Fluorescent tagged analysis of neural gene function using mosaics in zebrafish and Xenopus laevis.
An important question in the neurosciences is the role of specific gene expression in the control of neural morphology and connectivity. To address this question, methods are needed for expression of exogenous genes in a subset of neurons. This limited and mosaic expression allows the assessment of gene expression in a cell autonomous fashion without environmental contributions from neighboring expressing cells. These methods must also label neurons so that detailed morphology and neural connections can be evaluated. The labeling method should label only a subset of neurons so that neuronal morphology can be viewed upon a non-stained background, in a Golgi staining fashion. Here, we report methods using plasmids called pTAGUM (tagged analysis of genes using mosaics) that accomplish these goals. These methods should prove useful for the analysis of neural gene function in two important model organisms, the zebrafish and Xenopus laevis.
Conway
Greg
G
NASA Ames Research Center, Moffett Field, CA 94035, USA. gconway@mail.arc.nasa.gov
Torrejón
Marcela
M
Lin
Shuo
S
Reinsch
Sigrid
S
D000818:Animals; D030801:Animals, Genetically Modified; D001757:Blastomeres; D004247:DNA / Q000378:metabolism; D004270:DNA, Circular / Q000502:physiology; D015252:Deoxyribonucleases, Type II Site-Specific / Q000494:pharmacology; D004625:Embryo, Nonmammalian / Q000378:metabolism / Q000502:physiology; D047108:Embryonic Development; D005456:Fluorescent Dyes; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000235:genetics; D007267:Injections; D009030:Mosaicism; D009420:Nervous System / Q000196:embryology*; D009474:Neurons / Q000145:classification / Q000378:metabolism / Q000502:physiology; D009994:Osmolar Concentration; D029701:Saccharomyces cerevisiae Proteins; D013997:Time Factors; D014982:Xenopus laevis / Q000196:embryology* / Q000235:genetics*; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics*
D004270:DNA, Circular; D005456:Fluorescent Dyes; D029701:Saccharomyces cerevisiae Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D004247:DNA; C037725:SCEI protein, S cerevisiae; D015252:Deoxyribonucleases, Type II Site-Specific