Defense Date

1999

Document Type

Thesis

Degree Name

Master of Science

Department

Physiology

First Advisor

Raphael J. Witorsch

Abstract

The Nb2 lymphoma cell line is susceptible to glucocorticoid (GC)-induced apoptosis in the absence of mitogen and this effect is inhibited by the mitogen prolactin (PRL). This process is believed to be mediated by the glucocorticoid receptor (GR) and various downstream signals. The purpose of this study was to use the limiting dilution method to isolate subsets of Nb2 clones which expressed either highly enriched amounts of GR or an absence of GR and to relate the amount of GR to the magnitude of GC induced cytolysis. The presence of GR in these clones was determined by immunocytochemistry (ICC) using an affinity purified polyclonal rabbit antibody directed towards the GR. The presence of GR was then correlated to cytolysis in these cells produced by incubation with dexamethasone (Dex), a synthetic GC, in the presence and absence of PRL. Cytolysis was measured by two different methods, Trypan Blue exclusion counts, to determine the percentage of dead and the TUNEL (TdT-dependent dUTP-biotin Nick End Labeling) assay used to label apoptotic cells. The limiting dilution produced 28 clones with varying amounts of GR and all were responsive to Dex and this response was inhibited by PRL. A significant correlation (r2=0.419 and p

Comments

Scanned, with permission from the author, from the original print version, which resides in University Archives.

Rights

© The Author

Is Part Of

VCU University Archives

Is Part Of

VCU Theses and Dissertations

Date of Submission

2-21-2018

Included in

Physiology Commons

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