Vitamin D Pathway Status and the Identification of Target Genes in the Mouse Mammary Gland
Abstract
Mammary gland samples were isolated from wild type, vitamin D receptor knockout (VDRKO) and 1-alphahydroxylase knockout (1-alphaKO) female mice for whole mounts and paraffin embedding (inguinal) and for RNA and protein isolation (thoracic). Time points collected included 6-10wk old nulliparous, 9 and 16 days pregnancy, 5 and 10 days lactation and 3 and 6 days involution. All whole mounts were completed and showed increased branching during pregnancy in the VDRKO glands relative to wild type and 1-alphaKO glands. Paraffin embedded involution samples were stained with hematoxylin and eosin and showed an apparent decrease in alveolar breakdown in the first few days of involution in VDRKO and 1-alphaKO glands compared to wild type controls. Organ culture studies show that treatment of wild type and 1-alphaKO glands with 1,25 dihydroxyvitamin D decreases proliferation and retards tertiary branching. This is not the case in VDRKO glands. Gene expression analysis via microarray and qPCR provides only a glimpse into the complexities of the signaling involved in this process and is being complimented by ongoing protein analysis. These results suggest that 1-alphaKO mammary glands do not develop exclusively similar to VDRKO or wild type glands which verifies our need to complete the remainder of our studies to determine if the VDR is acting to control mammary gland development through the vitamin D pathway or through some other ligand or possibly without a ligand.
Document Details
- Document Type
- Technical Report
- Publication Date
- Nov 01, 2014
- Accession Number
- ADA613883
Entities
People
- Donald Matthews
- JoEllen Welsh