Ridet J L, Tamir H, Privat A
INSERM U. 336-EPHE, Université Montpellier II, France.
J Neurosci Res. 1994 May 1;38(1):109-21. doi: 10.1002/jnr.490380114.
In the present study, we performed the immunodetection of serotonergic (5-HT) receptor subtypes in the spinal cord by using an anti-idiotypic antiserum (TH8) at light and electron microscopic levels. This antibody has been shown to recognize 5-HT1B, 5-HT1C, and 5-HT2 receptor subtypes (Tamir et al.: J Neurochem 57:930-942, 1991). The TH8 immunoreactivity was observed in the dorsal and ventral horns of the gray matter. Light microscopy revealed that small cell bodies located in laminae I-III of the dorsal horn were intensely immunolabeled. A more homogenous and discrete staining was also observed throughout the entire dorsal horn. In the ventral horn, motoneurons were also immunoreactive (IR). Peroxidase deposits were observed as numerous patches covering the motoneuronal surface. Numerous interneurons were moderately and homogeneously immunostained. With the electron microscope, most of the labeled structures were identified as neurons (dendrites and perikarya) in both the dorsal and ventral horns. In the dorsal horn, immunoreactivity was present in dendrites and neuronal perikarya. A large majority of the immunoreactivity found in dendrites was not associated with synaptic differentiations. Indeed, the dendrites, in which peroxidase deposit was seen, were not locally involved in synapses. Very scarce synaptic varicosities were observed in close apposition with IR dendrites. In the ventral horn, TH8 immunoreactivity was present in dendrites, with an accumulation of peroxidase deposit on the active zone of synapses, facing presynaptic membranes. Both the postsynaptic membrane and the submembrane area were IR. In addition, a few astroglial fine processes were immunostained; most of them were observed in the dorsal horn. Scarce IR astroglial profiles were observed in the ventral horn. These observations show that such an antiserum constitutes a useful tool for the ultrastructural analysis of 5-HT receptor distribution. Finally, correlation between the immunocytochemical localization of 5-HT receptor subtypes and the modes of 5-HT transmission in the spinal cord (wiring and volume transmissions) is discussed in the present report.
在本研究中,我们使用抗独特型抗血清(TH8)在光镜和电镜水平对脊髓中的5-羟色胺能(5-HT)受体亚型进行了免疫检测。已证明该抗体可识别5-HT1B、5-HT1C和5-HT2受体亚型(塔米尔等人:《神经化学杂志》57:930 - 942,1991)。在灰质的背角和腹角观察到TH8免疫反应性。光镜显示,位于背角I - III层的小细胞体被强烈免疫标记。在整个背角也观察到更均匀且离散的染色。在腹角,运动神经元也具有免疫反应性(IR)。过氧化物酶沉积物表现为覆盖运动神经元表面的众多斑块。许多中间神经元呈中度且均匀的免疫染色。通过电子显微镜观察,在背角和腹角中,大多数标记结构被鉴定为神经元(树突和胞体)。在背角,免疫反应性存在于树突和神经元胞体中。在树突中发现的大部分免疫反应性与突触分化无关。实际上,可见过氧化物酶沉积物的树突并非局部参与突触形成。在与IR树突紧密相邻处观察到非常稀少的突触曲张。在腹角,TH8免疫反应性存在于树突中,在突触的活性区有过氧化物酶沉积物的积累,朝向突触前膜。突触后膜和膜下区域均为IR。此外,少数星形胶质细胞的细突起被免疫染色;其中大多数在背角观察到。在腹角观察到稀少的IR星形胶质细胞轮廓。这些观察结果表明,这种抗血清是用于5-HT受体分布超微结构分析的有用工具。最后,本报告讨论了5-HT受体亚型的免疫细胞化学定位与脊髓中5-HT传递模式(线路传递和容积传递)之间的相关性。