In JoVE (1)
Articles by Kimberly P. Keil in JoVE
A High Throughput in situ Hybridization Method to Characterize mRNA Expression Patterns in the Fetal Mouse Lower Urogenital Tract Lisa L. Abler1, Vatsal Mehta1, Kimberly P. Keil1, Pinak S. Joshi1, Chelsea-Leigh Flucus1, Heather A. Hardin1, Christopher T. Schmitz1, Chad M. Vezina1 1Department of Comparative Biosciences, School of Veterinary Medicine, University of Wisconsin-Madison Here, we describe an efficient high throughput in situ hybridization (ISH) method for visualizing patterns of mRNA expression in developing fetal mouse prostate tissue sections. The method can be easily adapted to visualize mRNA expression patterns in other mouse tissues or in tissues from other species.
Other articles by Kimberly P. Keil on PubMed
A High-resolution Molecular Atlas of the Fetal Mouse Lower Urogenital Tract Developmental Dynamics : an Official Publication of the American Association of Anatomists. Oct, 2011 | Pubmed ID: 21905163 Epithelial-stromal interactions in the lower urogenital tract (LUT) are integral to prostatic and seminal vesicle development in males, vaginal and uterine development in females, and urethral development in both sexes. Gene expression profiling of isolated LUT stroma and epithelium has unraveled mechanisms of LUT development, but such studies are confounded by heterogeneous and ill-defined cell sub-populations contained within each tissue compartment. We used in situ hybridization to synthesize a high-resolution molecular atlas of 17-day post-coitus fetal mouse LUT. We identified mRNAs that mark selective cell populations of the seminal vesicle, ejaculatory duct, prostate, urethra, and vagina, subdividing these tissues into 16 stromal and 8 epithelial sub-compartments. These results provide a powerful tool for mapping LUT gene expression patterns and also reveal previously uncharacterized sub-compartments that may play mechanistic roles in LUT development of which we were previously unaware.
Atlas of Wnt and R-spondin Gene Expression in the Developing Male Mouse Lower Urogenital Tract Developmental Dynamics : an Official Publication of the American Association of Anatomists. Nov, 2011 | Pubmed ID: 21936019 Prostate development is influenced by Î²-catenin signaling, but it is unclear which Î²-catenin activators are involved, where they are synthesized, and whether their mRNA abundance is influenced by androgens. We identified WNT/Î²-catenin-responsive Î²-galactosidase activity in the lower urogenital tract (LUT) of transgenic reporter mice, but Î²-galactosidase activity differed among the four mouse strains we examined. We used in situ hybridization to compare patterns of Wnts, r-spondins (Rspos, co-activators of Î²-catenin signaling), Î²-catenin-responsive mRNAs, and an androgen receptor-responsive mRNA in wild type fetal male, fetal female, and neonatal male LUT. Most Wnt and Rspo mRNAs were present in LUT during prostate development. Sexually dimorphic expression patterns were observed for WNT/Î²-catenin-responsive genes, and for Wnt2b, Wnt4, Wnt7a, Wnt9b, Wnt10b, Wnt11, Wnt16, and Rspo3 mRNAs. These results reveal sexual differences in WNT/Î²-catenin signaling in fetal LUT, supporting the idea that this pathway may be directly or indirectly responsive to androgens during prostate ductal development.