Genetic analysis of the homeodomain transcription factor Chx10 in the retina using a novel multifunctional BAC transgenic mouse reporter.
Chx10 is a homeobox-containing transcription factor critical for progenitor cell proliferation and bipolar cell determination in the developing retina. Its expression in the retina has been reported to be restricted to these cell populations. To further understand Chx10 regulation and function, a multifunctional reporter construct consisting of GFP, alkaline phosphatase, and Cre recombinase was integrated into a BAC encoding Chx10. Stable lines of transgenic mice expressing this BAC were generated and analyzed. The reporter expression was faithful to the endogenous retinal Chx10 expression pattern and revealed a previously unappreciated locus of Chx10 expression in a subset of Müller glial cells. In addition, Chx10 reporter activity was identified in mature orJ-Chx10 mutant retinas, although these retinas lack Chx10-expressing bipolar cells. Reporter and molecular analysis showed that the reporter-expressing cells in the mutant had hallmarks of progenitor cells or partially differentiated Müller glial cells. These results strongly suggest that Chx10 promotes bipolar fate by affecting differentiation of late progenitor cells. Crosses of the Chx10 BAC reporter mice to R26R mice for fate-mapping experiments revealed that Chx10 reporter-expressing progenitor cells contribute to all mature cell types of the retina. These results demonstrate the utility of these lines for generation of mosaic or complete genetic manipulations of the retina.
Rowan
Sheldon
S
Department of Genetics and Howard Hughes Medical Institute, Harvard Medical School, Boston, MA 02115, USA.
Cepko
Constance L
CL
D000469:Alkaline Phosphatase / Q000235:genetics / Q000378:metabolism; D000818:Animals; D015139:Blotting, Southern; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology*; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics / Q000378:metabolism; D003433:Crosses, Genetic; D017931:DNA Primers; D020869:Gene Expression Profiling; D049452:Green Fluorescent Proteins; D018398:Homeodomain Proteins / Q000235:genetics* / Q000378:metabolism; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D019426:Integrases / Q000235:genetics / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D009457:Neuroglia / Q000378:metabolism*; D012160:Retina / Q000378:metabolism* / Q000502:physiology; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D001616:beta-Galactosidase
D017931:DNA Primers; D018398:Homeodomain Proteins; D008164:Luminescent Proteins; D014157:Transcription Factors; C088975:Vsx2 protein, mouse; D049452:Green Fluorescent Proteins; C045073:Cre recombinase; D019426:Integrases; D000469:Alkaline Phosphatase; D001616:beta-Galactosidase