Deviant reporter expression and P2X4 passenger gene overexpression in the soluble EGFP BAC transgenic P2X7 reporter mouse model
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Deviant reporter expression and P2X4 passenger gene overexpression in the soluble EGFP BAC transgenic P2X7 reporter mouse model. / Ramírez-Fernández, Antonio; Urbina-Treviño, Lidia; Conte, Giorgia; Alves, Mariana; Rissiek, Björn; Durner, Anna; Scalbert, Nicolas; Zhang, Jiong; Magnus, Tim; Koch-Nolte, Friedrich; Plesnila, Nikolaus; Deussing, Jan M; Engel, Tobias; Kopp, Robin; Nicke, Annette.
In: SCI REP-UK, Vol. 10, No. 1, 16.11.2020, p. 19876.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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T1 - Deviant reporter expression and P2X4 passenger gene overexpression in the soluble EGFP BAC transgenic P2X7 reporter mouse model
AU - Ramírez-Fernández, Antonio
AU - Urbina-Treviño, Lidia
AU - Conte, Giorgia
AU - Alves, Mariana
AU - Rissiek, Björn
AU - Durner, Anna
AU - Scalbert, Nicolas
AU - Zhang, Jiong
AU - Magnus, Tim
AU - Koch-Nolte, Friedrich
AU - Plesnila, Nikolaus
AU - Deussing, Jan M
AU - Engel, Tobias
AU - Kopp, Robin
AU - Nicke, Annette
PY - 2020/11/16
Y1 - 2020/11/16
N2 - The ATP-gated P2X7 receptor is highly expressed in microglia and has been involved in diverse brain diseases. P2X7 effects were also described in neurons and astrocytes but its localisation and function in these cell types has been challenging to demonstrate in situ. BAC transgenic mouse lines have greatly advanced neuroscience research and two BAC-transgenic P2X7 reporter mouse models exist in which either a soluble EGFP (sEGFP) or an EGFP-tagged P2X7 receptor (P2X7-EGFP) is expressed under the control of a BAC-derived P2rx7 promoter. Here we evaluate both mouse models and find striking differences in both P2X expression levels and EGFP reporter expression patterns. Most remarkably, the sEGFP model overexpresses a P2X4 passenger gene and sEGFP shows clear neuronal localisation but appears to be absent in microglia. Preliminary functional analysis in a status epilepticus model suggests functional consequences of the observed P2X receptor overexpression. In summary, an aberrant EGFP reporter pattern and possible effects of P2X4 and/or P2X7 protein overexpression need to be considered when working with this model. We further discuss reasons for the observed differences and possible caveats in BAC transgenic approaches.
AB - The ATP-gated P2X7 receptor is highly expressed in microglia and has been involved in diverse brain diseases. P2X7 effects were also described in neurons and astrocytes but its localisation and function in these cell types has been challenging to demonstrate in situ. BAC transgenic mouse lines have greatly advanced neuroscience research and two BAC-transgenic P2X7 reporter mouse models exist in which either a soluble EGFP (sEGFP) or an EGFP-tagged P2X7 receptor (P2X7-EGFP) is expressed under the control of a BAC-derived P2rx7 promoter. Here we evaluate both mouse models and find striking differences in both P2X expression levels and EGFP reporter expression patterns. Most remarkably, the sEGFP model overexpresses a P2X4 passenger gene and sEGFP shows clear neuronal localisation but appears to be absent in microglia. Preliminary functional analysis in a status epilepticus model suggests functional consequences of the observed P2X receptor overexpression. In summary, an aberrant EGFP reporter pattern and possible effects of P2X4 and/or P2X7 protein overexpression need to be considered when working with this model. We further discuss reasons for the observed differences and possible caveats in BAC transgenic approaches.
U2 - 10.1038/s41598-020-76428-0
DO - 10.1038/s41598-020-76428-0
M3 - SCORING: Journal article
C2 - 33199725
VL - 10
SP - 19876
JO - SCI REP-UK
JF - SCI REP-UK
SN - 2045-2322
IS - 1
ER -