Biomedical subjects

G M Brown

Publications and source records attributed to G M Brown.

At least 73 records · Page 4Linked to original sources

PubMed1996

The hormonal response to intravenous 5-hydroxytryptophan in bulimia nervosa.

In recent years evidence has accumulated to implicate a disturbance in serotonin function in the eating disorder bulimia nervosa. This study employs a neuroendocrine technique to assess the effect of intravenous 5-hydroxytryptophan, the immediate precursor to serotonin, on peripheral hormones in bulimia nervosa subjects and controls. Blunted prolactin and growth hormone responses were observed among bulimia nervosa subjects; the possible pathophysiology and implications of the findings are discussed.

5-Hydroxytryptophan↗

PubMed1996

2-[125I]Iodomelatonin binding sites in the quail heart: characteristics, distribution and modulation by guanine nucleotides and cations.

To investigate whether melatonin has a direct action on the cardiovascular system, putative melatonin receptors were studied in quail heart membrane preparations using the specific melatonin agonist 2-[125I]iodomelatonin (125I]Mel, as the radioligand. The [125I]mel binding demonstrated in the mature quail heart was saturable, highly 5.2 pM; Bmax = 1.32 +/- 0.25 fmol/mg protein; n = 8). The linear Scatchard plots and the close to unity Hill coefficient indicated a single class of binding sites. The pharmacological profile was in the affinity order of 2-iodomelatonin = 2-phenylmelatonin > melatonin > 6-chloromelatonin >> 6-hydroxymelatonin > 6-sulphatoxymelatonin >> N-acetylserotonin>>>5-hydroxytryptamine. Guanosine 5'-triphosphate and guanosine 5'-O-(3-thiotriphosphate) (GTP gammaS) dose dependently inhibited the binding. Ten microM GTPgammaS lowered the binding affinity by 50% in saturation studies. The order of potency of inhibition by cations was: Ca2+ > Mg2+ > Li+ > Na+ > K+ > choline chloride. Contrary to most other melatonin binding sites, millimolar concentrations of Ca2+ and Mg2+ did not promote binding in the quail heart membranes. In vitro autoradiography indicated homogenous labeling in the heart. Our results demonstrated [125I]Mel binding sites in the quail heart. That guanine nucleotides and Na+ inhibited the binding indicated that these putative melatonin receptors are coupled to guanine nucleotide-binding proteins (G-proteins).

Animals↗

PubMed1996

The Drosophila developmental gene fat facets has a human homologue in Xp11.4 which escapes X-inactivation and has related sequences on Yq11.2.

EST 221 derived from human adult testis detects homology to the Drosophila fat facets gene (fat) and has related sequences on both the X and Y chromosomes mapping to Xp11.4 and Yq11.2 respectively. These two loci have been termed DFFRX and DFFRY for Drosophila fat facets related X and Y. The major transcript detected by EST 221 is-8 kb in size and is expressed widely in a range of 16 human adult tissues. RT-PCR analysis of 13 different human embryonic tissues with primers specific for the X and Y sequences demonstrates that both loci are expressed in developing tissues and quantitative RT-PCR of lymphoblastoid cell lines carrying different numbers of X chromosomes reveals that the X-linked gene escapes X-inactivation. The amino acid sequence (2547 residues) of the complete open reading frame of the X gene has 44% identity and 88% similarity to the Drosophila sequence and contains the conserved Cys and His domains characteristic of deubiquitinating enzymes, suggesting its biochemical function may be the hydrolysis of ubiquitin from protein-ubiquitin conjugates. The requirement of faf for normal oocyte development in Drosophila combined with the map location and escape from X-inactivation of DFFRX raises the possibility that the human homologue plays a role in the defects of oocyte proliferation and subsequent gonadal degeneration found in Turner syndrome.

Adult↗

PubMed1996

Melatonin receptors in the chicken kidney are up-regulated by pinealectomy and linked to adenylate cyclase.

