A STUDY OF CELLULAR NETWORKS WITHIN THE CENTRAL NERVOUS SYSTEM BY USING CONNEXIN45 GENE KNOCKOUT-LACZ GENE KNOCKIN MICE.

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We have investigated connexin45 (Cx45) gene knockout-lacZ gene knockin mice in order, to clarify Cx45 expressing cells in the central nervous system. 1. X-gal staining of those animal tissues enable us to visualize Cx45 expressing cells by the blue coloring. We performed X-gal staining on 1mm thick tissue slices and embedded in Technovit7100 resin. One to five μm thick sections were cut and observed after staining by either toluidine blue or neutral red. In the X-gal staining positive cells, bluish reaction products were observed around cell nucleus. 2. We observed those blueish reaction products by electron microscopy (EM). Slightly electron dense amorphous materials were identified as reaction products, which enable us to identify lacZ positive cells under EM. 3. Quantum dots worked well as marker for immunocytochemistry of light microscopic level for lacZ, however, they were not effective enough for EM level because of their small size. 4. By experiments in which bone marrow cells of GFP expressing transgenic mice were transplanted to wild type donor mice, we found bone marrow derived cells were distributed in tissues as fibroblast-like cells that connected by gap junctions. A part of c-Kit expressing cells were suggested to be bone marrow derived.