Targeted deletion of GD3 synthase protects against MPTP-induced neurodegeneration.

Parkinson's disease is a debilitating neurodegenerative condition for which there is no cure. Converging evidence implicates gangliosides in the pathogenesis of several neurodegenerative diseases, suggesting a potential new class of therapeutic targets. We have shown that interventions that simultaneously increase the neuroprotective GM1 ganglioside and decrease the pro-apoptotic GD3 ganglioside - such as inhibition of GD3 synthase (GD3S) or administration of sialidase - are neuroprotective in vitro and in a number of preclinical models. In this study, we investigated the effects of GD3S deletion on parkinsonism induced by 1-methyl-4phenyl-1,2,3,6-tetrahydropyridine (MPTP). MPTP was administered to GD3S-/- mice or controls using a subchronic regimen consisting of three series of low-dose injections (11 mg/kg/day × 5 days each, 3 weeks apart), and motor function was assessed after each. The typical battery of tests used to assess parkinsonism failed to detect deficits in MPTP-treated mice. More sensitive measures - such as the force-plate actimeter and treadmill gait parameters - detected subtle effects of MPTP, some of which were absent in mice lacking GD3S. In wild-type mice, MPTP destroyed 53% of the tyrosine-hydroxylase (TH)-positive neurons in the substantia nigra pars compacta (SNc) and reduced striatal dopamine 60.7%. In contrast, lesion size was only 22.5% in GD3S-/- mice and striatal dopamine was reduced by 37.2%. Stereological counts of Nissl-positive SNc neurons that did not express TH suggest that neuroprotection was complete but TH expression was suppressed in some cells. These results show that inhibition of GD3S has neuroprotective properties in the MPTP model and may warrant further investigation as a therapeutic target.

Dissimilar effects of subchronic clozapine and haloperidol on operant lever pressing in C57BL/6J, BALB/cJ, and LP/J mice.

The effects of the atypical antipsychotic, clozapine, and the typical antipsychotic, haloperidol, on operant behavior have been well described in the rat; however, little such work has been done with mice. In the present study, the effects of clozapine (2.0, 4.0, and 6.0 mg/kg) and haloperidol (0.06, 0.13, and 0.25 mg/kg) were evaluated in three inbred strains of mice (C57BL/6J, BALB/cJ, and LP/J mice) across six consecutive daily sessions at each dose, in which each lever press produced access to milk. Tolerance to the rate-reducing effects of clozapine was observed in the BALB/cJ strain, but not in the C57BL/6J and LP/J strains. In contrast, sensitization was observed in the three mouse strains treated with subchronic haloperidol. These results are at odds with the operant rat literature on the effects of clozapine, but are consistent with the idea that genetic variables contribute, at least in part, to some of the differences observed in response to antipsychotics.

Motor and associative deficits in D2 dopamine receptor knockout mice.

Behavioral abnormalities produced by D2 dopamine receptor gene deletion in mice have been attributed either to resulting Parkinson-like features (i.e. response slowing and response initiation difficulties) or to behavioral deficits contributed by alleles of the originating 129Sv strain. Three strategies were used to address these conflicting hypotheses: (1) we used mice congenic at n10 backcross into the C57BL/6 line to minimize the 129Sv contribution; (2) we compared mice that were wild-type (+/+), heterozygous (+/-), or homozygous (-/-) for the D2 gene with the two most relevant inbred lines (129Sv and C57BL/6) and (3) we used both conventional and novel behavioral assessment methods. Behavioral attributes were expressed in terms of locomotor activity, wall rearing, rotarod performance, operant response acquisition, operant response performance, lick dynamics (force, rhythm), grip strength, and tremor in response to harmaline challenge. Results showed that, compared to controls, the -/- mice exhibited longer duration wall rears, retarded operant response acquisition, increased latencies to move from the operandum to the reward well, and exaggerated response to harmaline. Age was investigated as a variable (10-11 weeks versus 41-44 weeks of age) in the locomotor activity and wall rear assessments. A gene dosage effect (deficits in the +/- mice) on these two variables became apparent in the older mice. Taken together, the results showed that mice without the D2 gene exhibited Parkinson-like behavioral features that were not easily attributed to alleles contributed by the 129Sv strain, but were consistent with basal ganglia dysfunction.

Low grip strength, impaired tongue force and hyperactivity induced by overexpression of neurotrophin-3 in mouse skeletal muscle.