The effect of pinealectomy on the characteristics of melatonin receptors in the chicken kidney was studied. One-day-old chicks were operated and kept under a 12 h/12 h light/dark photoperiod. Six weeks after operation, the animals were sacrificed at mid-light and mid-dark. Serum melatonin was determined by radioimmunoassay and kidney melatonin receptors were studied by radioreceptor assay using the melatonin agonist 2-[125I]iodomelatonin as the radioligand. Pinealectomy significantly reduced the mid-dark serum melatonin level and abolished the diurnal rhythm of 2-[125I]-iodomelatonin binding in the kidney. The density of 2-[125I]-Iodomelatonin binding sites in the kidney at mid-dark was increased significantly to a value comparable to the mid-light density after pineal ablation. Our results suggest that melatonin receptors in the chicken kidney are directly regulated by melatonin in the circulation. The coupling of kidney melatonin receptors to adenylate cyclase was investigated. The basal and forskolin-stimulated cAMP production in chicken kidney explants was studied following melatonin or melatonin plus pertussis toxin treatment. Levels of cAMP in chicken kidney explants were extracted and determined by radioimmunoassay. Melatonin had no effect on basal cAMP levels. However, melatonin significantly inhibited the forskolin-stimulated cAMP accumulation at a concentration of 10 pmol/l. Inhibitory effects of melatonin on the forskolin-stimulated cAMP increase in the chicken kidney were totally blocked by preincubating the kidney tissue with 1.0 micrograms/ml pertussis toxin. Our results suggest that kidney melatonin receptors may modulate the adenylate cyclase leading to biological responses in the renal system.

Adenylate Cyclase Toxin↗

PubMed1995

Molecular and functional characterization of a partial cDNA encoding a novel chicken brain melatonin receptor.

An approach based on homology probing was used to clone a partial cDNA encoding a novel melatonin (ML) receptor (MLR) from chicken (Gallus domesticus) brain. Based on available deduced amino-acid sequence, the chicken MLR (cMLR) displayed greater sequence homology to the frog (Xenopus) MLR than cloned human/mammalian receptors, with overall identities of 73% and 66%, respectively. In order to gain functional expression, a chimeric frog/chicken (flc)MLR was constructed in which the 5' end of the cMLR, including the N-terminus, TM1 and part of the first intracellular loop was substituted by fMLR sequence. [125I]Iodo-ML bound with high affinity (Kd of approximately 35 pM) to COS-7 cells transiently expressing the flcMLR in a saturable and guanine nucleotide-sensitive manner with the following rank order of potency: 2-iodo-ML > ML > 6-Cl-ML > S20750 > 6-OH-ML > S20642 > S20753 > N-acetyl-5HT >> 5-HT. Estimated Ki values for these compounds at the flcMLR correlated well to those obtained in native chicken brain membranes. In line with the observed structural similarity to the fMLR, the flcMLR exhibited affinities for ML, 6-Cl-ML and 6-OH-ML approximately 10-fold lower than mammalian receptors. Functionally, opposing interactions between ML and dopamine receptor signal transduction pathways were observed with ML potently inhibiting dopamine D1A-receptor-mediated cAMP accumulation in cells (HEK-293) transiently co-expressing these receptors. cMLR mRNAs were found expressed in chicken brain and kidney with trace levels observed in the lung. The availability of cloned vertebrate MLRs distinct at both the amino acid and pharmacological level from their mammalian counterparts may now allow for the identification of those amino-acid residues and structural motifs that regulate ML-binding specificity and affinity.

Adenylyl Cyclase Inhibitors↗

PubMed1995

Functional role of the prefrontal cortex in retrieval of memories: a PET study.

Retrieval of information from episodic memory involves the processes invoked by the attempt to remember (retrieval attempt) as well as processes associated with the successful retrieval of stored information (ecphory). Previous PET studies of memory have shown an activation of the prefrontal cortex in memory retrieval tasks, and we hypothesised that this activation represents retrieval attempt, not ecphory. This hypothesis was directly directed using [15O]H2 PET imaging in 19 healthy subjects who performed three matched tasks which involved different levels of retrieval attempt and ecphory. The results showed that retrieval attempt was associated with activation of the prefrontal cortex, right greater than left, while ecphory involved the posterior cortical regions. These findings illuminate the functional role of the different neuroanatomical regions involved in episodic remembering.

