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Biomedical subjects

M Wilkinson

Publications and source records attributed to M Wilkinson.

At least 253 records · Page 14Linked to original sources

Role of the E receptor in interferon-gamma expression: sheep erythrocytes augment interferon-gamma production by human lymphocytes.

The E receptor is found on virtually all human T cells and is defined by its ability to bind to sheep erythrocytes (SRBCs). It was recently shown that a monoclonal antibody (MoAb) binding to the E receptor regulates interferon-gamma (IFN-gamma) production and lymphocyte activation. Here the effects of SRBCs on IFN production is examined since these bind to the E receptor. IFN-gamma production induced by phytohemagglutinin (PHA) in both fresh peripheral blood mononuclear leukocytes ( PBML ) and T-lymphocyte lines was augmented by 5- to 10-fold when cultured with SRBCs. The SRBCs increased IFN by interacting with the E receptor since its augmenting effect was blocked by the E-receptor MoAb 9.6. The augmenting effect of SRBCs was independent of blastogenic events since DNA synthesis was not similarly augmented by SRBCs and because IFN-gamma production was only increased after blastogenesis had occurred (in already differentiated lymphoblasts or in PBML 2-3 days after induction). Since it was found that only a short pulse of SRBCs was required for their action, this suggests that the studies employing SRBC rosetting as a fractionation technique for investigating the cell source of IFN-gamma should be viewed with caution.

Animals↗

Brain slices in radioligand binding assays: quantification of opiate, benzodiazepines and beta-adrenergic ([3H] CGP-12177) receptors.

We have characterized and quantified specific binding of [3H]-flunitrazepam (FNZ; (benzodiazepine), [3H]-naloxone (NAL; (opiate) and [3H] CGP-12177(CGP; (beta-adrenergic) to thick slices (230-400 micron) of mouse and rat brain. The binding sites are stereospecific, saturable and of high affinity. In all cases, the binding of the ligands is readily reversible and demonstrates the appropriate drug specificity. In mouse brain [3H]-NAL binding is elevated by chronic treatment with naloxone (via capsules). We have been unsuccessful in quantifying beta adrenoreceptors with the archetypal ligand [3H]-dihydroalprenolol (DHA). However, the use of [3H]-CGP 12177 enabled us to detect high-affinity beta adrenoreceptors in brain slices. [3H]-CGP also permits the demonstration of rapid and reversible agonist-induced down-regulation (internalization) of beta binding sites. We have been successful in quantifying beta adrenergic sites in single pineal glands of rat and hamster.

Animals↗

Alterations in receptor number, affinity and laminar distribution in cat visual cortex during the critical period.

The number, affinity, and laminar distributions of various receptors in cat visual cortex were examined during postnatal development using homogenate and in vitro autoradiographic techniques. For all receptor populations examined, the total number of receptors (Bmax) increased from relatively low early values to peak values during the first three months of postnatal life followed by a drop or plateau in the number of receptors. This peak in Bmax occurred during the physiologically-defined period for cortical plasticity. For most receptors examined, the affinity (KD) was also altered during postnatal development. Many of the receptor populations examined exhibited changes in their initial laminar distributions during the first three months of postnatal development, although other did not. The results show a more complex picture of receptor ontogenesis than previously reported, and suggest that the observed receptor modifications affect the synaptic efficacy and the basic chemical circuitry of the visual cortex during the critical period.

Animals↗

Opiate ([3H]-naloxone) binding to hypothalamic and cerebral cortical slices of mouse brain.

We have characterized the binding of [3H]-naloxone to thick (400 microns) slices of hypothalamus and cerebral cortex from mouse brain. Binding is reversible, saturable, stereospecific, thermolabile, readily displaceable by opiates and sensitive to phenoxybenzamine and phentolamine. Values for KD and Bmax are very close to published figures obtained in brain homogenates. Metabolic inhibitors (ouabain and azide) have no effect on specific binding. The assay is rapid, simple and involves minimal tissue preparation.

Animals↗

Benzodiazepine ([3H])-flunitrazepam) binding sites in cerebellar and cerebral cortical slices of mouse brain.

We have characterized and quantified the specific binding of [3H]-flunitrazepam ( FNZ ) to thick (230 micron) slices of mouse brain. The binding site has the characteristics of a benzodiazepine receptor, i.e., binding of FNZ is reversible, stereospecific, saturable and of high affinity. Clonazepam, but not R05 -4864, readily displaces the label. In contrast to results from homogenate assays, neither GABA nor bicuculline has any effect on [3H]- FNZ binding. However, as previously reported, the slice assay confirms the lower number of benzodiazepine receptors in "emotional" mouse brain. In addition, we have confirmed that the neurotoxin DSP4 can modify [3H]- FNZ binding though in our hands this compound elevates rather than reduces binding. The speed, simplicity and minimal tissue preparation involved suggests that this slice assay could be a valuable addition to neurochemical studies of neurotransmitter receptors.

