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. 2012 Jan 16;16(1):213–233. doi: 10.1017/S1461145711001933

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© CINP 2012

The online version of this article is published within an Open Access environment subject to the conditions of the Creative Commons Attribution-NonCommercial-ShareAlike licence <http://creativecommons.org/licenses/by-nc-sa/2.5/>. The written permission of Cambridge University Press must be obtained for commercial re-use.

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Fig. 5. — Mood-stabilizing phenotype is not detectable in chimeric mice transplanted with the bone marrow of P2rx7^−/− mice. (a) General scheme of bone marrow transplantation. (b) Flow cytometric analysis of the expression of the donor-specific CD45.2 allele on peripheral blood granulocytes of an intact C57BL/6J mouse (donor), a CD45.1-expressing congenic mouse (recipient) and bone marrow chimeras engrafted with P2rx7^+/+ and P2rx7^−/− bone marrow cells (P2rx7^+/+ and P2rx7^−/− chimeras, respectively). (c–f) Immunocytochemical demonstration of the P2rx7 in mouse microglial cells. Co-localization of P2rx7s (labelled red) with microglial labelling CD11b (green) was found in microglia cells retrieved from P2rx7^+/+ mouse brain tissue (c) and in chimeras transplanted with P2rx7^+/+ bone marrow [(e) CD45.1/ P2rx7^+/+]. Microglial cells of P2rx7^−/− animals (d) did not show P2rx7 immunoreactivity. In chimeras transplanted with P2rx7^−/− bone marrow [(f), CD45.1/P2rx7^−/−], the P2rx7 (labelled red) was not found on microglial cells (labelled green) but was in some unidentified cells and cell debris in the cell suspension/fraction. Electron microscopy supported this finding. The cell membrane of morphologically characterized microglial cells – dark nuclei, either oval or bean shaped, electron dense cytoplasm, long cisternae of granular endoplasmic reticulum and large inclusions of phagocytosed material in the cell bodies that are commonly found in old animals – were covered by diaminobenzidine precipitates, demonstrating P2rx7 immunoreactivity (c and e rows). Cell processes of unidentified cells also showed P2rx7 immunoreactivity (e). Microglial cells of P2rx7 gene knockout animals (d) or microglial cells from CD45.1/P2rx7^−/− animals (f) were free of precipitate. (g, h) Basal immobility time in the tail suspension test (g) and amphetamine-induced hyperactivity (h) were not different in CD45.1/P2rx7^+/+ and CD45.1/P2rx7^−/− mice. Mice were submitted to behaviour tests 8 wk after engraftment (n = 13–14/group).