Decline in motor functions in aging is related to the loss of NMDA receptors.

The aim of the study was to assess the contribution of central dopaminergic and glutamatergic systems to the age-dependent loss of motor functions in rats. Rats of three age groups were compared: young (3-5-month-old), middle-aged (20-21-month-old) and old (29-31-month-old). The obtained results showed an age-dependent decline in the electromyographic (EMG) resting and reflex activities in the gastrocnemius and tibialis anterior muscles, as well as in the T-maze performance. Although these disturbances were accompanied with significant age-dependent decreases in the binding to NMDA, AMPA and dopamine D2 receptors, and a decline in the number of nigral dopamine neurons, they were significantly correlated with the loss of the binding to NMDA receptors only. The reduction in T-maze performance with aging was additionally correlated with a decrease in motor functions (EMG activity). The study suggests a crucial role of the loss of NMDA receptors in age-dependent motor disabilities, as well as in disturbances measured in the T-maze.

L-701,324, a selective antagonist at the glycine site of the NMDA receptor, counteracts haloperidol-induced muscle rigidity in rats.

RATIONALE: It has recently been suggested that the overactivity of glutamatergic neurotransmission may contribute to the pathophysiology of Parkinson's disease. Therefore, a search for new compounds which block glutamatergic receptors and show antiparkinsonian properties in animal models of this disease seems to be justified. OBJECTIVE: The aim of this study was to determine whether L-701,324 [7-chloro-4-hydroxy-3(3-phenoxy) phenylquinoline-2-(H)-one], a selective and full antagonist at the glycine site of the NMDA receptor, counteracts parkinsonian-like muscle rigidity and catalepsy induced by haloperidol in rats. METHODS: The muscle tone was measured as the resistance developed to passive flexion and extension of the hind limb. Electromyographic (EMG) activity was additionally recorded in the gastrocnemius and tibialis anterior muscles. RESULTS: L-701,324 (2.5-40 mg/kg IP) dose-dependently decreased the muscle tone enhanced by haloperidol (1-5 mg/kg IP). Likewise, the haloperidol-enhanced resting EMG activity and the EMG reflex response to passive movements were diminished by lower and almost abolished by higher doses of L-701,324. However, up to a dose of 20 mg/kg IP, L-701,324 did not influence haloperidol (0.5 mg/kg IP)-induced catalepsy. Moreover, L-701,324 (1.25-5 mg/kg IP) given alone or together with haloperidol (0.5-1 mg/kg IP) disturbed rotarod performance. Gross observation of behaviour indicated that rats injected with L-701,324 in doses equal to or higher than 5 mg/kg, alone or in combination with haloperidol, were markedly ataxic, i.e. rats showed signs of disturbed balance and loss of control over their hind limbs. CONCLUSIONS: The present study suggests that L-701,324 exhibits a beneficial action in the animal model of parkinsonian rigidity, but not that of parkinsonian akinesia. Nonetheless, this compound is not devoid of motor side-effects.

Muscle rigidity induced by fluphenazine in rats is antagonized by L-DOPA, an antiparkinsonian drug.

The aim of the present study was to find out whether the classic neuroleptic fluphenazine is a good model compound for inducing parkinsonian-like muscle rigidity in rats. The muscle tone was measured as resistance developed by the rat's hind foot to passive flexion and extension. Fluphenazine in doses of 0.4-3.0 mg/kg i.p. induced a dose-dependent increase in the hind foot resistance to passive movements. The muscle rigidity induced by fluphenazine 1.5 mg/kg i.p.) was counteracted in a dose-dependent manner by the main antiparkinsonian drug L-DOPA (25-75 mg/kg i.p.). The present results suggest that the fluphenazine-induced muscle rigidity may be a useful model of parkinsonian rigidity.

Age-related muscle stiffness: predominance of non-reflex factors.

This study was aimed at assessing the contribution of reflex and non-reflex factors to the muscle tone of old female Wistar rats. The hind foot of a rat was flexed or extended at the ankle joint by 25 degrees over 250 ms. The resistance of the foot to passive movements (torque, mechanomyogram), as well as the reflex electromyographic activity in the gastrocnemius and tibialis anterior muscles, were recorded simultaneously. Moreover, the impact of the blockade of the reflex activity caused by the local anesthetic lignocaine (1-2 ml of a 2% solution, injected in the vicinity of the sciatic nerve) on the muscle tone was investigated. Additionally, old rats' hind leg muscle samples were analysed using fluorescent microscopy for the expression of fibronectin, which is an early marker of connective tissue formation. It has been shown that old rats are characterized by (i) a substantially increased resistance of flexor muscle stiffness (measured during extension) and unchanged resistance of extensors (measured during flexion), (ii) the loss of a major part of the reflex electromyographic activity and (iii) the increased content of fibronectin in muscles. Moreover, it has been shown that lignocaine, which completely blocked the electromyographic reflex activity in the gastrocnemius and tibialis anterior muscles in young animals, was unable to counteract the resistance of these muscles to passive movements in old rats. The present results suggest that the muscle stiffness seen in old rats is not due to a reflex response, but depends mainly on non-reflex factors--chiefly on a large overgrowth of non-elastic connective tissue replacing degenerated active muscle fibers.

