Réthelyi M, Metz C B, Lund P K
Department of Physiology, University of North Carolina, Chapel Hill 27514.
Neuroscience. 1989;29(1):225-39. doi: 10.1016/0306-4522(89)90345-x.
In situ hybridization histochemistry was used to localize calcitonin gene-related peptide mRNAs in spinal cord, brain stem and dorsal root ganglion neurons of the rat and guinea-pig. A 32P-labeled 23-base-long (23mer) oligodeoxyribonucleotide (oligomer) complementary to calcitonin gene-related peptide mRNA sequences encoding residues 23-30 of calcitonin gene-related peptide was used primarily as a probe (CGRP I probe). A 32mer complementary to mRNA sequences for residues 10-20 of calcitonin gene-related peptide (CGRP II probe) was also used as a positive control for specificity of the 23mer for calcitonin gene-related peptide mRNA. In both the guinea-pig and rat calcitonin gene-related peptide mRNA was localized specifically to neurons of the dorsal root ganglion, to spinal motoneurons and to motoneurons of the hypoglossal, facial and accessory facial motor nuclei. Differences in the distribution of calcitonin gene-related peptide mRNA between the rat and guinea-pig included a higher proportion of rat dorsal root ganglion neurons containing calcitonin gene-related peptide mRNA and the localization of calcitonin gene-related peptide mRNA to motoneurons of the ambiguus motor nucleus, parabrachial and peripeduncular nucleus of the rat but not the guinea-pig. In the guinea-pig, in contrast, calcitonin gene-related peptide mRNA was localized also to motoneurons of the abducens, trigeminal, trochlear and oculomotor nerves. The neuronal groups in the intact rat found here to contain calcitonin gene-related mRNA have also been shown previously to contain calcitonin gene-related peptide immunoreactivity in colchicine-treated rats. Colchicine-treated rats, however, have been found to contain additional groups of calcitonin gene-related peptide immunoreactive neurons which, in the intact rats used in the present study, showed no detectable hybridization with the calcitonin gene-related peptide probe.
原位杂交组织化学法用于在大鼠和豚鼠的脊髓、脑干及背根神经节神经元中定位降钙素基因相关肽的信使核糖核酸。一种与编码降钙素基因相关肽第23 - 30位残基的信使核糖核酸序列互补的32P标记的23个碱基长的(23聚体)寡脱氧核糖核苷酸(寡聚物)主要用作探针(降钙素基因相关肽I探针)。一种与降钙素基因相关肽第10 - 20位残基的信使核糖核酸序列互补的32聚体(降钙素基因相关肽II探针)也用作23聚体对降钙素基因相关肽信使核糖核酸特异性的阳性对照。在豚鼠和大鼠中,降钙素基因相关肽信使核糖核酸均特异性定位于背根神经节神经元、脊髓运动神经元以及舌下神经、面神经和副面神经运动核的运动神经元。大鼠和豚鼠之间降钙素基因相关肽信使核糖核酸分布的差异包括:大鼠背根神经节中含降钙素基因相关肽信使核糖核酸的神经元比例较高,以及大鼠疑核、臂旁核和脚周核的运动神经元中有降钙素基因相关肽信使核糖核酸的定位,而豚鼠中没有。相反,在豚鼠中,降钙素基因相关肽信使核糖核酸还定位于展神经、三叉神经、滑车神经和动眼神经的运动神经元。在本研究中发现,完整大鼠中含降钙素基因相关信使核糖核酸的神经元群,在秋水仙碱处理的大鼠中先前也已显示含有降钙素基因相关肽免疫反应性。然而,已发现秋水仙碱处理的大鼠含有另外一些降钙素基因相关肽免疫反应性神经元群,在本研究中使用的完整大鼠中,这些神经元群与降钙素基因相关肽探针未显示可检测到的杂交信号。