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A new direct injection/on-line guard cartridge extraction-tandem mass spectrometry (DI/GCE-MS-MS) method has been validated for high-throughput evaluation of cytochrome P450 (CYP) 2C19 inhibition potential using human hepatic microsomes and 96-well microtiter plates. Microsomal incubations were quenched with formic acid, centrifuged, and the resulting supernatants were injected for DI/GCE-MS-MS analysis. Due to the use of a C(18) guard cartridge (4x2.0 mm I.D.), this method exhibits several advantages such as little sample preparation, rapid on-line extraction, short analysis time (2.5 min), and minimal source contamination and performance deterioration. The DI/GCE-MS-MS method demonstrates an inter-day accuracy ranged from 0.3 to 2.4% with precision ranging from 2.0 to 3.0% for the quantification of 4-hydroxymephenytoin, a marker metabolite of S-mephenytoin mediated by CYP2C19, in microsomal incubations. The CYP2C19 inhibition assay has been validated using tranylcypromine as a known inhibitor of the isoform. The IC(50) value (43.5 microM) measured by the new method is in agreement with a reported literature value (approximately 30 microM).

A highly efficient direct injection/on-line guard cartridge extraction–tandem mass spectrometry (DI/GCE–MS–MS) method has been validated for high-throughput evaluation of cytochrome P450 (CYP) 2D6 inhibition potential using human hepatic microsomes and 96-well microtiter plates. Microsomal incubations were terminated with formic acid, centrifuged, and the resulting supernatants were injected for DI/GCE–MS–MS analysis. Due to the novel use of an extremely short C 18 guard cartridge, this method exhibits several advantages, such as no sample preparation, excellent on-line extraction, short run time (2.5 min), and minimized source contamination and performance deterioration. The DI/GCE–MS–MS method demonstrates acceptable accuracy and precision for the quantification of dextrorphan, a marker metabolite of dextromethorphan mediated by CYP2D6, in microsomal incubations. The CYP2D6 inhibition assay has been validated using quinidine as a known selective inhibitor of the isoform. The IC 50 value (0.20 μ M ) measured by the new method is in good agreement with the literature value (0.22 μ M ).

An efficient direct injection/on-line guard cartridge extraction-tandem mass spectrometry (DI/GCE--MS--MS) method has been validated for high-throughput evaluation of cytochrome P450 (CYP) 1A2 inhibition potential using human hepatic microsomes and 96-well microtiter plates. Microsomal incubations were terminated with formic acid, centrifuged, and the resulting supernatants were injected for DI/GCE--MS--MS analysis. Due to the use of an extremely short C(18) guard cartridge, this method offers several advantages such as no sample preparation, excellent on-line extraction, short run time and minimal source contamination and performance deterioration. The DI/GCE--MS--MS method demonstrates acceptable accuracy and precision for the quantification of resorufin, a marker metabolite of ethoxyresorufin mediated by CYP1A2, in microsomal incubations. The inhibition potential of CYP1A2 has been evaluated using its selective inhibitors, alpha-naphthoflavone and furafylline. The IC(50) values (120 nM for alpha-naphthoflavone and 5.1 microM for furafylline) measured by the new method are in agreement with the literature values.

The inhibition potential of drugs towards five major human hepatic cytochrome P450 (CYP) isozymes (CYP2A6, 3A4, 2C9, 2D6, and 2E1) was investigated via cassette dosing of the five probe substrates (coumarin, midazolam, tolbutamide, dextromethorphan, and chlorzoxazone) in human liver microsomes using a 96-well plate format. After microsomal incubations had been terminated with formic acid, the five marker metabolites (7-hydroxycoumarin, 1'-hydroxymidazolam, 4-hydroxytolbutamide, dextrorphan, and 6-hydroxychlorzoxazone) were simultaneously quantified using direct injection/online guard cartridge extraction/tandem mass spectrometry (DI-GCE/MS/MS). Several advantages resulted from the use of a short C(18) guard cartridge (4 mm in length) for DI-GCE/MS/MS, including minimal sample preparation, fast online extraction, short analysis time (2.5 min), and minimal source contamination. In addition, this method demonstrated an inter-day accuracy range from -8.7 - 7.4% with a precision less than 8.3% for the quantification of all the marker metabolites. The inhibition assay for the five CYP isozymes was evaluated using their known selective inhibitors via individual and cassette dosing of the probe substrates. The IC(50) values measured via cassette dosing were consistent with those observed via individual dosing, which were all in agreement with the reported values. In addition, the validated assay was used to evaluate the inhibitory potential of 23 generic drugs (randomly selected) towards the five CYP isozymes. The results suggest the integration of the cassette dosing strategy and the DI-GCE/MS/MS method can provide a reliable in vitro approach to screening the inhibitory potential of new chemical entities, with maximal throughput and cost-effectiveness, in support of drug discovery and development.

Aims We investigated the pharmacokinetics and safety profile of oral ganciclovir coadministered with trimethoprim in HIV-and CMV-seropositive patients. Methods In an open-label, randomized, 3-way crossover study, 12 adult males received oral ganciclovir 1000 mg every 8h, oral trimethoprim 200 mg once daily, or both drugs concomitantly in a sequence of three 7-day treatment periods. Pharmacokinetic parameters were determined and adverse events recorded for each treatment. Results The presence of trimethoprim significantly decreased CLr (12.9%, P=0.0068) and increased t1/2 (18.1%, P=0.0378) of ganciclovir. However, these changes are unlikely to be clinically meaningful. There were no statistically significant changes in trimethoprim pharmacokinetic parameters in the presence of ganciclovir, with the exception of a 12.7% increase in Cmin. Ganciclovir was well tolerated when administered alone or in combination with trimethoprim. Conclusions There was no clinically significant pharmacokinetic interaction between oral ganciclovir and trimethoprim when coadministered.

