Lewis S. Seiden

Long-lasting depletions of striatal dopamine and loss of dopamine uptake sites following repeated administration of methamphetamine.

Repeated administration of high doses of methamphetamine produced long-term decreases in dopamine (DA) levels and in the number of DA uptake sites in the rat striatum. These two effects were dose-related and did not appear to be due to the continued presence of drug in striatal tissue. Long-lasting depletions induced by methamphetamine were selective for striatal DA neurons since norepinephrine (NE) levels in all of the rat brain regions examined were not changed on a long-term basis by methamphetamine treatments. Supersensitivity of DA receptors did not accompany the loss of striatal DA and its uptake sites.

Long-term effects of repeated methylamphetamine administration on dopamine and serotonin neurons in the rat brain: a regional study.

Repeated high doses (25 and 100 mg/kg) of methylamphetamine produce long-term depletions of both dopamine (DA) and serotonin (5-HT) in the rat brain. In the DA system, depletions are most pronounced in the neostriatum and substantia nigra, with decreased levels in these two regions being significantly correlated. Within the 5-HT system, levels are most reduced in the amygdala, frontal cortex and neostriatum. When both the DA and 5-HT depleting actions of methylamphetamine are considered, the hypothalamus stands out as one of the more resistant brain regions. The regional pattern of reduced 5-HT levels following methylamphetamine is similar to that seen after p-chloroamphetamine. After both methylamphetamine and p-chloroamphetamine, a loss of 5-HT synaptosomal uptake sites occurs. Serotonergic systems are more sensitive than DA systems to the apparent neurotoxic actions of methylamphetamine.

A rapid method for the regional dissection of the rat brain.

A method is described for the rapid dissection of seventeen areas of the rat brain. Regions from fresh unfrozen brain tissue are dissected from coronal brain slices obtained with use of a cutting block. This method is applicable to pharmacological and behavioral studies which require the dissection of numerous brains during short time intervals.

Dopamine nerve terminal degeneration produced by high doses of methylamphetamine in the rat brain

Numerous recent studies indicate that when amphetamines are administered continuously or in high doses, they exert long-lasting toxic effects on dopamine (DA) neurons in the central nervous system (CNS). Specifically, it has been shown that amphetamines can decrease the content of brain DA, reduce the number of synaptosomal DA uptake sites and selectively depress the in vitro activity of neostriatal tyrosine hydroxylase (TH). To date, however, anatomical evidence of DA neuronal destruction following amphetamines has not been reported. In this study, chemical methods were used in conjunction with the Fink-Heimer method which allows for the selective silver impregnation of degenerating nerve fibers, in order to determine whether methylamphetamine, a potent psychomotor stimulant often abused by man, causes actual DA neural degeneration. It has been found that methylamphetamine induces terminal degeneration along with correlative DA neurochemical deficits in the neostriatum and nucleus accumbens; furthermore, that in cresyl violet-stained sections of the substantia nigra (SN), pars compacta, and ventral tegmental area (VTA), there is no evidence of cell body loss in rats in which 50-60% of neostriatal DA terminals have been destroyed.

Small Changes in Ambient Temperature Cause Large Changes in 3,4-Methylenedioxymethamphetamine (MDMA)-Induced Serotonin Neurotoxicity and Core Body Temperature in the Rat

The amphetamine derivative 3,4-methylenedioxymethamphetamine (MDMA) is a drug of abuse and has been shown to be neurotoxic to 5-HT terminals in many species. MDMA-engendered neurotoxicity has been shown to be affected by both ambient temperature and core body temperature. We now report that small (2°C) changes in ambient temperature produce changes in core temperature in MDMA-treated rats, but the same changes in ambient temperature do not affect core temperature of saline-treated animals. Furthermore, increases in core temperature of MDMA-treated animals increase neurotoxicity. Rats were given MDMA (20 or 40 mg/kg) or saline and placed in an ambient temperature of 20, 22, 24, 26, 28, or 30°C using a novel temperature measurement apparatus that controls ambient temperature ±0.5°C. Two weeks after MDMA treatment, the rats were killed, and regional 5-HT and 5-hydroxyindole acetic acid levels were analyzed as a measure of neurotoxicity. Rats treated with MDMA at 20 and 22°C showed a hypothermic core temperature response. Treatment with MDMA at 28 and 30°C produced a hyperthermic response. At ambient temperatures of 20–24°C, neurotoxicity was not observed in the frontal cortex, somatosensory cortex, hippocampus, or striatum. At ambient temperatures of 26–30°C, neurotoxicity was seen and correlated with core temperature in all regions examined. These data indicate that ambient temperature has a significant affect on MDMA neurotoxicity, core temperature, and thermoregulation in rats. This finding has implications on both the temperature dependence of the mechanism of MDMA neurotoxicity and human use because fatal hyperthermia is associated with MDMA use in humans.

