The method of simultaneous detection of connexin 43 and astrocyte-like cells in the human pineal gland.

Abstract

ACKGROUND: Connexin 43 is one of the important gap junction proteins of astrocytes and is required for intercellular communication. To date, data on gap junctions in the human pineal gland are scarce, and connexin 43 has not been previously studied in this organ. AIM: The study was aimed to develop a method for the simultaneous detection of the gap junction marker connexin 43 (Cx43) and the astrocyte marker glial fibrillary acidic protein (GFAP) in the human pineal gland. METHODS: Fixed and paraffin-embedded samples of the human pineal gland (n = 4) were used in the study. For the simultaneous detection of Cx43 and GFAP in the human pineal gland, immunohistochemistry was used followed by analysis using a LSM800 confocal laser scanning microscope (Zeiss, Germany). RESULTS: Our immunohistochemical study for the first time showed the presence of the gap junction protein connexin 43 in the human pineal gland. The confocal microscopy with double immunolabeling of Cx43 and GFAP proteins visualized the individual clusters of connexin 43-containing structures that were undistinguishable under transmitted light microscopy, and also to show the localization of the connexin 43 on the membrane of astrocyte-like cells. CONCLUSION: The developed method makes it possible to determine Cx43-positive structures in human pineal tissue, which are localized mostly in the area of processes of astrocyte-like cells.