Animals↗

Periventricular and suprachiasmatic lesion effects on photoperiodic responses of the hamster hypophyseal-gonadal axis.

We examined the involvement of neural mechanisms within the suprachiasmatic nucleus (SCN) and periventricular area (PVA), and the role of prolactin (Prl) in control of endocrine function in short day-exposed Syrian hamsters. Hamsters bearing lesions of the SCN or PVA, hamsters implanted with an anterior pituitary under the kidney capsule to provide sustained Prl levels, and sham-operated hamsters were exposed to either 14L:10D or 8L:16D. After 9 wk, hamsters were sacrificed, and their testes and pituitaries were studied in vitro to assess their secretory capacity. SCN lesions and large periventricular lesions impinging on the paraventricular nucleus prevented testicular regression during short-day exposure. Small periventricular lesions and pituitary implants did not prevent gonadal regression in hamsters exposed to short days. Testis weights were positively correlated with basal and luteinizing hormone (LH)-stimulated androgen production in the control and lesioned groups; pituitary implants prevented the decline in androgen production in vitro in gonadally regressed animals. The relative in vitro pituitary response to gonadotropin-releasing hormone (GnRH) stimulation in control and lesioned groups was not reduced by short-day exposure. These data indicate that either axons coursing dorsally from the SCN or extra-SCN structures in the periventricular/paraventricular area are necessary for testicular regression in short photoperiods.

Androgens↗

Chronic treatment with [D-Ala6, des Gly-NH2(10)]-LHRH ethylamide reversibly delays puberty in the female rat.

Luteinizing hormone-releasing hormone (LHRH) agonist analogs have been suggested as a useful treatment for precocious puberty, though there is some concern that long-term treatment might be deleterious to normal sexual development. We have taken advantage of the very short maturation period of the female rat (approximately 35 days from birth) to examine the effects of chronic (daily) treatment with [D-Ala6, des Gly-NH2(10)]-LHRH ethylamide. We have observed that this treatment (either 1 or 2 micrograms/day) from day 5 after birth significantly delays sexual maturation but does not affect subsequent sexual cycles.

Animals↗

Short-day exposure eliminates the LH response to naloxone in golden hamsters.

Short-day-induced testicular regression in golden hamsters is accompanied by changes in opiatergic control of LH release. Serum LH in long-day-exposed hamsters is elevated by subcutaneous injections of naloxone. By day 42 of short-day exposure, naloxone is unable to stimulate LH secretion; responsiveness to naloxone is restored after reexposure to long days. The change in response to naloxone may reflect a loss of opiate regulation of LH release; an alternative, but not mutually exclusive, hypothesis is that additional opiate-independent mechanisms inhibit LH despite the blockade of opiate LH inhibition by naloxone.

Animals↗

Ergotamine toxicity and serum concentrations of ergotamine in migraine patients.

Twenty-five migraine patients (9 males and 16 females) aged 22-71 who had used between 7 and 60 mg ergotamine tartrate per week for 1.5-30 y volunteered to participate in the study. Side-effects attributable to ergotamine were wide ranging and included daily headache and pain in the limbs. Wide variation in sensitivity to the drug was observed and side-effects were not always proportional to the dose of ergotamine. Random serum ergotamine concentrations were estimated in all patients and 10 of them volunteered to take a 2 mg oral challenge of ergotamine tartrate. Ergotamine was not detected in 44% of the random estimations even though all patients exhibited clinical signs of ergotamine tartrate overdose. The remaining 56% estimations all showed low drug concentrations. After the 2 mg oral challenge of ergotamine in the 10 patients, significantly higher concentrations, but still within therapeutic range, were detected in the sera when compared with mean concentrations achieved after the same oral dose of the drug in healthy, non-migrainous subjects.

Adult↗

Prolonged elevation of hypothalamic opioid peptide activity in women taking oral contraceptives.

To examine the hypothesis that endogenous opioid peptide activity is chronically elevated by oral contraceptives, we infused either naloxone or saline into 10 women during the use of, or 5-6 and 9-10 days after stopping, combination birth control pills. A paradoxical increase in prolactin occurred with naloxone infusion during and 5-6 days after stopping the pills. Serum LH levels were not significantly elevated by naloxone until 9-10 days after cessation of pill use. These results suggest that hypothalamic opioid peptide activity is continuously elevated in women taking oral contraceptives.

Adult↗

Abnormal gonadotrophin release from pituitaries of muscular dystrophic mice and hamsters.

LHRH-stimulated LH and FSH secretion was studied in hemipituitaries, in vitro, obtained from several dystrophic mouse mutants (male: 129/ReJ-dy; 129B6F1/J-dy; C57BL/6J-dy and C57BL/6J-dy2J; female: 129B6F1/J-dy) and a dystrophic hamster mutant (male and female CHF-147). Without exception, pituitary tissue from dystrophic animals released significantly more FSH than did tissue obtained from controls. LH secretion was more variable; in the male mice released was inhibited, whereas in the male dystrophic hamsters secretion was elevated above normal. The female mouse mutant pituitary released more LH whereas in the female hamster LH secretion was normal. The reduction in body weight of the mutants studied could have contributed to the observations of impaired anterior pituitary function.