Long-term methamphetamine induced changes in brain catecholamines in tolerant rhesus monkeys

(Received September 1, 1975) : d-Amphetamine, methamphetamine and other phenethylamines cause decreases in food intake, stereotypic behavior, disruption of behavior that is under stimulus control, as well as general sympathomimetic effects such as hyperthermia, increased blood pressure and piloerection. The doses required to produce different effects vary. In addition, most psychomotor stimulants are subject to abuse in man and will be self-administered by animals [ 1]. Repeated administration of the amphetamines leads to the development of tolerance to many of its effects on behavior although the degree of tolerance depends to a large extent on the frequency of administration, the dose, the route of administration and the behavior in question. Several p investigators have presented evidence that amphetamines exert their effects upon behavior by increasing the concentration of catecholamines at the synaptic cleft by blocking re-uptake and/or promoting release. The overall purpose of this investigation was to determine the effect of long-term repeated intravenous administration of methamphetamine in increasing doses to rhesus monkeys. This study was part of a larger research program designed to determine if long-term administration of methampheta-mine results in irreversible effects in the rhesus monkey. To this end, measures were made on behavior, neuropathology and brain catecholamines. In this paper we shall report both short and long-term changes in brain cate-cholamines that were caused by prolonged administration of methampheta-mine. It was found that methamphetamine induced changes in regional brain catecholamines, some of which persisted even after the drug was discontinued. Rhesus monkeys were injected intravenously 8 times daily. The dose of methamphetamine was increased until a final dose was reached which could through observing diminished food intake and weight loss. Doses were incremented only when signs of behavioral tolerance appeared. Two studies were carried out: in the first, monkeys were treated for 3-6 months {final dose 3.0-6.5 mg/kg/inj) eight times per day. Monkeys were either killed 24 hours or 3-6 months after the last injection. In the

Formation of 6-hydroxydopamine in caudate nucleus of the rat brain after a single large dose of methylamphetamine ☆

We now report that 6-hydroxydopamine (0.39 +/- 0.31 nanograms/mg of tissue at 2 hr) is formed in the rat caudate nucleus after a single injection of methylamphetamine (100 mg/kg). The same dose of methylamphetamine causes approximately 50% depletion of caudate dopamine 2 weeks after the injection. We suggest that the formation of 6-hydroxydopamine from endogenous dopamine is responsible for the neurotoxicity to dopamine terminals seen after methylamphetamine administration.

Further evidence that amphetamines produce long-lasting dopamine neurochemical deficits by destroying dopamine nerve fibers

Methamphetamine and amphetamine were continuously administered to rats for 3 days by means of subcutaneously implanted osmotic minipumps. The total daily dose of each drug was approximately 4 mg/day. Dopamine, norepinephrine and serotonin determinations two weeks later indicated that both amphetamines produced a selective striatal dopamine depletion. Anatomical studies indicated that this depletion was associated with striatal nerve fiber degeneration. To determine whether this fiber degeneration induced by amphetamines was dopaminergic, the long-lasting dopamine depletion produced by methamphetamine was antagonized with alpha-methyl-para-tyrosine. This prevented the appearance of nerve fiber degeneration after methamphetamine. These findings suggest that amphetamines produce a long-term striatal dopamine depletion by destroying striatal dopamine nerve fibers.

Long-term effects of chronic methamphetamine administration in rhesus monkeys

Biochemical and neuropathological effects of exposure to a high dose regimen of methamphetamine were evaluated in rhesus monkeys approximately 4 years after the last drug injection. Concentrations of dopamine and serotonin in caudate were below control levels as were concentrations of serotonin in several other brain regions. These changes were more severe in a monkey that was exposed twice to the drug regimen. A decrease in caudate synaptosomal uptake of both neurotransmitters as well as neuropathological changes were evident in that monkey. Although it is possible that partial recovery occurred, these results strongly suggest that methamphetamine-induced neurotoxicity may be permanent.

Effects of repeated injections of cocaine on D1 and D2 dopamine receptors in rat brain

In order to determine if chronic administration of cocaine produced long-lasting alterations in dopamine receptor binding, rats were treated with single daily injections of cocaine (0, 10, or 20 mg/kg) for 15 consecutive days and killed either 20 min or 2 weeks after the last injection. The density of D1 binding sites in frontal cortex was either unchanged (10 mg/kg) or slightly increased (20 mg/kg) 20 min after the last daily injection, but was decreased 2 weeks later. D1 sites in striatum were decreased both immediately and 2 weeks after the injection regimen. Decreases in D1 binding site density in nucleus accumbens were observed only immediately after the last injection. In contrast to these effects on D1 binding sites, D2 binding sites were decreased in striatum and frontal cortex and increased in the nucleus accumbens 20 min after repeated cocaine, but were unaffected 2 weeks after repeated cocaine. Computer-assisted analysis of the saturation isotherms revealed that chronic administration of cocaine did not affect the affinity (Kd) of the radioligands used to label D1 or D2 sites. These findings suggest that repeated administration of cocaine results in long-term decreases in D1 binding sites in striatum and frontal cortex and transient decreases in D2 binding sites. Furthermore, cocaine caused opposite, transient effects on D1 and D2 site density in nucleus accumbens.