Biomedical subjects

Michael Wilkinson

Publications and source records attributed to Michael Wilkinson.

16 recordsLinked to original sources

PubMed2006

Increased expression of the adipokine genes resistin and fasting-induced adipose factor in hypoxic/ischaemic mouse brain.

Adipose tissue is the primary source of the adipokines resistin and fasting-induced adipose factor (FIAF). We reported that the brain is also a site of adipokine expression, although their function there is unknown. Peripheral resistin and fasting-induced adipose factor are reported to be inflammatory markers, and we hypothesized that they would be induced in the brain by hypoxia/ischaemia. We show that neonatal hypoxia/ischaemia rapidly increased fiaf mRNA in the injured cortex and hippocampus at 2 and 7 days after hypoxia/ischaemia. In contrast, resistin (retn) mRNA was increased in the cortex only at 21 days after hypoxia/ischaemia. As a lipopolysaccharide-induced inflammatory response did not increase brain fiaf and retn mRNA levels, we conclude that brain injury may be responsible for the novel hypoxia/ischaemia-induced changes in adipokine gene expression. In summary, our results indicate that brain injury, or an inflammatory stimulus, regulates the central expression of two genes normally considered to be adipose tissue-specific. These observations add to our previous evidence that the brain is an important site of adipokine gene expression.

Angiopoietin-Like Protein 4↗

PubMed2006

Caustic activation of rain showers.

We show quantitatively how the collision rate of droplets of visible moisture in turbulent air increases very abruptly as the intensity of the turbulence passes a threshold, due to the formation of fold caustics in their velocity field. The formation of caustics is an activated process, in which a measure of the intensity of the turbulence, termed the Stokes number St, is analogous to temperature in a chemical reaction: the rate of collision contains a factor exp(-C/St). Our results are relevant to the long-standing problem of explaining the rapid onset of rainfall from convecting clouds. Our theory does not involve spatial clustering of particles.

Causality↗

PubMed2006

Is there an optimal training intensity for enhancing the maximal oxygen uptake of distance runners?: empirical research findings, current opinions, physiological rationale and practical recommendations.

The maximal oxygen uptake (V-dotO(2max)) is considered an important physiological determinant of middle- and long-distance running performance. Little information exists in the scientific literature relating to the most effective training intensity for the enhancement of V-dotO(2max) in well trained distance runners. Training intensities of 40-50% V-dotO(2max) can increase V-dotO(2max) substantially in untrained individuals. The minimum training intensity that elicits the enhancement of V-dotO(2max) is highly dependent on the initial V-dotO(2max), however, and well trained distance runners probably need to train at relative high percentages of V-dotO(2max) to elicit further increments. Some authors have suggested that training at 70-80% V-dotO(2max) is optimal. Many studies have investigated the maximum amount of time runners can maintain 95-100% V-dotO(2max) with the assertion that this intensity is optimal in enhancing V-dotO(2max). Presently, there have been no well controlled training studies to support this premise. Myocardial morphological changes that increase maximal stroke volume, increased capillarisation of skeletal muscle, increased myoglobin concentration, and increased oxidative capacity of type II skeletal muscle fibres are adaptations associated with the enhancement of V-dotO(2max). The strength of stimuli that elicit adaptation is exercise intensity dependent up to V-dotO(2max), indicating that training at or near V-dotO(2max) may be the most effective intensity to enhance V-dotO(2max) in well trained distance runners. Lower training intensities may induce similar adaptation because the physiological stress can be imposed for longer periods. This is probably only true for moderately trained runners, however, because all cardiorespiratory adaptations elicited by submaximal training have probably already been elicited in distance runners competing at a relatively high level.Well trained distance runners have been reported to reach a plateau in V-dotO(2max) enhancement; however, many studies have demonstrated that the V-dotO(2max) of well trained runners can be enhanced when training protocols known to elicit 95-100% V-dotO(2max) are included in their training programmes. This supports the premise that high-intensity training may be effective or even necessary for well trained distance runners to enhance V-dotO(2max). However, the efficacy of optimised protocols for enhancing V-dotO(2max) needs to be established with well controlled studies in which they are compared with protocols involving other training intensities typically used by distance runners to enhance V-dotO(2max).

Adaptation, Physiological↗

PubMed2005

Clustering by mixing flows.

We calculate the Lyapunov exponents for particles suspended in a random three-dimensional flow, concentrating on the limit where the viscous damping rate is small compared to the inverse correlation time. In this limit Lyapunov exponents are obtained as a power series in epsilon, a dimensionless measure of the particle inertia. Although the perturbation generates an asymptotic series, we obtain accurate results from a Padé-Borel summation. Our results prove that particles suspended in an incompressible random mixing flow can show pronounced clustering when the Stokes number is large and we characterize two distinct clustering effects which occur in that limit.

Journal Article↗

PubMed2005

RNAi-mediated silencing of leptin gene expression increases cell death in C6 glioblastoma cells.

