Electron microscopic immunocytochemical evidence that the calmodulin-dependent cyclic nucleotide phosphodiesterase is...

The calmodulin-dependent cyclic nucleotide phosphodiesterase represents an important junction between the Ca2+ and the cyclic AMP/cyclic GMP second messenger systems. In brain it is a major cyclic nucleotide-degrading activity and is selectively expressed in the soma and dendrites of regional output neurons [Kincaid et al. (1987) Proc. natn. Acad. Sci. U.S.A. 84, 1118-1122]. In this study the subcellular localization of this enzyme in cerebral cortex, hippocampus and inferior colliculus of rat brain was analysed by electron microscopic immunocytochemical methods using affinity-purified antibodies. The immunoreactivity was found exclusively within neurons whereas glial cells were unstained; preabsorption of antibody with phosphodiesterase eliminated this reactivity, demonstrating the specificity of immunostaining. In the neuronal cell bodies, deposits of immunoreaction product occurred as sparse patches in the cytoplasm and were often associated with organelles such as mitochondria, Golgi-complex and endoplasmic reticulum; nuclei, however, were free from immunoreaction product. In the neuronal processes immunoreactivity was found within dendrites and dendritic spines, whereas the myelinated axons and axon terminals were immunonegative. The postsynaptic densities of asymmetric synapses were associated with especially high concentrations of immunoreaction product. However, the immunopositive synaptic profiles appeared to be quite selective, comprising only a small percentage of the total number of synapses in the neuropil. Our results indicate that the calmodulin-dependent cyclic nucleotide phosphodiesterase is concentrated at postsynaptic sites in specific classes of neurons. This finding supports other morphological evidence indicating a primary role for cyclic nucleotide action in postsynaptic and not presynaptic structures. Furthermore, since this enzyme is regulated by Ca2+, this interface between second messenger systems seems to play a significant role in the postsynaptic integration of Ca(2+)-mediated neuronal inputs.