Imipramine affects circadian leaf movements in Oxalis regnellii

The sensitivity of the circadian leaf movement of Oxalis regnellii Mig. to imipramine (a tricyclic dibenzazepine) was investigated. Imipramine, like Li⁺, is used as a therapeutic agent against depressive disorders in man. The therapeutic effects of the two substances might be mediated via effects on basic circadian rhythms and the cellular level. It was indeed possible to influence the circadian movement of Oxalis by imipramine; pulses (10⁻³ M , 4h) phase shifted the rhythms and caused advances. A phase response curve is presented. No period change of the movements was caused by permanent presence of imipramine (5 - 10⁻⁵ or 10⁻⁵ M ). The nature of the imipramine-induced phase shift is discussed and compared with Li⁻¹ effects on the same circadian system.     

Characteristics of the combined action of melatonin and imizin on the structure of forced swimming and the circadian rhythm

Acute administration of melatonin (10 mg/kg) produced a depressed effect on the mice behavior. But it's injection after chronic imipramine (10 mg/kg/day, 2 weeks) potentiated antidepressant ability and shortened the duration of immobility in the structure of swimming. In rats, which had a high amplitude of circadian rest-activity rhythm, melatonin produced imipramine effect on circadian mobility, but potentiated antidepressant action in animals with low initial level of locomotor activity.     

Circadian rhythm of circulating corticosterone and urinary excretion of catecholamines, serotonin and their catabolites in rats treated with imipramine

The circadian rhythm of serum corticosterone was assessed in rats entrained to a 12:12 LD cycle and treated with tricyclic imipramine (25 mg/kg/day) via osmotic pumps for a period of 14 days; urinary excretion of catecholamines, serotonin and their catabolites was also assessed. We observed that imipramine did not modify the phase position of the corticosterone rhythm but rather lowered the animal's responsiveness as shown by the lower peak of corticosterone at 2000 and by the smaller amplitude of its circadian rhythm; moreover imipramine had no effect on urinary excretion of catecholamines, serotonin and their catabolites during LD cycle.     

Phase shifting of the circadian conidiation rhythm in Neurospora crassa by calmodulin antagonists

The effects of chemicals capable of antagonizing the functions of calmodulin, such as trifluoperazine, chlorpromazine, imipramine, alprenolol, W7, and W13, on the circadian conidiation rhythm of Neurospora crassa were examined. Trifluoperazine, at a 30-microM concentration, was most effective in shifting the phase of the conidiation rhythm and caused a maximum phase delay at circadian time (CT) 6 and maximum phase advance at CT 9. Chlorpromazine was less effective than trifluoperazine, and a 300-microM concentration of chlorpromazine was required for a similar phase shift. Imipramine, at a 1-mM concentration, caused only a small phase shift, while alprenolol had little effect on biological clock function. W7 and W13 caused phase delays longer than 10 hr at CT 6 and caused a phase advance of about 5 hr at CT 10 when present at a 200-microM concentration. However, W5 and W12, the dechlorinated homologues of W7 and W13, had no effects on clock function at the same concentration. Calmodulin was assayed by measurements of stimulation of cyclic nucleotide diphosphodiesterase activity. Calmodulin content remained constant in trifluoperazine-sensitive and trifluoperazine-insensitive phases for two cycles following the light-dark transition.     

Chronic Administration of Imipramine and Lithium Changes the Phase-Angle Relationship Between the Activity and Core Body Temperature Circadian Rhythms in Rats

Evidence suggests that there is an association between the pathophysiology of depression and a disturbance of circadian rhythms. Accordingly, attention has focused on the possible effects of antidepressants on circadian rhythms. In the present study, we examined the effects of chronic administration of two clinically effective antidepressant agents, imipramine and lithium, on several circadian rhythms in the rat. Activity, core body temperature, and drinking rhythms were assessed in constant darkness (DD) and light-dark (LD) conditions. In DD, lithium significantly lengthened the circadian period of the activity, temperature, and drinking rhythms, while imipramine had no effect. In LD, both drugs significantly delayed the phase of the activity rhythm, but did not change that of the other two rhythms. As a result, the phase-angle differences between the activity and temperature rhythms significantly increased. Neither lithium nor imipramine produced any effect on the resynchronization of these rhythms after an 8-h delay in the LD cycle. These results indicate that although both drugs produced different effects on the circadian period of individual rhythms, both caused a relative phase advance of the temperature rhythm as compared to the activity rhythm, and this effect may be related to the similarity in their antidepressant effects. (Chronobiology International, 13(4), 251-259, 1996)     