Impaired plasticity of neurons in aging. Biochemical, biophysical, and behavioral studies.

Age-related correlation of impaired plasticity of neurons (biochemical and biophysical aspects) and behavioral alterations were investigated in young (3.5 months) and extremely aged (approximately 40 months) female Wistar rats. Age-dependent significant differences in second messenger (cAMP and Ins (1,4,5)P3) concentration and signal transduction via muscarinic and dopaminergic receptors were found. The results point to the specifically impaired coupling between dopamine D1 receptor and GS protein, which underlies normal brain aging. However, cholinergic neurotransmission may be modulated at another level in extremely aged rats. Thus, it appears that the site of affection in coupling of receptor and G protein and/or G protein-dependent signal transduction in aging cannot be generalized. This indicates that alterations in the coupling of signal transduction depend on diverse neurotransmitter receptors with advanced age. The age-dependent alterations in the cAMP and PI signal pathways could be due to changes in the physical properties of the membranes. To support this hypothesis, age-dependent changes in the physical state and the biochemical composition of synaptosomal membranes from the cortex, cerebellum, and striatum were examined by measuring the steady-state fluorescence amisotropy of the membrane probes 1,6-diphenyl-1,3,5-hexatriene (DPH), trimethylammonium-DPH (TMA-DPH), and trimethylammoniumpropyl-DPH (TMAP-DPH). Significant differences in the physical properties of the synaptosomal membranes existed between young and very aged rats, expressed by a higher anisotropy in the 40-month-old rat brain tissue. The changes in the physical properties of the membranes were in line with the determined age-dependent alterations in the chemical composition, e.g., the increase in cholesterol content of the aged membranes.

The role of reflex activity in the regulation of muscle tone in rats.

The aim of the study was to assess the contribution of reflex activity to the regulation of muscle tone in rats. The experiment was carried out on young Wistar male and female rats. The hindfoot of a rat was flexed or extended at the ankle joint by 25 deg over 250 ms. The resistance of the foot to passive movements, as well as the electromyographic (EMG) activity in the gastrocnemius and the tibialis anterior muscles, were recorded simultaneously. During passive movements, reflex EMG activity developed simultaneously in both antagonistic muscles of the foot. Three components were distinguished: a short-latency EMG-A (within the first 0-20 ms of a movement), long-latency EMG-B (within 60-160 ms), and EMG-C (within 220-340 ms). When the amplitudes of EMG-B and EMG-C components of the gastrocnemius muscle reflex response were greater than 50 microV, a significant correlation was found between them and the maximum resistance of the hindfoot (MMGmax) during flexion, whereas no such correlation was observed for the tibialis anterior muscle. No correlation was found when the amplitudes of the log-latency components of the gastrocnemius muscle were less than 50 microV. Moreover, no correlation was observed between the EMG-A and the MMG(max). The above results suggest that: (1) the muscle tone of the gastrocnemius muscle in rats seems to be regulated by long-latency (supraspinal) reflexes only when the level of EMG activity exceeds a critical threshold of ca 50 microV; (2) when the level of EMG activity is lower, a major role in the resistance of hindlimb muscles is played by some non-neuronal factors; and (3) the proposed animal model emphasizes new aspects of the reflex which may be useful in a search for basic mechanisms underlying changes in the muscle tone.

The effects of prenatal exposure to hypoxia on the behavior of rats during their life span.

The aim of this study was to investigate the influence of moderate prenatal damage on adaptability during the juvenile, adult, and senile phases. Pregnant rats were exposed to a 12% normobaric hypoxia from day 1 to 17 postconception. Pregnancy was normal in both the treated animals and the controls. Erythrocytes, hemoglobin, and hematocrit did not increase in the treated pregnant animals. During the first 3 weeks, the F1 generation showed developmental deviations in physiological characteristics. Throughout subsequent ontogeny, motor performance, cognitive ability, and adaptability to physical stress were determined with a test battery of varying demands. Some of the differences (e.g., locomotor activity, learning ability) between juvenile untreated and treated rats disappeared during the adult phase. Motor and coordinative abilities, however, remained partially impaired in the old rats, especially under high demands. This study, and previous findings with alcohol (37), indicate that prenatal exposure to a noxa may result in a highly differentiated brain injury pattern. Depending on the different functions, damage may intensify age-dependent adaptive disorders or provoke impairment without influencing the course of development.

