Histological and neurochemical effects of fetal treatment with methylazoxymethanol on rat neocortex in adulthood.

Forebrain microencephaly results when developing rats are exposed to methylazoxymethanol acetate (MAM) at 15 days of gestation (DG). This potent alkylating agent is selectively cytotoxic for dividing cells. Since distinct neuronal populations in neocortex vary greatly with respect to timing of mitotic activity during gestation, it was predicted that some groups would be differentially reduced by treatment. Histological examination of neocortex from treated rats grown to adulthood revealed major losses of laminae II--IV with relative preservation of deeper layers. The atrophic adult neocortex was further characterized by assay of several defined pre- and postsynaptic neurochemical markers. Total markers for GABAergic neurons were greatly reduced (glutamate decarboxylase -71%, [3H]GABA synaptosomal uptake -63% and endogenous GABA -59%). Total [3H]GABA binding to cortical membranes was reduced 67%. Total [3H]glutamate synaptosomal uptake and endogenous glutamate were reduced 71% and 65% respectively. In contrast, total presynaptic markers for noradrenergic innervation were minimally altered but concentration of tyrosine hydroxylase, [3H]norepinephrine synaptosomal uptake and endogenous norepinephrine were increased by 275%, 130% and 133%, respectively. Concentration of cholinergic presynaptic markers was also increased (choline acetyltransferase +97%, endogenous acetylcholine +64%) in atrophic cortex, but to a lesser degree than for noradrenergic innervation. Specific binding of muscarinic cholinergic antagonist [3H]quinuclidinyl benzilate and the beta-adrenergic receptor antagonist [3H]dihydroalprenolol was reduced 25% and 29% respectively in treated cortex. Thus, MAM treatment at 15 DG severely reduces intrinsic neuronal populations including GABAergic and glutamatergic neurons, and produces a shrunken cortex relatively hyperinnervated by noradrenergic and cholinergic neurons. MAM-induced microencephaly is a useful model system for producing relatively selective lesions of telencephalic neurons and for study of altered neurochemical relationships following developmentally incurred brain damage.

Role of dopamine storage function in the control of rat striatal tyrosine hydroxylase activity.

Pretreatment of rats with reserpine prevents and post-treatment with RO4-1284 depletes the gamma-butyrolactone (GBL)-induced increase of striatal dopamine (DA) levels. This suggests that the accumulation of DA in striatal nerve endings that normally follows GBL-induced cessation of nigrostriatal impulse flow is in reserpine-sensitive sites. Three days after a single injection of reserpine, the ability of either haloperidol, a DA receptor blocker, or GBL to enhance DA synthesis is greatly reduced and these responses recover slowly over a two week period. Similarly, the ability of haloperidol to elevate striatal DA metabolite concentrations shows a similar pattern of inhibition. The rate of recovery after reserpine of haloperidol effects on DA metabolite concentrations and the activation of striatal tyrosine hydroxylase (measured in vivo by the 30 min L-DOPA accumulation after decarboxylase inhibition with NSD-1015) after either haloperidol or GBL parallels the rate of recovery of basal DA levels. The accumulation of DA after GBL proceeds for 60 min before beginning to plateau in normal rats, but 3 days after reserpine the DA elevation stops after 15 min and lasts for only 30 min in 10 day reserpinized animals. The initial 15 min accumulation of DA after GBL is the same in normal, 3 day and 10 day reserpinized rats, indicating that the initial enzymic rate of activity is the same, but the duration of activation is less. Thus, inhibition of DA storage function by reserpine alters the coupling of DA autoreceptor activity with tyrosine hydroxylase activity. It is suggested that DA storage function modulates tyrosine hydroxylase activity by controlling the amount of DA available for attachment to and inhibition of tyrosine hydroxylase enzyme. This hypothesis is consistent with recent immunocytochemical observations which suggest an association of tyrosine hydroxylase with synaptic vesicles in DA neuronal terminal areas.

Acetylcholinesterase from Wistar rat brain.

Acetylcholinesterase (E.C.3.1.1.7) was partially purified from rat brains stored in toluene. Extraction was performed using buffers containing non-ionic tensoactive detergents. Some properties of the enzyme were affected by the use of different activity measurement methods, such as the short-time radiometric or the long-time colorimetric method. There were two zones of maximum activity in the range pH 7.5-8.0 and 8.0-8.6, respectively. There seems to be a histidine residue in the enzyme that participates in the catalytic process. Thermal denuration presented first order kinetics and different thermodynamic parameters were obtained on using different incubation periods. On using the short-time activity measurement method there was activation at high substrate concentration, but with the long time method there was a marked inhibition produced by excess of substrate. However, if the enzyme was extracted from fresh rat brain, toluene untreated, these differences dissapeared. Gel filtration and disc electrophoresis showed the presence of multiple and interconvertible forms of the enzyme.

Light effect on the synaptic organ of the rat.

The photoreceptor synapses of albino rats show considerable pathologic changes following fluorescent light exposure. The changes in the synapses and in the lamellar membranes of the outer segments occur and progress simultaneously. Membranes proliferate in the paramitochondrial zone of the rod synaptic spherule and fine budding of the smooth endoplasmic reticulum occurs in the cone pedicle within one hour's exposure to the brightness of 500 foot candles. Proliferating paramitochondrial membranes have no cytochrome c oxidase activity and degenerate together with mitochondria by further exposure. The cone pedicles remain relatively intact in photically damaged retina.

[Effect of butaclamol enantiomers on hypothalamic tyrosine hydroxylase in the rat brain]. / Vliianie énantiomerov butaklamola na tirozingidroksilazu gipotalamusa mozga krys.

The authors demonstrate stereospecificity of the action of butaclamol enantiomers on substrate inhibition of hypothalamic tyrosine hydroxylase (TH) and regulation of the tyrosine hydroxylase response by the presynaptic membrane (presynaptic receptors) of rat hypothalamus synaptosomes under membrane activation with dopamine. The effect of (+)-butaclamol on the substrate inhibition of TH was noticeable at a concentration of 10(-8)M, reaching a maximum at 10(-5)M. (-)-Butaclamol administered at the same concentrations did not influence the substrate inhibition of the enzyme. (+)-Butaclamol added to the incubation medium containing hypothalamic synaptosomes concurrently with dopamine (10(-5)M) completely blocked the regulatory action of the latter on TH, with this action mediated via presynaptic receptors. (-)-Butaclamol (10(-5)M) antagonized the action of dopamine under the same conditions. The data obtained indicate high stereo-specificity of butaclamol enantiomers as regards their effect on presynaptic regulation of TH, suggesting that elimination of the substrate inhibition of hypothalamic TH is a stereoselective effect of neuroleptics and can be a prognostically important criterion in the appraisal of compounds with potential neuroleptic activity.