Heteroatom substitution within the zileuton nucleus led to a congener series which was evaluated for potency and rate of glucuronidation, a major route for elimination of zileuton. Subtle changes in structure were found to significantly effect glucuronidation rate.

The single dose pharmacokinetic characteristics of four aspirin formulations in humans were compared with their in vitro dissolution properties. The pharmacokinetic parameters were determined by measuring salicylate concentrations in the plasma. Dissolution was measured by using the USP XX single time point dissolution test. The four aspirin products were a commercial tablet, a commercial capsule, a capsule filled with a commercial granulation, and a slow dissolving capsule formulation that failed the USP dissolution specification. It was found that there was a poor correlation between the in vitro dissolution results and the in vivo computed pharmacokinetic parameter statistics. In vivo testing in humans showed that all of the formulations were bioequivalent in terms of half-life and AUC, and the capsule that failed to pass the dissolution specification was bioequivalent to two of the three products that did pass the specification with respect to the maximum plasma concentration. None of the prod...

This paper is based on my experiences in 40 years of research in behavioral neuroscience. It is aimed at giving help to beginning graduate students with advice for how to do their research.

The word “synthetic” in the title of this paper has three meanings, each of which highlights a fundamental problem with physiological psychology, and. at the same time, offers a solution. (1) Synthetic means artificial—made by man, not by nature. To separate physiological psychology from medicine, and to build a science of pure function in humans (which is what psychology is), apart from a science of the mechanics of living human tissue (which is what medicine is), synthetic physiological psychology adopts a physically achieved analytic approach to the design, not of people, but of robots that behave like people. This avoids the misuse of reductionism. (2) Analysis by itself is not adequate; every analysis must be validated immediately by synthesis. Some forms of physically achieved analysis and resynthesis, particularly appropriate for synthetic physiological psychology, are contrasted with the hypothetico-deductive method, as a complementary scientific method of arriving at simple facts about complex living systems. (3) Synthetic also means integrative—at present, the various subfields of psychology are not united by principles based on the action of common units of function, derived by a physically achieved analysis. A simple procedure is described that can provide them.

The current explanation of air righting in animals is that when falling supine in the air, labyrinthine stimulation triggers head rotation. The head rotation involves neck rotation which, via the cervical righting reflex, triggers rotation of the body. (In cats and monkeys, when the labyrinths are absent, visual stimulation when falling supine can also trigger this righting sequence.) In the present paper, a descriptive analysis of air righting in the rat shows that the shoulders rotate, carrying the unmoving head and neck passively along. Thus, for this species, labyrinthine input appears to trigger shoulder rotation directly, independently of the cervical righting reflex. This suggests that at least two physiological mechanisms exist for labyrinthine control of head rotation during air righting, one via the neck and the other via the shoulder girdle.

The development of contact-righting by rats was studied from birth to weaning. Such righting involved both vestibular and tactile forms, which matured at different rates. At birth, only righting triggered by snout contact with the ground (i.e., trigeminal righting) had the adult form of cephalocaudal axial rotation in which the limbs flex to accommodate placement on the ground while the body is rotated to prone. Vestibularly triggerd righting from the supine position initially only activated head and neck rotation, and failed to recruit the shoulders and pelvis to rotate to prone. In the fully adult form, achieved by about 12 days, vestibularly triggerd righting activated shoulder-led roration which passively carried the head and neck to prone, and then recruited the pelvis to rotate to prone. Asymmetrical contact of the body surface with the ground triggered both body-on-head (forequarter-led) and body-on-body (hindquarter-led) forms of righting. At birth, both involved limb righting movements (foreleg and hindleg, respectively), without rotation of the axial musculature: For both, the fully mature forms involved axial rotation by the shoulders and pelvis, respectively. In the adult, the body-on-body form of righting only occurs when the head is restrained by the experimenter; whereas in the infant it also occurs when the head is unrestrained, and thus simultaneously with those forms of righting which involve the forequarters. The simultaneous occurrence and differential rate of maturation of these forms of righting produce complex, seemingly bizarre, patterns of righting during development, which are unlike those present in adults. This study clearly shows that the postnatal development of contactrighting involves a complex interaction of righting subsystems.

Based on the fact that thalidomide, at a certain point in human pregnancy, produces autism, we propose administering thalidomide to pregnant monkeys at an appropriate point after conception. The infant monkeys born after thalidomide treatment of the pregnant mothers should manifest aberrations in social vocalization and in socialization behavior. Histological analysis of their brains should reveal areas whose damage will lead to autism. This can then be produced stereotaxically in infant monkeys to allow the better determination of the relation of degree of damage in these areas to the severity of autism.

