Department of Molecular Biology and Endocrinology, VINČA Institute of Nuclear Sciences - National Institute of thе Republic of Serbia, University of Belgrade, Belgrade, Serbia.
Department for General Physiology and Biophysics, Faculty of Biology, University of Belgrade, Belgrade, Serbia.
Histol Histopathol. 2022 Oct;37(10):919-936. doi: 10.14670/HH-18-471. Epub 2022 May 16.
Adenosine 5'-triphosphate (ATP) and other nucleotides and nucleosides, such as adenosine, are versatile signaling molecules involved in many physiological processes and pathological conditions in the nervous system, especially those with an inflammatory component. They can be released from nerve cells, glial cells, and vascular cells into the extracellular space where they exert their function via ionotropic (P2X) or metabotropic (P2Y) receptors. Signaling via extracellular nucleotides and adenosine is regulated by cell-surface located enzymes ectonucleotidases that hydrolyze the nucleotide to the respective nucleoside. This review summarizes a histochemical approach for detection of ectonucleotidase activities in the cryo-sections of brain tissue. The enzyme histochemistry (EHC) might be used as suitable replacement for immunohistochemistry, since it gives information about both localization and activity, thus adding a functional component to a classical histological approach. With this technique, it is possible to visualize spatial distribution and cell-specific localization of ecto-nucleoside triphosphate diphosphohydrolases (NTPDases) and ecto-5'-nucleotidase (eN/CD73) activities during brain development, after different hormonal manipulations, during neurodegeneration, etc. EHC is also suitable for investigation of microglial morphology in different (patho)physiological conditions. Furthermore, the review describes how to quantify EHC results.
三磷酸腺苷（ATP）和其他核苷酸和核苷，如腺苷，是多功能信号分子，参与神经系统中的许多生理过程和病理状况，特别是那些具有炎症成分的过程和状况。它们可以从神经元、神经胶质细胞和血管细胞释放到细胞外空间，在那里通过离子型（P2X）或代谢型（P2Y）受体发挥作用。细胞外核苷酸和腺苷的信号转导受位于细胞膜表面的酶——核苷酸酶的调节，这些酶将核苷酸水解为相应的核苷。这篇综述总结了一种用于检测脑组织冷冻切片中核苷酸酶活性的组织化学方法。酶组织化学（EHC）可以作为免疫组织化学的合适替代品，因为它提供了关于定位和活性的信息，从而为经典的组织学方法添加了一个功能成分。通过该技术，可以在脑发育过程中、在不同的激素处理后、在神经退行性变等过程中，可视化核苷酸三磷酸二磷酸水解酶（NTPDases）和 5'-核苷酸酶（eN/CD73）的空间分布和细胞特异性定位。EHC 也适用于研究不同（病理）生理条件下的小胶质细胞形态。此外，该综述还描述了如何量化 EHC 结果。
