A noninvasive genetic/pharmacologic strategy for visualizing cell morphology and clonal relationships in the mouse.
Analysis of cellular morphology is the most general approach to neuronal classification. With the increased use of genetically engineered mice, there is a growing need for methods that can selectively visualize the morphologies of specified subsets of neurons. This capability is needed both to define cell morphologic phenotypes and to mark cells in a noninvasive manner for lineage studies. To this end, we describe a bipartite genetic system based on a Cre-estrogen receptor (ER) fusion protein that irreversibly activates a plasma membrane-bound alkaline phosphatase reporter gene by site-specific recombination. Because the efficiency and timing of gene rearrangement is controlled pharmacologically, a sparse subset of labeled cells can be generated from the set of CreER-expressing cells at any time during development. Histochemical visualization of alkaline phosphatase activity reveals neuronal morphology with strong and uniform labeling of all processes.
Badea
Tudor C
TC
Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
Wang
Yanshu
Y
Nathans
Jeremy
J
D000469:Alkaline Phosphatase / Q000032:analysis / Q000378:metabolism; D000818:Animals; D001921:Brain / Q000166:cytology / Q000254:growth & development; D019070:Cell Lineage / Q000502:physiology; D002999:Clone Cells / Q000145:classification* / Q000166:cytology* / Q000378:metabolism; D018390:Gene Targeting; D017930:Genes, Reporter; D006651:Histocytochemistry / Q000379:methods*; D019426:Integrases / Q000235:genetics; D051379:Mice; D009474:Neurons / Q000145:classification* / Q000166:cytology* / Q000378:metabolism; D010641:Phenotype; D011960:Receptors, Estrogen / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic; D015203:Reproducibility of Results; D020845:Selective Estrogen Receptor Modulators / Q000494:pharmacology; D013629:Tamoxifen / Q000494:pharmacology; D019076:Transgenes; D014764:Viral Proteins / Q000235:genetics
D011960:Receptors, Estrogen; D011993:Recombinant Fusion Proteins; D020845:Selective Estrogen Receptor Modulators; D014764:Viral Proteins; D013629:Tamoxifen; C045073:Cre recombinase; D019426:Integrases; D000469:Alkaline Phosphatase