A highly efficient direct injection on-line guard cartridge extraction/tandem mass spectrometry (DI-GCE/MS/MS) method has been validated for high-throughput evaluation of cytochrome P450 (CYP) 3A4, 2D6 and 2E1 inhibition potential via cassette dosing of midazolam, dextromethorphan and chlorzoxazone using human hepatic microsomes and 96-well microtiter plates. Microsomal incubations were terminated with formic acid, centrifuged, and the resulting supernatants were injected for analysis by DI-GCE/MS/MS. Due to the novel use of an extremely short C18 guard cartridge (4 mm in length), this method exhibits several advantages such as no sample preparation, excellent on-line extraction, short run time (2.5 min), and minimized source contamination and performance deterioration. The DI-GCE/MS/MS method demonstrates acceptable accuracy and precision for the simultaneous quantification of 1′-hydroxymidazolam, dextrorphan and 6-hydroxychlorzoxazone in microsomal incubations. The inhibition potential of CYP3A4, 2D6 and 2E1 has been evaluated using their known selective inhibitors. The IC50 values measured by the cassette dosing approach (high-throughput) are consistent with those observed by an individual dosing regimen (conventional) and are all in good agreement with the literature values. The results suggest that the cassette probe-dosing strategy may provide an in vitro approach to minimize cost while maximizing throughput of CYP inhibition evaluation of new chemical entities in support of drug discovery and development. Copyright © 2000 John Wiley & Sons, Ltd.

A highly efficient direct injection/on-line guard cartridge extraction/tandem mass spectrometry (DI-GCE/MS/MS) method utilizing electrospray polarity switching was developed for the simultaneous detection of probe substrates and marker metabolites of seven human hepatic cytochrome P450 (CYP) isozymes: CYP1A2, 2A6, 3A4, 2C9, 2C19, 2D6 and 2E1. Microsomal incubations were terminated with formic acid, centrifuged, and the resulting supernatants were injected for analysis by DI-GCE/MS/MS. This method employed an extremely short C18 cartridge (4 mm in length) which allowed rapid cleanup of sample matrices while retaining the analytes an appropriate time (2.0–2.2 min). From 1.5 to 2.7 min the effluent was directed to the mass spectrometer for detection otherwise diverted to waste. As a result of the efficient on-line extraction, matrix (e.g., salts and proteins) suppression was minimized. In addition, no visible source contamination was observed and system performance (chromatographic and mass spectrometric) did not significantly deteriorate after 500 consecutive injections. Electrospray polarity switching was strategically executed on a Micromass Quattro II mass spectrometer by establishing dummy ion transitions to protect the analytes from the interference of the overwhelming noise which was unavoidable for the first transition scanned following each polarity switch. This unique strategy led to the simultaneous detection of seven CYP probe substrates and seven corresponding marker metabolites (12 by positive mode and 2 by negative mode). Copyright © 2000 John Wiley & Sons, Ltd.

Two open-label, randomized, multiple-dose, three-way crossover studies were performed to assess the pharmacokinetics and safety of oral ganciclovir 1000 mg q8h in asymptomatic patients seropositive for human immunodeficiency virus and cytomegalovirus. Ganciclovir was administered alone and in combination with zalcitabine 0.75 mg q8h (study 1) or stavudine 40 mg q12h (study 2). In the presence of zalcitabine, the only statistically significant change in the pharmacokinetic parameters of ganciclovir was a 22.2% mean increase in AUC0-8. However, there was no significant change in the renal clearance of ganciclovir when coadministered with zalcitabine, suggesting that the increase in serum ganciclovir concentration cannot be attributed to competition for active renal tubular secretion. No change in zalcitabine pharmacokinetics was observed in combination with ganciclovir. There were no significant changes in the pharmacokinetics of ganciclovir or stavudine when coadministered. Ganciclovir was well tolerated when given alone and in combination with either zalcitabine or stavudine.

Gait in Parkinson's disease is characterised by slowed velocity; shuffling, small steps; and absent arm swing. Drug therapy intervention is beneficial in improving mobility, though with prolonged use its effects may diminish. The purpose of this study was to examine whether Parkinsonian patients could improve their gait patterns in response to five instructional sets: natural walking; walking while deliberately swinging the arms; walking with large steps; fast walking; and walking while counting aloud. Eight subjects with idiopathic Parkinson's disease and eight age matched control subjects were tested using motion analysis. The findings indicated that parkinsonian patients followed the instructions which immediately altered a series of single walking variables. Simultaneously, automatically activated changes occurred in other gait variables producing more normal gait. The instructional set is a strategy which can aid normalisation of Parkinsonian gait although its benefits may depend on the stage of disease progression and the degree of attention to the instructions.

During unconstrained locomotor behavior, rats move in and out of a straight posture of the body (including the head). In the present study, the stability of maintaining a straight body was examined in untreated rats and in rats treated with saline (SAL) or with 1 of 3 dopamine stimulants (n = 4 rats per group). The stability of maintaining a straight body can range from very high (with 0.5 mg/kg quinpirole [QUIN]), to high (first half-session with 5 mg/kg (+)-amphetamine [AMPH]), to very low (second half-session with 5 mg/kg AMPH), or can be maintained at a level similar to that observed in untreated rats (with 1.25mg/kg apomorphine [APO]). Stability was assessed by videotaping the rats and, then, by using frame-by-frame analysis, scoring the cumulative proportion of time spent in a straight posture, the frequency of transitions from one hemisphere to the other without being trapped in the midline plane, and the degree of lateral bending during turning and during walking on a curved path. The present study is one in a series identifying key variables that constrain as many degrees of freedom as possible in rat locomotor behavior. The uncovering of such variables is an indispensible step that precedes dynamic systems stability analysis and provides candidates for key variables for the modeling of motor coordination.

