Immunochemical Identification of 2′, 3′‐Cyclic Nucleotide 3′‐Phosphodiesterase in Central and Peripheral Nervous System Myelin, the Wolfgram Protein Fraction, and Bovine Oligodendrocytes

Abstract

Myelin proteins and the total Wolfgram protein fraction were isolated from the CNS of several mammalian species and characterized with rabbit anti-bovine 2', 3'-cyclic nucleotide 3'-phosphodiesterase (CNP) antisera after sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electrophoretic transfer to nitrocellulose membranes. The corresponding CNP proteins cross-reacted across all species examined, suggesting that the CNP amino acid sequence was fairly well conserved in all six species. The same corresponding proteins were also identified immunochemically in the crude total Wolfgram protein fraction in the region of the W1 myelin protein, thus further supporting and extending two different previous reports indicating a relationship between CNP and the W1 protein. In addition to these CNS enzyme sources, peripheral nervous system CNP (rabbit and rat sciatic nerve) was also recognized by these same rabbit anti-bovine (CNS) CNP antisera. CNP was also detected in freshly isolated delipidated bovine oligodendrocyte membranes. These results suggest that rabbit anti-bovine CNP antisera may be of use in localization and structural studies of this enzyme in several different species and will permit clear identification of CNP in oligodendrocytes and their isolated membrane fractions.

Differential Ultrastructural Localization of Myelin Basic Protein, Myelin/Oligodendroglial Glycoprotein, and 2′,3′‐Cyclic Nucleotide 3′‐Phosphodiesterase in the CNS of Adult Rats

“…CNP is an enzyme involved in the hydrolysis of 2',3'cyclic nucleotide phosphates (Drummond et al, 1962) and is found by biochemical and immunocytochemical methods in highest activities in central myelin and oligodendrocytes (Kurihara and Tsukada, 1 967;Roussel et al, 1978;Roussel and Nussbaum, 1981;Sheedlo et ULTRASTRUCTURAL DISTRIBUTION OF MYELIN ANTIGENS al., 1983;Sprinkle et al, 1983;McMoms et al, 1984). Work by Braun et al (1988) in normal and myelindeficient (mld) mice also shows, that immunoreactive CNP is particularly concentrated at the periphery of the oligodendrocyte, generally adjacent to the Plasma membrane but not especially associated with membranes.…”

Identification of oligodendrocytes in experimental disease models

“…While CC1 is directed against APC, the adenomatous polyposis coli, a tumor suppressor protein, it also binds to a nonspecific antigen in oligodendrocytes (Bhat et al, 1996;Brakeman et al, 1999). CC1 is (Sprinkle et al, 1983). Although CNPase is expressed early in oligodendrocyte development, its expression is greatly increased in mature, myelinating cells.…”

A Rabbit Autoantibody Specific for the 46‐kDa Form of 2′,3′‐Cyclic Nucleotide 3′‐Phosphodiesterase

“…The higher band in the PNS homogenates that was stained with the autoantibody also stained with known anti-CNP antiserum (not shown). Although the reason for this reactive component with a higher apparent molecular weight in the PNS that has also been observed by Sprinkle et al (1983) is not known, it may be a CNP-lipid complex because the relative amount of it appeared to be less when the samples were delipidated with ether-ethanol or chloroform-methanol before electrophoresis. Panel C shows that purified PNS myelin from rabbit and human nerve also contained the 46-kDa antigen and the higher molecular weight band.…”