2',3'-Cyclic nucleotide 3'-phosphodiesterase is associated with mitochondria in diverse adrenal cell types.
In this study, we have examined the expression and intracellular localisation of the myelin protein 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNP) in bovine adrenal medullary chromaffin cell cultures. By immunoblotting, using two distinct anti-CNP monoclonal antibodies, CNP was detected in medullary cell cultures and expression of CNP was confirmed by reverse transcription and PCR amplification. CNP did not leak from digitonin-permeabilised chromaffin cells, suggesting that there is no cytosolic pool of this protein. Immunofluorescence studies with both antibodies showed that all cells in the medullary chromaffin cell culture were stained with a punctate appearance consistent with an intracellular localisation for CNP. More specifically it was demonstrated that CNP is co-localised with mitochondria. Various cell types in chromaffin cell cultures were stained with a mitochondrial pattern and CNP staining was co-localised with mitochondrial staining. These results show that CNP is a widely expressed protein that is associated with mitochondria and provides new clues as to its cellular function outside of myelin structures.
McFerran
B
B
The Physiological Laboratory, University of Liverpool, Liverpool L69 3BX, UK.
Burgoyne
R
R
D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases / Q000032:analysis* / Q000096:biosynthesis*; D000313:Adrenal Medulla / Q000166:cytology / Q000201:enzymology*; D000818:Animals; D000911:Antibodies, Monoclonal; D001921:Brain / Q000201:enzymology; D002417:Cattle; D002478:Cells, Cultured; D019439:Chromaffin Cells / Q000145:classification / Q000166:cytology / Q000201:enzymology*; D007150:Immunohistochemistry; D008928:Mitochondria / Q000201:enzymology* / Q000648:ultrastructure; D016133:Polymerase Chain Reaction; D051381:Rats; D017208:Rats, Wistar
D000911:Antibodies, Monoclonal; D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases