Diurnal and circadian variation of protein kinase C immunoreactivity in the rat retina

We studied the dependence of the expression of protein kinase C immunoreact ivity (PKC-IR) in the rat retina on the light:dark (LD) cycle and on circad ian rhythmicity in complete darkness (DD). Two anti-PKC alpha antibodies we re employed: One, which we call PKC alpha beta recognized the hinge region; the other, here termed PKC alpha, recognized the regulatory region of the molecule. Western blots showed that both anti-PKC antibodies stained an ide ntical single band at approximately 80 kD. The retinal neurons showing PKC- IR were rod bipolar cells and a variety of amacrine neurons. After 3 weeks on an LD cycle, PKC alpha beta -IR in both rod bipolar and certain amacrine cells manifested a clear rhythm with a peak at zeitgeber time (ZT) of 06-1 0 hours and a minimum at ZT 18. No rhythm in total PKC-IR was observed when using the PKC alpha antibody, but, at ZT 06-10 hours, rod bipolar axon ter minals showed increased immunostaining. After 48 hours in DD, with either a ntibody, rod bipolar cells showed increased PKC-IR. The PKC alpha antibody alone revealed that, after 48 hours, AII amacrine neurons, which lacked PKC -IR in an LD cycle, manifested marked PKC-IR, which became stronger after 7 2 hours. Light administered early in the dark period greatly increased PKC alpha beta -IR in rod bipolar and some amacrine neurons. Our data indicate that light and darkness exert a strong regulatory influence on PKC synthesi s, activation, and transport in retinal neurons. (C) 2001 Wiley-Liss, Inc.