Central and peripheral norepinephrine concentrations in rat strains selectively bred for differences in response to stress: confirmation and extension

Inbred rats from the Maudsley Reactive (MR) strain had lower concentrations of norepinephrine in hypothalamus, heart, and spleen, and lower total catecholamines in adrenal than inbred Maudsley Non-Reactive rats (MNRA line). In contrast, they had a higher concentration of telencephalic NE than MNRA rats. These results confirmed previous findings obtained on rats maintained by non-systematic breeding within the two lines. Comparisons were also made between MR and a second Maudsley Non-Reactive strain (MNR). Rats of the two Non-Reactive lines (MNRA, MNR) have been bred from the same foundation population and selected for the same behavioral characteristics, but have been genetically isolated from each other for many generations. It was found that MR rats showed differences from MNR rats in hypothalamic and peripheral (but not telencephalic) NE concentrations similar to those seen in MR/MNRA comparisons. Since rats of the two Non-Reactive lines differ appreciably from MR rats in open-field defecation (the criterion on which they were selected), their differences from MR rats in a neurochemical system involved in sympathetic function suggests that this system may be functionally related to well established behavioral and physiological differences between the lines.     

An examination of the effects of stimulant and depressant drugs on escape/avoidance conditioning in strains of rats selectively bred for emotionality/non-emotionality

Summary Two of the main hypotheses concerning the role of emotionality (as an excitatory state) in the determination of escape/avoidance conditioning are juxtaposed, and their explanatory force tested by the use of constitutionally differentiated levels of reactivity in rats and the interaction these constitutional states may have with different drugs (assumed to affect different neural structures) and different doses of such drugs. From an analysis of these main effects and their interactions a complex set of relationships emerge which suggest that all the variables employed, with the exception of sex, are likely to be relevant to a consideration of those factors subserving differences in conditioning and learning.This experiment is part of the thesis submitted to the University of London in 1967 in partial fulfilment of the degree of Ph.D. in Psychology under the guidance of Professor H. J. Eysenck and Dr. H. C. Holland.     

Differences in the hypothermic response to ethanol in rats selectively bred for oral ethanol preference and nonpreference

The duration of retention of tolerance to ethanol was tested in the alcohol-preferring (P) and alcohol-nonpreferring (NP) rats lines, using ethanol-induced hypothermia as a measure of tolerance. Rats received two injections of ethanol (3.5 g/kg) body wt, IP) and the time between the injections was 1, 2, or 3 days. When one day separated the two injections, tolerance to the hypothermic effect of a second “test” injection was found in both lines. When 2 or 3 days separated the two injections, the P line showed a loss of tolerance and the NP line showed sensitization to ethanol. Sensitization in the NP line grew stronger when the interval between injections was increased from 2 to 3 days. The duration of retention of tolerance to ethanol-induced hypothermia in the P line was shorter than has previously been reported for motor impairment in this line. It appears that the duration of tolerance retention in the P line depends on the test used to measure tolerance. Sensitization to ethanol in the NP line may be associated with low oral ethanol intake. This research was supported, in part, by grants AA08312, AA03243, and AA07611 from the PHS     

Exaggerated response to restraint stress in rats congenic for the chromosome 1 blood pressure quantitative trait locus

1. To understand the roles of a putative hypertension gene in the chromosome 1 quantitative trait locus (QTL) region, the response to restraint stress was studied in strains congenic for this QTL. 2. To establish congenic strains, the QTL region was introgressed from stroke-prone spontaneously hypertensive rats (SHRSP)/Izm to Wistar-Kyoto/Izm (WKY/Izm) rats by repeated backcrossing. Two congenic strains (WKYpch1.0 and WKYpch1.1) were established to cover the whole QTL region between D1Wox29 and D1Arb21 (approximately 40 cM) and a smaller region between D1Smu11 and D1Arb21 (approximately 10 cM), respectively. After telemetry probes were implanted, rats were exposed to restraint stress to investigate the blood pressure response. 3. Basal blood pressure measured by radiotelemetry differed significantly between WKY rats and WKYpch1.0 (103 +/- 10 and 116 +/- 4 mmHg, respectively; P = 0.002 by anova). When exposed to restraint stress, WKYpch1.0 showed a greater increase in blood pressre than did WKY rats. The exaggerated response in the WKYpch1.0 strain was abolished by chemical sympathectomy using guanethidine. The WKYpch1.1 rats did not differ significantly from WKY rats either in basal blood pressure or in the response to restraint stress. 4. In conclusion, a QTL for high blood pressure was successfully introgressed in the established congenic strain, WKYpch1.0. A gene (or genes) in the chromosome 1 QTL region modulates the cardiovascular responses to restraint stress in these congenic rats, probably through the sympathetic nervous system.     