Acute and subchronic benzodiazepine-barbiturate-interactions on behaviour and physiological responses of the mouse.

Female NMRI mice were pretreated for 2 weeks with diazepam (D: 20 mg/kg/day), secobarbital (S: 23 mg/kg/day), or combination (D+S: 19 mg/kg/day, each) by means of the drinking fluid. A fourth group remained untreated. One day after this period the mice received an i.p. injection of one out of 16 drug combinations (crossover design: 0, 2, 4, 6 mg/kg D combined with 0, 6, 12, 18 mg/kg S). Open field behaviour, motor performance, and rectal body temperature were measured. In non-pretreated animals, D and S induced immobility, impairment of coordination and hypothermia in a dose-dependent manner. Excitation appeared after low doses of D (2 mg/kg) and high doses of S (12-18 mg/kg). Acute interactions between D and S were studied by means of isobolographic analysis. Dose-additivity indicating a common mechanism of action was confirmed for immobility, impairment of coordination, and hypothermia whereas excitation revealed a non-additive interaction and was reduced after combined administrations. After chronic pretreatment, the mode of acute drug interaction (dose-additive and non-additive, resp.) remained unchanged. Shifts of the isoboles indicated tolerance, cross-tolerance or sensitization. There was an asymmetry concerning the pretreatment with D and S. Chronic administration of D induced a tolerance to D in regard to all responses and a sensitization to S-effected motor incordination. Chronic administration of S sensitized the sedative and hypothermic responses to acute D. Metabolic tolerance could not account for the subchronic effects since distinct functional responses were concerned in different ways.(ABSTRACT TRUNCATED AT 250 WORDS)

The effects of prenatal alcohol exposure on the behavior of rats during their life span.

Ontogenesis is closely related to the ability to adapt to endogenous and exogenous stimuli. However, this precondition for viability does not remain stable; it changes in correlation to the phases of life. In this long-term study with rats, the extent to which moderate prenatal damage influences the adaptability in the juvenile, adult and senile phases was examined. For this purpose rat fetuses were exposed to alcohol (6 g/kg bw/day) by treating the pregnant rats via drinking water from gestation days 7 to 17. In the juvenile phase (0-3 months), no substantial differences in comparison to controls were recorded. Exceptions were a delayed development in the negative geotaxis (posture reflex) and a transiently higher exploratory activity in the open field test. In the adult phase (> 3-27 months), reduced performance was observed in basic functions such as body temperature regulation. Additionally, diminished performance was seen in response to high demands in motor-coordination (rotorod). During the senile phase (> 27 months), impairment of nearly all tested functions occurred earlier and was more pronounced.

Muscle stiffness and continuous electromyographic activity in old rats; an animal model for spasticity?

A mechanomyographic response of the hind foot to passive straightening and bending, as well as an electromyographic activity of the gastrocnemius and tibialis anterior muscles were recorded in old (35-44-month-old) and young female rats. In old rats, spontaneous, tonic electromyographic activity patterns were concurrently observed in both antagonistic muscles; they were low-amplitude, dense tonic activity and continuous, high-amplitude, sparse electromyographic activity. The tonic electromyographic activity was correlated with a decline in the strength and mass of muscles, as well as with motor disturbances, including paresis of the rigidly straightened backward hind legs, dragged behind by an animal. In muscles of old rats, morphological features of a chronic denervation atrophy were found. Baclofen (10 and 15 mg/kg, i.p.) diminished the spontaneous tonic electromyographic activity and potently decreased the whole body muscle tone, whereas Madopar (50 mg/kg of L-DOPA+12.5 mg/kg of benerazide) was ineffective. It is suggested that old rats in which the above-described pathologic alterations are observed might be a useful animal model in the search for basic etiopathological mechanisms of spasticity and similar disturbances found in humans.

Effects of age and drugs on food and fluid intake.