Adult↗

PubMed1995

Comparison of [125I]iodomelatonin binding sites in infant cerebellum of sudden infant death syndrome and non-sudden infant death syndrome.

[125I]Iodomelatonin bindings sites in human infant cerebellum were studied by radioligand receptor binding assay and in vitro quantitative autoradiography. The binding sites characterized in membrane preparations revealed saturable, reversible and highly specific binding sites with a Kd value of 21.2 +/- 8.33 pM and a Bmax value of 2.02 +/- 0.52 fmol/mg protein. 10 microM of GTP gamma S significantly reduced the binding capacity, suggesting the possible G-protein coupling of the binding sites. Autoradiographic study showed that the labelling was mainly located in the cerebellar cortex. On comparison of the binding parameters from cerebellum of sudden infant death syndrome (SIDS) and non-SIDS infants, no significant change in binding capacity and binding affinity was detected. These findings suggest that the binding sites in cerebellum may not be related to the etiology of SIDS.

Analysis of Variance↗

PubMed1995

13C-NMR spectroscopy of keratan sulphates. Assignments for four poly(N-acetyllactosamine)-repeat-sequence tetrasaccharides derived from bovine articular cartilage keratan sulphate by keratanase II digestion.

Skeletal keratan sulphate has been fragmented using the enzyme keratanase II, and the complete 13C chemical shift data are reported for the four tetrasaccharides which derived from the repeat region. They have the structures: Gal beta(1-4)GlcNAc(6S)beta(1-3)Gal beta(1-4)GlcNAc(6S)-ol, Gal(6S)beta(1-4)GlcNAc(6S)beta(1-3)Gal(1-4)GlcNAc(6S)-ol, Gal beta(1-4)GlcNAc(6S)beta(1-3)Gal(6S)beta(1-4)GlcNAc(6S)-ol, and Gal(6S)beta(1-4)GlcNAc(6S)(1-3)Gal(6S)beta(1-4)GlcNAc(6S)-ol, where GlcNAc(6S)-ol represents N-acetyl-glucosaminitol-6-O-sulphate. The value of these data for microstructural analysis of undegraded keratan sulphate samples using 13C-NMR spectroscopy is discussed with respect to galactose sulphation levels and the block sulphation structure.

Animals↗

PubMed1995

Keratanase digestion of keratan sulphates: characterization of large oligosaccharides from the N-acetyllactosamine repeat sequence and from the non-reducing terminal chain caps.

Keratan sulfate (KS) chains prepared from both bovine tracheal rings and bovine femoral head cartilage were digested with the enzyme keratanase from Pseudomonas species; large repeat-sequence and non-reducing terminal oligosaccharides were fractionated and purified using high-performance ion-exchange chromatography. The main beta-linked pentasulfated hexasaccharide repeat segment, [R6], GlcNAc(6S)1-1-3Gal(6S)1-4GlcNAc(6S)1-3Gal(6S)1-4GlcNAc(6S)1-3Gal-ol and the asialo beta-linked capping pentasulfated heptasaccharide, [C7], Gal1-4GlcNAc(6S)1-3Gal(6S)1-4GlcNAc(6S)1-3Gal(6S)1-4GlcNAc(6S) 1-3Gal-ol have been completely characterized by high-field NMR spectroscopy using one- and two-dimensional methods. Partial 1H assignments are summarized for three homologous series of higher oligosaccharides: GlcNAc(6S)[1-3Gal(6S)1-4GlcNAc(6S)]2-5(1-3)Gal-0l [R8,R10,R12] Gal1-4GlcNAc(6S)[1-3Gal(6S)1-4GlcNAc(6S)]3-5(1-3)Gal-ol [C9,C11,C13] NeuAc alpha 2-3Gal1-4GlcNAc(6S)[1-3Gal(6S)1-4GlcNAc(6S)]2-4(1-3)Gal-ol [C8,C10,C12] obtained from keratan sulfate by keratanase cleavage. The first shows that the unsulfated galactose residues within the repeat sequence region of KS may be separated by fully sulfated segments which have a wide distribution of lengths. The others, viz. those with sialylated caps, and the related galactose capped asialo-segments (derived from a KS digestion in which the keratanase also exhibited sialidase activity) represent an homologous series of epitopes in which the first internal unsulfated galactose is located at a position which may be up to five or more fully sulfated N-acetyllactosamine disaccharide repeat units along from the non-reducing terminus of the KS polymer.

