Hurbin A, Boissin-Agasse L, Orcel H, Rabié A, Joux N, Desarménien M G, Richard P, Moos F C
Centre National de la Recherche Scientifique, Unité Propre de Recherche 9055, Centre de Pharmacologie-Endocrinologie, Montpellier, France.
Endocrinology. 1998 Nov;139(11):4701-7. doi: 10.1210/endo.139.11.6320.
We have identified and visualized the vasopressin (VP) receptors expressed by hypothalamic magnocellular neurons in supraoptic and paraventricular nuclei. To do this, we used RT-PCR on total RNA extracts from supraoptic nuclei or on single freshly dissociated supraoptic neurons, and in situ hybridization on frontal sections of hypothalamus of Wistar rats. The RT-PCR on supraoptic RNA extracts revealed that mainly V1a, but also V1b, subtypes of VP receptors are expressed from birth to adulthood. No V2 receptor messenger RNA (mRNA) was detected. Furthermore, the single-cell RT-nested PCR indicated that the V1a receptor mRNA is present in vasopressinergic magnocellular neurons. In light of these results, in situ hybridization was performed to visualize the V1a and V1b receptor mRNAs in supraoptic and paraventricular nuclei. Simultaneously, we coupled this approach to: 1) in situ hybridization detection of oxytocin or VP mRNAs; or 2) immunocytochemistry to detect the neuropeptides. This provided a way of identifying the neurons expressing perceptible amounts of V1a or V1b receptor mRNAs as vasopressinergic neurons. Here, we suggest that the autocontrol exerted specifically by VP on vasopressinergic neurons is mediated through, at least, V1a and V1b subtype receptors.
我们已经鉴定并可视化了视上核和室旁核中下丘脑大细胞神经元表达的血管加压素(VP)受体。为此,我们对视上核的总RNA提取物或单个新鲜解离的视上神经元进行了逆转录聚合酶链反应(RT-PCR),并对Wistar大鼠下丘脑额叶切片进行了原位杂交。对视上核RNA提取物进行的RT-PCR显示,从出生到成年,主要表达V1a亚型的VP受体,同时也表达V1b亚型的VP受体。未检测到V2受体信使核糖核酸(mRNA)。此外,单细胞RT巢式PCR表明,V1a受体mRNA存在于血管加压素能大细胞神经元中。鉴于这些结果,我们进行了原位杂交,以可视化视上核和室旁核中的V1a和V1b受体mRNA。同时,我们将这种方法与以下方法相结合:1)催产素或VP mRNA的原位杂交检测;或2)免疫细胞化学检测神经肽。这提供了一种将表达可感知量的V1a或V1b受体mRNA的神经元鉴定为血管加压素能神经元的方法。在此,我们认为VP对血管加压素能神经元的特异性自身调控至少是通过V1a和V1b亚型受体介导的。