In young adults rats (5-month-old) d-amphetamine (2 mg/kg/d), administered on a long-term basis via drinking water, caused a moderate reduction in the intake of nutriments, which in part normalized within three weeks. Self-administration of a daily dose of 20 mg per kg diazepam over a period of 26 days led neither to hypodipsia nor to anorexia. Pentylenetetrazol (70 mg/kg/d) primarily produced a hypodipsia. The three drugs did not influence body weight. In 27-month-old rats d-amphetamine and pentylenetetrazol had the same qualitative effects. Intake of nutriments and the development of body weight were influenced more strongly than in young rats. Diazepam also had a marked effect in old animals. Nootropics (piracetam, pyrithinol, hydergin, centrophenoxin, aniracetam) had no effects on the parameters observed. When the agents were given in combination in both age groups the nootropic piracetam (230 mg/kg/d) weakened the effects induced by d-amphetamine, pentylenetetrazol or diazepam alone. The benzodiazepine, however, enhanced the loss of body weight and fluid intake in old rats caused by the stimulant or analeptic, whereas food intake remained unaffected. The results support the hypothesis that an organism's adaptivity to external and internal stimuli is reduced in later life. The behavior of young and old rats in the open field was not affected by any drug medication.

Learning abilities of rats in multiple T-mazes of two degrees of complexity under the influence of d-amphetamine.

The effect of d-amphetamine on the learning capacity of male Wistar rats was investigated in multiple T-mazes in two experiments of increasing or decreasing degree of difficulty. Running speed, distance covered and the number of errors were scored to indicate proficiency and success of learning. These parameters, as well as the distribution of errors (goal-directed orientation), correction of errors (situational orientation) and latency at the decision points (discrimination time) were considered to represent cognitive components. The results demonstrated an experiment effect in that the rats showed more difficulty in learning, as exhibited by a slower running speed and more errors, in the maze with successively increasing demands than in the one with decreasing demands. Oral self-administered d-amphetamine in a dosage of 3-4 mg/kg/day or 7-8 mg/kg/day significantly increased the running speed in a dose-dependent manner. By contrast, success of learning and goal-directed orientation decreased. Situational orientation was, however, dose-dependently improved, at least in the experiment with the increasing demand.

Adaptation capacity of biogenic amines turnover to hypoxia in different brain areas of old rats.

Catecholamines and indoleamines serve in the CNS as neurotransmitters in a great number of functional pathways. In order to contribute findings which might help to understand differences in functioning and behavioral performances with aging, the concentrations of dopa, dopamine, noradrenaline, 5-hydroxytryptophane (5-HTP), 5-hydroxytryptamine (5-HT), 5-hydroxyindoleacetic acid (5-HIAA) in four brain regions (hypothalamus, c. striatum, hippocampus, cortex) of young and old rats were determined 30 min after i.p. injection of NSD1015 (3-hydroxybenzylhydrazine hydrochloride). The influence of 12 h hypoxia of 10 and 8% O(2) as well as 36 h hypoxia of 10% was investigated. Accumulation of dopa and 5-HTP under normoxic conditions is reduced in old rats compared to young rats; the concentration of dopamine, noradrenaline and 5-HT is not significantly different between the age groups. After 12 h of 10% O(2) in young rats a drop of dopa accumulation occurs, only at 8% O(2) in both groups can a significant reduction be observed; dopamine and noradrenaline do not show a uniform tendency. Under the same conditions 5-HTP accumulation is reduced in both groups, 5-HT and 5-HIAA decrease at 10% O(2) but are in the range of controls at 8% O(2). After 36 h 10% O(2) hypoxia dopa accumulation in young rats returns to normal whereas in the striatum of old rats the decrease continues, but in the hypothalamus an increase above normal occurs. Dopamine and noradrenaline return to normal. Besides, in the hypothalamus of young rats 5-HTP accumulation is compensated. 5-HT and 5-HIAA rise even above control values.

P-hydroxy-norephedrine as a possible mediator causing the reduction of oral intake of D-amphetamine in rats.

In rats D-amphetamine is predominantly metabolized by hydroxylation to p-hydroxy-norephedrine (p-HNE); in guinea pigs, however, by deamination to benzoic acid. After 2-3 days on dosages of 1 mg/kg per day and more rats begin to reduce their oral intake of the stimulant whereas guinea pigs do not. In the present study we examined the hypothesis that the formation of p-HNE in the CNS is partially responsible for this aversion. To determine the elimination of D-amphetamine and the increase in p-HNE, groups of male Wistar rats were given various doses (0.5-5 mg/kg per day) of D-amphetamine in their drinking water intragastrically and intravenously. D-Amphetamine in the brain was determined by radioimmunoassay, p-HNE by high performance liquid chromatography followed by electrochemical detection. In contrast to the concentration of D-amphetamine, the p-HNE-content is independent of the route of administration; after oral treatment it showed a linear increase. The results reveal that p-HNE induces the aversion to the stimulant and that the ratio of D-amphetamine to its metabolite determines the onset of this aversion. No p-HNE was found in the brain of guinea pigs. Guinea pigs do not show any aversion to drinking D-amphetamine solutions, even in high dosages.

