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J G Mercer

Publications and source records attributed to J G Mercer.

At least 73 records · Page 4Linked to original sources
PubMed1996

Coexpression of leptin receptor and preproneuropeptide Y mRNA in arcuate nucleus of mouse hypothalamus.

Leptin, the protein product of the adipose tissue-specific ob (obese) gene (1), reduces the body weight, adiposity and food intake of obese ob/ob mice on peripheral or central injection (2, 3, 4). [125I]leptin binding has been detected in mouse choroid plexus (5), from which a leptin receptor gene was expression cloned (5). The gene has at least 6 splice variants (6, 7). Leptin receptor mRNA was localized in the hypothalamus by in situ hybridization being particularly abundantly expressed in the arcuate nucleus (8). There is evidence linking the physiological effects of injected leptin with hypothalamic neuropeptide Y (9, 10) (NPY), which has potent central effects on food intake and energy balance (11), and is also expressed in the arcuate nucleus. Here we report dual in situ hybridization studies for leptin receptor and NPY gene expression in the mouse arcuate nucleus, where the majority of cells examined expressed both genes. This provides the first direct evidence that leptin acts on cells that express NPY mRNA.

Animals↗
PubMed1996

The ovine pars tuberalis secretes a factor(s) that regulates gene expression in both lactotropic and nonlactotropic pituitary cells.

The purpose of this study was to determine whether the cells of the ovine pars tuberalis (PT) secrete a factor(s) that can influence the activity of cells in the pars distalis (PD). By Northern blotting of total RNA isolated from PD cells that had been stimulated in the presence of cycloheximide (10 micrograms/ml), PT cell-conditioned medium was shown to induce a significant increase in the expression of the early response gene, c-fos, above both PD cell-conditioned and nonconditioned medium control levels (P < 0.05). Although forskolin (5 microM) induced a weak increase in c-fos expression in PD cells, the effect of PT medium conditioned in the presence of forskolin enhanced this expression more than additively (P < 0.05); furthermore, this effect was reversed by melatonin. These results are consistent with the release of a factor(s) from the PT, which for simplicity we have called tuberalin. This factor was released from PT cells in a time-dependent and cycloheximide-sensitive manner and was resistant to heating at 100 C for 10 min. Tuberalin activity could be size-fractionated using molecular size cut-off filters to produce activity in both the 1- to 10-kDa and more than 10-kDa size ranges. The activities in both of these fractions were sensitive to trypsin degradation and, therefore, appeared to be peptidergic. However, it was not clarified whether the biological activities were due to one or two components. Tuberalin also induced c-fos expression in other cell types, including GH3 and NIH3T3 cells. Dual labelling of PD cells by in situ hybridization using riboprobes for c-fos and PRL demonstrated that both the less than and more than 10-kDa fractions of tuberalin activated c-fos expression in some, but not all, lactotrophs in PD cell cultures, suggesting that a primary function of the PT is to regulate the activity of lactotrophs. This was supported further by enhanced secretion of PRL from PD cells in the presence of either PT-conditioned medium or PT cells in coculture. In addition, PT-conditioned medium was found to increase c-fos in a second cell type, which did not hybridize positively for PRL, indicating the existence of other endocrine interactions between the PT and PD.

3T3 Cells↗
PubMed1995

Fat mobilisation in short days is not associated with altered noradrenergic sensitivity of adipocytes in Djungarian hamsters.

One of the primary physiological responses of the Djungarian hamster to short photoperiods is a reduction in body weight with fat mobilisation. The depletion of fat to a minimum level may be regulated either in the periphery, through the sensitivity of adipocytes to hormonal stimulation, or centrally, via adjustments in efferent activity. To investigate this, we examined the lipolytic pathway in fat cells from animals at various stages of entrainment to long or short photoperiod. Short photoperiod exposure of up to a 10-week duration was without effect on basal glycerol release by unstimulated cells or on the ability of norepinephrine or an adenosine analogue to stimulate or inhibit lipolysis, respectively. Prolonged exposure to short photoperiod reduced basal glycerol release, but adipocytes retained their sensitivity to hormonal stimulation. Short photoperiod had no effect on the density or affinity of membrane-bound beta-adrenergic or adenosine receptors, or upon the ability of isoproterenol or forskolin to stimulate adenylate cyclase in adipocyte membranes. This suggests that the regulation of fat depletion in short photoperiod is determined centrally and does not involve alterations in adipocyte sensitivity and, in particular, the desensitisation of the adipocyte beta-adrenergic receptor-linked adenylate cyclase pathway.