We previously demonstrated that the brain, pituitary, and C6 glioblastoma cells express leptin. To determine the physiological role of brain-derived leptin, we sought to selectively silence its expression using RNA interference (RNAi) in vitro. One of four potential targets, siRNA L7, reduced leptin mRNA by 50% (P < 0.05) and protein by 55% (P < 0.0001) in C6 cells. RNAi also induced a twofold increase in cell death as seen by ethidium homodimer-1 (P < 0.015) and TUNEL (P < 0.005) staining. These data suggest that endogenous leptin may be a critical factor promoting cell survival in the brain.

Analysis of Variance↗

PubMed2005

Hypothalamic resistin immunoreactivity is reduced by obesity in the mouse: co-localization with alpha-melanostimulating hormone.

Resistin is a new adipokine expressed in mouse, rat and human adipose tissue. Resistin may be an important link between obesity and insulin resistance, though this controversial view is complicated by the discovery of multiple sites of resistin expression, including human macrophages, placenta and pancreas. In previous studies we demonstrated that the mouse hypothalamo-pituitary system was also a site of resistin production. Pituitary resistin is developmentally regulated, reduced in the ob/ob mouse and severely down-regulated by food deprivation (24 h). An unexpected finding was that hypothalamic resistin mRNA remained unaffected by fasting. The present experiments examined the localization and possible regulation of hypothalamic resistin protein. Using immunohistochemistry we observed a complex network of resistin+ fibres extending rostrally from the arcuate nucleus of the hypothalamus (ARC) to the preoptic area. Labelled cell bodies occurred only in the ARC and in a periventricular region of the dorsal hypothalamus. Hypothalamic resistin immunoreactivity (ir) was unaffected by fasting (48 h) or by a high fat diet, but the periventricular staining was greatly increased in the lactating mouse. Marked reductions in resistin+ fibres were seen in brain tissue from: (a) ob/ob mice, (b) young mice made underweight for their age by raising them in large litters (20 pups per litter) and (c) mice with hypothalamic lesions induced by monosodium glutamate (MSG) or gold thioglucose (GTG). We speculate that the resistin-ir deficit in genetically obese mice, and in severely underweight mice, could be due to low or absent leptin. In contrast, though MSG- and GTG-treated mice have high levels of circulating leptin, in the presence of excessive visceral fat deposits, we hypothesize that damage to the ARC destroys the resistin+ cell bodies. This latter supposition led us to an additional hypothesis, that resistin-ir would be contained in neurons expressing the proopiomelanocortin (POMC) gene. This proved to be correct. Double label immunofluorescence histochemistry revealed that alpha-MSH-ir, a marker for POMC neurons, was co-localized with resistin-ir. In conclusion, our data reveal a second example of an adipocytokine co-localized with a hypothalamic neuropeptide. We reported previously that leptin was co-localized with oxytocin and vasopressin. RT-PCR analysis confirmed that resistin mRNA is readily detectable in ARC, but further work is required to determine whether the resistin gene is expressed in POMC neurons or if resistin is specifically accumulated by these cells. Nonetheless, our data suggest that the hypothalamus is a target tissue for resistin.

Age Factors↗

PubMed2005

Pituitary resistin gene expression is upregulated in vitro and in vivo by dexamethasone but is unaffected by rosiglitazone.

A physiological role for the adipose-derived hormone, resistin, remains unsolved and its putative involvement in insulin resistance is largely controversial. Like leptin and other adipokines, we detected resistin in the rodent hypothalamic-pituitary system. In addition, the pituitary corticotrope cell line, AtT20, is also a source of resistin. These data suggested that resistin could be involved in diverse physiological processes in non-adipose tissue. Initial studies indicated that pituitary resistin gene expression was regulated in a nutritional (fed>fasted), age- (young>old) and gender-specific manner (male>female). In the present work we hypothesized that pituitary resistin expression should be regulated through signalling pathways similar to those reported for adipose tissue. For example, dexamethasone (DEX) potently stimulates production of resistin in murine adipose tissue, whereas thiazolidinediones such as rosiglitazone (ROSI), acting via peroxisome proliferator-activated receptor (PPAR)gamma, reportedly inhibit or stimulate resistin expression. Using quantitative real-time RT-PCR we determined that injection of DEX (10 and 50 microg) yielded 7- and 9-fold increases in pituitary resistin gene expression in prepubertal, but not adult mice. In addition, adrenalectomy attenuated pituitary resistin gene expression, which was restored by DEX (10 microg). In AtT20 cells, exposure to corticosterone (10(-7) and 10(-6)M; 24 h) and DEX (10(-9)-10(-6)M) stimulated resistin mRNA more than 65% (p<0.05) and 115% (p<0.0001), respectively. In contrast, ROSI, injected (5 and 20 mg/kg s.c. for 14 days) or given orally (3 mg/kg/day to 10 mg/kg/day for up to 7 weeks), failed to alter pituitary resistin gene expression in healthy male CD1 mice. Treatment of AtT20 cells with ROSI (10(-5)-10(-10)M; 24 h or 96 h) or the PPARgamma agonist GW 1929 (10(-9) and 10(-5)M; 24 h) had no effect on resistin mRNA. The PPARgamma antagonist GW 9662 (10(-6) and 10(-5)M) was also ineffective. In conclusion, pituitary resistin mRNA levels are robustly stimulated by corticosteroids, particularly at the time of puberty. This is consistent with our previous suggestion that resistin may be involved in maturation of the hypothalamic-pituitary axis. In contrast, pituitary resistin gene expression appears to be PPARgamma-independent and therefore different from the situation in adipose tissue.

