Tetrahymena, a protozoan eucaryotic ciliate, has been utilized in many basic research because of its structural similarities to mammalian cells. The overall general objectives of this project are: (1) To better understand the mechanism of alphaprodines hydrochloride and Benzodiazepines on the cellular level; (2) To study the drugs-cell receptors activity; (3) To document structural changes in Tetrahymena Cells induced to conjugate while exposed to these drugs; (4) To provide an environment where minority students will develop an interest in biomedical research and gain experience in all aspects of this research. The PI has carried out extensive study on the ciliates, their structure, mating mechanism and growth studies that are relevant to this project. We intend to investigate the mechanism of actions of the drugs using the ciliates. Part I of this project is designed to define the molecular process involved in the drugs' action at the cellular level. I intend to study the drugs' uptake by the cells, follow the pathway from cell membrane into the cytoplasm. Part II will involve isolation of binding sites on the cell membrane and in the cytoplasm. Such data will provide more information on the drugs' actions. Part III will involve the studying of cell development and differentiation in the ciliate. The methods of approach for this project will include a combination of biochemical and morphological procedures: Light microscopy, Electron Microscopy (SEM/TEM), Growth Studies, Autoradiography and Physiochemical analysis of conjugation. This project will serve as important step for further experiments on the actions of narcotic drugs on the cellular level. Knowledge of the biology of Tetrahymena and effects of drugs on the cell-cell interaction (conjugation) is essential to the understanding of narcotic drugs' action and treatment of drug abuse. This research will also provide a better understanding of morbidity and mortality with alphaprodine (sometimes involving misuse) and its' route of administration; and can ultimately result in an increase in the number of undergraduate minority students who develop interest in cellular pharmacological research.