In his classic article, Stellar (1954) proposed that diverse motivated behaviors reflected the activity of excitatory and inhibitory centers in the hypothalamus. His specific and testable ideas provided the theoretical focus for a great deal of fruitful research on the biological bases of behavior for 2 decades. Subsequently, new findings and technical developments again changed the perspective and experimental approaches in behavioral neuroscience. The authors suggest that the modern emphasis on the anatomy and chemical function of neuronal systems has come at the expense of understanding the subcomponents of behavior and the hierarchical levels of integration involved in transforming reflexes into operant acts. Increased attention in the future to the infrastructure of the behaviors being elucidated, when combined with reductionistic studies of neurons, will fulfill the potential contribution to behavioral neuroscience that is implicit in Stellar's article.

Rats made immobile and cataleptic by haloperidol, a dopamine receptor blocker, maintain their static stable equilibrium by employing a variety of allied postural support reflexes. Under some test conditions, competition between such reflexes occurs, and in haloperidol-treated rats, unlike undrugged controls, proprioceptive-tactile stimuli appear to be dominant over vestibular stimuli. We investigated this relationship in rats by testing their air-righting with and without simultaneous contact of the tail on a wooden platform. The rats were lightly held in a supine position by the shoulders and pelvis, with or without tail contact on a small wooden platform 47 cm above the ground. Undrugged rats showed the normal pattern of righting which involves axial rotation with cephalocaudal recruitment whether the tail is contacting the platform or not. Upon release, the haloperidol-treated rats (2.5 mg/kg) gripped the platform with their tail, which interfered with the air-righting reflex. This demonstrates that in haloperidol-treated rats, the dominance of tactile-proprioceptive postural support reflexes over those triggered vestibularly.

The vestibular head righting reflex can be demonstrated by holding an adult rat vertically downward, so that the snout points downward. In this situation, the animal dorsiflexes its head and neck, bringing the head towards its normal orientation in space. Bilateral labyrinthectomy not only blocks this response, but releases an actively maintained ventroflexion of the head and neck. Bilateral electrolytic lesions of the lateral hypothalamus (LH) exaggerate such ventroflexion in labyrinthectomized rats. By themselves, LH lesions had no such effect. Therefore, it is argued that there are vestibular and supraspinal inhibitory mechanisms which, in the intact adult animal, keep this ventroflexion response in check. In addition, when the rats were held with their heads down, and with gentle paw contact with the ground, they did not ventroflex. However, they ventroflexed immediately upon releasing this paw contact. These observations suggest that there are tactile mechanisms which can also inhibit this exaggerated ventroflexion released by labyrinthectomy.

The dorsal immobility response (DIR) is typically seen in the infants of many altricial mammalian species. Lifting the animal into the air by the nape of the neck is the primary releasing stimulus. Functionally, this response appears to facilitate carrying of the infants by the adults. When grasped by the nape and lifted into the air, adult rats will also exhibit the DIR. In this paper, the role of the labyrinths in the DIR of adult male rats was examined. Vestibular stimulation produced by vertical circular acceleration increased the duration of the DIR, while labyrinthectomy greatly diminished the DIR. In rats with intact labyrinths, visual occlusion greatly potentiated the DIR, whereas, in labyrinthectomized rats, visual occlusion had little effect. These data indicate that the vestibular system plays a major role in mediating the DIR of adult rats. The retention of the DIR into adulthood and the possible increased role of the labyrinths in the control of the adult DIR, are discussed with respect to the possible role of the DIR as an anti-predator mechanism.

Compensatory head movements as responses to passive angular acceleration are obvious in lower vertebrates, but they are usually small and difficult to observe in mammals. The may simply be cut short very early by an inhibitory mechanism linked to the initiation of anticompensatory head movements (quick phases of head nystagmus). After electrolytic lesions of the pontine reticular formation (PRF), rats made no spontaneous lateral head movements, but exhibited large compensatory lateral head movements and abnormal postrotatory circus movements, both of which can be regarded as consequences of a loss of the quick phase of head nystagmus. The quick phase of ocular nystagmus was also lost, corroborating earlier findings of others in primates, cats, and rabbits, and supporting the view that saccadic movements of the head and eyes are generated by closely related neural substrates. Rats with bilateral PRF lesions were unable to right themselves in the air. Together with additional observations, this indicates involvement of the PRF in some functions of the otolith organ.

Localized lesions or local applications of gamma-aminobutyric acid (GABA) in the nucleus reticularis tegmenti pontis (NRTP) of rats cause rapidly accelerating forward locomotion. Such "festination" can coexist with blockade of the dopamine system. We suggest that (i) the akinesia produced by dopamine deficiency results at least in part from release of excessive inhibition of locomotion by a neural system whose final common inhibitory path includes the region of the NRTP and (ii) when it occurs in addition to nigrostriatal damage, destruction in the region of the NRTP might be the cause of a form of festination seen in some patients suffering from Parkinsonism.