Transgenic mice overexpressing neurotrophin-3 (NT-3) in skeletal muscle (mlc/NT-3 mice) develop abnormal muscle spindles in skeletal muscle and display abnormal motor function in the form of gait and locomotive disturbances. The purpose of this work was to characterize the functional consequences of NT-3 overexpression in skeletal muscle with further behavioral assessments that permitted inferences about muscle weakness in the tongue or forelimbs as well as potential central nervous system (CNS) abnormalities compared to wild-type controls. Wild-type (n=12) and mlc/NT-3 (n=12) male mice were tested in five procedures (in chronological order): lick dynamics, locomotor activity, grid ataxia, go-no-go discrimination procedure, and grip strength. Relative to wild-type mice, the mlc/NT-3 mice exhibited lower tongue force, hyperactivity, slowed limb retrieval in the grid ataxia test, similar discrimination performance, and lower grip strength. Overall, the data suggest that chronically elevated levels of NT-3 in mouse skeletal muscle cause muscle weakness in the mlc/NT-3 mice. Surprisingly, mlc/NT-3 mice also exhibited significant hyperactivity, suggesting that NT-3 overexpression in the periphery may have caused abnormalities in the CNS that are related to the cortical processing of proprioceptive afferent information.

Concurrent quantification of tremor and depression of locomotor activity induced in rats by harmaline and physostigmine.

RATIONALE: Both harmaline and physostigmine are known to produce whole-body tremors in rodents, yet there is little quantitative information on the differential frequency characteristics of these tremors or on other possible motor effects of these drugs. In addition, the degree to which tolerance develops to their tremorogenic effects is uncertain. OBJECTIVES: The purpose of this work was to use a new kind of instrument (the force-plate actometer) to describe quantitatively the frequency characteristics of tremor induced by the two drugs and simultaneously to measure locomotor activity effects. Another aim was to detect any tolerance effects on the tremor or locomotor measurements. METHODS: Sprague-Dawley rats in four separate groups received harmaline (0, 4.0, 8.0, and 16.0 mg/kg) while another four groups received physostigmine (0, 0.10, 0.25, and 0.50 mg/kg). Dosing continued for 4 consecutive days. Each day, every rat's whole-body tremor and locomotor activity were recorded for 30 min. Fourier analysis of the force-plate recordings was used to quantify tremor and characterize its frequency components. RESULTS: Harmaline induced a dose-related tremor that peaked in the 10 Hz region of the spectrum, while the dose related-tremor of physostigmine was demonstrably of a broader-band frequency composition. While harmaline tremor exhibited pronounced tolerance, physostigmine tremor remained substantial across the 4 days of dosing. Both drugs suppressed locomotor activity, and there was no tolerance on this measure for either drug. CONCLUSIONS: In regard to tremor, harmaline and physostigmine induced quantitatively different kinds of tremor, and these differences probably reflect the drugs' different sites of action in the brain (harmaline: inferior olive; physostigmine: basal ganglia). For harmaline, tolerance to the tremorogenic effect was concurrent with a lack of tolerance to locomotor suppression, and this finding suggested the presence of a strong motor effect of harmaline that continued to be expressed despite the absence of tremor.

Comparison of two intracranial self-stimulation (ICSS) paradigms in C57BL/6 mice: head-dipping and place-learning.

A variety of intracranial self-stimulation (ICSS) paradigms have been utilized for investigations of reward. Among them, nose-poking and spatial-preference paradigms are known to be relatively more resistant to the effects of drug-induced motor-deficits in rat studies, although these two ICSS paradigms have not been directly compared in previous studies. In the present study, head-dipping and place-learning (forms of nose-poking and spatial-preference tasks, respectively) paradigms with lateral hypothalamus stimulation were systematically analyzed using C57BL/6 mice in the presence and absence of two motor-deficit-inducing drugs: tolperisone and harmaline. Rapid acquisition and rapid extinction patterns of ICSS responding were observed in the head-dipping and place-learning paradigms. In contrast to these pre-drug similarities in responding, dramatic differences were noted after drug administration. Tolperisone significantly reduced head-dipping but not place-learning ICSS responding. Similarly, reduction of ICSS responding after harmaline was more pronounced in the head-dipping task. Therefore, the place-learning paradigm may be superior for the assessment of reward values under motor-deficit-inducing conditions in C57BL/6 mice. The relative benefits and disadvantages of both ICSS paradigms are discussed. Combinations of complementary ICSS paradigms using mice may be useful for further investigations of the molecular bases of reward.

