Assembly and Maintenance of the Axonal Cytoskeleton in      Cultured Neurons
{'Code': '08090200', 'Directorate': {'Abbreviation': 'BIO', 'LongName': 'Direct For Biological Sciences'}, 'Division': {'Abbreviation': 'IOS', 'LongName': 'Division Of Integrative Organismal Systems'}}
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NB2a/dl neuroblastoma cells provide a model system for the study                of axonal cytoskeletal dynamics.  Following differentiation with                dbcAMP, NB2a/dl cells express and phosphorylate all three                       neurofilament protein subunit (NFPs) and assemble them into                     neurofilaments in a sequence of events closely paralleling that                 observed in vivo.  Axonal microtubules of these cells contain                   acetylated alpha.tubulin, a post.translational modification of                  tubulin that confers stability to microtubules.  NB2a/dl cells                  also express calpain, the calcium.activated protease that                       preferentially degrades cytoskeletal proteins and its regulatory                molecule, calpastatin.  In this project, the sequence of events                 of NFP and alpha.tubulin synthesis, modification, assembly and                  degradation, and the synthesis and activity of calpain and                      calpastatin will be defined during differentiation by: 1.                       immunoblot analysis of steady.state levels present in                           cytoskeletal and soluble fractions utilizing antisera specific                  for unmodified subunits: 2. immunoprecipitation of radiolabeled                 .                                                                               J                                                                               H.subunits; and 3. proteolysis of isolated subunits by endogenous               calpain.  The hypothesis that alterations in one or more of the                 events is responsible for axonal atrophy will be tested by                      withdrawal of dbcAMP (which induces resorption of axons)                        followed by analysis of cultures by the above methods.  Direct                  introduction into cells of calpain inhibitory molecules will be                 utilized to further examine the role of proteolysis in axonal                   atrophy.
{'FirstName': 'Thomas', 'LastName': 'Shea', 'PI_MID_INIT': 'B', 'PI_SUFX_NAME': None, 'PI_FULL_NAME': 'Thomas B Shea', 'EmailAddress': 'Thomas_Shea@uml.edu', 'NSF_ID': '000099882', 'StartDate': '03/15/1988', 'EndDate': None, 'RoleCode': 'Principal Investigator'}
{'Name': 'McLean Hospital', 'CityName': 'BELMONT', 'ZipCode': '024781064', 'PhoneNumber': '6178552868', 'StreetAddress': '115 MILL ST', 'StreetAddress2': None, 'CountryName': 'United States', 'StateName': 'Massachusetts', 'StateCode': 'MA', 'CONGRESSDISTRICT': '05', 'CONGRESS_DISTRICT_ORG': 'MA05', 'ORG_UEI_NUM': 'MCKWJYCWNVN3', 'ORG_LGL_BUS_NAME': 'THE MCLEAN HOSPITAL CORPORATION', 'ORG_PRNT_UEI_NUM': None}
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{'Code': '1192', 'Text': 'NEURONAL AND GLIAL MECHANISMS'}
{'url': 'https://www.nsf.gov/awardsearch/download?DownloadFileName=1987&All=true', 'xml': '8719823.xml'}