Transgenic mice for conditional gene manipulation in astroglial cells.
Astrocytes are thought to exert diverse functions in the brain, but it has been difficult to prove this in vivo because of a scarcity of tools to manipulate these cells. Here, we report the generation of new transgenic mouse lines that allow for conditional gene ablation in astrocytes using the tamoxifen- (TAM-) inducible CreER(T2)/loxP system and bacterial artificial chromosome (BAC)-based transgenesis. In adult transgenic mice, where CreER(T2) expression is driven by the promoter of the sodium-dependent glutamate/aspartate transporter (Glast/Slc1a3) or of connexin 30 (Cx30/Gjb6), intraperitoneal TAM-injection induced Cre-mediated recombination in astroglial cells throughout the brain. Targeting efficacies varied in a region-specific manner from 20 to 90% as indicated by enzyme-based reporter lines and immunohistochemical staining. In addition, the Glast-line allowed to target retinal Müller cells and adult neural stem/progenitor cells in neurogenic regions of the adult brain. Transgenic mice expressing CreER(T2) under the control of the apolipoprotein e (ApoE) or aquaporin 4 (Aqp4) promoter showed inducible recombination in different areas of the central nervous system (CNS) albeit at low levels. Transgenic lines showed TAM-induced recombination in specific peripheral organs. These new mouse lines should help to further explore the relevance of astrocytes for brain function, as well as their contribution to pathological conditions because of aging, disease or injury.
Slezak
Michal
M
Department of Neurotransmission/Neuroendocrine Secretion, Institute of Cellular and Integrative Neuroscience (INCI), UMR 7168/LC2 CNRS/Université Louis Pasteur, Strasbourg, France.
Göritz
Christian
C
Niemiec
Aurore
A
Frisén
Jonas
J
Chambon
Pierre
P
Metzger
Daniel
D
Pfrieger
Frank W
FW
D000818:Animals; D001057:Apolipoproteins E / Q000235:genetics; D051401:Aquaporin 4 / Q000235:genetics; D001253:Astrocytes / Q000502:physiology*; D002460:Cell Line; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics; D000075302:Connexin 30; D017630:Connexins / Q000235:genetics / Q000502:physiology; D004247:DNA / Q000235:genetics; D004965:Estrogen Antagonists / Q000494:pharmacology; D027341:Excitatory Amino Acid Transporter 1 / Q000235:genetics / Q000502:physiology; D018014:Gene Transfer Techniques; D007150:Immunohistochemistry; D019426:Integrases / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009474:Neurons / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics / Q000502:physiology; D012160:Retina / Q000166:cytology / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013629:Tamoxifen / Q000494:pharmacology