3-acetylpyridine reduces tongue protrusion force but does not abolish lick rhythm in the rat.

Data from other laboratories suggest that neurons in the inferior olivary nucleus (IO) may play a role in the modulation of rhythmic tongue movements in rats. Because of its known harmful effects on neurons of the IO, it was suspected that administration of the neurotoxin 3-acetylpyridine (3AP) would affect subsequent tongue dynamics during rat licking. In the present study, the task of licking water from a force-transducing disk was investigated in water-restricted rats that received systemic administration of 3AP (12.5, 25, and 50 mg/kg). After recovery from the acute toxic effects of 3AP, tongue dynamics were assessed by measuring lick force, lick rhythm, variability of timing within bursts of licking, and number of licks per 2-min session. At 50 mg/kg, 3AP resulted in: (1) reduced lick force; (2) reduced number of licks; and (3) increased variance in the timing within bursts. Lick rhythm was not significantly affected by any dose of 3AP. All 3AP treatment groups and the vehicle control group displayed slowing of lick rhythm after harmaline challenge. Compared to vehicle controls, rats receiving lower and mid-range doses of 3AP displayed indistinguishable lick behaviors, with one exception--when the lick task was made incrementally more difficult by extending the distance required to make contact with the lick-disk, rats that had received 25 mg/kg 3AP persevered at the task more than all other rats. The various changes in lick dynamics may be due to the detrimental effects of 3AP at the IO, and possibly at the hypoglossal nucleus and other sites.

Digital measurement of operant disk press force maintained in CD-1, BALB/c, and C57BL/6 mice.

Force of disk press responses by inbred (C57BL/6 and BALB/c) and outbred (CD-1) mice were measured with a PC/DOS-based system, which allowed for continuous measurement of pressing, as well as for control of reinforcer presentation on the basis of response-force dimensions. Photobeam-based measurement of mouse entry into a hopper where reinforcers were presented provided additional information about anticipatory and consumatory behavior in relation to environmental stimuli leading up to reinforcer delivery. Disk pressing was generated and measured with the use of an analog-to-digital interface with all three mouse strains. The strains differed in the physical and temporal characteristics of the disk press, as well as in the behavioral chain leading to reinforcer presentation. These measurement methods appear well suited for quantitating functional behavioral differences occasioned by genetic variations in mice.

Haloperiodol-induced microcatalepsy differs in CD-1, BALB/c, and C57BL/6 mice.

The degree of arrest of movement (microcatalepsy) induced by haloperidol at doses equipotent for operant rate suppression was measured with computerized instrumentation. The inbred C57BL/6 mouse strain displayed more susceptibility to microcatalepsy than the CD-1 and BALB/c strains. In addition, the C57BL/6 strain exhibited a greater degree of sensitization to repeated dosing than did the other 2 strains. The results were consistent with the C57BL/6 mouse's hypodopaminergic profile reported in the literature but were at odds with results reported for conventional catalepsy testing. The C57BL/6 mouse may serve as a model for genetic vulnerability to extrapyramidal motor side effects and may be useful in quantifying the mild extrapyramidal motor side effects of atypical antipsychotic drugs.

The inebri-actometer: a device for measuring the locomotor activity of Drosophila exposed to ethanol vapor.

Drosophila melanogaster can be used as a model organism for probing the genetic basis for alcohol sensitivity. In this paper, we describe a new device, the inebri-actometer, which measures the locomotor activity of up to 128 individual flies simultaneously. The device consists of 128 pairs of emitter/detector photodiodes connected in series through a computer interlink. A single fly is placed in each of the 128 chambers and humidified air or air containing variable amounts of ethanol vapor is pumped through the chambers. When a fly blocks the infrared signal transmitted by an emitter photodiode, the computer records one movement for that fly. We present preliminary results showing the effect of ethanol on the activity of wild-type Oregon R Drosophila. Five preliminary runs with 95% ethanol vapor revealed that this concentration induces an approximately 3- to 4-fold increase in locomotor activity which peaks at about 5 min after the addition of ethanol vapor. This is followed by a gradual decrease in activity leading to a nearly total cessation of movement after 30 min. Statistically significant dose-related activity increases were obtained for ethanol concentrations of 8, 19, 50, and 100% of maximum, assessed in two replications at each dose. Unlike the complete suppression of locomotion seen in the last 10 min of the session at maximum ethanol exposure, the initial stimulation effect at the 19% concentration was maintained across the 30-min session.

