Chronic Treatment Of Mice Research Articles

Effect of the synthetic cannabinoid nabilone on spermatogenesis in mice

Chronic treatment of mice with the synthetic cannabinoid nabilone (50 mg/kg, 3 times per week) reduced the number of pachytene spermatocytes. Nabilone did not affect other cell types in the testis or the sex organ weights. Nabilone tended to increase the number of abnormal spermatozoa, but this did not reach statistical significance. It is concluded that nabilone causes less testicular toxicity than the natural cannabinoids.

Differential regulation of σ and PCP receptors after chronic administration of haloperidol and phencyclidine in mice

The existence of multiple receptor sites for the psychotomimetic agents phencyclidine (PCP) and some opiate-benzomorphans such as (+)N-allylnormetazocine ([+]SKF 10,047) in the mammalian central nervous system is well documented. These are: 1) sigma/PCP (sigma p) site, which binds both PCP and psychotomimetic opiates but not antipsychotics such as haloperidol, 2) PCP site, which selectively binds PCP analogs, and 3) sigma/haloperidol (sigma h) site, for which certain antipsychotics and (+)SKF 10,047, but not PCP analogs, display high affinity. In this study we examined the regulation of these receptor sites after chronic treatment of mice with either PCP or haloperidol. The following radiolabeled ligands were used to assess binding to the various receptor subtypes: [3H]-1-[1-[3-hydroxyphenyl)cyclohexyl]piperidine ([3H]PCP-3-OH; sigma p and PCP sites), [3H]thienyl-phencyclidine ([3H]TCP; PCP site), (+)-[3H]SKF 10,047 (sigma p and sigma h sites), and [3H]haloperidol (sigma h and D-2 dopamine receptors). Treatment of mice for 1, 7, 14, and 21 days with PCP (10 mg.kg-1.day-1) failed to induce variations in sigma p, sigma h, and PCP receptor binding. However, similar treatment with haloperidol (4 mg.kg-1.day-1) induced: 1) complete elimination of the binding to sigma h sites, 2) up-regulation of D-2 dopamine receptors, and 3) no change in sigma p and PCP receptor binding after 14 or 21 days of treatment. However, a single day of haloperidol treatment or in vitro incubation of mouse brain membranes with haloperidol failed to alter receptor binding. This study suggests that prolonged treatment of mice with haloperidol induces a loss in sigma h receptors that are presumably associated with certain psychotomimetic effects. This phenomenon is accompanied by an up-regulation of D-2 dopamine receptors.

The effects of central dopamine function of chronic L-DOPA (methyl ester hydrochloride) treatment of mice

Mice were treated for 28 days with drinking water containing L-DOPA methyl ester hydrochloride (DME) plus carbidopa, carbidopa alone, or with the vehicle. All mice were then given the vehicle for 1 day and behavioural and biochemical assessments made on the 29th day. On average, mice consumed between 181 and 302 mg/kg of DME (expressed as the base) each day. In behavioural experiments DME- and carbidopa-treated mice were subsensitive to the locomotor stimulating effects of apomorphine, after their pretreatment with reserpine plus α-methyl-p-tyrosine to remove endogenous stores of dopamine and to stop its synthesis. Even mice pretreated for only one day with chronic DME or carbidopa displayed some subsensitivity to apomorphine challenge, but the effect was more marked the longer the chronic treatment. Other mice were chronically treated for 28 days with α-methylDOPA or vehicle, and these mice when challenged with apomorphine after dopamine depletion (as described above), were also markedly subsensitive to the locomotor activating effects of apomorphine. There were no changes in sensitivity of drug-treated mice to the hypothermic effects of apomorphine, to the stereotypy-inducing effects of apomorphine or d-amphetamine, or to the locomotor activating effects of L-DOPA itself or to bromocriptine. There were, however, some changes in the basal grooming behaviour of both DME- and carbidopa-treated mice, and in their response to SKF38393 challenge. Striatal binding studies with [ 3H]-spiperone and [ 3H]-SCH23390 indicated that there were no marked changes in K d or B max of either D-1 or D-2 receptors. The data indicate that chronic treatment of mice with L-DOPA (as the methyl ester hydrochloride) produces a subsensitivity to one particular behavioural effect of apomorphine, in the absence of any change in D-1 and D-2 receptors. Furthermore, the behavioural subsensitivity to apomorphine would appear to be a result of the chronic treatment with carbidopa. Carbidopa, while not crossing the blood-brain barrier, can be metabolized in part to α-methylDOPA, which can cross teh blood-brain barrier.