Animals↗

PubMed1995

Spectroscopic characterisation of disaccharides derived from keratan sulfates.

Skeletal keratan sulfates have been degraded by three independent techniques and the resultant, borohydride-reduced, disaccharides have been characterised by NMR spectroscopy. The 1H and 13C (where available) chemical shifts are reported for the following substances, where GalNAc-ol represents N-acetyl-galactosaminitol, GlcNAc-ol represents N-acetyl-glucosaminitol, GlcNAc(6S)-ol represents N-acetyl-glucosaminitol 6-O-sulfate and 2,5AnMan(6S)-ol represents 2,5-anhydro-D-mannitol 6-O-sulfate. (a) GlcNAc(6S)beta(1-3)Gal-ol, isolated after keratanase (from Pseudomonas sp.) digestion. (b) Gal beta(1-4)GlcNAc(6S)-ol and Gal(6S)beta(1-4) GlcNAc(6S)-ol, the 1H chemical shifts have been reported previously [Brown, G. M., Huckerby, T. N., Morris, H. G., Abram, B. L. & Nieduszynski, I. A. (1994) Biochemistry 33, 4836-4846; Brown, G. M., Huckerby, T. N. & Nieduszynski, I. A. (1994) Eur. J. Biochem. 224, 281-308], GlcNAc(6S)beta(1-6)GalNAc-ol, [formula: see text], [formula: see text], all isolated after keratanase II digestion. (c) Gal beta(1-4)2,5AnMan(6S)-ol and Gal(6S)beta(1-4)2,5AnMan(6S)-ol, isolated after hydrazinolysis and nitrous acid digestion. In addition, the model compounds Gal beta(1-4)GlcNAc-ol and Gal beta(1-6)GlcNAc-ol have also been examined by 1H and 13C NMR spectroscopy. The value of these data for microstructural analysis of keratan sulfate samples is discussed.

Animals↗

PubMed1995

Effects of exogenous arginine vasotocin on circulating levels of thyroid hormones and melatonin in the pigeon, Columbia livia.

The effects of intravenous injections of two different preparations of synthetic arginine vasotocin (AVT), deamino-dicarba-arginine vasotocin and arginine vasotocin diacetate tetrahydrate on serum levels of thyroxine (T4), triiodothyronine (T3) and melatonin were studied in the resting pigeon. Although variations depending on factors such as the type of AVT preparation and the dose used and also the duration of treatment were apparent, both preparations of AVT produced an increase in serum levels of T4 and a decrease in that of T3. The T3:T4 ratio was also significantly lower in response to AVT administration. However, no statistically significant changes in serum levels of melatonin were discernible with the administration of either of the two AVT preparations, even though melatonin levels in most AVT-treated groups showed a trend (P > 0.05) to be higher than that in the controls. The rise in T4 levels is indicative of increased release of T4 and/or the inhibition of its conversion to the active hormone, T3. The drop in T3 may account for the AVT-induced reduction in the metabolic rate of the pigeon observed in an earlier study. The absence of a significant increase in melatonin indicates that AVT could not produce such a change in the daytime in resting pigeons, in contrast to that in flown homing pigeons, suggesting that the modulating effect of AVT on melatonin levels is dependent on the physiological state of the animal.