A force-plate actometer for quantitating rodent behaviors: illustrative data on locomotion, rotation, spatial patterning, stereotypies, and tremor.

This report describes a new kind of actometer for recording the behavior of rodents or other small animals. The instrument, a force-plate actometer, uses a stiff, low-mass horizontal plate coupled to four supporting force transducers positioned at the corners of the plate. When an animal moves on the plate, its movements are sensed by the transducers whose signals are processed by computer to yield measurements of a wide range of behaviors or behavioral attributes, such as locomotor activity, rotation around the center, whole-body tremor, and amphetamine-induced stereotypies. Spatial resolution is less than 1 mm, and temporal resolution is 0.02 s. Sample data were presented comparing the locomotor activity of CD-1, BALB/c, and C57BL/6 mice before and after treatment with D-amphetamine sulfate. Rotational behavior was recorded in an amphetamine-treated rat that had sustained a unilateral 6-hydroxydopamine-induced lesion of the nigrostriatal system. In the C57BL/6 mouse, harmaline-induced tremor was quantified. With rats as subjects, the force-plate actometer was used to quantify amphetamine-induced stereotypies, to demonstrate the development of sensitization to amphetamine's effects, and to quantitate the consistent 11-12 Hz rhythmicities that underlie the sterotypies. The performance of the force-plate actometer was compared with that of a variety of instruments reported in the literature on behavioral instrumentation. Finally, potential applications in neuroscience research other than those illustrated in this report were discussed.

Behavioral tolerance to the force differentiation effects of diazepam and midazolam in rats.

RATIONALE: Several benzodiazepines (BZs) have been shown to increase the peak force of operant responses at doses that increased, decreased, or had no effect on response rate, suggesting that operant response force may be a sensitive index of BZs' effects rather than solely a correlate of rate-dependent effects. In addition, contingent tolerance to the rate-dependent effects of BZs has been reported, but the degree of contingent tolerance that develops when the critical variable of the task is force of the response has not been explored. OBJECTIVES: These experiments examined the effects of acute and repeated oral administration of diazepam (DZ) and midazolam (MZ) on a force-differentiation task to explore the importance of task requirements on the development of contingent tolerance. METHODS: Two groups of rats were trained to press a force-sensing operandum, and responses having peak forces falling within fixed lower and upper limits [low force (8-10 g) or high force (40-50 g)] were reinforced with water. Acute effects of the oral administration of DZ (0.3, 1.0, 3.0, 10.0, 30.0 mg/kg) and MZ (same doses) were determined for the discriminated-force task before and after a repeated-administration procedure. RESULTS: When administered acutely, both drugs increased the peak force of responses in a dose-related manner and concomitantly reduced the proportion of reinforced responses, with MZ exhibiting greater potency. For the next 36 days, one group received drug before experimental sessions and the other group received drug after the experimental session. A second dose-effect determination demonstrated that rats chronically dosed with DZ or MZ pre-session displayed more contingent tolerance to alterations in peak force than rats that had received 36 drug injections postsession, where there was no opportunity to practice the force-discrimination response while under the drug state. CONCLUSIONS: These results suggest that perceptual motor difficulty of the task rather than effort may be an important variable in predicting the degree of contingent tolerance that develops. Additionally, these results suggest that both behavioral and pharmacological mechanisms are involved in the development of drug tolerance to the BZs.

The relationship between isometric force requirement and forelimb tremor in the rat.