Analysis of the circadian rhythm of motor activity in white rats by using several pharmacologic agents which act on cerebral monoamines]

Three pharmacological agents, (disulfiram, imipramine and reserpine) influencing the brain monoamine transmitters have been studied to explain the mechanism involved in the motor circadian rhythm. The new results corroborate our previous ones : the norepinephrine transmitter responsible for the paradoxical sleep is unable to explain the depressed metabolic phase of the circadian rhythm; the negative results obtained with disulfiram and imipramine corroborate the previous results with nialamide. On the contrary, the agents which reduce the serotonine transmitter mechanisms (p-chlorophenylalanine) decrease the difference between active and sleep phases by their action involved in the non-activated sleep. Reserpine, supposed to reduce both transmitter mechanisms (serotonine and norepinephrine) shows a p-chlorophenylalanine-like effect, perhaps more strong. It thus appears that the non-activated sleep is responsible for the two essential sleep functions : the restitution function and the depressed phase of the circadian rhythm.     

The temporal dynamics of swimming in rats as a possible index of the circadian mobility of the animals

By the time course of forced swimming all rats may be separated in several groups with typical patterns of circadian motility. In case when active swimming movements and reduced immobility predominated, invert type of daily chronogram with low activity at the night hours and high sensitivity to chronic imipramine treatment were observed. Considerable immobility correlates with clear circadian rhythm and weak response to antidepressant. It is suggested that rhythmical pattern of the forced swimming may be used as predicator of circadian motility type.     

Effects of N-acetylcysteine and imipramine in a model of acute rhythm disruption in BALB/c mice

Circadian rhythm disturbances are among the risk factors for depression, but specific animal models are lacking. This study aimed to characterize the effects of acute rhythm disruption in mice and investigate the effects of imipramine and N-acetylcysteine (NAC) on rhythm disruption-induced changes. Mice were exposed to 12:12-hour followed by 10:10-hour light:dark cycles (LD); under the latter, mice were treated with saline, imipramine or NAC. Rhythms of rest/activity and temperature were assessed with actigraphs and iButtons, respectively. Hole-board and social preference tests were performed at the beginning of the experiment and again at the 8th 10:10 LD, when plasma corticosterone and IL-6 levels were also assessed. Actograms showed that the 10:10 LD schedule prevents the entrainment of temperature and activity rhythms for at least 13 cycles. Subsequent light regimen change activity and temperature amplitudes showed similar patterns of decline followed by recovery attempts. During the 10:10 LD schedule, activity and temperature amplitudes were significantly decreased (paired t test), an effect exacerbated by imipramine (ANOVA/SNK). The 10:10 LD schedule increased anxiety (paired t test), an effect prevented by NAC (30?mg/kg). This study identified mild but significant behavioral changes at specific time points after light regimen change. We suggest that if repeated overtime, these subtle changes may contribute to lasting behavioral disturbancess relevant to anxiety and mood disorders. Data suggest that imipramine may contribute to sustained rhythm disturbances, while NAC appears to prevent rhythm disruption-induced anxiety. Associations between sleep/circadian disturbances and the recurrence of depressive episodes underscore the relevance of potential drug-induced maintenance of disturbed rhythms.     

Hemodynamic and cardiac actions of trazodone and imipramine in the anesthetized dog.

