microdialysis in blood and brain, shortly after a single acute antisense infusion. We also employed chronic antisense infusion via osmotic minipumps or by repeated local infusion into the targeted brain region; for example, septal vasopressin receptor downregulation impairs the ability of male rats to discriminate between juvenile rats. Further, reduction of the amount of available CRH, vasopressin, and oxytocin within the hypothalamic paraventricular nuclei alters the neuroendocrine stress response of the HPA axis.
脑内神经肽或神经肽受体表达可能存在序列依赖性、短暂性和局部抑制，这使得反义靶向对于那些对脑内神经肽系统参与行为和神经内分泌调节感兴趣的人来说是一种有吸引力的方法。在此，我描述了我们尝试通过反义寡脱氧核苷酸来操纵下丘脑 - 神经垂体系统（即催产素和加压素）以及下丘脑 - 垂体 - 肾上腺（HPA）轴的肽能系统的合成活性。文中提供了详细的实验方案，包括在麻醉或清醒大鼠中进行脑内反义应用的不同方法。由于在下丘脑视上核内进行局部催产素或加压素反义治疗，数小时后神经元活动的各个方面就已发生改变。因此，在单次急性反义输注后不久，我们监测了催产素能和加压素能神经元的电生理参数、催产素神经元中刺激诱导的Fos蛋白表达，以及通过血液和脑内同时进行微透析测量的催产素或加压素向血液以及下丘脑内通过树突和细胞体的刺激释放。我们还通过渗透微型泵或通过反复局部输注到靶向脑区进行慢性反义输注；例如，隔区加压素受体下调会损害雄性大鼠区分幼鼠的能力。此外，下丘脑室旁核内可用的促肾上腺皮质激素释放激素（CRH）、加压素和催产素量的减少会改变HPA轴的神经内分泌应激反应。
