Deviant reporter expression and P2X4 passenger gene overexpression in the soluble EGFP BAC transgenic P2X7 reporter mouse model.

Scientific Reports
Antonio Ramírez-FernándezAnnette Nicke

Abstract

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.

References

Apr 16, 1996·Proceedings of the National Academy of Sciences of the United States of America·F SotoW Stühmer
Dec 6, 2001·Nature Reviews. Neuroscience·N Heintz
Oct 18, 2007·Journal of Cell Science·Omar S QureshiRuth D Murrell-Lagnado
Dec 18, 2007·Neuroscience·P Marín-GarcíaM T Miras-Portugal
Mar 14, 2008·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Jessica A ShuenNicole Calakos
Apr 23, 2008·Current Protocols in Neuroscience·X William Yang, Shiaoching Gong
Jan 28, 2009·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Miguel Díaz-HernándezJosé J Lucas
Oct 5, 2010·Nature Protocols·Shiaoching GongNathaniel Heintz
Dec 27, 2011·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Tobias EngelMiguel Diaz-Hernandez
Jun 6, 2012·Nature Structural & Molecular Biology·Laure WeillRaúl Méndez
Aug 18, 2012·Journal of Biomedicine & Biotechnology·J BeilT Buch
Nov 10, 2012·The Journal of Biological Chemistry·Paula García-HuertaRosa Gómez-Villafuertes
Jan 31, 2013·Brain : a Journal of Neurology·Tobias EngelDavid C Henshall
Mar 5, 2013·Cold Spring Harbor Protocols·Eric F SchmidtNathaniel Heintz
Mar 6, 2015·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Yuri HirayamaSchuichi Koizumi
Aug 16, 2015·Journal of Medical Genetics·Isabelle PerraultFriedhelm Hildebrandt
Jun 3, 2016·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Alba Jimenez-PachecoDavid C Henshall
Aug 26, 2016·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Ji XuBaljit S Khakh
Feb 18, 2017·Frontiers in Neuroscience·Edward BeamerTobias Engel
Mar 3, 2017·Genome Research·Prashant Kumar SrivastavaMichael R Johnson
Jul 28, 2017·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Peter IllesPatrizia Rubini
Jul 28, 2017·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·M Teresa Miras-PortugalMiguel Díaz-Hernández
Apr 10, 2018·Current Opinion in Immunology·Francesco Di VirgilioFabio Grassi
Aug 11, 2018·Trends in Molecular Medicine·Jan M Deussing, Eduardo Arzt
Apr 4, 2019·The EMBO Journal·Stefanie WachterEsben Lorentzen
Apr 16, 2019·Current Opinion in Pharmacology·Friedrich Koch-NolteSahil Adriouch
Apr 25, 2019·Current Opinion in Pharmacology·María Domercq, C Matute
Apr 30, 2019·Frontiers in Cellular Neuroscience·Carlos Martínez-FrailesMiguel Díaz-Hernández
May 1, 2019·Current Opinion in Pharmacology·Ruth D Murrell-Lagnado, Manfred Frick
Jun 16, 2019·ENeuro·Xiao-Feng ZhaoYunfei Huang
Aug 24, 2019·Frontiers in Molecular Neuroscience·Robin KoppAnnette Nicke
Sep 12, 2019·Frontiers in Pharmacology·Knut BiberThomas Möller
Sep 26, 2019·Frontiers in Cellular Neuroscience·Jean M Kanellopoulos, Cécile Delarasse
Feb 6, 2020·Frontiers in Molecular Neuroscience·Björn RissiekFriedrich Koch-Nolte

❮ Previous
Next ❯

Citations

Feb 16, 2021·Biochemical Pharmacology·María Teresa Miras-PortugalEsmerilda G Delicado
Mar 2, 2021·Frontiers in Molecular Neuroscience·Ya-Fei ZhaoPeter Illes
Jul 27, 2021·Journal of Inflammation Research·Tobias EngelMariana Alves

❮ Previous
Next ❯

Methods Mentioned

BETA
transgenic
PCR
FACS
flow cytometry
gene modification
genotyping
lavage

Software Mentioned

Primer3
Zeiss Axiovision
Treestar
GraphPad Prism
ImageJ
LightCycler
StrataQuest
Labchart
FlowJo
ImageStudio

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.