- Friday, September 23, 2016
- 1:30 PM–2:30 PM
- Science Building 010
Title: Mechanical and Chemical Isolation of Mouse RPE Layer
The retinal pigment epithelium (RPE) is a single layer of cuboidal cells that composes the dorsal-most part of the retina and participates in the blood brain barrier. As a barrier tissue, the RPE is replete with GLUT1. RNA analysis and paraffin sample staining demonstrate that GLUT1 is present on the apical and basal RPE membrane while TXNIP is only present on the apical RPE membrane. In an attempt to co-localize the apical GLUT1 and TXNIP, we established and optimized protocols for murine RPE whole mount preparation and confocal imaging. Despite successful isolation and mounting, difficulties with antibody specificity and tissue morphology have prevented quantification of the difference in TXNIP immunofluorescence between WT and TXNIP KO mice.
Title: Unlocking the secrets of urine: assaying urinary extracellular mRNA in healthy volunteers
The admixture of molecules in urine is directly affected by renal physiology and pathophysiology. Though extracellular messenger RNA in urine might be informative about renal physiology, a gold standard clinically applicable approach for isolating extracellular messenger RNA (mRNA) from urine has not been established. Key knowledge gaps include the limits of detection of urinary mRNA transcripts, whether mRNA degradation can be prevented, and the variability in urinary mRNA transcripts in healthy volunteers. The goal of this pilot study was to determine which method best allowed detection of renally expressed RNA targets in urine, for use in future studies in patients.