Expression of α-, β- and γ-subspecies of protein kinase C in the motor neurons in the embryonic and postnatal rat spinal cord

Abstract

Using polyclonal antibodies against alpha-, beta- and gamma-subspecies of protein kinase C, developmental changes in expression of these subspecies in the motor neurons in the rat cervical spinal cord were immunohistochemically investigated. On embryonic day-12, the motor neurons began to differentiate from undifferentiated neuroepithelial cells. On embryonic day-13, they began to express weak immunoreactivity for alpha- and beta-protein kinase C and slightly more evident immunoreactivity for gamma-protein kinase C. Immunoreactivity for protein kinase C in these neurons gradually became stronger, as the development progressed. Between embryonic day-18 and postnatal day-7, the motor neurons showed distinct immunoreactivity in the nucleus, perikaryal cytoplasm, axon and dendrites. At these stages, distribution and intensity of immunoreactivity for alpha-, beta- and gamma-protein kinase C were very similar. Thereafter, the expression of this enzyme in the nucleus gradually declined, while in the other structures, expression of each subspecies changed independently. On postnatal day-28 and 35, expression of beta-protein kinase C in the axons was stronger than that of alpha- and gamma-protein kinase C, and immunoreactivity for gamma-protein kinase C in the perikaryal cytoplasm and dendrites was slightly weaker than that for alpha- and beta-protein kinase C. Expression of this enzyme in the motor neurons at these stages was almost the same as in the adult animal. Electron microscopically, immunoreactivity for protein kinase C was randomly distributed in the nucleus, and in the perikaryal cytoplasm, often near the cisterns of the endoplasmic reticulum. Expression of protein kinase C in the growing axons was quite different from that in the mature axons. In the dendrites, immunoreactivity for protein kinase C was distributed randomly in the cytoplasm and at the postsynaptic densities. These findings suggest that protein kinase C might regulate not only the neural functions, but also several aspects of the differentiation process in the motor neurons.