Approaches to Identifying Approaches to Identifying Possible - PowerPoint PPT Presentation

Approaches to Identifying Approaches to Identifying Possible Mechanisms of Kidney Possible Mechanisms of Kidney Damage Damage Mellisa Manson Mellisa Manson Virginia Comonwealth University Virginia Comonwealth University

Goal of Research Goal of Research � Groundwork for finding a mechanism of � Groundwork for finding a mechanism of kidney damage kidney damage � Aid in understanding hypertension � Aid in understanding hypertension � Aid in understanding problems in kidney � Aid in understanding problems in kidney function function

Background Background � Function of the kidney � Function of the kidney � Normal � Normal � Hypertension � Hypertension � Possible Markers � Possible Markers � S1P1 � S1P1 � Hypoxyprobe � Hypoxyprobe � Caspase-3 � Caspase-3

Methods Methods � Immunohistochemistry � Immunohistochemistry � Preservation and Slide Preparation � Preservation and Slide Preparation � Deparaffination � Deparaffination � Antigen Retrieval � Antigen Retrieval � Staining � Staining � Block Serum � Block Serum � Primary Antibody � Primary Antibody � Secondary Antibody � Secondary Antibody � Hematoxylin � Hematoxylin

S1P1 S1P1 � Endothelial Marker � Endothelial Marker � Expressed all over body � Expressed all over body � Role at kidney � Role at kidney � Possible Mechanisms � Possible Mechanisms

Hypoxyprobe Hypoxyprobe � Marker for hypoxia � Marker for hypoxia � Distribution � Distribution � Role at kidney � Role at kidney � Possible Mechanism � Possible Mechanism

100x Low Salt Hypoxyprobe Hypoxyprobe Hypoxyprobe Negative 1:16000 1:32000 1:64000 Cortex Medulla Papilla

Caspase-3 Caspase-3 � Marker for substance essential to apoptosis � Marker for substance essential to apoptosis � Distribution � Distribution � Role at kidney � Role at kidney � Possible Mechanism � Possible Mechanism

Cortex 100x 400x 1000x 1000x Control Low Salt Normal Diet High Salt

Conclusion Conclusion � Substances and the trends in their presence � Substances and the trends in their presence � Ways to combat hypertension � Ways to combat hypertension � S1P1 receptors � S1P1 receptors � Hypoxia � Hypoxia � Apoptosis � Apoptosis