Adenylyl Cyclase Inhibitors↗
PubMed1995

Anorexia induced by the parasitic nematode, Nippostrongylus brasiliensis: effects on NPY and CRF gene expression in the rat hypothalamus.

Infections of the gastrointestinal nematode, Nippostrongylus brasiliensis, in the laboratory rat result in a characteristic biphasic anorexia which is followed by hyperphagia once the worm burden has been cleared. Despite the importance of parasite-induced anorexia, relatively little is known of the underlying mechanisms. We have investigated the involvement of the central appetite drive in this anorexia by studying the gene expression of two neuropeptides with opposing actions on energy balance, NPY and CRF. Gene expression was assessed by in situ hybridization at 2, 8 and 16 days post-infection (p.i.) in infected rats, in uninfected controls, and in a group with food intake restricted to match that taken voluntarily by the parasitize animals. The sampling intervals corresponded to each of the two phases of maximum anorexia and the period of compensatory hyperphagia. Surprisingly, we found that increases in NPY gene expression in the hypothalamic arcuate nucleus (ARC) accompany anorexia in rats infected with N. brasiliensis; there was a significant relationship between degree of anorexia and induction of NPY mRNA after 8 days of infection. Furthermore, ARC NPY mRNA levels in parasitized animals were similar to those in pair-fed individuals with food intake restricted to match the infected rats. The number of larvae used to establish the infection affected both the degree of anorexia and the level of NPY mRNA at 8 days p.i. in a dose-dependent manner. NPY gene expression remained elevated in infected rats during at least the initial stages of compensatory hyperphagia. This suggests that animals detect a state of energy deficit during the early stages of the infection, yet do not feed, but become hyperphagic coincident with worm loss. The failure of anorectic parasitized animals to feed in response to activation of the NPYergic system makes this a novel system in which to study the regulation of hypothalamic NPY by physiological challenge. There were no significant differences in CRF gene expression between the groups at any of the sampling intervals.

Animals↗
PubMed1995

Hypothalamic NPY and prepro-NPY mRNA in Djungarian hamsters: effects of food deprivation and photoperiod.

Two catabolic states leading to loss of body weight were compared in the Djungarian hamster (Phodopus sungorus campbelli). Hypothalamic neuropeptide Y (NPY) and gene expression for NPY and corticotropin-releasing factor (CRF) were examined after withdrawal of food for 48 h or exposure to short photoperiod for 10 or 20 wk. Food deprivation was accompanied by increases in both NPY and prepro-NPY mRNA in the hypothalamic arcuate nucleus (ARC). Increases in gene expression were limited compared with published data from the rat and were inversely related to predeprivation body weight. Exposure to short photoperiod for 20 wk reduced body weight by 39%, but the activity of the NPY-ergic system was not affected; peptide concentration and gene expression were similar in short photoperiod hamsters and long photoperiod controls. The hypothalamic NPY-ergic system of the Djungarian hamster is sensitive to weight loss due to imposed manipulations of energy balance, but the catabolism observed in short photoperiod gives rise to a body weight that is appropriate to the season encoded by the photoperiod. CRF gene expression was not affected by food deprivation or short photoperiod.

Animals↗
PubMed1994

Effect of photoperiod on mitochondrial GDP binding and adenylate cyclase activity in brown adipose tissue of Djungarian hamsters.

Experiments were designed to investigate the involvement of brown adipose tissue (BAT) thermogenesis in the weight loss exhibited by Djungarian hamsters (Phodopus sungorus campbelli) in response to a short photoperiod. Significant decreases in body weight preceded reductions in food intake, suggesting a photoperiod-induced change in energy expenditure. Sixteen weeks exposure to short photoperiod resulted in large decreases in body weight and interscapular BAT mass that were accompanied by an increase in the thermogenic activity of BAT (estimated by mitochondrial GDP binding). However, exposure to short photoperiod for 8 weeks, that induced smaller but significant reductions in body weight, was without effect on the BAT parameters measured. This suggests that increased BAT thermogenesis is unlikely to initiate, or contribute to, the early stages of photoperiod-induced weight loss. In addition, short photoperiod failed to induce any change in the specific activity or sensitivity of adenylate cyclase in BAT membranes, in contrast to the downregulation of catecholamine-stimulated cAMP production observed in BAT following cold exposure.

