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A novel technique for temporally regulated cell type-specific Cre expression and recombination in the pituitary gonadotroph.
J Mol Endocrinol. 2006 Aug; 37(1):63-9.JM

Abstract

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.

Authors+Show Affiliations

Department of Medicine, University of Chicago, Chicago, IL 60637, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural

Language

eng

PubMed ID

16901924

Citation

Naik, Karuna, et al. "A Novel Technique for Temporally Regulated Cell Type-specific Cre Expression and Recombination in the Pituitary Gonadotroph." Journal of Molecular Endocrinology, vol. 37, no. 1, 2006, pp. 63-9.
Naik K, Pittman I, Wolfe A, et al. A novel technique for temporally regulated cell type-specific Cre expression and recombination in the pituitary gonadotroph. J Mol Endocrinol. 2006;37(1):63-9.
Naik, K., Pittman, I., Wolfe, A., Miller, R. S., Radovick, S., & Wondisford, F. E. (2006). A novel technique for temporally regulated cell type-specific Cre expression and recombination in the pituitary gonadotroph. Journal of Molecular Endocrinology, 37(1), 63-9.
Naik K, et al. A Novel Technique for Temporally Regulated Cell Type-specific Cre Expression and Recombination in the Pituitary Gonadotroph. J Mol Endocrinol. 2006;37(1):63-9. PubMed PMID: 16901924.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A novel technique for temporally regulated cell type-specific Cre expression and recombination in the pituitary gonadotroph. AU - Naik,Karuna, AU - Pittman,Isaiah,4th AU - Wolfe,Andrew, AU - Miller,Ryan S, AU - Radovick,Sally, AU - Wondisford,Fredric E, PY - 2006/8/12/pubmed PY - 2007/4/11/medline PY - 2006/8/12/entrez SP - 63 EP - 9 JF - Journal of molecular endocrinology JO - J Mol Endocrinol VL - 37 IS - 1 N2 - 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. SN - 0952-5041 UR - https://www.unboundmedicine.com/medline/citation/16901924/A_novel_technique_for_temporally_regulated_cell_type_specific_Cre_expression_and_recombination_in_the_pituitary_gonadotroph_ L2 - https://jme.bioscientifica.com/doi/10.1677/jme.1.02053 DB - PRIME DP - Unbound Medicine ER -