The relative cardiovascular effects of trazodone and imipramine were compared in two open-chest, anesthetized dog models. Trazodone lowered arterial blood pressure (0.3 mg/kg), slowed heart rate (3 mg/kg) and reduced myocardial contractile force (3–10 mg/kg) following i.v. administration. Low i.v. doses (0.05–0.15 mg/kg) of imipramine increased arterial blood pressure and heart rate, presumably as a consequence of its known anticholinergic properties and/or effects on neuronal catecholamine re-uptake mechanisms. Subsequent to administration of 1.5 and 5 mg/kg, however, the vascular and myocardial depressant effects of imipramine were evident. Trazodone (1–10 mg/kg, i.v.), unlike imipramine, effected a substantial level of $\text{alpha}$-adrenergic blockade $\text{vs.}$ a fixed challenge dose of norepinephrine, although less than that associated with phentolamine. Both trazodone and imipramine reduced aortic flow although $\text{via}$ different mechanisms. The reduction following administration of trazodone resulted from a decrease in heart rate whereas imipramine depressed aortic flow by lowering stroke volume.     

The effect of the deprivation of the paradoxical sleep stage on the temporal dynamics of swimming in rats with antidepressive and depressant exposures]

After REM sleep deprivation the time-course of the forced swimming was reorganized. As shown, reduction of rhythmical index of depression, such effect has an antidepressive nature. In this model potentiation of specific activity of antidepressant imipramine and attenuation of depressive properties of clonidine were observed. These results suggest that shifts in sleep phase structure may be a source of restriction of circadian desynchronosis, upon which depression is based.     

Tricyclic Imipramine Modification of the Circadian Rhythms of Hypothalamic Serotonin, its Precursors and Acid Catabolite in Individually Housed Rats

Circadian rhythms of serotonin (5HT), its precursors tryptophan (TP) and 5-hydroxy-tryptophan (5HTP) and its acid catabolite 5-hydroxy-indoleacetic acid (5HIAA), were determined in the hypothalamus of control rats and rats which had been treated continuously with subcutaneous imipramine (10 mg/kg/day) for 2 weeks.

Rats were individually housed and entrained to LD12:12. Controls showed the 5HT and TP peaks in the light and dark periods respectively, as reported in the literature, but no inverted correlation (antiphase) between SHT and 5HIAA rhythms.

Imipramine significantly modified circadian rhythm characteristics: the 5HT acrophase was advanced, that of TP and 5HIAA was delayed. Imipramine also significantly increased hypothalamic SHT and TP concentrations.     

A biorhythmologic approach to evaluating forced swimming as an experimental model of a "depressive" state

Rhythmical structure of forced swimming was studied on rats. Reserpine (1 mg/kg, 24 h before testing), clonidine (150 mkg/kg) and prolonged repeated striatal stimulation induced behavioural depression with reorganization of swimming rhythm and increase of short cycles (less than 6 s) of immobility. After chronic administration of antidepressants (imipramine, amitriptyline, niamid, 10 mg/kg/day, during 14 days), on the contrary, the number of these cycles diminished, while the number of active swimming cycles increased. Chrono-biological "index of depression" is suggested to express more exactly behavioural depression and specific activity of antidepressants than usual registration of immobility time.     

Determination of imipramine and seven of its metabolites in human liver microsomes by a high-performance liquid chromatographic method

The metabolism of the widely used antidepressant drug imipramine is subject to marked interindividual variation. A sensitive and specific reversed-phase high-performance liquid chromatography method for the simultaneous determination of imipramine and seven of its metabolites in human liver microsomal preparations was developed. These metabolites include 10-hydroxy-desipramine, 10-hydroxyimipramine, 2-hydroxydesipramine, 2-hydroxyimipramine, desipramine, didesmethylimipramine, and imipramine N-oxide. The detection limit for imipramine and the metabolites was approximately 20 pmol. At concentrations of 100 and 500 pmol per tube, the reproducibility showed a coefficient of variation less than 10%, except for the 2-hydroxy-desipramine (16%), 2-hydroxyimipramine (15%), and imipramine N-oxide (17%), all three at 100 pmol per tube. Linear standard curves were obtained for all the compounds within a concentration range of 50 to 1000 pmol per tube. This assay will provide a tool to assess the contribution of different enzymes to the formation of imipramine metabolites.     