Animals↗

The inhibitory effect of opiates on gonadotrophin secretion is dependent upon gonadal steroids.

We have attempted to clarify the physiological involvement of endogenous opiates in the steroid-mediated control of gonadotrophin release. Our studies showed that there was an acute reduction in the inhibitory effects of endogenous opiates on LH and FSH release following gonadectomy in the rat. This was indicated by a significant reduction in the ability of naloxone to stimulate serum LH/FSH levels (sampled at 15 min) in 26-day-old female rats 48 h after ovariectomy. Luteinizing hormone was highly sensitive to the inhibitory effects of the synthetic met-enkephalin analogue, FK 33-824, at this time (sampled at 90 min). An unexpected observation was that long-term absence of gonadal steroids also disrupted the ability of exogenous opiates, FK 33-824 and morphine, to influence LH release. This was seen as an inability of FK 33-824 (1.0 or 3.0 mg/kg) to inhibit LH secretion. The effects of gonadectomy on opiate control of LH occurred at all developmental stages and were not due to a disruption of sexual maturation. Opiate involvement in prolactin secretion did not appear to be adversely affected by an absence of gonadal steroids. Another novel aspect of this work was that the opiatergic component in the control of gonadotrophin secretion could be reinstated in long-term gonadectomized rats by treatment with oestradiol benzoate or testosterone propionate. Similarly, priming with increasing dosages of oestradiol benzoate which resulted in progressively lower LH levels gave larger naloxone in progressively lower LH levels gave larger naloxone responses.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Interferon with novel characteristics produced by human mononuclear leukocytes.

Treatment of human peripheral blood mononuclear leukocytes with phytohaemmaglutinin and the tumour promoter teleocidin, results in the production of large amounts of interferon-gamma and significant amounts of a novel interferon-like substance which we tentatively class as interferon-delta. This novel interferon type possesses all the important characteristics of classical interferon but, of various cell types tested, has antiviral activity only in trisomy-21 human fibroblasts. It differs decisively from previously identified interferon types in its antigenic, biological and physicochemical properties.

Animals↗

Strain differences in brain alpha2- and beta-adrenergic receptor binding in dystrophic mice.

We have examined alpha 2-adrenergic [( 3H]-clonidine) and beta-adrenergic [( 3H]-dihydroalprenolol; DHA) binding in brain membranes from four different dystrophic mouse mutants: 129/ReJ-dy; 129 B6F1/J-dy; C57BL/6J-dy and C57BL/6J-dy2J. As shown previously, male 129/ReJ-dy mice have significantly fewer brain [3H]-DHA binding sites than their littermate controls. In marked contrast, the remaining three mutants showed no differences in [3H]-DHA binding when compared to their respective controls although there were inter-strain differences, e.g., C57BL/6J-dy2J mice have significantly fewer binding sites than the 129B6F1/J-dy mice. On the other hand, we saw no strain differences in alpha 2-binding. In one mutant, however, (129 B6F1/J-dy) we observed a 50% increase in alpha 2-binding sites compared to littermate controls. We conclude that weight loss or undernutrition may be an important factor in determining brain levels of adrenergic binding sites in dystrophic mouse brain.

Animals↗

alpha- and beta-adrenergic binding sites in sheep cerebral cortex: characterisation, effects of photoperiod and treatment with estrogen/progesterone.

The radioligands [3H]-dihydroergocryptine and [3H]-dihydroalprenolol were used to characterise alpha- and beta-adrenergic binding sites, respectively, in membrane fractions of sheep cerebral cortex. In terms of affinity, density and specificity these sites possess properties similar to those previously characterised in rat brain. Further, in preliminary studies, these sites also appear to be responsive to treatment with estradiol/progesterone as well as to photoperiod. Thus, estrogen treatment can elevate both alpha- and beta-adrenergic binding sites in cortical tissue of sheep kept in natural light. In contrast, artificial light either has no effect or inhibits binding in response to estrogen.

Animals↗

Benzodiazepine receptors in fish brain: [3H]-flunitrazepam binding and modulatory effects of GABA in rainbow trout.

We have identified and partially characterised benzodiazepine binding sites in whole brain membranes of male rainbow trout. In terms of Bmax and KD values trout brain receptors are remarkably similar to those in rat and human brain. The Hill coefficient was 0.98, indicating a single binding site. GABA (10(-4) M) was able to significantly elevate binding of [3H]-FNZ through a change in KD rather than Bmax. This effect was prevented by the GABA receptor antagonist bicuculline methiodide.

Animals↗