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去甲肾上腺素能神经元中GAP - 43和p38(突触素)水平的去中心化效应:一项使用免疫荧光和共聚焦激光扫描显微镜的半定量研究

Effects of decentralization on the levels of GAP-43 and p38 (synaptophysin) in sympathetic adrenergic neurons: a semi-quantitative study using immunofluorescence and confocal laser scanning microscopy.

作者信息

Hou X E, Dahlström A

机构信息

Department of Anatomy and Cell Biology, Göteborg University, Sweden.

出版信息

Brain Res. 1995 May 8;679(1):49-63. doi: 10.1016/0006-8993(95)00219-g.

Abstract

The distribution of GAP-43 in superior cervical ganglion (SCG) and iris were studied in normal animals and following decentralization using immunofluorescence and confocal laser scanning microscopy (CLSM). GAP-43-like immunoreactivity (LI) was compared with p38 (synaptophysin)-LI, and tyrosin hydroxylase (TH)-LI. In the control SCG, GAP-43-LI and p38-LI were mainly localized in nerve terminals around the principal neurons. The neuronal perikarya were negative for GAP-43, but positive for p38 in a perinuclear zone, as well as positive for TH. SIF cells (Small Intensely Fluorescent cells, ganglionic interneurons) were positive for GAP-43, TH and p38. One day after decentralization, GAP-43-LI and p38-LI in nerve terminals around principal neurons had disappeared. Some of the principal neurons showed a weak GAP-43-immunoreactivity. Three days post-decentralization, GAP-43- and p38-positive nerve terminals around the neurons had reappeared in considerable numbers and the intra-ganglionic nerve bundles were positive for both antibodies. In the control irides, GAP-43-LI and p38-LI were distributed in a varicose pattern in the nerve bundles, around blood vessels and in the network of terminals. Double labelling studies showed that GAP-43-LI was colocalized with TH-LI and p38-LI. The network of terminals in the dilator plate of the irides was quantified by measuring the fluorescence intensity of randomly selected areas, using CLSM. Three days after decentralization the intensity of GAP-43-LI and p38-LI had significantly increased. TH-LI had decreased 8 days after decentralization. The results indicate that GAP-43-LI and p38-LI are normally present in the nerve fibers and terminals of both pre- and post-ganglionic neurons in adult rats. The expression of GAP-43-LI and p38-LI in post-ganglionic neurons is preganglionically regulated, as indicated by the increased expression after decentralization. The expression of p38 in these neurons is probably regulated via mechanisms that are separate from those which regulate GAP-43, since it showed a different time course than that of GAP-43-LI.

摘要

采用免疫荧光和共聚焦激光扫描显微镜(CLSM)技术,研究正常动物及去神经支配后颈上神经节(SCG)和虹膜中生长相关蛋白43(GAP - 43)的分布。将GAP - 43样免疫反应性(LI)与突触素(p38)-LI以及酪氨酸羟化酶(TH)-LI进行比较。在对照SCG中,GAP - 43-LI和p38-LI主要定位于主神经元周围的神经终末。神经元胞体GAP - 43呈阴性,但在核周区p38呈阳性,TH也呈阳性。小而强荧光细胞(SIF细胞,神经节中间神经元)GAP - 43、TH和p38均呈阳性。去神经支配1天后,主神经元周围神经终末的GAP - 43-LI和p38-LI消失。部分主神经元显示出微弱的GAP - 43免疫反应性。去神经支配3天后,神经元周围GAP - 43和p38阳性神经终末大量重新出现,神经节内神经束两种抗体均呈阳性。在对照虹膜中,GAP - 43-LI和p38-LI以曲张形式分布于神经束、血管周围及终末网络中。双重标记研究显示GAP - 43-LI与TH-LI和p38-LI共定位。利用CLSM通过测量随机选择区域的荧光强度对虹膜瞳孔开大肌中的终末网络进行定量分析。去神经支配3天后,GAP - 43-LI和p38-LI强度显著增加。去神经支配8天后TH-LI降低。结果表明,成年大鼠中GAP - 43-LI和p38-LI正常存在于节前和节后神经元的神经纤维和终末中。去神经支配后表达增加表明节后神经元中GAP - 4

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