To determine the properties of the @entity268 -specific, @entity7428 -sensitive phosphodiesterases ( @entity268 -PDEs) that are expressed in different organs, monoclonal and polyclonal antibodies were raised against different epitopes present in the @entity268 -PDE sequences. Of the several antibodies generated against peptides and fusion proteins, one monoclonal and four polyclonal antibodies recognized both the native @entity268 -PDEs as well as the denatured proteins on Western immunoblot analysis. An immunoprecipitation assay demonstrated that these antibodies recognized the recombinant @entity35 @entity53398 , @entity53399 , and @entity14686 proteins with different avidity. The polyclonal antibody K118 and the monoclonal M3S1 were most specific for @entity35 @entity53399 and @entity14686 forms, respectively, whereas the AC55 antiserum displayed the highest affinity for @entity53398 forms. This selectivity was confirmed by Western blot analysis using recombinant @entity35 @entity53398 , @entity53399 , and @entity14686 proteins expressed in a heterologous system. These antibodies were used to characterize the @entity268 -PDEs expressed in the @entity35 brain. An immunoblot of extract of cortex and cerebellum demonstrated that at least seven different polypeptides specifically cross-reacted with the different antibodies, indicating that multiple @entity268 -PDEs are expressed in this tissue. On the basis of cross-reactivity with @entity14686 but not @entity53398 or @entity53399 antibodies, 93- and 105-kDa @entity14686 species were detected in the cortex and cerebellum extract. These forms are different from the 68-kDa @entity14686 form expressed in endocrine cells after hormonal stimulation. Although the 93-kDa form was recovered in both the soluble and particulate fractions, the 105-kDa polypeptide was mostly particulate in the cortex and cerebellum extracts. @entity53399 forms of 90-87 kDa were recovered in both soluble and particulate compartments of the brain extract. These forms were different from the previously identified @entity53398 variants of 110 and 75 kDa. These data demonstrate that the presence of multiple @entity268 -PDE genes is translated into @entity268 -PDE proteins of different sizes and distinct immunological properties and that multiple variants derived from these @entity268 -PDE genes are expressed in different regions of the brain and different subcellular compartments. These immunological tools will be useful to identify different @entity268 -PDE forms expressed in organs targeted for pharmacological intervention with PDE4 inhibitors.

Characterization of the XXXX -sensitive, cyclic AMP-specific phosphodiesterases: identification and differential expression of immunologically distinct forms in the @entity35 brain.

@entity268 :: ('CHEBI:17489', 'Chemical') :: ['cAMP'] @entity7428 :: ('MESH:D020889', 'Chemical') :: ['rolipram'] @entity53398 :: ('25638(Tax:10116)', 'Gene') :: ['PDE4A'] @entity53399 :: ('24626(Tax:10116)', 'Gene') :: ['PDE4B'] @entity35 :: ('10116', 'Species') :: ['rat'] @entity14686 :: ('24627(Tax:10116)', 'Gene') :: ['PDE4D']
@entity7428 :: (MESH:D020889,Chemical) :: ['rolipram']