The Eshkol-Wachmann movement notation is used to analyze and describe neurological recovery from the akinesia caused by severe bilateral lateral hypothalamic (LH) damage in rats. Exploratory movement recovers along several relatively independent dimensions which appear successively. First, lateral head scanning movements recover. At about the same time or later, longitudinal (backward-forward) head scans appear. After movements along these two dimensions increase in amplitude and involve the whole body, vertical (dorsal-ventral) head scans with snout contact (along vertical surfaces) typically appear, and increase gradually in amplitude. Later, vertical rearing without snout contact emerges. Recovery proceeds cephalocaudally, as more caudal limb and body segments are recruited along each of the above dimensions separately. LH rats show delayed recruitment of caudal limb and body segments (‘strait-jacket ohenomenon’). Support of the body and management of limb and body segments' contact with the ground also recover relatively independently, in a proximodistal fashion. In recovery, arrests between bouts of activity become shorter. Movement first becomes organized in relation to the animals' own body, and only much later, in relation to the environment. In each sequence of movement after pronounced immobility, the rat recapitulates the process of recovery; and, any time it starts to move, it repeats the movements at a particular amplitude several times until there is an increase to the next larger size movement (‘warm-up’ phenomenon). These regularities explain the apparently bizarre stereotypes behavior in partial enclosures (behavioral traps) seen in LH rats recovering from akinesia. They also explain some aspects of exploration in rats and normal social behavior of wild animals, particularly in situations involving fear and conflict.

Apomorphine, at doses greater than or equal to 10 mg/kg (intraperitoneally), produced two patterns of stereotypy. In rats from one supplier it induced predominantly gnawing while in those from another predominantly climbing, suggesting that the response to the drug is influenced by genetic and/or experimental factors. At lower doses, apomorphine induced climbing in both groups (ED50 = 1.4 mg/kg in each group) but oral behavior in only one of them (ED50 = 1.3 mg/kg in one, and 8 mg/kg in the second group). Thus, at a given dose of apomorphine, different patterns of stereotypy may result from an interaction between two phenomena: the relative setting of the thresholds to mouth and to climb, and an inverse relation between oral activity and climbing. Analysis of climbing suggests that this response is comprised of two (previously unidentified) fundamental effects of apomorphine: snout contact fixation and bodywise forward progression.

Excessive, abnormal locomotion occurs after a high dose (25-50 mg/kg) of atropine sulfate to rats already akinetic due to catecholamine deficiency from intraventricular administration of 6-hydroxydopamine. This abnormal locomotion involves an abnormal gait sequence [right (R) hindleg (H), left (L) foreleg (F), LH, RF] instead of the normal gait sequence (RH, RF, LH, LF). In such animals atropine progressively (i) decreases hindleg step size, (ii) decreases arching of the trunk, and (iii) increases foreleg step size. These factors combine to change the ratio of front/hind body support. If the body stretches too far and the hindleg step is too small, a given hindleg step supports insufficient weight to remove weight from the ipsilateral foreleg; consequently, the opposite foreleg must execute the next step, producing the abnormal gait sequence. Thus, atropine affects gait sequence indirectly; it acts on at least three variables that affect how body weight is distributed and shifted during locomotion. To maintain stability during such locomotion, gait sequence is appropriately altered.

The seemingly unrelated stereotyped locomotor “acts” reported in the literature to be produced by apomorphine in rats are shown to be composites, whose form and sequence are determined by the particular values of a few component variables which form a common denominator in each of the behaviors. Three variables, continuous snout contact, forward progression and turning, account for much of the behavior. In the course of the drug's action these emerge in succession and vary in amount, the latter two successively reaching a peak and subsiding. The interaction between forward progression and turning yields in sequence, forward walking, circling, revolving, tight pivoting and finally side-to-side movements of the forequarters around the relatively stationary hindquarters. Later behaviors in this list are gradually incorporated into the sequence as earlier ones are eliminated. The course of change in forward progression and turning is also reflected in changes in the sequence and in the direction of stepping of each of the four legs. The order in which the behavior unfolds under the drug is opposite to that manifested in ontogeny and in recovery from lateral hypothalamic damage, suggesting that at the particular high dose used, apomorphine is acting not only to activate the behavior but also to shut it down.

In haloperidol-treated rats, bracing, i.e., resistance to displacement along a horizontal surface, was found to involve four components: gripping by the digits, extension by the limbs, stiffening of the body axis and arching the vertebral column towards the displacing force. Labyrinthectomy weakened the bracing of the forequarters as did application of a head bandage. Labyrinthectomy, when combined with head bandage, completely abolished all forequarter bracing responses. Neither manipulation affected the bracing responses of the hindquarters. The hindquarter bracing reaction was abolished by application of an abdominal bandage, which left most of the forequarter responses intact. Isolation of fore- and hindquarter bracing responses revealed that whereas gripping by the digits and extension by the limbs could occur independently in either part of the body, stiffening of the body axis and arching of the vetebral column originated in the hindquarters. However, although these latter components of bracing originate in the hindquarters, as evidenced by their abolition with application of an abdominal bandage, their recruitment into the anterior of the body only occurred when the bracing of the forequarters was unimpaired. Either labyrinthectomy or head bandage prevented stiffening and arching of the vertebral column from occurring in the forequarters, even though these procedures had no effect on the hindquarters. Labyrinthectomy, head bandage and abdominal bandage all fractionated the bracing response over a wide range of haloperidol dosages (0.5, 1.0, 2,5, 5,0 and 7.5 mg/kg).

