Date of Award

Spring 1992

Document Type

Thesis

Department

Life & Environmental Sciences

First Advisor

John Addis

Second Advisor

Rev. Joseph Harrington

Third Advisor

Robert Swartout

Abstract

Means of localizing molecules are essential in contemporary biological research attempting to unlock the mysteries of how cells function, interact, and go wrong. Enzyme cytochemistry and immunohistochemistry are two methods which utilize different characteristics of given molecules to localize them in tissue. The objective of this study was to localize two proteins, Na,K-ATPase and vinculin, in frog and mouse liver sections using these methods. Na,KATPase activity contributes to overall p-nitrophenylphosphatase (jpNPPase) activity, and sites of p-NPPase activity were determined using an enzyme cytochemical method. Sites of p-NPPase activity were thus extrapolated to indicate regions of probable Na,K-ATPase activity. Vinculin was localized immunohistochemically using an antibody prepared specifically against the vinculin molecule. Liver sections mounted on glass slides were subjected to appropriate reaction conditions to carry out either the enzyme cytochemical method for p-NPPase, or the immunohistochemical method for vinculin. Results of the vinculin localization were inconclusive, but results of the p-NPPase localization enabled probable identification of Na,K-ATPase at lateral hepatocyte membranes, along the lining of the sinusoids, and associated with the bile canalicular membranes.

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