Dose-response effects of alcohol upon rat strains bred for differences in reactivity to alcohol

Two rat strains, designated LA and MA, selectively bred for differential impairment of motor activity following an injection of alcohol, were tested in stabilmeters and compared over a range of ethanol doses. As expected, increasing doses of ethanol produced progressively greater activity decrements in both strains; however, the same dose of ethanol induced a more pronounced decrement in the MA strain than in LA strain at all doses. At the highest alcohol dose (2.25 g/kg), the LA animals were twice as active as were the MA strain at the 1.5 g/kg dose. This strain difference in impairment was evident within 3 min postinjection and remained throughout the 30 min test session. The results are discussed in terms of differential neural and behavioral toelrance to ethanol in the two strains.     

Digoxin increases peripheral chemosensitivity and the ventilatory response to exercise in normal subjects

1. The contribution of peripheral chemoreceptors to the regulation of ventilation during exercise remains incompletely understood. Digoxin has been reported to increase chemoreflex sensitivity in humans. In the present randomized, cross‐over, double‐blind study, we tested the hypothesis that this increases the ventilatory response to exercise in normal subjects, as assessed by changes in minute ventilation (V_E) in response to the rate of CO₂ production (Vco ₂). 2. Minute ventilation, end‐tidal Pco ₂, pulse oximetric O₂ saturation (S_po ₂), heart rate and blood pressure (BP) were measured in 11 healthy young male untrained subjects after intravenous infusion of digoxin (0.01 mg/kg) or placebo during normoxia, isocapnic hypoxia and hyperoxic hypercapnoea. All participants underwent a maximum cardiopulmonary exercise test. 3. During normoxia, digoxin increased systolic BP only. During hypoxia, digoxin increased V_E compared with placebo (P = 0.009) for the same fall in S_po ₂ (P = NS). Moreover, no significant effects on ventilation and haemodynamic responses were recorded during hypercapnoea. Digoxin increased the V_E /Vco ₂ slope above the anaerobic threshold from 30.4 ± 2.9 to 32.8 ± 3.7 (P < 0.05), but did not affect Vo _2max. 4. In conclusion, enhanced peripheral chemosensitivity with digoxin increases the ventilatory response to CO₂ production above the anaerobic threshold, but does not affect exercise capacity in healthy humans.     

Central and peripheral tissue distribution of diclofenac after subcutaneous injection in the rat

Nonsteroidal anti-inflammatory drugs (NSAIDs) are viewed as peripherally acting analgesics; however, evidence is accruing that they also act centrally. To help clarify the regional actions of diclofenac, we studied its plasma and tissue concentrations after subcutaneous injection in a model of acute pain, the rat tail ischaemia-reperfusion acute hyperalgesia model. After antinociceptive doses (20 and 40 mg kg^-1), diclofenac plasma C _max occurred at 20 min and neither plasma or tissue concentrations were affected by application of the pain model. Brain, spinal cord, heart, skeletal muscle, (injured) tail cartilaginous tissue, stomach and fat had similar distribution coefficients (0.05-0.2). Distribution coefficients for liver (0.5-1.4) and kidney (0.3-0.5) increased with time, presumably reflecting their roles in the elimination of diclofenac. Subcutaneous injection was found a pharmacokinetically reliable method for administering diclofenac for antinociception studies in the rat. The finding that CNS and peripheral tissue distribution coefficients of diclofenac were similar provides indirect support for a potential central action of NSAIDs.     

Dermatotoxicity of oral cadmium is strain-dependent and related to differences in skin stress response and inflammatory/immune activity

Adverse effects of non-occupational exposure to cadmium (Cd) are increasingly acknowledged. Since our previous study has showed that orally acquired Cd affects skin, the contribution of genetic background to dermatotoxicity of oral cadmium was examined in two rat strains, Albino Oxford (AO) and Dark Agouti (DA), which differed in response to chemicals. While similar accumulation of Cd in the skin of both strains was noted, the skin response to the metal differed. DA rat individuals mounted antioxidant enzyme defense in the skin already at lower Cd dose, in contrast to AO rats which reacted to higher metal dose solely (and less pronounced), implying higher susceptibility of DA strain to Cd dermatotoxicity. Epidermal cells from both strains developed stress response, but higher intensity of antioxidant response in AO rats implied this strain`s better ability to defend against Cd insult. Cd induced epidermal cells’ proinflammatory cytokine response only in DA rats. Increased IL-10 seems responsible for the lack of response in AO rats. Differences in the pattern of skin/epidermal cell responsiveness to cadmium give a new insight into repercussion of genetic variability to dermatotoxicity of orally acquired cadmium, bearing relevance for variations in the link between dietary cadmium and inflammation-based skin pathologies.     