Role of the amygdala in the sedation induced by low doses of apomorphine

In the present study the role of amygdala in the antidepressant action of imipramine is discussed. An animal model of depression is induced, in rats, by systemic injection of low doses of apomorphine. Systemic administration of imipramine prevents, as already reported, apomorphine-induced sedation. The same effect is observed following intra-amygdaloid imipramine administration. On the contrary, local injection of imipramine in frontal cortex or caudate nucleus does not affect apomorphine-induced sedation.     

Pineal-independent regulation of photo-nonresponsiveness in the Siberian hamster (Phodopus sungorus)

The pineal hormone melatonin influences circadian rhythms and also mediates reproductive responses to photoperiod. The authors tested whether pinealectomy influences circadian oscillators responsible for induction of nonresponsiveness to short day lengths by preventing normal short-day patterns of circadian entrainment. Adult male Siberian hamsters were pinealectomized or sham operated, maintained in either 18 h light per day (18L) or 15L for 10 weeks, and then tested for responsiveness to 10L. Because pinealectomized hamsters do not show gonadal regression in short day lengths, responsiveness was assessed by measuring phase angle of entrainment and the length of the nightly activity period following transfer to 10L. The incidence of nonresponsiveness was significantly higher in 18L hamsters than in 15L hamsters but was unaffected by pineal status. Fully 88% of 18L hamsters failed to entrain to 10L in the normal short-day manner; the duration of nightly activity remained compressed, and the phase angle of entrainment was large and negative relative to lights off. The 15L hamsters entrained normally to 10L. Exposure to constant light after 10L treatment was equally effective in inducing arrhythmicity in pinealectomized and intact hamsters. Changes in the period of morning and evening circadian oscillators subsequent to 18L treatment did not predict circadian responsiveness to short photoperiod. Long-day induction of photo-nonresponsiveness, which prevents winter responses to short day lengths, occurs independently of pineal melatonin feedback on the circadian system.     

Relationship between disruption of rhythmicity and reentrainment in surgical patients

Urine samples for assay, temperature, heart rate, and blood pressure were collected daily at 2-h intervals from 11 consenting subjects undergoing abdominal surgery, as well as 10 age-and sex-matched control subjects. Alterations in level, and timing of circadian excretion of catecholamine metabolites, adrenal cortical hormones, sodium, potassium, creatinine, and vital signs following surgery were measured. Data were examined to determine if a relationship exists between the degree of circadian alteration and the subject's return to typical circadian profiles. The data suggest that certain circadian rhythms of hospitalized subjects were altered and uncoupled from external stimuli. In addition, subjects with less disruption in some variables following surgery regained rhythmicity more quickly than more disrupted subjects. These findings suggest that health professionals should individualize patient care to promote rhythmicity. In addition, patient assessment should consider individual circadian patterns and disruption following surgery.     

Testosterone has antidepressant-like efficacy and facilitates imipramine-induced neuroplasticity in male rats exposed to chronic unpredictable stress

Hypogonadal men are more likely to develop depression, while testosterone supplementation shows antidepressant-like effects in hypogonadal men and facilitates antidepressant efficacy. Depression is associated with hypothalamic–pituitary–adrenal (HPA) axis hyperactivity and testosterone exerts suppressive effects on the HPA axis. The hippocampus also plays a role in the feedback regulation of the HPA axis, and depressed patients show reduced hippocampal neuroplasticity. We assessed the antidepressant-like effects of testosterone with, or without, imipramine on behavioral and neural endophenotypes of depression in a chronic unpredictable stress (CUS) model of depression. A 21-day CUS protocol was used on gonadectomized male Sprague–Dawley rats treated with vehicle, 1 mg of testosterone propionate, 10 mg/kg of imipramine, or testosterone and imipramine in tandem. Testosterone treatment reduced novelty-induced hypophagia following CUS exposure, but not under non-stress conditions, representing state-dependent effects. Further, testosterone increased the latency to immobility in the forced swim test (FST), reduced basal corticosterone, and reduced adrenal mass in CUS-exposed rats. Testosterone also facilitated the effects of imipramine by reducing the latency to immobility in the FST and increasing sucrose preference. Testosterone treatment had no significant effect on neurogenesis, though the combination of testosterone and imipramine increased PSA-NCAM expression in the ventral dentate gyrus. These findings demonstrate the antidepressant- and anxiolytic-like effects of testosterone within a CUS model of depression, and provide insight into the mechanism of action, which appears to be independent of enhanced hippocampal neurogenesis.     