Animals↗

PubMed1995

Skeletal keratan sulphate structural analysis using keratanase II digestion followed by high-performance anion-exchange chromatography.

A semi-quantitative fingerprinting method has been developed for the structural analysis of skeletal keratan sulphates. This involves the digestion of the parent keratan sulphate chains with the enzyme keratanase II (Bacillus sp.), followed by reduction of the resulting oligosaccharides with sodium borohydride and chromatography on a Dionex AS4A-SC column. This column has been calibrated for the elution positions of 26 previously characterized oligosaccharides (Brown et al., Biochemistry, 33, 4836-4846, 1994; Brown et al., Eur. J. Biochem., 224, 281-308, 1994). The technique permits sample analysis with pulsed electrochemical detection (sensitive to approximately 5 ng of oligosaccharide) or by monitoring [3H] or [35S] radiolabel (potentially sensitive to approximately 1 pg or less of an oligosaccharide); thus permitting the study of sub-microgram amounts of keratan sulphates. Skeletal keratan sulphates from a number of sources have been examined in this chromatographic system and their structural features are discussed.

Acetylglucosaminidase↗

PubMed1995

Melatonin and steroids in human pre-ovulatory follicular fluid: seasonal variations and granulosa cell steroid production.

Follicular fluid samples were obtained from the largest pre-ovulatory follicle of 120 women undergoing in-vitro fertilization and were examined for melatonin by enzyme-linked immunosorbent assay and the steroids oestradiol and progesterone by radioimmunoassay. The concentrations (mean +/- SE) of melatonin (213.4 +/- 18.9 pmol/l) and progesterone (20.1 +/- 1.1 mumol/l) in follicular fluid during the autumn and winter (dark) months were significantly higher than during the spring and summer (light) months, melatonin (138.4 +/- 12.5 pmol/l) and progesterone (11.6 +/- 0.8 mumol/l). By contrast, oestradiol concentrations were significantly lower during the dark months than during the light months (264.7 +/- 44.1 and 661.8 +/- 55.1 nmol/l respectively). There was a positive correlation between follicular fluid melatonin and progesterone concentrations (r = 0.271, P < 0.05, n = 120) and a negative relationship between melatonin and oestradiol (r = -0.254, P < 0.05, n = 120). The effects of melatonin alone and in combination with human chorionic gonadotrophin (HCG) or follicle stimulating hormone (FSH) on steroidogenesis by human granulosa cell culture were also investigated. Melatonin had minimal effects on oestradiol or progesterone production by granulosa cells. Interestingly, the oestradiol response in culture appeared to be different according to the time of the year when harvested. During the light period oestradiol production was enhanced. Melatonin also synergized with HCG in increasing progesterone production on days 6 and 7 after treatment during both light and dark periods. FSH stimulated oestradiol production by the cells on day 2 of culture. Melatonin had no effect on FSH stimulation of oestradiol production. The results of this study suggest that melatonin may be involved in the regulation of steroidogenesis by the human ovaries.

Cells, Cultured↗

PubMed1995

Responsiveness of Keitel functional index compared with laboratory measures of disease activity in rheumatoid arthritis.

This study compares functional changes to change in measures of disease activity following the introduction of slow-acting anti-rheumatic drugs (SAARD) in patients with active rheumatoid arthritis (RA). Clinical and laboratory variables were simultaneously monitored at 6-monthly intervals, over approximately 18 months. Function was measured by a performance testing, the Keitel function index (KFI), which was divided into sections representing small and large joints [hand (HFI); wrist (WFI) and limb function index (LFI)]. One-hundred-and-fifteen patients were studied, of whom 21 were male. The mean age of the subjects was 49 yr (S.D. +/- 12) and mean duration of disease 7 yr (S.D. +/- 7). The mean KFI at entry was 38 (S.D. +/- 18) while at the end of the study it was 31 (S.D. +/- 17) (P < 0.0001). The change in KFI following therapy correlated with the change in Ritchie articular index (RAI) (r = 0.4; P < 0.0001), early morning stiffness (EMS) (r = 0.3; P = 0.004), swollen joint count (JC) (r = 0.4; P = 0.0005), C-reactive protein (CRP) (r = 0.2; P < 0.05) and Lansbury systemic index (LSI) (r = 0.35; P = 0.002), but not with change in Westergren erythrocyte sedimentation rate (ESR) or change in time to onset of fatigue. Multiple regression analysis showed that 32% of the variation in KFI at the end of the study could be predicted by a combination of ESR, sulphasalazine therapy, RAI, disease duration and chloroquine treatment at onset (P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Anti-Inflammatory Agents↗

