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Purinergic Signal 2021 Sep 01;173:449-465. doi: 10.1007/s11302-021-09792-9.
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BAC transgenic mice to study the expression of P2X2 and P2Y1 receptors.

Grohmann M , Schumacher M , Günther J , Singheiser SM , Nußbaum T , Wildner F , Gerevich Z , Jabs R , Hirnet D , Lohr C , Illes P , Schmalzing G , Franke H , Hausmann R .

Extracellular purines are important signaling molecules involved in numerous physiological and pathological processes via the activation of P2 receptors. Information about the spatial and temporal P2 receptor (P2R) expression and its regulation remains crucial for the understanding of the role of P2Rs in health and disease. To identify cells carrying P2X2Rs in situ, we have generated BAC transgenic mice that express the P2X2R subunits as fluorescent fusion protein (P2X2-TagRFP). In addition, we generated a BAC P2Y1R TagRFP reporter mouse expressing a TagRFP reporter for the P2RY1 gene expression. We demonstrate expression of the P2X2R in a subset of DRG neurons, the brain stem, the hippocampus, as well as on Purkinje neurons of the cerebellum. However, the weak fluorescence intensity in our P2X2R-TagRFP mouse precluded tracking of living cells. Our P2Y1R reporter mice confirmed the widespread expression of the P2RY1 gene in the CNS and indicate for the first time P2RY1 gene expression in mouse Purkinje cells, which so far has only been described in rats and humans. Our P2R transgenic models have advanced the understanding of purinergic transmission, but BAC transgenic models appeared not always to be straightforward and permanent reliable. We noticed a loss of fluorescence intensity, which depended on the number of progeny generations. These problems are discussed and may help to provide more successful animal models, even if in future more versatile and adaptable nuclease-mediated genome-editing techniques will be the methods of choice.

PubMed ID: 34050505
Article link: Purinergic Signal

Species referenced: Xenopus laevis
Genes referenced: drg1 p2rx2 p2ry1

References [+] :
Alexander, Effect of transgene copy number on survival in the G93A SOD1 transgenic mouse model of ALS. 2004, Pubmed