The neuroleptic drug haloperidol, which interferes with catecholaminergic activity in the brain, was used to produce akinesia and catalepsy in rats. Such akinesia and catalepsy can be viewed as one in which a subsystem of integrated motor functions (e.g., righting, standing still, clinging, and bracing), is operating to achieve and defend static stable equilibrium, whereas other movement subsystems such as scanning, orienting, and quadrupedal locomotion are meanwhile inoperative. Thus, the catecholamine-deficient rat is a useful preparation to study the isolated components of static stable equilibrium and their interaction, without the complexities encountered by the operation of alternative movement subsystems. Righting in the air was completely inhibited by clinging, no matter how ineffective such clinging was in actually providing physical support. High speed film analysis and lightweight wire-mesh frames of varying stability were used to determine the minimal stimuli that inhibit righting. We suggest that an antagonistic relationship exists between clinging and righting, in which the inputs involved in clinging are dominant over (and may be potentiated by) the vestibular inputs involved in labyrinthine righting. In addition, the bandage-backfall phenomenon, a hitherto bizarre reaction pattern that can be elicited in cataleptic adult animals and normal infants, can be explained by a related antagonistic interaction.

Mild tail-pinch induces eating in animals which are aphagic following lateral hypothalamic lesions. This study compares the specificity of the behavior produced by tail-pinch to that produced by vestibular stimulation in cats which are akinetic and aphagic following lateral hypothalamic lesions. In these cats, tail-pinch elicited licking and biting at food while vestibular stimulation preferentially elicited forward locomotion. These results suggest that such stimuli activate specific movement subsystems rather than producing general, nonspecific arousal.

Electromyographic recordings from antagonistic flexor and extensor muscles in the forelegs (biceps and triceps) and hind legs (tibialis and gastrocnemius-soleus) of freely moving rats demonstrated that haloperidol (5 and 10 mg/kg, i.p.) and morphine (20 and 40 mg/kg, i.p.) produced contrasting patterns of rigidity. Haloperidol catalepsy was characterized by increases in frequency, intensity, and duration of simultaneous tonic cocontractions in antagonistic flexor and extensor muscles of the limbs. Such synergistic rigidity suggested the release of an adaptive mechanism involved in static support and the maintenance of stable static equilibrium (the positive supporting reaction of Schoen and Magnus), at the expense of locomotor mechanisms. In contrast, morphine produced antagonistic/reciprocal rigidity, which was insensitive to challenges to static equilibrium, and was compatible with locomotion. Contrary to the haloperidol-induced limb postures, which were enhanced supporting reactions, those induced by morphine were “frozen” phases of the step cycle. Haloperidol-induced synergistic rigidity and morphine-induced antagonistic/reciprocal rigidity are discussed as manifestations of contrasting movement subsystems underlying these functionally opposite immobility states. In addition, we present hypotheses concerning supraspinal and spinal mechanisms underlying cataleptic rigidity states, and their relevance as models of parkinsonian rigidity and akinesia.

Electrolytic lesions in the caudal part of the medial pontine reticular formation (PRF) in the rat abolished apomorphine- or amphetamine-induced stereotypic lateral head movements and turning to the damaged side. Rats with unilateral PRF lesions turned and circled only to the intact side, and rats with bilateral lesions did not turn at all. PRF lesions were also effective in abolishing or reducing amphetamine-induced circling in rats with unilateral nigrostriatal bundle damage. Thus, head movements induced by dopamine agonists are added to the class of head movements mediated by the PRF. It is proposed that the decussations of the motor pathways for drug-induced turning are located between the midbrain and caudal pons.

A characteristic of catecholamine-depletion-induced catalepsy is that such animals resist horizontal displacement, forward, backward or sideward, by bracing, i.e., pushing against the displacing force rather than stepping away as do normal animals [16]. In this report the bracing responses of two cataleptic preparations were compared: (1) intact rats given haloperidol, a dopamine antagonist, and (2) rats with large lesions of the lateral hypothalamus (LH). Although both preparations exhibited exaggerated bracing responses when pushed, the LH rats acted in a non-cataleptic manner when pushed backward by the head; they retreated backward. The backward walking was a head-elicited response, because pushing the body backwards by the shoulders elicited bracing, not walking. Other forms of head displacement elicited body bracing if the body's stability was challenged: as when the head (and thus the body) was pulled forward; or no strong bracing responses if the head was displaced without affecting the body's stability, i.e., when as the head was pushed laterally. Therefore, retreating rather than bracing was elicited by one specific form of head displacement: backward. In contrast, haloperidol-treated rats braced whenever the body's stability was challenged, including when the head was pushed backward.

An analysis was made of the movement subsystems involved in the stereotyped behaviors and electroencephalographic (EEG) activity that appear in rats given the anticholinergic drug atropine sulfate (50-75 mg/kg). One form of stereotypy occurred when the drugged faced the closed end of an alleyway. Instead of scanning briefly and then turning around to face the open end, as undrugged rats do, they typically "trapped", i.e., they continued to engage in rapid, repetitive scanning by the snout up and down or side to side along the surfaces of the closed end for long periods of time. Atropine appears to produce an exaggerated snout thigmotaxis. Accordingly, the behavioral pattern can be manipulated simply by changing the configuration of the environment, which alters sensory input to the snout. Therefore, such stereotyped behavior is not a motor automatism but rather a circular chain of reflexive reactions to surfaces which trap the animal. During rapid shifts in direction of movement in undrugged rats, there were often extremely brief periods of hippocampal theta that could be blocked by atropine. The presence of theta during such shifts might be important for normal behavioral sequencing. This could partially account for the fact that atropine-treated rats failed to change from their initial reactions in the alleyway to more adaptive behaviors. When a low roof was placed over the alleway, head scanning was greatly limited, and immobility in a lying posture rapidly became the more frequent behavior. This immobility was accompanied by a sleeplike (synchronized) neocortical EEG pattern rather than the activated (desynchronized) neocortical pattern that occurs during repetitive thigmotactic scanning. It is hypothesized that scanning stereotypy and activated EEG are maintained through movement-concurrent positive feedback.

