Transgenic mice expressing Cre-recombinase specifically in retinal rod bipolar neurons.
OBJECTIVE
To establish a transgenic mouse line that expresses Cre-recombinase in retinal rod bipolar cells for the generation of rod bipolar cell-specific knockout mutants.


METHODS
The IRES-Cre-cDNA fragment was inserted into a 173-kb bacterial artificial chromosome (BAC) carrying the intact Pcp2 gene, by using red-mediated recombineering. Transgenic mice were generated with the modified BAC and identified. The Cre-transgenic mice were crossed with ROSA26 and Z/EG reporter mice to detect Cre-recombinase activity.


RESULTS
X-gal staining showed that strong Cre-recombinase activities were present in retinal inner nuclear layers and cerebellar Purkinje cells. Double staining with an anti-GFP antibody and an anti-PKCalpha antibody (specific for retinal rod bipolar cells) revealed that Cre-recombinase activity localized exclusively to the rod bipolar cells in the retina.


CONCLUSIONS
A mouse BAC-Pcp2-IRES-Cre transgenic line that expresses Cre-recombinase in retinal rod bipolar neurons has been established. Because mutations in some ubiquitously expressed genes may result in retinal degenerative diseases, the mouse strain BAC-Pcp2-IRES-Cre will be a useful new tool for investigating the effects of retinal rod bipolar cell-specific gene inactivation.
Zhang
Xin-Mei
XM
Department of Biochemistry, University of Hong Kong, SAR, China.
Chen
Bai-Yu
BY
Ng
Alam Hoi-Lam
AH
Tanner
Julian A
JA
Tay
David
D
So
Kwok-Fai
KF
Rachel
Rivka A
RA
Copeland
Neal G
NG
Jenkins
Nancy A
NA
Huang
Jian-Dong
JD
D000818:Animals; D022202:Chromosomes, Artificial, Bacterial; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D005697:Galactosides / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D020662:Guanine Nucleotide Exchange Factors; D007211:Indoles / Q000378:metabolism; D019426:Integrases / Q000235:genetics / Q000378:metabolism*; D007395:Interneurons / Q000201:enzymology*; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008822:Mice, Transgenic; D009479:Neuropeptides / Q000235:genetics; D011247:Pregnancy; D011689:Purkinje Cells / Q000201:enzymology; D017948:Retinal Rod Photoreceptor Cells / Q000166:cytology* / Q000196:embryology; D001616:beta-Galactosidase / Q000378:metabolism
D005697:Galactosides; D020662:Guanine Nucleotide Exchange Factors; D007211:Indoles; D009479:Neuropeptides; C104971:Pcp2 protein, mouse; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; C045073:Cre recombinase; D019426:Integrases; D001616:beta-Galactosidase; C044888:5-bromo-4-chloro-3-indolyl beta-galactoside