Immunocytochemical localization of calmodulin in PC12 cells and its possible interaction with histones.
The subcellular localization of calmodulin, a multi-functional calcium-binding regulatory protein, was examined immunocytochemically in undifferentiated PC12 rat pheochromocytoma cells and cells differentiated with nerve growth factor (NGF) and dibutyryl cyclic AMP. In undifferentiated PC12 cells, diffuse immunostaining for calmodulin was observed in the cytoplasm, and weak, patch-like staining was found in the nucleus. In differentiated cells, intense immunostaining for calmodulin was observed in the cytoplasm, while nuclear immunostaining was still evident. Immunoreactivity for calmodulin was also observed along newly-formed neuritic processes, with strong staining in varicosity-like structures and growth cones. Using double-label immunochemistry, the relative intensity of immunostaining for calmodulin among the nuclei was found to correlate with the relative intensity of immunostaining for histones in the same nuclei. A comparison of a profile of 125I-calmodulin binding in the nuclear fraction from PC12 cells to that of immunoblotting for histones in the same fraction indicated that some of the histones are calmodulin-binding proteins in PC12 cells. These results show that the level and subcellular distribution of calmodulin are altered during the course of nerve cell differentiation and suggest the possibility that histones may function as major nuclear binding proteins for calmodulin.
D000818:Animals; D000918:Antibody Specificity; D003994:Bucladesine / Q000494:pharmacology*; D002147:Calmodulin / Q000032:analysis*; D002454:Cell Differentiation / Q000187:drug effects; D004396:Coloring Agents; D006657:Histones / Q000032:analysis*; D015151:Immunoblotting; D007150:Immunohistochemistry; D009414:Nerve Growth Factors / Q000494:pharmacology*; D016716:PC12 Cells; D051381:Rats; D013347:Subcellular Fractions / Q000737:chemistry*; D014404:Tubulin / Q000032:analysis
D002147:Calmodulin; D004396:Coloring Agents; D006657:Histones; D009414:Nerve Growth Factors; D014404:Tubulin; D003994:Bucladesine