Inhibition of growth and linoleate-enhanced metastasis of a transplantable mouse mammary tumor by indomethacin

The influence of the cyclo-oxygenase inhibitor, indomethacin (IM), on the metastasis, development and prostaglandin E (PGE) levels of line 4526 mammary tumors grown in mice fed high fat (HF, 20%, w w ) diets containing various levels of linoleic acid (18:2) was investigated. Control mice that grew primary tumors and were fed HF diets containing 12% 18:2 ( w w ) had 2–3 times the number of lung metastases than mice fed 1%, 4%, or 8% 18:2. Chronic treatment of mice with 10 μg/ml IM in drinking water reduced metastasis in 1% and 4% 18:2-fed mice compared to controls and completely inhibited the increased metastasis of mice fed the 12% 18:2 diet. Treatment with IM also increased the latency and decreased the growth rates of primary 4526 tumors of all dietary groups. Treatment of mice with a higher dosage of IM (20 μg/ml), decreased tumor metastasis even further compared to controls, but did not decrease tumor growth rate compared to the lower dosage of IM (10 μg/ml). Tumor PGE levels, measured by radioimmunoassay (RIA), were decreased by IM treatment. These data provide evidence that arachidonic acid metabolites such as PGE may be involved in the metastasis of 4526 mammary tumors.

Co-existence of kindling induced by the β-carboline, FG 7142, and tolerance to diazepam following chronic treatment in mice

The effect of chronic treatment with the β-carboline, FG 7142, followed by chronic treatment with diazepam (DZP) on the acute effects of DZP and FG 7142 were studied. Mice were treated for 16 days with FG 7142 (40 mg/kg i.p.) followed by treatment with DZP (5 or 20 mg/kg i.p.) for 9 days. At the end of this period, the anticonvulsant, antipunishment and locomotor sedative properties of a test dose of DZP were assessed, as were the convulsant properties of FG 7142. During the chronic treatment with FG 7142, 80% of the mice developed clonic convulsions in response to the β-carboline, and this increased sensitivity to FG 7142 (kindling) remained following the chronic DZP treatment. Thus long-term treatment with DZP does not reverse the changes which occur during FG 7142-induced kindling. Chronic treatment with DZP for 9 days gave rise to tolerance to its pharmacological effects as assessed in the 4-plate test of antipunishment activity, in a test of locomotor sedation, and by its ability to increase the convulsant threshold of intravenously administered pentylenetetrazol. The development of tolerance to DZP was not affected by a prior chronic treatment with FG 7142. Nor were the acute effects induced by DZP altered by a prior chronic treatment with FG 7142. Apart from reducing its own convulsant threshold, chronic treatment with FG 7142 had no effect in any of the experiments. These results suggest that kindling induced by FG 7142 and tolerance to DZP depend on different mechanisms. In neither case were the pharmacological changes induced by chronic administration reflected by changes in the biochemical measures of the coupling between benzodiazepine binding sites and γ-aminobutyric acid (GABA) receptors.

Effects of ethanol on ouabain inhibition of mouse brain (Na +,K +)ATPase activity

Plots of ouabain inhibition of mouse cerebral cortical (Na +K +ATPase activity fitted a two-site model significantly better than a one-site model, consistent with the presence of two forms of the enzyme with different affinities for ouabain. The fraction of enzyme activity with high affinity for ouabain (HAO: K i = 500 nM), suggested to be localized neuronally, constituted the major portion (60–70%) of activity. Ouabain inhibition of both components of enzyme activity was reduced as KC1 concentrations were increased. In vitro, only high concentrations of ethanol affected (Na +,K +)ATPase activity and ouabain inhibition of activity. Ethanol (500 mM) selectively reduced the activity, and increased the sensitivity to ouabain inhibition, of the HAO component, with no significant effect on the low-affinity (LAO) component. On the other hand, following chronic treatment of mice with ethanol in vivo, in a paradigm that produced tolerance and physical dependence, the sensitivity to ouabain of the HAO form of the enzyme was selectively increased. The relative proportions, and the activities of the HAO and LAO components, were not altered. The effects of ethanol, added in vitro, on the HAO component were decreased in ethanol-tolerant animals. The selective effect of chronic ethanol ingestion on (Na +,K +)ATPase activity indicates the specificity of action of ethanol in the CNS.