Adenylyl Cyclases↗
PubMed1993

Satiety signals in sheep: involvement of CCK, propionate, and vagal CCK binding sites.

The satiety effects of the hormone, cholecystokinin (CCK), and propionate, an important gluconeogenic substrate, were studied in ad lib-fed sheep. Hepatic portal infusion of either sodium propionate (1.2 mmol/min) or sulphated CCK-8 (sCCK-8; 18.3 pmol/kg/min) had no effect on food intake. However, together, they decreased intake by 44%; similar to the effect of 2.4 mmol/min propionate alone. CCK infusions reduced the frequency of reticular contractions in the presence or absence of propionate. The effects of infusions on motility and food intake were, therefore, dissociated. Further studies demonstrated axonal transport of CCK binding sites in the ovine vagus. Binding sites accumulated to a similar extent on both sides of a ligature indicating the existence of both anterograde and retrograde transport which was limited to a small proportion of fibres. Binding incubations carried out in the presence of the CCK receptor antagonists, MK-329 and L-365,260, indicated that the majority of binding sites, if not the total population, possessed pharmacology typical of type B CCK receptors.

Animals↗
PubMed1993

G-protein coupling of vagal CCK binding sites and comparisons of transport rates.

The ability of nanomolar concentrations of guanine, but not adenine, nucleotides to inhibit specific 125I-Bolton-Hunter CCK binding to ligated rat vagus nerve demonstrated that vagal CCK binding sites were linked to G-proteins during axonal transport. The GTP analogue, GTP[S], reduced specific binding to both anterogradely and retrogradely transported binding sites by more than 90% at 1 microM. Transport of these putative receptor-G-protein complexes was examined under conditions of food deprivation or physiological hyperphagia induced by either lactation or genetic obesity. None of the physiological or imposed manipulations of food intake had any effect on the axonal transport of CCK binding sites. Transection of the cervical vagus resulted in an accumulation of binding sites at the lesion site that was indistinguishable from that seen following ligation for the same period.

Animals↗
PubMed1992

Selectivity of cholecystokinin (CCK) receptor antagonists, MK-329 and L-365,260, for axonally-transported CCK binding sites on the rat vagus nerve.

The ability of the cholecystokinin (CCK) receptor antagonists, MK-329 and L-365,260, to selectively inhibit 125I-Bolton-Hunter-CCK8 binding to ligated rat vagus nerve in vitro was examined at concentrations ranging from 10(-10) M to 10(-6) M. Both antagonists inhibited binding to CCK binding sites accumulating proximal to ligatures on the cervical vagus. Incubation of nerve sections in the presence of both antagonists produced an additive effect, indicating that both CCK-A and CCK-B binding sites are transported towards the periphery. In contrast, CCK binding sites accumulating distal to the ligature possessed the pharmacological characteristics of the CCK-B receptor sub-type only.

Animals↗
PubMed1992

Effect of neonatal capsaicin treatment on cholecystokinin-(CCK8) satiety and axonal transport of CCK binding sites in the rat vagus nerve.

Cholecystokinin (CCK) binding sites which accumulate at ligatures placed on the rat vagus nerve may mediate the satiety actions of CCK. Treatment of neonatal rats with capsaicin attenuated the satiety effect of injected CCK in adult life. Capsaicin pretreatment also reduced, but did not eliminate, the accumulation of CCK binding sites proximal and distal to ligatures on either cervical trunk. A similar effect was observed following ligation of subdiaphragmatic vagal trunks. The CCK receptor antagonists, MK-329 and L-365,260, inhibited binding to capsaicin- and vehicle-treated nerves to a similar degree. Densities of CCK binding sites in the nucleus tractus solitarius and area postrema were also markedly affected by neonatal capsaicin treatment.

Animals↗
PubMed1992

Analysis of ecdysteroids in the trematodes, Schistosoma mansoni and Fasciola hepatica.

