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Histamine Receptors in Guinea Pig Trachea and Their Relations to Cyclic Nucleotides

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Histamine Receptors in Guinea Pig Trachea and Their Relations to Cyclic Nucleotides INFORMATION TO USERS This was produced from a copy of a document sent to us for microfilming. While the most advanced technological means to photograph and reproduce this document have been used, the quality is heavily dependent upon the quality of the material submitted. The following explanation of techniques is provided to help you understand markings or notations which may appear on this reproduction. 1. The sign or “target” for pages apparently lacking from the document photographed is “ Missing Pagc(s)”. If it was possible to obtain the missing pagc(s) or section, they arc spliced into the film along with adjacent pages. This may have necessitated cutting through an image and duplicating adjacent pages to assure you of complete continuity. 2. When an image on the film is obliterated with a round black mark it is an indication that the film Inspector noticed cither blurred copy because of movement during exposure, or duplicate copy. Unless we meant to delete copyrighted materials that should not have been filmed, you will find a good image of the page in the adjacent frame. If copyrighted materials were deleted you will find a target note listing the pages In the adjacent frame. 3. When a map, drawing or chart, etc., is part of the material being photo­ graphed the photographer has followed a definite method in “sectioning” the material. It is customary to begin filming at the upper left hand corner of a large sheet and to continue from left to right in equal sections with small overlaps. If necessary, sectioning is continued again—beginning below the first row and continuing on until complete. 4. For any illustrations that cannot be reproduced satisfactorily by xerography, photographic prints can be purchased at additional cost and tipped into your xerographic copy. Requests can be made to our Dissertations Customer Services Department. 5. Some pages In any document may have indistinct print. In all cases we have filmed the best available copy. University Moorilms International 300 N. ZEEB RD., ANN ARBOR, Ml 48106 8121805 Ja c k so n , G a y l e L a t r ic u M a r t in THE EXISTENCE OF MULTIPLE HISTAMINE RECEPTORS IN GUINEA PIG TRACHEA AND THEIR RELATIONS TO CYCLIC NUCLEOTIDES The Ohio Slate University PH.D. 1981 University Microfilms International 300 N. Z«b Road, Aon Arbor. MI 48106 THE EXISTENCE OF MULTIPLE HISTAMINE RECEPTORS IN GUINEA PIG TRACHEA AND THEIR RELATIONS TO CYCLIC NUCLEOTIDES DISSERTATION Presented In Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in The Graduate School of The Ohio State University By Gayle Latrlda Martin Jackson, B.S. * * * * * The Ohio State University 1981 Reading Committee: Approved By Richard Fertel, Ph.D Joseph Blanchlne, M.D., Ph.D Jack Rail, Ph.D. Advisor Department of Pharmacology Gopl A. Tejwani, Ph.D ACKNOWLEDGEMENTS I thank Dr. Richard Fertel, my adviser, for his support, guidance, criticism, patience and most of all his understanding. 1 wish to thank my Reading Committee: Drs. J. Blanchlne, R. Fertel, J. Rail and G. Tejwanl for their critical evaluation of this thesis. There have been so many generous and kind people who have helped me to reach this goal. Let me say a special "Thank You" to: Dr. G. Tejwani for his assistance In biochemical techniques; Carol Oravec, Novecia Custls and Christine Albrlghtson for their technical assistance; Janet Rice and Jill Williams for statistical assistance; James Greenwald, photography; Nancy Sally for graphic illustrations, and Reece Smith for Xeroxing. My friends, Ricardo Wright, Richard and Fannie Watson, Lee Erby, Crosby and Florence Christian and the Kwaanza family, who in their own special ways inspired me; To my babysitters, Judy and Carol, who were always available in my times of greatest need; To Carol Jones for her patience in typing this manuscript and her assistance in my years as a graduate student; ii To my parentsr Abraham and Mattie who always encouraged me to reach a little higher. To my brother, Abraham; my sisters Frances, Mamie and Felicia; my sister-in-law, Theresa and my inlaws Hazel and Charlie Allen, who were always supportive; To my children, Nkenge and Atlba, who have shown the greatest patience and understanding during this long ordeal; Finally, to my husband Alvin, to whom I dedicate this work* My love and thanks for all the encouragement and assistance you have given me, and understanding the importance of this accomplishment to me* ill VITA October 27 1950 Born - Baltimore, Maryland 1972 B.S., Morgan State University Baltimore, Maryland 1972-1973 University Fellow The Graduate School The Ohio State University Columbus, Ohio 1973-1974 Graduate Student Representative to