Twenty years of dopamine research: individual differences in the response of accumbal dopamine to environmental and pharmacological challenges

Individual differences in the dopaminergic system of the nucleus accumbens of rats have extensively been reported. These individual differences have frequently been used to explain individual differences in response to environmental and pharmacological challenges. Remarkably, only little attention is paid to the factors that underlie these individual differences. This review gives an overview of the studies that have been performed in our institute during the last 20 years to investigate individual differences in accumbal dopamine release. Data are summarised demonstrating that individual differences in accumbal dopamine release are due to individual differences in: the functional reactivity of the noradrenergic system, the accumbal concentration of vesicular monoamine transporters and tyrosine hydroxylase as well as in the quantal size of the presynaptic pools of dopamine. Our data are embedded in the available literature to create a model that illustrates the putative hardware giving rise to the individual-specific release of accumbal dopamine. An important role is contributed to individual differences in the reactivity of the: hypothalamic–pituitary–adrenal axes, the reactivity of second messenger systems as well in the aminergic reactivity of the accumbens shell and core. The consequences of the individual-specific make-up and reactivity of the nucleus accumbens on the regulation of behaviour and the response to drugs of abuse will also be discussed. Apart from agents that interact with dopaminergic receptors, re-uptake or breakdown, noradrenergic agents as well as agents that interact with vesicular monoamine transporters or tyrosine hydroxylase are suggested to have therapeutic effects in subjects that are suffering from diseases in which the dopaminergic system is disturbed.     

Characterization of testosterone metabolism and 7-hydroxycoumarin conjugation by rat and human liver slices after storage in liquid nitrogen for 1 h up to 6 months

1. Slices of human and rat liver were cryopreserved in 18% dimethyl sulphoxide (DMSO) and subsequently stored in liquid nitrogen for periods up to as long as 6 months. After thawing, the metabolism of testosterone to hydroxylated products and conjugation of 7-hydroxycoumarin were investigated. 2. Rat liver slices stored in liquid nitrogen for 6 months exhibited rates of formation of 7α-, 6β- 16α- and 2β-hydroxytestosterone, and of androstenedione that did not differ significantly from those observed with fresh slices. 3. No formation of 2β-hydroxytestosterone was detected with slices of human liver. However, in contrast with the rat, human slices produced 2β-hydroxytestosterone. The rates of formation of 7α-, 6β-, 16α- and 2β-hydroxytestosterone and of androstenedione by human liver slices after 6 months of storage in liquid nitrogen were 82, 71, 236, 66 and 92%, respectively, of the corresponding rates by fresh slices. 4. The rates of sulphation and glucuronidation of 7-hydroxycoumarin by slices from rat liver were 97 and 119%, respectively, of the corresponding fresh values after 6 months of storage in liquid nitrogen. 5. 7-Hydroxycoumarin glucuronidation by human liver slices was 53% of the corresponding fresh values after 6 months of storage. However, human slices showed little or no capacity to conjugate 7-hydroxycoumarin with sulphate. 6. It was demonstrated that slices of both human and rat liver can be cryopreserved and stored in liquid nitrogen for at least 6 months without major changes in their rates of metabolism of testosterone to its hydroxylated products and of 7-hydroxycoumarin conjugation. These findings further emphasize that cryopreservation of liver slices can be an effective tool in the use of biological material of limited availability.     

Monoacylglycerol lipase inhibitors: modulators for lipid metabolism in cancer malignancy,neurological and metabolic disorders

Monoacylglycerol lipase (MAGL) is a serine hydrolase that plays a crucial role catalysing the hydrolysis of monoglycerides into glycerol and fatty acids. It links the endocannabinoid and eicosanoid systems together by degradation of the abundant endocannabinoid 2-arachidaoylglycerol into arachidonic acid, the precursor of prostaglandins and other inflammatory mediators. MAGL inhibitors have been considered as important agents in many therapeutic fields, including anti-nociceptive, anxiolytic, anti-inflammatory, and even anti-cancer. Currently, ABX-1431, a first-in-class inhibitor of MAGL, is entering clinical phase 2 studies for neurological disorders and other diseases. This review summarizes the diverse (patho)physiological roles of MAGL and will provide an overview on the development of MAGL inhibitors. Although a large number of MAGL inhibitors have been reported, novel inhibitors are still required, particularly reversible ones.