PubMed1995

Neutrophil sequestration in rat lungs.

BACKGROUND: The transit of neutrophils through the pulmonary microvasculature is prolonged compared with red blood cells and is increased further during cigarette smoking and in exacerbations of chronic obstructive pulmonary disease. The increased residence time (sequestration) of neutrophils in the pulmonary capillaries in these conditions may be the first step leading to the accumulation of cells within the lung interstitium and in the bronchoalveolar space, so potentiating lung damage. A rat model has been developed to investigate the factors which may influence neutrophil transit through the lung microvasculature. METHODS: Intratracheal instillation of the heat killed organism Corynebacterium parvum was used to induce an acute neutrophil alveolitis. Neutrophils and red blood cells were isolated from donor rats, labelled with two distinct radioisotopes, and then reinjected into recipient rats to assess their transit through the pulmonary circulation. To ascertain whether peripheral blood neutrophils were minimally altered by the isolation procedure their functional status in vitro was compared with that of inflammatory neutrophils in a number of assays commonly used as descriptors of neutrophil activation. The influence of neutrophil activation on the accumulation of cells in the lungs was assessed by comparing the lung sequestration of control neutrophils, isolated from peripheral blood, with that of inflammatory neutrophils obtained from bronchoalveolar lavage of inflamed rat lungs. Lung sequestration of neutrophils was defined as the fold increase in the ratio of neutrophils labelled with chromium-51 to red blood cells labelled with technetium-99m in lung tissue compared with the same ratio in peripheral blood. RESULTS: Sequestration of peripheral blood neutrophils occurred in control rat lungs as shown by a 17.5 (2.1) fold increase in the ratio of neutrophils to red blood cells in the pulmonary circulation compared with the ratio of these cells in the peripheral circulation. When inflammatory neutrophils, obtained by bronchoalveolar lavage from C parvum-treated animals, were injected into control rats, the increase was 90.6 (11.0) fold. Induction of an inflammatory response in the lung tissue of the recipient rat also caused an increase in the sequestration of control neutrophils compared with the same cells in control rat lungs which was, however, less marked than when inflammatory neutrophils were used (34.7 (4.7) fold). The mean (SE) pressure developed on filtration of inflammatory neutrophils in vitro through a millipore filter (7.53 (0.2) cm H2O) was greater than that of peripheral blood neutrophils (1.18 (0.2) cm H2O). Increased filtration pressure indicates a decrease in cell deformability and suggests that this may be a contributory factor to the increased sequestration of inflammatory neutrophils in the pulmonary vasculature. CONCLUSIONS: This study shows that there is sequestration of neutrophils in the pulmonary vasculature in normal rat lungs which increases in acute lung inflammation and when inflammatory neutrophils are injected into control animals. In this model changes in the neutrophil, such as cell deformability, may have a more important role in inducing increased neutrophil sequestration than the inflammatory response in the lungs.

Animals↗

PubMed1995

Ontogeny of 2-[125I]iodomelatonin binding sites in the chicken (Gallus domesticus) kidney and spleen.

