Baldino F, Fitzpatrick-McElligott S, Gozes I, Card J P
Medical Products Department, E.I. Dupont de Nemours and Co., Wilmington, Delaware.
J Mol Neurosci. 1989;1(4):199-207.
Messenger RNA (mRNA) coding for vasoactive intestinal polypeptide (VIP) and peptide histidine isoleucine (PHI-27) were localized in cortical and thalamic neurons with synthetic DNA probes complementary to the PHI-27 and VIP exon coding sequence of the rat VIP precursor gene. Hybridization signal with these probes was found widely distributed in the thalamus, neocortex, and pyriform cortex, and the distribution of hybridization signal for each probe was identical. Furthermore, the distribution of each message was correlated closely with peptide distribution demonstrated immunohistochemically. Labeling of individual neurons with in situ hybridization histochemistry was characterized by a dense accumulation of silver grains in the cytoplasm of these cells with little or no label in the nucleus. Labeled neurons in the thalamus were observed in the ventrolateral, ventromedial, ventrobasal, and lateral reticular nuclei. In the neocortex, the distribution of labeled neurons was concentrated in layers II and III with scattered cells also apparent in deeper cortical layers. Hybridization signal was limited to nonpyramidal neurons in both the neo- and pyriform cortex. The coextensive distribution of immunoreactivity and mRNA coding regions for VIP and PHI-27 establishes that these peptides are synthesized from the same precursor mRNA in the same thalamic and cortical cell groups. Although the physiological role of these peptides in thalamocortical function remains unknown, these data provide an anatomical substrate which suggests that VIP and PHI-27 may be cotransmitters in thalamic and cortical neurons.
利用与大鼠血管活性肠肽(VIP)前体基因的PHI-27和VIP外显子编码序列互补的合成DNA探针,对编码血管活性肠肽(VIP)和肽组氨酸异亮氨酸(PHI-27)的信使核糖核酸(mRNA)在皮质和丘脑神经元中进行定位。发现与这些探针的杂交信号广泛分布于丘脑、新皮质和梨状皮质,且每个探针的杂交信号分布相同。此外,每种信使核糖核酸的分布与免疫组织化学显示的肽分布密切相关。原位杂交组织化学对单个神经元的标记特征是,这些细胞的细胞质中有密集的银颗粒聚集,而细胞核中几乎没有标记或没有标记。在丘脑的腹外侧、腹内侧、腹基底和外侧网状核中观察到标记的神经元。在新皮质中,标记神经元的分布集中在II层和III层,在更深的皮质层中也可见散在的细胞。杂交信号仅限于新皮质和梨状皮质中的非锥体神经元。VIP和PHI-27的免疫反应性和mRNA编码区域的共同分布表明,这些肽是在相同的丘脑和皮质细胞群中由相同的前体mRNA合成的。尽管这些肽在丘脑皮质功能中的生理作用尚不清楚,但这些数据提供了一种解剖学基础,表明VIP和PHI-27可能是丘脑和皮质神经元中的共同递质。