ABSTRACT
The 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNP), a protein of unknown function in vivo, is abundantly expressed in myelinating glia in two isoforms, CNP1 and CNP2. In this study, immunoblot analysis showed that CNP1 is the major isoform in adult forebrain, and that both isoforms are included in the postsynaptic density (PSD) fraction and tyrosine-phosphorylated at the basal level. However, subcellular distribution and detergent extraction data showed that CNP is nonspecifically associated with the PSD fraction. Immunocytochemistry revealed that CNP is detected, in a weak but punctate pattern, in dissociated rat hippocampal neurons of 3 days to 2 weeks in vitro. The CNP-positive punctae were distributed throughout soma and dendrites, and distinct from PSD95-positive ones. Immunoblot analysis indicated that CNP is also expressed in neuronal stem cell lines, HiB5 and F11. Interestingly, in addition to the known two isoforms, a new CNP isoform of MW 45 kDa was expressed in these cell lines and was the major type of isoform in F11 cells. Taken together, our data suggest that CNP is expressed in the early stage of in vitro development and nonspecifically included in the adult rat PSD fraction.
Subject(s)
Animals , Rats , 2',3'-Cyclic-Nucleotide Phosphodiesterases/metabolism , Aging/physiology , Cells, Cultured , Hippocampus/cytology , Immunohistochemistry , Nerve Tissue Proteins/metabolism , Neurons/metabolism , Phosphotyrosine/metabolism , Prosencephalon/cytology , Rats, Sprague-Dawley , Substrate SpecificityABSTRACT
Classical studies of macroglial proliferation in muride rodents have provided conflicting evidence concerning the proliferating capabilities of oligodendrocytes and microglia. Furthermore, little information has been obtained in other mammalian orders and very little is known about glial cell proliferation and differentiation in the subclass Metatheria although valuable knowledge may be obtained from the protracted period of central nervous system maturation in these forms. Thus, we have studied the proliferative capacity of phenotypically identified brain stem oligodendrocytes by tritiated thymidine radioautography and have compared it with known features of oligodentroglial differentation as well as with proliferation of microglia in the opossum Didelphis marsupialis. We have detected a previously undescribed ephemeral, regionally heterogenous proliferation of oligodendrocytes expressing the actin-binding, ensheathment-related protein 2' 3'- cyclic nucleotide 3' -phosphodiesterase (CNPase), that is not necessarily related to the known regional and temporal heterogeneity of expression of CNPase in cell bodies. On the other hand, proliferation of microglia tagged by the binding of Griffonia simplicifolia B4 isolectin, which recognizes an alpha-D-galactosyl-bearing glycoprotein of the plasma membrane of macrophages/microglia, is known to be long lasting, showing no regional heterogeneity and being found amongst both ameboid and differentiated ramified cells, although at different rates. The functional significance of the proliferative behavior of these differentiated cells is unknown but may provide a lowgrade cell renewal in the normal brain and may be augmented under pathological conditions.
Subject(s)
Animals , Brain Stem/physiology , Cell Division , Microglia/physiology , Neuroglia/physiology , Oligodendroglia/physiology , Opossums/physiology , 2',3'-Cyclic-Nucleotide Phosphodiesterases , Autoradiography , Biomarkers , LectinsABSTRACT
During the last years there have been major advances in the knowledge of cyclic nucleotide phosphodiesterases. Particularly, referred to the presence of multiple different isozymes. Seven different phosphodiesterase gene families, have been described in mammalian tissues, containing several distinct genes, most of them expressed in different tissues as functionally unique splice variants. This article includes various aspects of the currently accepted nomenclature, structure and function of each family of phosphodiesterases. Finally, a brief discussion of the presence and role of these enzymes in the cell proliferation and differentiation processes, in parasites of the Trypanosmatidae family, is provided