Tolerance to effects of clonidine and morphine on sulfobromophthalein in disposition in mice

Chronic treatment of mice with clonidine or morphine caused tolerance to the analgesic and thermoregulatory effects of these drugs. After chronic morphine, mice also became tolerant to the analgesic and thermoregulatory effects of clonidine. Cross tolerance to the hypothermic effect of morphine was demonstrated after chronic clonidine administration, but no diminution of morphine-induced analgesia could be shown. Morphine and clonidine acutely increased the retention of sulfobromophthalein (BSP) in plasma and liver. Chronic dosing with morphine or clonidine caused partial tolerance and cross-tolerance to the rise in hepatic BSP caused by an acute challenge with either agonist. However, both drugs elevated plasma BSP levels similarly in tolerant and non-tolerant mice. Thus, regimens which readily induced tolerance to the analgesic and hypothermic effects of morphine or clonidine were only partially effective in modifying the acute hepatobiliary effects of these drugs.

BM-5, a centrally active partial muscarinic agonist with low tremorogenic activity. In vivo and in vitro studies.

The acute and chronic effects of the centrally active oxotremorine analog, BM-5, [N-methyl-N-(1-methyl-4-pyrrolidino-2-butynyl)-acetamide] were examined in rats and mice. In vivo studies in mice and rats indicated that this compound is a partial muscarinic agonist with large regional differences in its efficacy: BM-5 produced low tremor in doses which evoke full salivary response. The maximal tremor response to BM-5 was much smaller than that produced by oxotremorine, while the maximal salivary response to BM-5 was greater than that evoked by oxotremorine. The tremor response to BM-5 was bell-shaped, the peak dose being around 2 mg/kg. In contrast, the salivary response increased with increasing doses of BM-5. The apparent muscarinic antagonist properties of higher doses of BM-5 were specific to the striatum in which BM-5 (0.05-10 mg/kg) caused significant decreases in the level of acetylcholine while these levels were unaltered in the cerebral cortex, hippocampus, and brainstem. Pretreatment of rats with BM-5 (5 mg/kg) also prevented the increase in striatal acetylcholine induced by oxotremorine (0.75 mg/kg). Chronic treatment of mice with BM-5 (0.2-2 mg/kg) for 14 days also showed that BM-5 at higher doses, behaved as an antagonist, since it caused supersensitivity to oxotremorine on the tremor response. In addition, the number of receptor sites, as measured by binding of 3H-3-quinuclidinyl benzilate (3H-3-QNB), was increased in the striatum while no similar increase was observed in other brain areas.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of chronic drug treatment on the sensitivity of mouse vas deferens to drugs

Chronic treatment of mice with clonidine or morphine reduced the sensitivity of the isolated, field-stimulated, vas deferens to the inhibitory effects of acutely administered clonidine and morphine. Chronic treatment of mice with clonidine produced a postsynaptic non-specific supersensitivity to noradrenaline and carbachol, but not to potassium chloride. Chronic treatment with morphine produced a specific supersensitivity to noradrenaline. Chronic treatment of mice with naloxone increased the sensitivity of the isolated, field stimulated, vas deferens to the inhibitory effects of acutely administered morphine, but not to clonidine. Naloxone treatment of mice did not affect the sensitivity of the isolated vas deferens to noradrenaline, carbachol or potassium chloride. These results suggest a physiological role for the opioid receptors in the mouse vas deferens and an apparent cross-tolerance between clonidine and morphine. The possible mechanisms are discussed.

SUBACUTE BENZODIAZEPINE TREATMENT: OBSERVATIONS ON BEHAVIOURAL TOLERANCE AND WITHDRAWAL

The acute and subchronic actions of the benzodiazepines triazolam, oxazepam and diazepam have been examined using three behavioural tests. In the accelerating rotarod motor coordination test both diazepam and oxazepam on acute administration produced dose-related discoordination or motor impairment in mice. After chronic administration (50 mg/kg per day i.p. for 14 days) the corresponding dose-response lines were shifted to the right suggesting tolerance development. In a test for conflict behaviour in rats (conditioned suppression of drinking) oxazepam (20 mg/kg i.p.), triazolam (1 mg/kg i.p.) and diazepam (10 mg/kg i.p.) all augmented punished responding rates with no effect on unpunished responses and this has been speculated to reflect anxiolytic activity. These initial elevations in punished responding displayed a gradual decline during repeated daily administration over 20 days, there being little alteration in the unpunished response levels. In separate studies, chronic treatment of mice with triazolam in the drinking water over 30 days showed diminished pentylenetetrazol-induced seizure latencies compared to the level of protection afforded by triazolam at the beginning of the schedule. In additional experiments, mice injected daily with triazolam (1 mg/kg i.p. for 14 days) exhibited a higher seizure susceptibility than their corresponding controls. The results are discussed in relation to tolerance to the anxiolytic, anticonvulsant and neurological impairment properties of benzodiazepines.

