Laurence L. Brunton - US grants

In purified ventricular myocytes from adult rat heart, alpha adrenergic stimulation accelerates cAMP degradation. In rabbit cardiocytes, phosphodiesterase (PDE) inhibitors abolish apparent compartmentation of cAMP. We propose that these are two examples of intracellular modulation of cAMP metabolism to achieve specificity within the cell. We will investigate the mechanism of hormonal activation of PDE activity and the role of PDE in maintaining compartmentation of cAMP in adult cardiac myocytes. Using a new HPLC micromethod, isozymes of PDE will be resolved and characterized in terms of substrates, modulators and inhibitors, and subcellular localization. Such information will help identify PDEs involved when inhibitors are applied to whole cells to prevent alpha-adrenergic activation of PDE. The mechanism by which alpha- adrenergic stimuli activate PDE will be investigated (direct coupling to G-protein? cAMP stimulation of cAMP hydrolysis? any effect of electrical stimulation?, etc.). Attempts will be made to isolate stably altered PDE following alpha-adrenergic stimulation, and to reproduce activation in broken cell- reconstituted systems. Using fluorescent protein kinase inhibitor, regions of altered activation of cAMP-protein kinase will be mapped by fluorescent and light microscopy, and effects of hormones, modulators, and specific inhibitors studied to indicate localization of compartments created by PDE activities. Similar techniques will be applied to analysis of compartmentation of PGE1 response in cardiocytes. The cardiac myocyte alpha-1 receptor will be analyzed with respect to subtype and coupling to second messengers, especially cell Ca++ (assessed by FURA-2 fluorescence) and phosphatidylinositol metabolism. These studies will demonstrate cell specific localization, and possibly sub-cellular localization, of PDE isozymes, as well as the means by which hormones can alter these activities. This work will increase our understanding of how cells inegrate their responses to multiple hormonal signals, of how hormonal signals are transduced, and of how specificity and different responses may be achieved in response to elevation of cAMP, a general signal. The studies will also delineate the role played by PDE in maintenance of sub-cellular compartments of cAMP. Knowledge of these matters will be helpful in controlling specific aspects of cardiovascular function independently or coordinately of others using PDE inhibitors.

tissue /cell culture; heart failure; biomedical facility;

DESCRIPTION (provided by applicant): The University of California at San Diego (UCSD) and San Diego State University (SDSU) propose an institutional research and academic career development award (IRACDA) program for postdoctoral research fellows. The purpose of the UCSD/SDSU IRACDA will be to recruit and train top under-represented postdoctoral fellows in the best research labs and to couple this mentored training with evaluated training in teaching. The immediate goal is to place IRACDA Fellows in tenure track jobs within the UC and California State University systems. The long-term goal is to increase the number of under-represented faculty in biomedical sciences nation-wide. The program combines the facilities and selected biomedical research faculty of a large and well-known research university, the pool of candidates from the UC system (largest producer of minority Ph.D. graduates in the US), and the facilities and research professors from a large minority-serving institution with specialists in science education. In this three year training program, research training will occur at both institutions but primarily at UCSD, and will include weekly lab meetings, weekly meeting of the research division or department (postdocs present at least once a year), student journal club with evaluated presentations, a course in research ethics, courses in grant and paper writing and annual attendance at a national meeting (poster presentations in years 2 and 3). Fellows will deliver one lecture per semester in a beginning biology course throughout the training period, with training in classroom management, advising, assessment and Lecture skills and substantial development and analysis of individual teaching styles by one-on-one and group evaluations with a specialist in science education. The expectation is substantial esprit de corps amongst Fellows, a first publication by end of year two, a research grant in draft form by the second half of year three, a portfolio of six exemplary lectures by the end of year three and a job offer at the end of year three. Fellows will be followed for ten years from the start of the program (roughly to the time of a tenure decision) and their progress compared to that of the general pool of postdoctoral fellows on the two campuses.

[unreadable] DESCRIPTION (provided by applicant): In response to the RFA for a short course in integrative organ systems pharmacology, we propose a 3- week summer course, Systems Pharmacology and Translational Biology, for up to 20 participants. Our premise is that interpretation of complex phenotypes using molecular, genetic and proteomic information requires a familiarity with organ level function and its assessment. The course will focus on the CMS (nociception; neuropsychiatric disease), the cardiovascular system (autonomic regulation; hypertrophy; heart failure), and advanced imaging techniques. The aims of the Course are to provide: [unreadable] 1. For specific target cardiac and CMS phenotypes and for the process of nociception: [unreadable] a. review of basic molecular and cell biology, physiology, pathophysiology and pharmacology [unreadable] b. review of preclinical models used in current research in these areas [unreadable] c. hands-on training in the use of these models, ranging from mouse (KO, transgenic) to dog 2. Training in mouse imaging (PET, MRI, optical, ultrasound), radiopharmaceuticals & interpreting the data [unreadable] 3. Training in the assessment of phenotype in genetically altered animals [unreadable] 4. Familiarization with the use of animal and organ systems in the development of new drugs [unreadable] 5. Instruction and experience in the responsible and ethical use of animal subjects [unreadable] 6. Opportunities to analyze and present quantitative laboratory findings [unreadable] 7. Long-term tracking of and communication with and amongst participants [unreadable] Faculty are from UCSD and San Diego biotech/pharmaceutical companies. Students will be drawn from West Coast NIGMS Pharmacological Sciences training programs, regional biotech firms and research institutes, IRACDA fellows, and a national applicant pool attracted by web-based advertising. [unreadable] [unreadable]

[unreadable] DESCRIPTION (provided by applicant): The University of California at San Diego (UCSD) and San Diego State University (SDSU) propose to continue a successful institutional research and academic career development award (IRACDA) program for postdoctoral scholars. Six trainees will enter each year for a 3-year training period. The San Diego IRACDA Program will recruit and train under-represented postdoctoral fellows in top research labs, coupling mentored research with focused training in teaching, research ethics, grant writing, lab management, and a variety of [unreadable] professorial skills. The immediate goals are to help these postdocs begin independent research careers with appropriate skills to succeed, and to place IRACDA Fellows in tenure track jobs within the UC and California State University systems. The long-term goal is to increase the number of under-represented faculty and faculty committed to the education of URMs nation-wide. [unreadable] Research training will occur primarily at UCSD, and will include weekly lab meetings, departmental seminars, evaluated research presentations, monthly meetings with the PI, a course in research ethics, courses in grant and paper writing, and annual attendance at a national meeting (poster presentations in years 2 and 3) and the National IRACDA meeting. These activities will be coordinated by Drs. Brunton and Villarreal. Teaching will occur at SDSU, under the coordination of Dr. Pozos, in a design that keeps the emphasis on continual research progress. Following a six month period in the lab to establish their research, fellows will prepare & deliver 3 lectures per semester in a beginning biology course for 5 semesters, with training in classroom management, advising, assessment, e-curriculum development, and development of individual teaching styles. The academic emphasis is on mammalian systems biology, which also expands the Fellows' [unreadable] knowledge, since many of them have specialized without comprehensive training in biology. [unreadable] Expectations are substantial esprit de corps amongst Fellows, a major publication during year two, a research grant in draft form by the second half of year three, a portfolio of exemplary lectures by the end of year three, and a more advanced position or job offer at the end of year three. Fellows will be followed for ten years from the start of the program (roughly to the time of a tenure decision) and their progress compared to that of the general pool of postdoctoral fellows on the two campuses, majority postdocs in the San Diego IRACDA, other postdoctoral fellows in the same labs, and, as data become available, the national pool of NIH-supported postdocs. [unreadable] [unreadable] [unreadable]