To explore the effects of isometric force of rodent forelimb contraction on forelimb tremor, rats were trained to press downward on an isometric force transducer to raise a water-filled dipper cup and maintain force to keep the dipper in the raised position while licking. Force requirements were then manipulated parametrically to measure the effects of escalating force output on forelimb tremor and other variables. In the Peak-Force greater than Hold-Force (PF > HF) manipulation, the forces required to raise the dipper were 20, 40, and 60 g (each condition for about 2 weeks), while the force required to maintain the dipper in the raised position remained 6.7 g for all three conditions. In the Peak-Force equal to the Hold-Force (PF = HF) manipulation, rats were required to maintain the "dipper-raising" force throughout the response. The forces required were 20 g, 40 g, and 60 g (each for 2 weeks). For all force requirement manipulations, data were analyzed within and across conditions. As expected, force output increased with increased force requirements. Spectral analysis of force-time records revealed that during all manipulations, high-frequency (>10 Hz) forelimb tremor increased with increased force output, an effect that is consistent with human studies, and that may reflect increases in the number of motor units firing at higher rates. Additionally, with the exception of the 60-g PF = HF condition, there were within-condition decreases in tremor and increases in task engagement, evidence suggesting increased muscle strength as a function of experience (i.e., "physical training"). Taken together, the results suggest that the rodent-based method may provide a valuable, noninvasive functional assay for animal models of disorders that affect skeletal muscle control in humans.

Clozapine-like motor effects of the atypical antipsychotic risperidone in rats.

A behavioral preparation that affords concurrent measurement of forelimb force, lick rhythm, and forelimb tremor in rats was used to assess the effects of the atypical antipsychotic risperidone. Rats that were trained to press downward on an isometric force transducer while simultaneously licking water reinforcement were administered risperidone (0.08, 0.12, and 0.16 mg/kg). Risperidone suppressed task engagement and increased the duration of individual press-hold-release bouts, effects shared with both typical and atypical antipsychotic drugs in this task. Although risperidone did not significantly affect forelimb force output as clozapine does, it did significantly decrease tremor power in the high-frequency (10-25 Hz) band of the power spectrum. Risperidone dose-dependently decreased the dominant 6 Hz frequency of the power spectrum, a reflection of slowed lick rhythm which is an effect that is shared by other atypical antipsychotic drugs in this task. The results reported in the present study suggest that, although risperidone may not possess the exceptionally low extrapyramidal side-effect profile that clozapine does, its effects are more similar to clozapine than to the extrapyramidal side-effect-producing haloperidol in this task.

Effects of haloperidol and clozapine on tongue dynamics during licking in CD-1, BALB/c and C57BL/6 mice.

RATIONALE: In an initial effort to describe how genetic background influences the differential motor effects of haloperidol, a drug with high extrapyramidal side effect (EPS) liability, and clozapine, an antipsychotic low in EPS, both drugs were studied in inbred strains of mice (BALB/c and C57BL/6) previously shown to have differential sensitivities to haloperidol. OBJECTIVES: Behavioral differences in lick dynamics for male BALB/c, C57BL/6 and CD-1 (an outbred strain) were characterized. Effects of dose ranges of haloperidol and clozapine were then evaluated in the three strains. METHODS: The mice learned to lick milk from a force-sensing disk during daily 2-min sessions, while a computer counted the number of licks and measured lick peak force and lick rhythm. After training, acute doses of haloperidol (0.08-2.0 mg/kg) or clozapine (0.5-8.0 mg/kg) were administered i.p. 45 min before sessions. RESULTS: Prior to drug treatment, substantial quantitative strain differences in licking behavior were observed: C57BL/6 mice made fewer licks, licked with lower peak force per lick, and had a slower lick rhythm than the BALB/c and CD-1 mice. As in rats, clozapine slowed the lick rhythm in all three mouse strains much more than haloperidol did. Haloperidol produced a 50% greater suppression of number of licks in BALB/c than in C57BL/6 mice (ED50 values were 0.82 mg/kg and 1.22 mg/kg, respectively). For clozapine, lick suppression was greater in the C57BL/6 than in the BALB/c strain (ED50 values were 1.88 mg/kg and 2.65 mg/kg, respectively). Among the three strains examined, CD-1 was the most sensitive to haloperidol's suppression of licking, while its sensitivity to clozapine's lick-suppressing effect was similar to C57BL/6 mice. Clozapine lowered the lick peak force in the CD-1 and BALB/c strains more than in the C57BL/6 strain. CONCLUSIONS: Overall, the results suggest that genetic variables may influence both mice's tongue dynamics and their alteration by both typical and atypical antipsychotic drugs. In addition, while the BALB/c strain was more sensitive to haloperidol's lick-disruptive effects than the C57BL/6 strain, the size of the difference between strains was much smaller than the reported difference between the strains in the catalepsy test.

Effects of unilateral striatal dopamine depletion on tongue force and rhythm during licking in rats.