Faculty Department of Pharmacology The Ohio State University Columbus, Ohio 1973-1974 Member of Council of Graduate Students The Ohio State University Columbus, Ohio 1973-1975 National Institute of Health Trainee Department of Pharmacology The Ohio State University Columbus, Ohio 1975-1976 University Fellow The Graduate School The Ohio State University Columbus, Ohio 1976-1979 Fellow, National Fellowship Fund (Ford Foundation) 1977-1978 Coordinated and participated in teaching Pharmacology7 400 Department of Pharmacology The Ohio State University Columbus, Ohio iv 1977 Minority Recruiter Department of Pharmacology The Ohio State University Columbus, Ohio 1979 Participated In teaching Pharma* cology 600 Department of Pharmacology The Ohio State University Columbus, Ohio HONORS University Fellowship, The Ohio State University* 1972-1973, 1975-1976* National Fellowship Fund Fellow (Ford Foundation)* 1976-1979* Awarded Fellowship, American Association of University Women* 1976* The ICSABER Society Award, The Ohio State University* 1978* The Clayton S* Smith Memorial Award, The Ohio State University* 1979. v PUBLICATIONS AND PRESENTATIONS Martin, G.L., Lindower, J.O. and O'Neill, J.J. Barbiturate Induced . fine structure changes In guinea pig brain using an Improved terminal fixation procedure* Pharmacologist 16: 331, 1974 (abstract) Martin, G*L. and Fertel, R. The effect of histamine on the cyclic AMP to cyclic GMP ratio In guinea pig lung* Pharmacologist 19: 284, 1977 (abstract) Martin, G.L. and Fertel, R. Guinea pig lung may have both Hi and H2 receptors. Federation Proceedings 37: 393, 1978 (abstract) Martin, G. Role of Histamine In Asthma. Research Seminar, October 1977. Morgan State University, Baltimore, MD and Coppln State College, Baltimore, MD FIELDS OF STUDY Major Field: Biochemical Pharmacology Autonomic Pharmacology Drs. B.H. Marks and J.J. O'Neill Biochemical Pharmacology Dr. J.J. O'Neill Cardiovascular Pharmacology Dr. S. Dutta Drug Metabolism Dr. D. Courl Neuroanatomy Dr. A. Humbertsen Neurochemistry Dr. L. Horrocks Physiological Biochemistry Dr. A.J. Merola Physiology Department of Physiology Staff vi TABLE OF CONTENTS PAGE ACKNOWLEDGEMENTS ........................................ 11 VITA .................................................... iv LIST OF‘FI G U R E S ............................ ; ........... xlll INTRODUCTION ............................................ 1 I. THE ROLE OF HISTAMINE IN THE CONTROL OF RESPIRATORY SMOOTH MUSCLE ......................... 2 A. Historical Background ................. 1 Br Asthma and Anaphylaxis.................. 2 C. The Normal Physiologic Control of 3 Airway Tone ............................ II. THE MECHANISM OF ACTION OF H I S T A M I N E .............. 4 A. Evidence for the Existence of Two Histamine Receptors.....................* ......... 4 B. Histamine Receptors In Respiratory Smooth Muscle .................................. 5 1. Evidence for an H^ Receptor......... 5 2. Evidence for an H2 Receptor......... 7 3. Evidence for an H3 Receptor......... 9 III. CORRELATION BETWEEN AIRWAY TONE AND CYCLIC NUCLEOTIDES.................................... 10 A. Cyclic Nucleotides aa Second Messengers • . 10 1. Historical Background .............. 10 vil PAGE 2. Cyclic AMP- formation, metabolism and action........... 12 3* Cyclic GMP - formation, metabolism and action..................... ...... 13 B. The Mechanism of Action of Cyclic Nucleotides in Tracheal Smooth Muscle • • • .............. 14 1. Cyclic A M P ............................ 14 2. Cyclic G M P ............................ 15 C. The Role of Cyclic Nucleotides in the Actions of Histamine ............. 16 1. Cyclic A M P ............................ 16 2. Cyclic G M P ............................ 17 STATEMENT OF THE PROBLEM .................................... 19 MATERIALS AND M E T H O D S ........................................ 21 I. ANIMALS............................................. 21 II. DRUGS AND SOLUTIONS ............................... 21 A* Physiological Salt Solution (PSS)............ 22 III. PHARMACOLOGICAL AS S A Y ............................... 23 A. Preparation of Tracheal Rings ........ 23 B. Passive Force VersuB Active Force.............. 24 C. Equilibration of the Tissue.................. 27 D. Monitoring of Tissue Sensitivity... ........... 30 E. Histamine Tachyphylaxis .................... 30 F. Assay of Tissue ............................ 33 1. Dose Response ........ ........ 33 2. Time Response............. 33 3. Evaluation of Histamine Antagonist Activity . .............. 33 4. WaBhout of Drug •*..». ..... 34 viii PAGE G. Expression of Data • 34 IV. BIOCHEMICAL ASSAY ............................ 34 A. Preparation of the Tracheal Smooth Muscle . 34 •B. Incubation of the T i s s u e ................ 35 C. Measurement of Cyclic AMP and Cyclic GMP . 35 1. Preparation of Tracheal
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