Time‐dependent changes in protein phosphorylation patterns in rat brain synaptosomes caused by deltamethrin

Abstract

AbstractEffects of deltamethrin, a powerful pyrethroid insecticide, on the protein phosphorylation and dephosphorylation processes during depolarization in rat brain synaptosomes were studied by using [32P]phosphoric acid as a starting radiotracer and high external concentration of potassium ions or veratridine (10−‐5 M) as depolarizing agents. At the onset of depolarization there was a quick rise in phosphorylation in various synaptic proteins for about 15–30 s followed by a gradual decline in levels of phosphorylation. The effect of deltamethrin (10−‐7 M) on this system was found to be dependent on the length of preincubation of the synaptosome with the pesticide prior to depolarization. At an early stage (0–3 min preincubation period) it caused a modest suppression of protein phosphorylation activities. When the period of deltamethrin preincubation was extended to 5–20 min, however, it caused a significant increase in protein phosphorylation throughout the depolarization period. At the later stage of the action of deltamethrin (e.g. preincubation period of 30–40 min), deltamethrin‐treated synaptosomes no longer responded to the depolarization signal to raise the level of phosphorylation on many proteins. These results indicate that deltamethrin's actions on the synaptic process are complex. Depending on the length of exposure, its effects on protein phosphorylation responses in intact synaptosomes could be either stimulatory or inhibitory. To study the cause of deltamethrin‐induced synaptic block at the later stage, effects of deltamethrin on protein kinases were studied by using lysed synaptic membranes with [gamma‐32P]ATP. Deltamethrin was shown to inhibit calcium–calmodulin‐dependent protein phosphorylation activities at 10−‐7 M when given directly to the enzyme source 10 min prior to the addition of [32P]ATP. Such an observation helps to explain the inhibitory action of deltamethrin on protein phosphorylation which occurs at the late stage of its action (i.e. preincubation time > 20 min).