To quantitatively assess the orolingual dysfunctions produced by unilateral striatal dopamine depletions, rats first received 6-hydroxydopamine injections into the nigrostriatal bundle and were then trained to lap water from a force-sensing disk in 2-min sessions. Compared with controls and rats with moderate (<75%) dopamine depletions, subjects with substantial (>75%) dopamine depletions showed decreases in number of licks, lick rhythm, and lick peak force. Rats with substantial lesions were also impaired in making initial, within-session adjustments in lick peak force but not in lick rhythm. The results confirm the presence of Parkinson-like deficits in tongue dynamics during consummatory licking behavior in rats. The methods used here should prove useful in providing quantitative measures of the efficacy of experimental therapies in this rodent model of Parkinson's disease.

Haloperidol, raclopride, and eticlopride induce microcatalepsy during operant performance in rats, but clozapine and SCH 23390 do not.

The purpose of this work was (1) to assess the ability of selected antipsychotic and comparison drugs to induce arrest of movement phenomena during operant responding and (2) to evaluate the capacity of muscarinic anitcholinergics to block such effects. The effects of haloperidol (0.02-0.12 mg/kg, i.p., 45 min), raclopride (0.05-0.80 mg/kg, i.p., 30 min) eticlopride (0.02-0.16 mg/kg, i.p., 45 min), clozapine (1.0-8.0 mg/kg, i.p., 60 min) and SCH 23390 (0.01-0.16 mg/kg, i.p., 30 min) were administered to rats for 4 weeks in a between-groups dosing design. Operant responses in 15 min and the maximum duration of the rat's muzzle entry into the reinforcement dipper well (the measure of arrest of movement that reflected microcatalepsy) were the quantitative measures of behavior. The D2 antagonists dose-relatedly decreased operant responding and increased maximum muzzle duration, effects that were significantly reversed by the anticholinergic scopolamine (0.1 mg/kg) or atropine (6.0 mg/kg). Although the atypical antipsychotic drug clozapine and the selective D1 antagonist SCH 23390 both significantly reduced operant responding, these drugs did not produce microcatalepsy. The results suggested that microcatalepsy expressed in the context of ongoing operant behavior may model low-dose extrapyramidal side effects.

Chronic haloperidol produces a time- and dose-related slowing of lick rhythm in rats: implications for rodent models of tardive dyskinesia and neuroleptic-induced parkinsonism.

In order to characterize the development of orolingual motor effects of chronic haloperidol treatment in rats, this typical neuroleptic was administered for 102 days while daily measurements of tongue movement dynamics (peak force, lick rhythm, number of licks) during water licking were recorded. After chronic haloperidol dosing (vehicle, 0.06. 0.12, 0.24 mg/kg for four separate groups) for 32 days and continuing every second or third day of the chronic dosing period, the effects of cholinergic (scopolamine: 0.05-0.20 mg/kg; trihexyphenidyl: 0.15-1.0 mg/kg) or serotonergic (ritanserin: 0.5-4.0 mg/kg; quipazine: 0.5-4.0 mg/kg) probe drugs were examined for their capacity to antagonize the alterations in licking behavior induced by haloperidol. Haloperidol dose-dependently reduced peak force and number of licks, effects which were apparent within 2 or 3 days of the start of treatment. Significant effects of haloperidol on lick rhythm first emerged on day 13 and gradually increased in magnitude through the remaining treatment period. Scopolamine, trihexyphenidyl, and quipazine reduced haloperidol's effects on at least one measure of licking behavior. During a 7-day haloperidol withdrawal period, the four dosage groups were similar on all measures of tongue dynamics. Overall, the results exhibited features suggesting the co-occurrence of Parkinson-like and tardive dyskinesia-like effects.

Caffeine, but not time of day, increases whole-arm physiological tremor in non-smoking moderate users.

1. Consumptive levels of caffeine significantly increased whole-arm physiological tremor in young adult males at 3 but not 1 mg caffeine/kg bodyweight. 2. Physiological tremor was not affected by time of day (TOD) or the interaction of caffeine and TOD. 3. Findings suggest that ingestion of a single oral dose of caffeine at typical consumptive levels results in measurable changes in physiological functioning and that whole-arm physiological tremor is a sensitive measure of physiological change due to drug effects. 4. However, the results suggest that whole-arm physiological tremor may not detect reliably TOD changes in functioning.