The early stages of recovery from lateral hypothalamic lesions were analyzed in 30 adult cats. In addition to aphagia and adipsia, neurological examination revealed deficits suggestive of deficient endogenous arousal, including somnolence, catalepsy, akinesia, and sensory neglect. Manipulations (tail pinch and injection of amphetamine) that counteracted these deficits also restored feeding. During recovery from aphagia, feeding gradually became activated by sensory stimuli (sight, feel, and smell) associated with food. These data suggest that activation is an important component in the control of normal feeding.

This paper discusses the course of recovery from the thermoregulatory deficits produced in rats by electrolytic lesions in the preoptic/anterior hypothalamic area. Severe damage rendered rats ectothermic, that is unable to maintain their body temperatures at normal levels unless they were incubated at an ambient temperature of 30 degrees C. Less severe damage produced rats that maintained subnormal but stable body temperatures at 23 degrees C, but that did not increase metabolic rate or shiver and whose body temperatures dropped drastically in the cold (5 degrees C). As the animals recovered, nonshivering thermogenesis returned. Eventually the rats became excessively hyperthermic in normal room temperatures, due to very high metabolic rates. They were still unable to shiver or increase metabolic rate further in the cold and were therefore still unable to prevent a large drop in body temperature. Muscle tonus and shivering recovered gradually, and oxygen consumption returned to near normal levels. The data are described in terms of levels of integration of the nervous control of thermoregulation.

Cortical and hippocampal EEG patterns that persist after atropine sulfate administration are defined as atropine-resistant. In the rat, such patterns are movement-related, cortical low-voltage fast activity (LVFA) and movement-related, hippocampal rhythmic slow activity (RSA or “theta”). A close relationship exists between head movement and activation of atropine-resistant EEG. In the present study, we asked whether manipulation of vestibular sensory input would affect atropine-resistant EEG. To allow experimenter control, 6 vestibular-intact rats were made cataleptic with haloperidol and were then injected with atropine sulfate. Movement of the rats* heads by the experimenter (passive head movement) failed to activate atropine-resistant EEG. These animals were then rotated on a turntable. Such vestibular stimulation, the effectiveness of which was reflected by postrotatory ocular nystagmus, &lso failed to activate atropine-resistant EEG. These and 5 other rats were then subjected to surgical or chemical deafferentation of the labyrinth. They were tested for EEG atropine resistance before and after this surgery. Although atropine-resistant EEG patterns in response to active bodily movements still appeared after vestibular deafferentation, the frequency of hippocampal RSA during active movement was reduced with or without atropine. In contrast, &n atropine-sensitive component in the hippocampal EEG during immobility was not changed by vestibular deafferentation. We conclude that vestibular reafferent feedback is neither necessary nor sufficient for the atropine-resistant EEG, but that such feedback from active movement has a modulatory effect on hippocampal RSA.

Our experiments demonstrate that morphine and haloperidol produce two distinct and contrasting behavioral states, which can be thought of as exaggerated, isolated, and simplified forms of organized adaptive behavioral states functioning as components of normal motivated behavior. Haloperidol catalepsy constitutes an organized state in which tonic reactions subserving the maintenance of stable static equilibrium prevail, at the expense of phasic locomotor reactions. In contrast, morphine produces an immobility state characterized by inhibition of the postural support subsystem, and compatible with or preparatory to locomotor rather than static postural reactions. haloperidol-treated rats (1, 2.5, 5, 10 mg/kg i.p.) display exaggeraged bracing reactions to passive displacement as well as to stimuli which do not actively challenge stable equilibrium. In contrast, rats treated with morphine sulfate (10, 20, 40, 80 mg/kg i.p.) show a dose-dependent suppression of bracing and an exaggerated tendency to run in response to stimuli which produce bracing in haloperidol-treated rats. Further evidence that haloperidol-treated rats are organized to stand still in stable equilibrium includes their typical posture during akinesia (i.e. broad-based support), bradykinesia, tonic grasping and enhanced postural components of contact- and air-righting. Under morphine, however, the postural support subsystem is dispensed with, as evidenced by the posture of akinesia (i.e. a frozen phase of the locomotor step cycle associated with loss of limb support), absence of tonic grasping, and nature of the deficits in contact- and air-righting. Furthermore, the opiate-induced immobility state is accompanied by an increased readiness to locomote. Morphine produces an alternation between two extreme behavioral states: complete immobility (inhibition of the postural support subsystem) versus locomotor paroxysms (varying degrees of 'explosive motor behavior'). We suggest that the postures or actions adopted by morphine-treated rats involve movement subsystems concerned with the adaptive behavioral state known as the 'immobility reflex' ('tonic immobility', 'animal hypnosis').