To understand the possible role of melatonin receptors in the development of renal and immune functions, age-related variations of 2-[125I]iodomelatonin binding sites in the chicken kidney and spleen were investigated by radioreceptor assay. Chickens at embryonic day 20, as well as 2 days, 9 days, 2 weeks, 6 weeks, 12 weeks, and 16 weeks after hatching, were kept under a 12 h light:12 h dark photoperiod and killed at the middle of the light period. Binding sites for 2-[125I]iodomelatonin in membrane preparations of the chicken kidney and spleen were present on embryonic day 20. The maximum binding densities (Bmax) in the kidney increased to a peak between 9 days and 2 weeks of age, then progressively decreased. Bmax values of 2-[125I]iodomelatonin binding sites in the chicken spleen were lower than in the kidney. The peak density in the chicken spleen was recorded at day 2 after hatching and decreased significantly after 6 weeks of age. There were no significant differences in binding affinities (Kd) in kidney and spleen of chicken in the different age groups studied. The unity of Hill coefficients of 2-[125I]iodomelatonin binding sites of the chicken kidney and spleen in all age groups tested suggested that only a single class of binding sites was present in these tissues during development. It is proposed that the developmental changes in 2-[125I]iodomelatonin binding sites in the chicken kidney and spleen may be pertinent to the development of diurnal rhythms of kidney functions and the post-pubertal decline in immune functions of the chicken.

Age Factors↗

PubMed1995

Effects of sex hormones on the pineal response to isoproterenol and on pineal beta-adrenergic receptors.

Several studies have shown sex hormone effects on pineal function. In order to clarify the role of adrenergic mechanisms in these effects, we investigated the pineal response to adrenergic stimulation and pineal beta-adrenergic receptors following castration and/or sex hormone treatment in 2-month-old male and female Sprague-Dawley rats. The urinary 6-sulphatoxymelatonin (aMT6s) response to isoproterenol (ISO) was compared in castrated and sham-operated animals. Ovariectomy caused an increase and orchiectomy a decrease in ISO-induced urinary aMT6s excretion. The melatonin response to ISO was examined in pineal glands and serum samples obtained from castrated, sex-hormone-treated castrated and sham-operated rats. Consistently, ovariectomy increased pineal and serum melatonin responses to ISO, while orchiectomy decreased the responses; oestradiol and testosterone treatments, respectively, reversed these effects. 3H-dihydroalprenolol binding was measured in single pineal glands from castrated, sex-hormone-treated castrated and sham-operated rats. Ovariectomy increased the density of beta-adrenoceptors, whereas orchiectomy decreased the density; oestradiol and testosterone replacement, respectively, blocked these effects. No significant effect on receptor Kd values was found. These data suggest that sex hormones regulate pineal melatonin production by modifying beta-adrenergic mechanisms.

Animals↗

PubMed1995

2-[125I]iodomelatonin-binding sites in the human kidney and the effect of guanosine 5'-O-(3-thiotriphosphate).

Putative melatonin receptors in normal kidney cortical tissues of patients with transitional cell carcinoma or renal cell carcinoma were characterized using a melatonin agonist, 2-[125I]iodomelatonin, as the radioligand. 2-[125I]Iodomelatonin-binding sites in the human kidney were stable, saturable, reversible, and of high affinity. The binding affinity was 15.2 +/- 2.5 pmol/L, and the binding density was 1.79 +/- 0.19 fmol/mg protein. The unity of the Hill coefficients and linearity of the Scatchard plots suggested that 2-[125I]iodomelatonin was bound to a single class of binding sites. Pharmacological characterization showed that these binding sites were highly specific to 2-iodomelatonin, melatonin, 6-hydroxymelatonin, and 6-chloromelatonin. Guanosine 5'-O-(3-thiotriphosphate) decreased the binding affinity and density of 2-[125I]iodomelatonin-binding sites in the kidney, suggesting that these binding sites are coupled to a G-protein. The characterization of 2-[125I]iodomelatonin-binding sites in normal kidney tissues taken from patients with transitional cell carcinoma or renal cell carcinoma suggests the existence of 2-[125I]iodomelatonin-binding sites in the human kidney cortex, which is in line with the findings of 2-[125I]iodomelatonin-binding sites in kidneys of other mammals and birds. The implication of a direct melatonin action on renal function in the human is proposed.

Adult↗