Subcellular Site of Action of Imipramine in Rodent Brain

To determine the site of action of imipramine, the subcellular distribution of [3H]imipramine in rodents was followed after both in vivo administration and in vitro incubation with tissue slices under "physiological" conditions. Total [3H]imipramine (10-1,000 nM) binding was associated with all primary fractions, but in particular with the nuclear (P1) and mitochondrial (P2) pellets and the synaptosomal (P2B) and myelin (P2A) fractions. Using an excess of imipramine to define any nonspecific interactions, a specific association was observed mainly in those fractions containing isolated nerve terminals and to a lesser extent with the purified myelin fraction. Preparation of subsynaptosomal fractions by osmotic lysis indicated that [3H]imipramine was associated with the synaptic vesicle and microsomal fractions and also with synaptosomal membranes. The degree of binding to the vesicular and microsomal fractions was increased with the length of preparation time, whereas there was an inverse relationship between the length of preparation and the amount bound to the synaptosomal membrane fraction. There was no evidence of an intrasynaptosomal accumulation of [3H]imipramine at concentrations up to 1,000 nM. [3H]2-Nitroimipramine, a slowly dissociating imipramine derivative, was exclusively located in synaptic membrane fractions. Prior treatment of rats with a combination of 5,7-dihydroxytryptamine and desipramine reduced 5-hydroxytryptamine levels and the levels of [3H]imipramine associated with the synaptosomal fractions to the same extent. It is concluded that imipramine is associated with a binding site localised on 5-hydroxytryptaminergic nerve terminals and that there is a redistribution to other sites (vesicular and microsomal) during the isolation procedure.     

Assessment of rat brain alpha1-adrenoceptor binding and activation of inositol phospholipid turnover following chronic imipramine treatment

Chronic (21 days) treatment of rats with imipramine (10 mg/kg) did not change the density or affinity of alpha₁-adrenoceptors as measured by the specific binding of [³H]prazosin in rat cortical membranes, but produced the expected significant decrease in the density of beta-adrenoceptors labeled by [¹²⁵I]iodocyanopindolol. The functional status of brain alpha₁-adrenoceptors was also assessed by measuring the noradrenaline (NA)-induced accumulation of [³H]inositol 1-phosphate (IP₁) in brain slices from these animals. No apparent change was observed in the concentration-response relationship between NA and [³H]IP₁ accumulation in rat cerebral cortex after chronic treatment with imipramine. At concentrations higher than 1 M in vitro, imipramine and its metabolite, desipramine, produced a concentration-dependent decrease in the [³H]IP₁ accumulation elicited by NA. This inhibitory effect is likely mediated by direct blockade of alpha₁-adrenoceptors by these drugs. As the endogenous drug concentration would not reach 1 M in our preparation, the lack of changes in alpha₁-adrenoceptor response following chronic imipramine treatment are not likely attributable to residual imipramine or desipramine retained in the tissues. In conclusion, the above findings do not support previous suggestions that brain alpha₁-adrenoceptors are upregulated following chronic imipramine administration.     

Similarities and Differences Between High-Affinity Binding Sites for Cocaine and Imipramine in Mouse Cerebral Cortex

Previously we found close similarities between high-affinity binding sites for [3H]cocaine and those for [3H]imipramine in the mouse cerebral cortex in regard to their association with neuronal uptake of serotonin. In the present study we investigated whether the two ligands bind to the same site. The two ligands had the following high-affinity binding properties in common: localization in both synaptosomal and microsomal fractions; vulnerability to treatment with N-ethylmaleimide, trypsin, and phospholipase A2; and resistance to exposure to dithiothreitol. In contrast, cocaine binding in the cerebral cortex was more sensitive to heat inactivation than imipramine binding. In addition, the mechanism by which cocaine inhibited [3H]imipramine binding differed from that by which imipramine inhibited [3H]cocaine binding. These data suggest that the high-affinity binding sites for [3H]cocaine and [3H]imipramine in the cerebral cortex are distinct entities.