A novel technique for temporally regulated cell type-specific Cre expression and recombination in the pituitary gonadotroph.
Inducing tissue-specific genetic alterations under temporal control allows for the analysis of gene function in particular cell types at specified points in time. We have generated a system for tetracycline-controlled expression of Cre recombinase in mice using the unique CreTeR vector. The gonadotroph-specific bovine alpha-subunit (Balpha) promoter fragment was subcloned into the CreTeR vector, creating a technique for highly regulated expression of Cre recombinase exclusively in pituitary gonadotrophs. Control of Cre recombinase in the CreTeR vector was demonstrated in LbetaT2 pituitary cell lines, where Cre protein was detected in cells treated with doxycycline, but not in untreated cells. In transgenic mice, Cre was expressed in pituitary gonadotrophs of mice treated with doxycycline, but not in non-pituitary tissues or in transgenic mice not treated with doxycycline. We demonstrated Cre expression in the gonadotroph by immunostaining showing co-localization of Cre recombinase with the beta-subunit of LH (LH-beta). Furthermore, by crossing Balpha/CreTeR with R26R mice, we were able to demonstrate functional recombination within pituitary gonadotrophs, detected by lacZ expression. The Balpha/CreTeR mice described here can be used to study the function of virtually any gene in the gonadotroph; in particular, this will be useful in studying genes, which may have distinct roles in development and in the adult.
Naik
Karuna
K
Department of Medicine, University of Chicago, Chicago, IL 60637, USA.
Pittman
Isaiah
I
Wolfe
Andrew
A
Miller
Ryan S
RS
Radovick
Sally
S
Wondisford
Fredric E
FE
D000818:Animals; D002417:Cattle; D002460:Cell Line; D004318:Doxycycline / Q000378:metabolism; D005786:Gene Expression Regulation; D005821:Genetic Techniques; D052681:Gonadotrophs / Q000166:cytology / Q000502:physiology*; D019426:Integrases / Q000235:genetics / Q000378:metabolism*; D037101:Luteinizing Hormone, beta Subunit / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D010902:Pituitary Gland / Q000166:cytology; D011401:Promoter Regions, Genetic; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism; D011995:Recombination, Genetic; D013997:Time Factors; D019076:Transgenes
D037101:Luteinizing Hormone, beta Subunit; D021122:Protein Subunits; C045073:Cre recombinase; D019426:Integrases; D004318:Doxycycline