Chan-Palay V, Palay S L
Proc Natl Acad Sci U S A. 1979 Mar;76(3):1485-8. doi: 10.1073/pnas.76.3.1485.
Guanosine 3',5'-cyclic monophosphate (cGMP) immunoreactivity in the rat's cerebellum was studied with light and electron microscopy by the indirect fluorescence method and the peroxidase-antiperoxidase method. Labeled cells included neuroglial cells in the cerebellar cortex, white matter, and deep nuclei; some stellate and basket cells in the cortex; and some large neurons in the deep nuclei. No evidence was found for sagittal microzonation in the cGMP distribution. In the labeled cells, cGMP immunoreactive sites were localized to surface membranes, organelles, and the cytoplasmic matrix. Specificity was indicated by the same pattern of labeling after treatment with cGMP immunoglobulin that had been adsorbed with adenosine 3',5'-cyclic monophosphate (cAMP) and by the failure to label after treatment with normal rabbit sera or with cGMP immunoglobulin that had been adsorbed with 1 mM cGMP. Cerebella treated with cAMP antisera, however, showed immunoreactivity in Purkinje cells, granule cells, and Golgi cells in addition to neuroglia in cortex and deep nuclei. Sequential norepinephrine and glutamate superfusions generally intensified cGMP immunoreactivity, not only in neuroglial cells but also in the background. Under these conditions some Purkinje cells and some granule cells were also labeled. Increased cGMP immunoreactivity was also obtained by treatment with harmaline, gamma-aminobutyric acid and aminooxyacetic acid, muscimol, gamma-aminobutyric acid, or apomorphine in order of decreasing effectiveness. Serotonin and colchicine produced no detectable increase of cGMP immunoreactivity above normal, and diazepam and sodium pentobarbital decreased it. In these experiments, diethyl ether was preferable to sodium pentobarbital for anesthesia on account of the depressive action of the latter on cGMP immunoreactivity. Thus, drugs that increase cerebellar activity enhance cGMP levels, whereas those that decrease cerebellar activity decrease cGMP levels. However, it is not clear whether these fluctuations in cGMP levels are a direct consequence of neurotransmitter function or are sequelae to other related events. The present study suggests that some neurons and many neuroglial cells are the major sites of cGMP in the cerebellum.
采用间接荧光法和过氧化物酶-抗过氧化物酶法，通过光学显微镜和电子显微镜对大鼠小脑鸟苷3',5'-环磷酸（cGMP）免疫反应性进行了研究。标记细胞包括小脑皮质、白质和深部核团中的神经胶质细胞；皮质中的一些星状细胞和篮状细胞；以及深部核团中的一些大神经元。未发现cGMP分布存在矢状微区划分的证据。在标记细胞中，cGMP免疫反应位点定位于表面膜、细胞器和细胞质基质。用已被腺苷3',5'-环磷酸（cAMP）吸附的cGMP免疫球蛋白处理后呈现相同的标记模式，以及用正常兔血清或已被1 mM cGMP吸附的cGMP免疫球蛋白处理后未出现标记，均表明了特异性。然而，用cAMP抗血清处理的小脑，除了皮质和深部核团中的神经胶质细胞外，浦肯野细胞、颗粒细胞和高尔基细胞也显示出免疫反应性。去甲肾上腺素和谷氨酸的顺序灌注通常会增强cGMP免疫反应性，不仅在神经胶质细胞中，而且在背景中也是如此。在这些条件下，一些浦肯野细胞和一些颗粒细胞也被标记。用哈马灵、γ-氨基丁酸和氨氧基乙酸、蝇蕈醇、γ-氨基丁酸或阿扑吗啡处理也可增加cGMP免疫反应性，其效果依次递减。5-羟色胺和秋水仙碱未使cGMP免疫反应性高于正常水平出现可检测到的增加，而地西泮和戊巴比妥钠则使其降低。在这些实验中，由于戊巴比妥钠对cGMP免疫反应性有抑制作用，因此在麻醉时乙醚比戊巴比妥钠更可取。因此，增加小脑活动的药物会提高cGMP水平，而降低小脑活动的药物会降低cGMP水平。然而，尚不清楚这些cGMP水平的波动是神经递质功能的直接结果还是其他相关事件的后遗症。本研究表明，一些神经元和许多神经胶质细胞是小脑中cGMP的主要位点。
