Immunocytochemistry of cGMP in the Cerebellum of the Immature, Adult, and Aged Rat: the Involvement of Nitric Oxide. A Micropharmacological Study
Immunocytochemistry of cGMP in the Cerebellum of the Immature, Adult, and Aged Rat: the Involvement of Nitric Oxide. A Micropharmacological Study
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未成熟、成年和老年大鼠小脑中 cGMP 的免疫细胞化学：一氧化氮的参与。
DOI：
10.1111/j.1460-9568.1990.tb00396.x
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发表时间：
1990
影响因子：
3.4
通讯作者：
H. Steinbusch
中科院分区：
文献类型：
--
作者：
J. Vente;J. Bol;H. S. Berkelmans;J. Schipper;H. Steinbusch
在这项研究中，我们描述了甲醛固定的cGMP免疫反应性（cGMP-IR）在体外培养的大鼠小脑组织切片中的定位。在不存在磷酸二酯酶抑制的情况下，在从未成熟小脑制备的组织切片中，cGMP免疫荧光强度较低。向孵育培养基中添加异丁基甲基黄嘌呤（IBMX）导致在内部颗粒层中的星形胶质细胞簇中出现cGMP-IR。加入N-甲基-D-天冬氨酸（NMDA）、红藻氨酸、心房利钠因子（ANF）或硝普钠（SNP）后，Bergmann纤维、Bergmann细胞体和内部颗粒层中的星形胶质细胞中的cGMP-IR增强。在ANF或硝普钠存在下孵育切片后，白色物质中的星形胶质细胞显示cGMP-IR，但在NMDA或红藻氨酸存在下未显示。此外，在SNP刺激cGMP产生后，在胶质细胞酸性蛋白（GFAP）不阳性的纤维中发现cGMP-IR。在成人小脑切片中，在Bergmann纤维、Bergmann细胞体和颗粒层中的星形胶质细胞中观察到强烈的基础cGMP免疫染色。在浦肯野细胞中未观察到cGMP-IR。免疫细胞化学结果表明，NMDA、ANF或SNP刺激神经胶质结构中的cGMP含量。然而，当测量生化，只有SNP的影响是统计学上显着的，和免疫细胞化学表明，SNP明显刺激cGMP合成的神经元细胞结构。在老年大鼠的小脑中，与成年大鼠相比，发现cGMP-IR降低，在成年大鼠中显示cGMP-IR的相同结构中。在存在血红蛋白、亚甲蓝的情况下，通过用NG-单甲基-L-精氨酸（NGMAr）抑制一氧化氮合成，或通过消耗外部Ca 2+，降低基础cGMP-免疫染色。此外，NMDA和ANF（部分）对cGMP-IR的刺激作用也被这些化合物抑制。SNP刺激鸟苷酸环化酶后的cGMP-IR因血红蛋白或亚甲蓝的同时存在而降低，但NGMAr或Ca 2+的缺失则不降低。我们的研究结果表明，cGMP在小脑神经胶质组织的功能中发挥着重要作用，也表明一氧化氮在小脑谷氨酸和ANF的功能中作为细胞间介质的作用。
In this study we describe the localization of formaldehyde‐fixed cGMP‐immunoreactivity (cGMP‐IR) in rat cerebellar tissue slices incubated in vitro. In the absence of phosphodiesterase inhibition, cGMP‐immunofluorescence was of low intensity in tissue slices prepared from immature cerebella. Addition of isobutylmethylxanthine (IBMX) to the incubation medium resulted in the appearance of cGMP‐IR in clusters of astrocytes in the internal granular layer. Addition of N‐methyl‐D‐aspartate (NMDA), kainic acid, atrial natriuretic factor (ANF), or sodium nitroprusside (SNP) gave an intense cGMP‐IR in Bergmann fibres, Bergmann cell bodies, and astrocytes in the internal granular layer. Astrocytes in the white matter showed cGMP‐IR after incubation of the slice in the presence of ANF or nitroprusside, but not after NMDA or kainic acid. In addition, after SNP stimulation of cGMP production, cGMP‐IR was found in fibres which were not positive for glial fibrillary acidic protein (GFAP). In the adult cerebellar slice, intense basal cGMP‐immunostaining was observed in Bergmann fibres, Bergmann cell bodies, and astrocytes in the granular layer. No cGMP‐IR was observed in Purkinje cells. Stimulation of the cGMP‐content in the glial structures by NMDA, ANF, or SNP, was suggested by the immunocytochemical results. However, when measured biochemically, only the effect of SNP was statistically significant, and immunocytochemistry showed that SNP clearly stimulated cGMP synthesis in neuronal cell structures. In the cerebellum of the aged rat a reduced cGMP‐IR was found compared to the adult, in the same structures which showed cGMP‐IR in the adult. Basal cGMP‐immunostaining was reduced in the presence of haemoglobin, methylene blue, by inhibiting nitric oxide synthesis with NG‐monomethyl‐L‐arginine (NGMAr), or by depletion of external Ca2+. Also the stimulatory effect of NMDA and of ANF (partly) on the cGMP‐IR was inhibited by these compounds. cGMP‐IR after stimulation of guanylate cyclase by SNP was reduced by the concomitant presence of haemoglobin or methylene blue, but not by NGMAr, or by omission of Ca2+. Our results point to an important role for cGMP in the functioning of glial tissue in the cerebellum and also suggest a role for nitric oxide as an intercellular mediator in the functioning of glutamate and ANF in the cerebellum.
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DOI：
10.1016/0167-4889(89)90060-8
发表时间：
1989-03-06
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA
影响因子：
--
作者：
HIRATA, M;CHANG, CH;MURAD, F
通讯作者：
MURAD, F
影响因子：
15.9
作者：
MURAD, F
通讯作者：
MURAD, F
影响因子：
2.9
作者：
MARLETTA, MA;YOON, PS;WISHNOK, JS
通讯作者：
WISHNOK, JS
DOI：
10.1073/pnas.85.22.8664
发表时间：
1988-11-01
影响因子：
11.1
作者：
SAKUMA, I;STUEHR, DJ;LEVI, R
通讯作者：
LEVI, R
鸟苷酸环化酶，一种细胞表面受体。
DOI：
--
发表时间：
1989
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Garbers,DL
通讯作者：
Garbers,DL