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钙调神经磷酸酶(钙神经素)催化亚基的神经亚型在中枢神经系统神经元中的亚细胞定位差异:利用微波辐射进行抗原修复的福尔马林固定石蜡切片免疫组织化学研究

Differential subcellular localization of neural isoforms of the catalytic subunit of calmodulin-dependent protein phosphatase (calcineurin) in central nervous system neurons: immunohistochemistry on formalin-fixed paraffin sections employing antigen retrieval by microwave irradiation.

作者信息

Usuda N, Arai H, Sasaki H, Hanai T, Nagata T, Muramatsu T, Kincaid R L, Higuchi S

机构信息

Department of Anatomy and Cell Biology, Shinshu University School of Medicine, Matsumoto, Japan.

出版信息

J Histochem Cytochem. 1996 Jan;44(1):13-8. doi: 10.1177/44.1.8543776.

Abstract

We examined the immunohistochemical distribution of the two mammalian isoforms of calcineurin catalyic subunits, A alpha and A beta, in central nervous system (CNS) tissues of cows, rats, and humans. Cryostat sections and paraffin sections of parformaldehyde-fixed tissues were stained with antipeptide antibodies for each isoform. The same localization pattern was observed in both cryostat and paraffin sections. In the latter, the intensity of the staining was dramatically enhanced by microwave irradiation. Calcineurin isoforms were localized in a variety of nerve cells but not in neuroglial cells. Their differential expression as the A alpha isoform in the nucleus and the A beta isoform in the cytoplasm was present in a variety of CNS nerve cells, most distinctively in Purkinje cells of the cerebellum and pyramidal cells of the cerebrum, irrespective of species. These results suggest that each isform has distinct intracellular sites of action in CNS neurons and that the phenomenon has been conserved during mammalian evolution.

摘要

我们研究了钙调神经磷酸酶催化亚基的两种哺乳动物同工型Aα和Aβ在牛、大鼠和人类中枢神经系统(CNS)组织中的免疫组化分布。用针对每种同工型的抗肽抗体对多聚甲醛固定组织的冰冻切片和石蜡切片进行染色。在冰冻切片和石蜡切片中均观察到相同的定位模式。在石蜡切片中,微波照射可显著增强染色强度。钙调神经磷酸酶同工型定位于多种神经细胞,但不在神经胶质细胞中。它们在细胞核中以Aα同工型、在细胞质中以Aβ同工型的差异表达存在于多种中枢神经系统神经细胞中,最明显的是在小脑的浦肯野细胞和大脑的锥体细胞中,且与物种无关。这些结果表明,每种同工型在中枢神经系统神经元中具有不同的细胞内作用位点,并且这种现象在哺乳动物进化过程中得以保留。

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