The influence of brain catecholamines on 'drug taking behaviour' relative to oral self-administration of d-amphetamine by rats.

Oral administration of 5 mg/kg of d-amphetamine to adult Wistar rats caused brain NE to decrease to approx. 80% of the control level during 4-24 h after acute treatment and slowly further to 65% after 24 days of self-administration via drinking water. The norepinephrine (NE)-reducing effect was first recognized at 1 mg/kg and appeared to peak at 5 mg/kg of d-amphetamine. Brain dopamine (DA) and 3,4-dihydroxyphenylacetic acid (DOPAC) were only shortly affected. Neither iprindole nor desipramine altered the effect of amphetamine on brain NE. DA was decreased by both inhibitors depending on the duration of pre-treatment. Iproniazid and alpha-methyl-p-tyrosine antagonized and potentiated respectively the amphetamine effect on NE- and DA-concentration after 4 days of simultaneous treatment. In the free choice experiment (water vs. 0.005% d-amphetamine solution) rats developed an aversion to amphetamine. The number of rats taking the drug and the consumption by rats still drinking it declined gradually from 100% and approx. 3 mg/kg/day to 50% and approx. 1.5 mg/kg/day, respectively, during 18 days. The time course of the developing aversive reaction to oral amphetamine ran approximately parallel to that of NE-depletion. Iprindole and desipramine intensified, iproniazid and propranolol weakened, while alpha-methyl-p-tyrosine and haloperidol hardly influenced the aversive effect of amphetamine. It is concluded that the development of aversive behaviour in response to oral d-amphetamine is mediated not only through the depleting effect of amphetamine on NE stores but also by its direct stimulation at beta-adrenergic receptors in the CNS.

Psychostimulants, analeptics, nootropics: an attempt to differentiate and assess drugs designed for the treatment of impaired brain functions.

The common characteristic properties of psychostimulants, analeptics, and nootropics are excitatory and disinhibitory effects on the central nervous system. The differences lie in the type of excitatory effect. Psychostimulants produce a general, yet nonphysiologic, activation with subsequent sedation. They generally act in a destabilising manner, disturbing the homoeostatic functions of centrally regulated reactions. Nootropics produce a physiological activation of disturbed or reduced adaptation functions. They have a stabilising effect, increasing the homoeostatic functions of the centrally regulated reactions that have become susceptible to disturbances. Analeptics differ from psychostimulants and nootropics. The effects of neuronal excitation or disinhibition are mainly restricted to the respiratory and circulatory systems. In high dosages they produce convulsions and corresponding motor reactions. No conclusive evidence for a general efficacy in the treatment of organic mental disorders has been furnished for any of the three drug classes. Yet there is sufficient proof that nootropics, unlike psychostimulants and analeptics, can produce therapeutic results in at least some patients, even if it is not yet clear under what conditions they can be meaningfully applied. There is a fundamental difference between the three groups with regard to the potential for abuse. While tolerance and extreme physiological dependence can occur rapidly under treatment with psychostimulants, such risks are not a typical feature of nootropics or analeptics.

Interaction of two barbiturates and an antihistamine on body temperature and motor performance of mice.

As an example for a drug combination used as a sedative (Vesparax) the actions of secobarbital (S), brallobarbital (B) and the antihistamine etodroxizine (E) were studied in mice after single and concomitant oral application. S and B caused a dose-dependent increase of exploratory locomotor performance enclosing a short period of locomotor depression. Such excitatory effect was suppressed by concomitant application. E produced a decrease in locomotion and--when added to S and B--enhanced their sedative effects. Explorative rearing was suppressed by low doses of S and B, by B, by E, and by all the combinations, whereas high single doses of S and B (20 mg/kg and 25 mg/kg, resp.) caused a triphasic temporal pattern. Motor coordination was investigated by means of fixed-bar and rotarod procedures. Increasing doses of S and B impaired motor performance within a small dose range. In concomitant applications rotarod performance revealed a dose-additive synergism whereas the effect was moderately superadditive in the fixed-bar test. Neither in single applications nor in combinations E showed any effects. Rectal temperature was poorly affected by the single drugs, but hypothermia was strongly enhanced by concomitant application of S and B and further potentiated by addition of E. The results suggest that responses to combinations of drugs even belonging to the same category cannot sufficiently be deduced from the single components alone.