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
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