An ultrastructural study into the effects of pentobarbitone on synaptic organization

A series of adult male rats was analyzed to test the effects of varying doses of pentobarbitone sodium on the ultrastructure of synapses in the molecular layer of the parietal cortex. The rats were divided into the following categories: unanaesthetized stunned, unanaesthetized cannulated and those subjected to 40, 80, 160, 300 or 400 mg/kg pentobarbitone. All material was examined both qualitatively and quantitatively after aldehyde-OsO4 or ethanolic phosphotungstic acid (E-PTA) treatment. The principal qualitative observations were: the preponderance in the unanaesthetized stunned and 160–400 mg/kg material of a variety of intraterminal profiles including synaptic vesicles, mitochondria, coated vesicles, tubular profiles, vacuoles, cisterns and double membrane profiles; the presence of exocytotic sites along the presynaptic membrane in the unanaesthetized stunned and cannulated material; the presence of endocytotic sites over the limiting membrane of the terminal away from the cleft in the unanaesthetized stunned and 160–400 mg/kg material; the prominence of the presynaptic network in the unanaesthetized and 40 mg/kg E-PTA material; discontinuity of the cleft material in the 40–160 mg/kg material. Findings to emerge from the quantitative aspect of the study show that pentobarbitone influences synaptic curvature, with a marked increase in curvature negativity over the 0–80 mg/kg dose range and a decrease in negativity at higher dose levels. The increase in curvature negativity is accompanied by an increase in synaptic length and dense projection numbers, with a consonant increase in the perimeter and area of the presynaptic terminal. Reversal of the negativity trend at higher dose levels is parallelled by reversal of these accompanying trends. Both sets of findings can be accounted for by membrane recycling within the terminal, supporting a membrane redistribution hypothesis.