Adult Schistosoma mansoni from experimentally infected mice and Fasciola hepatica recovered from ovine livers post mortem were analyzed for free and conjugated ecdysteroids by radioimmunoassay, high-performance liquid chromatography monitoring fractions by radioimmunoassay, and by capillary gas chromatography/mass spectrometry (selected ion monitoring). Both species contained ecdysone and 20-hydroxyecdysone as free ecdysteroids and as polar conjugates. F. hepatica also contained a polar conjugate of 2-deoxyecdysone. Evidence of apolar ecdysteroid conjugates was only obtained for F. hepatica. The free ecdysteroid-containing fraction of S. mansoni contained small amounts of unidentified immunoreactive material.

Animals↗
PubMed1991

The effect of ecdysteroids on the microfilarial production of Brugia pahangi and the control of meiotic reinitiation in the oocytes of Dirofilaria immitis.

The effects of the ecdysteroids ecdysone and 20-hydroxyecdysone on microfilarial release in Brugia pahangi and on meiotic reinitiation in the oocytes of Dirofilaria immitis were studied. Ecdysone was found to stimulate microfilarial release at 2 x 10(-6) M, but this effect was reduced at higher and lower concentrations. 20-Hydroxyecdysone was found to have no such effect. In D. immitis, ecdysone at 10(-5) M was also found to overcome the period of meiotic arrest that occurs during the pachytene stage of prophase 1. This effect was reduced when 10(-6) M ecdysone was used. These results help support the theory that the ecdysteroids play a hormonal role in filarial worms similar to that found in insects.

Animals↗
PubMed1991

Metabolism and fate of ecdysteroids in the nematodes Ascaris suum and Parascaris equorum.

When injected with [3H]ecdysone and maintained in vitro, the parasitic nematodes, Ascaris suum and Parascaris equorum each produced a series of polar and relatively apolar metabolites. A. suum metabolised the compound into ecdysonoic acid ([3H]EOIC), ecdysone 25-glucoside ([3H]E25gluc), putative ecdysone 22-phosphate ([3H]E22P) and a series of at least six relatively apolar metabolites. All of these, except ecdysonoic acid, were hydrolysed by a crude enzyme preparation from Helix pomatia, releasing ecdysone. In a similar study, P. equorum produced ecdysone 25-glucoside, putative ecdysone 22-phosphate and a series of relatively apolar compounds all of which were hydrolysed by H. pomatia enzymes, releasing ecdysone. [3H]Ecdysone 25-glucoside was the most abundant single metabolite in both species, and in P. equorum, at least, was released into the culture medium in relatively large amounts. Apolar metabolites were present in worm samples and were the major, if not the only radiolabelled compounds detected in eggs of both species. Data indicated a metabolic relationship between some of the apolar conjugates found in both nematode species and ecdysone 25-glucoside.

Animals↗
PubMed1990

Investigation of ecdysteroid excretion by adult Dirofilaria immitis and Brugia pahangi.

The excretion of ecdysteroids by the filarial nematode species, Dirofilaria immitis and Brugia pahangi, was examined both in vitro, by the analysis of culture medium, and in vivo, through analysis of serum samples from experimentally infected hosts. There was no evidence of ecdysteroid excretion by intact parasites of either species in vitro. Free ecdysteroids were detected in the serum of ferrets and dogs infected with D. immitis, but concentrations would be at or below the limit of detection in sub-millilitre serum samples. The detection of ecdysteroids in the serum of potential hosts is unlikely to be of value in the diagnosis of filarial infections due to a combination of low titre in the presence of current infection and measurable titre in its absence. Ecdysteroids of dietary origin may contribute to the latter.

Animal Feed↗
PubMed1989

Effects of potential inhibitors on Brugia pahangi in vitro: macrofilaricidal action and inhibition of microfilarial production.

A series of compounds that apparently disrupt hormonally regulated processes in insects have been examined for effects on the viability and microfilarial production of adult Brugia pahangi cultured in vitro. The azasteroids, 25-azacoprostane and 25-azacholestane, inhibited the production of microfilariae at 5 ppm, the former also exhibiting macrofilaricidal activity at this concentration. The brassinosteroids examined inhibited microfilarial production at 5 ppm but did not affect worm viability. Azadirachtin also proved to be a significant inhibitor of microfilarial release without effect on worm motility or viability. Of all the compounds tested, the non-steroidal amines appeared to be the most promising as potential filaricides, several of them proving to be macrofilaricidal at 1 ppm and affecting microfilarial production at even lower concentrations.

Amines↗