Protein kinase C mediates transient spinule-type neurite outgrowth in the retina during light adaptation

Light and dark adaptation of the teleost retina is accompanied by a remarkable morphological rearrangement of the synaptic connections between photoreceptors and second-order neurons: during light adaptation, numerous new neurites, the so-called spinules, arise from the terminal dendrites of horizontal cells invaginating the cone pedicle, and during dark adaptation, these spinules are retracted. The formation of these spinules is paralleled by the appearance of color opponency in horizontal and ganglion cells, which led to the suggestion that these spinules are the site of the inhibitory synapses in the negative feedback loop between cones and horizontal cells. The formation of the spinules in the light and their disappearance in darkness have a time course of minutes and are modulated by the neurotransmitters dopamine and glutamate, respectively. Neurotransmitters can modulate neuronal processing through a variety of second messengers that activate protein kinases, resulting most commonly in protein phosphorylation. Herein we report that activation of protein kinase C by phorbol esters promotes the formation of new horizontal-cell spinules in animals kept in the dark. Partial inhibition of protein kinase C activation with sphingosines prevents the formation of new spinules during light adaptation but does not affect established spinules. The spinule-forming effect of phorbol esters is not mediated by dopaminergic neurons, since the effect is also seen in retinas depleted of dopaminergic neurons. Phorbol esters also initiate the formation of spinules in synaptically isolated horizontal cells, demonstrating that they have a direct action on these cells. In addition, isolated horizontal cells have substrate proteins that are phosphorylated in a protein kinase C-dependent manner.