Contact righting, that is, turning from a recumbent position to prone, is abolished for a few days after large electrolytic lesions of the lateral hypothalamus. With recovery, contact righting reappears, but does so in a distinct manner. At first the body is righted by backleg movements, in the absence of any active axial rotation. Later, righting switches from back to front, so that righting begins in the shoulders and then proceeds to the pelvis. Such righting is achieved by axial rotation, that is, the limbs are carried by the torso, rather than vice versa. Labyrinthectomy, when combined with lateral hypothalamic (LH) damage, slows this recovery (now taking as long as 3 weeks), and reveals many intermediate stages of contact-righting. The absence of axial rotation in the early stages of recovery from combined LH damage and labyrinthectomy is compared to the ‘axial apraxia’ seen in some parkinsonian patients.

A cataleptic animal clings in a vertical position, unmoving, for abnormally long periods by supporting some of its weight on its hindlegs, grasping with the forepaws, flexing its forelimbs, and holding the head horizontal. When the head is snugly wrapped with a bandage, the head slowly falls backward, the neck hyperextends, the forelimbs extend and the grasp is released, resulting in the animal falling backward to the ground. It was earlier suggested that in cataleptic animals, the bandage inhibits vestibular and kinesthetic mechanisms of head support, yielding the backfall sequence [35]. However, preliminary experiments showed that labyrinthectomized rats made cataleptic by haloperidol fall backwards when placed in a vertical clinging position, even without a bandage, suggesting that in the rat the bandage-backfall reaction depends only on the vestibular system. In the present paper, this result is verified but, by additional experiments, the latter conclusion is shown to be incorrect. In labyrinthectomized rats made cataleptic by other means (lateral hypothalamic damage, or bulbocapnine), backfall from clinging did not occur unless a bandage was applied. Therefore, the bandage does indeed appear to inhibit the kinesthetic mechanisms that maintain head support in labyrinthectomized cataleptic rats. Haloperidol, particularly in high doses, greatly weakens postural support in labyrinthectomized rats (causing the animal to sag down and fall back when clinging), although the effect is not detectable in rats with labyrinths intact. However, labyrinthectomy reveals that the bandage can trigger an active dorsiflexion of the neck which in itself appears to inhibit clinging and righting. Bandage-induced dorsiflexion is present to a much lesser degree in intact animals, indicating that labyrinthine mechanisms inhibit the dorsiflexion reflex. Therefore, in the intact, cataleptic rat the bandage backfall reaction appears to be produced by the combined effects of a passive component (inhibition of kinesthetic support mechanisms), and an active component (elicitation of dorsiflexion of the neck).

After lateral hypothalamic damage in rats, somnolence, akinesia, and sensory neglect combine to produce complete aphagia. Only simple automatisms (such as grooming, chewing, licking) are present, but intense stimuli can activate more complex actions (walking, orientation, swimming). In the anorexic stage, tactile stimuli dominate in steering locomotion and "spontaneous" locomotion depends on activation from the empty stomach.

Following extensive bilateral lateral hypothalamic damage, rats appear “somnolent.” Cortical EEG shows persistent high voltage delta, reinforcing the impression of sleep. Preoperatively and postoperatively, we simultaneously measured cortical and subcortical (hippocampal and pontine) EEG, muscular events (neck muscle EMG and eye movement EOG), and behavior, which, as aggregates, differentially define quiet sleep, active sleep, and waking. Postoperatively, though cortical activity was persistently slow, subcortical EEG, muscular events, and behavior, as aggregates, revealed quiet sleep, active sleep, and waking, organized subcortically, intact and alternating, but disconnected from the persistent slow cortical activity. For example, preoperatively, active sleep included cortical low voltage fast activity, hippocampal theta, episodic pontine spike bursts, flat EMG, and rapid eye movements, without any organized behavior. Postoperatively, the same aggregate of subcortical and muscular events indicated the presence of active sleep. Similarly so, for subcortically organized quiet sleep and spontaneous waking. Such waking, termed “drowsy-wakefulness,” is a low-arousal form, perhaps related to drowsiness in other species, and to human hypersomnia.

In cataleptic clinging, produced either by catecholamine-blocking drugs or lateral hypothalamic damage in adult cats, rats, or monkeys, bandaging the face and neck causes the head to fall backwards.Early in development, a similar reaction can be seen in normal undrugged infants.

In earlier work, we showed that in rats, proprioceptive-tactile information is sufficient for contact-righting on the ground (from lying on one side to prone). Thus, axial rotation, starting with the shoulders and followed by the pelvis, occurs normally in labyrinthectomized animals with eyes occluded. After damage to the lateral hypothalamus, even with labyrinths intact, contact-righting is at first abolished (1-2 days postoperatively), and when it reappears, involves pushing by the hindlegs. Rostrocaudal contact-righting, involving axial rotation, takes 3-4 days to recover. If labyrinthectomy is combined with lateral hypothalamic damage, the deficit is exaggerated and recovery is greatly slowed down, now requiring 2-3 weeks. The present paper shows that during this prolonged period of recovery several transitional forms of righting are present, each produced by a different combination of limb and body axis movements. At first, axial rotation is absent, and righting is achieved only by pushing with the limbs. This is followed by a transitional form in which, even though axial rotation cannot be triggered directly by contact with the ground, it can be triggered indirectly as an allied reflex when the paw places on the ground. Eventually the body axis actively initiates the rotation to proneness (at first, in the pelvis, later in recovery, in the shoulders), with the limbs being carried. Recovery of axial rotation overlaps with the recovery of cephalic dominance, yielding complex intermediate forms of righting.