The effects of chronic morphine administration to mice on the contractile activity in vitro of the vas deferens without and with morphine

Chronic treatment of mice with morphine enhances maximal responses to noradreneline in isolated vasa deferentia. Addition of morphine to the bath further increases the responses to noradrenaline (facilitatory effect). Vasa deferentia from reserpine-pretreated mice showed increased responses to noradrenaline, but upon addition of morphine to the bath no further facilitatory response to noradrenaline. The facilitatory response was not inhibited by naloxone and it was not elicited by narcotic analgesics other than morphine. High calcium in the medium (3.8 mM) suppressed the enhanced maximal responses to noradrenaline of vasa deferentia from mice chronically treated with morphine and also suppressed the facilitatory response to noradrenaline. Low calcium in the medium (1.43 mM) had the opposite effects. Reserpine-induced changes of noradrenaline responses were not affected by alteration of the calcium concentration in the medium. In the K-depolarized vas deferens, a reduction of the depressant effect of morphine on the calcium-induced contractions was observed after chronic administration of morphine.

The direction of opioid agonists towards μ-, δ- and ε-receptors in the vas deferens of the mouse and the rat

Chronic treatment of mice with specific opioids results in the development of tolerance of particular opiate receptors in the mouse vas deferens (MVD). Accordingly, the infusion of animals with the specific δ-receptor ligand [D-Ala 2,D-Leu 5]-enkephalin (DADL) or the potent μ-agonist sufentanyl (SUF) produces MVD highly tolerant to δ- and μ-agonists, respectively. Investigating a series of opioids in these preparations provides unequivocal evidence for the simultaneous existence of δ- and μ-receptors in the MVD. Thus, the possibility exists to obtain vasa deferentia, which almost exclusively contain either μ- or δ-opiate receptors. In combination with the rat vas deferens (RVD), a supposedly selective ε-receptor preparation, useful tools are provided for the classification of opioids according to their preference for the μ-, δ- and ε-type of the opiate receptors.

Effect of Treatment with Phenformin, Diphenylhydantoin or <i>L</i>-Dòpa on Life Span and Tumour Incidence in C3H/Sn Mice

The chronic treatment of female C3H/Sn mice with phenformin (2 mg/day) and diphenylhydantoin (2 mg/day) prolonged mean life span by 23 and 25%, respectively, and decreased spontaneous tumour incidence by 4.0 and 2.3 times, respectively. The chronic treatment of mice with L-dopa (2 mg/day) did not change these parameters and decreased the multiplicity of mammary tumours. The mechanisms of the drug action on mouse life span and tumour incidence are discussed.

Effects of chronic guanethidine on the sympathetic nervous system of mouse and toad

1. Chronic treatment of mice and toads with low (5–10 mg/kg per day) or high (25–150 mg/kg per day) doses of guanethidine depleted neuronal catecholamine stores, reduced adrenergic nervemediated responses and produced supersensitivity to noradrenaline. 2. No ultrastructural changes were seen in toad adrenergic cell bodies and only minor changes were seen in mouse noradrenergic cell bodies after treatment with high doses. Cells in later stages of degeneration were not observed. 3. After cessation of treatment, catecholamine levels and nerve-mediated responses increased considerably in both species. 4. Thus, the cytotoxic effects of chronically administered guanethidine vary considerably between species. Widespread degeneration of adrenergic neurons has so far been observed only in rats.

Mutagenicity of DDT in mice, Drosophila melanogaster and Neurospora crassa.

The pesticide DDT was examined for possible mutagenicity in mice, D. melanogaster and N. crassa. Through the use of the dominant-lethal assay it was found that acute oral doses of DDT (2 x 150 mg/kg body weight) in male mice induced dominant lethal mutations in early spermatid and spermato-cyte stages. Chronic oral doses of DDT (2 x 100 mg/kg body weight per week for 10 weeks) in male mice caused a persistent increase in the number of dominant lethal mutations. Histological sections showed that chronic treatment of mice with DDT caused changes in seminiferous tubule morphology and degeneration of B-type spermatogonia. Acute treatment of mice with DDT caused an increase in spermatocyte chromosome breakage, stickiness and precocious separation of the X and Y bivalent.

The effect on the thyroid gland and the gonads following chronic treatment of mice with low doses of radioiodine (I131)

The effect on the thyroid gland and the gonads following chronic treatment of mice with low doses of radioiodine (I¹³¹)