Adrenalectomy↗

PubMed2004

Coagulation by random velocity fields as a Kramers problem.

We analyze the motion of a system of particles suspended in a fluid which has a random velocity field. There are coagulating and noncoagulating phases. We show that the phase transition is related to a Kramers problem, and we use this to determine the phase diagram in two dimensions, as a function of the dimensionless inertia of the particles, epsilon, and a measure of the relative intensities of potential and solenoidal components of the velocity field, Gamma. We find that the phase line is described by a function which is nonanalytic at epsilon=0, and which is related to escape over a barrier in the Kramers problem. We discuss the physical realizations of this phase transition.

Journal Article↗

PubMed2004

Long-term methylphenidate treatment down-regulates c-fos in the striatum of male CD-1 mice.

Methylphenidate (Ritalin) is routinely prescribed to children with attention deficit hyperactivity disorder, but little is known about its long-term consequences on brain development. We treated pre- and peri-pubertal male CD-1 mice with repeated injections of methylphenidate hydrochloride (MPH) and quantified the expression of the immediate early gene c-fos in the striatum. A single injection of MPH (5 or 40 mg/kg) significantly elevated FOS immunoreactivity in the striatum in a dose-dependent manner, compared with saline. Repeated MPH treatment attenuated the effect of a single challenge dose of MPH on striatal c-fos expression. These results replicate those observed with rats and indicate that long-term use of MPH may alter neural activity by down-regulation of gene expression in the striatum.

Animals↗

PubMed2004

Pituitary resistin gene expression: effects of age, gender and obesity.

Resistin is a new adipocytokine which is expressed in rat, mouse and possibly human adipose tissue. Its putative role as a mediator of insulin resistance is controversial. We hypothesized that resistin, like leptin, would have multiple roles in non-adipose tissues and we reported that resistin is expressed in mouse brain and pituitary. Moreover, resistin expression in female mouse pituitary is developmentally regulated and maximal expression occurs peripubertally. Although the role of endogenous resistin in mouse brain and pituitary has not been determined, our data suggest that resistin could be important in the postnatal maturation of the hypothalamic-pituitary system. In the present study we compared the ontogeny of resistin gene expression in the pituitary of male and female mice using semi-quantitative RT-PCR analysis. We show that resistin expression is developmentally regulated in the pituitary of male and female CD1 mice. However, significant gender differences were evident (male > female at postnatal day 28 and 42) and this was not modified by neonatal treatment of female pups with testosterone. Since resistin expression in adipose tissue is also influenced by obesity, we evaluated resistin expression in fat, brain and pituitary of the obese ob/ob mouse. Resistin mRNA was significantly increased in both visceral and subcutaneous adipose depots in postnatal day 28 ob/ob mice compared to controls, but pituitary resistin expression was significantly reduced. In contrast to the prepubertal levels, and in agreement with other reports, adipose resistin expression was reduced in adult ob/ob mice. In a third set of experiments we examined the influence of food deprivation on pituitary and fat resistin mRNA. Resistin gene expression was severely down-regulated by a 24-hour fast in adipose and pituitary tissue but not in hypothalamus. In conclusion, pituitary resistin expression is age- and gender-dependent. In ob/ob mice, and in fasted mice, resistin is regulated in a tissue-specific manner. Thus in visceral fat obesity increases but starvation decreases resistin mRNA. In contrast, pituitary levels are decreased in the presence of both high (ob/ob) and low (fasting) adipose stores. Further studies are required to define the unexpected hormonal regulation of resistin gene expression in the pituitary.

Adipose Tissue↗

PubMed2004

Food restriction regulates adipose-specific cytokines in pituitary gland but not in hypothalamus.

White adipose tissue is now recognized as the source of a growing list of novel adipocyte-specific factors, or adipokines. These factors regulate energy homeostasis, including the response to food deprivation. We hypothesized that the brain and pituitary gland would also express adipokines and their regulatory factors and subsequently demonstrated that the rodent brain-pituitary system expresses mRNA and protein for leptin and resistin. We now report that the adipokines FIAF and adiponutrin, as well as the nuclear hormone receptor PPAR gamma, are expressed in pituitary, brain and adipose tissue. In pituitary gland, 24 h of food restriction reduced PPAR gamma expression by 54% whereas both adiponutrin and FIAF were increased 1.7 and 2.3 fold, respectively. These changes in expression were similar to those observed in fat, except for adiponutrin, which by contrast is dramatically reduced 95% by fasting. Furthermore, whereas PPAR gamma 2 is the main isoform affected by fasting in adipose tissue, our data suggest that only PPAR gamma 1 is present and downregulated by fasting in pituitary tissue. In contrast to the sensitivity of pituitary tissue to the effects of fasting, no significant change in expression was observed in basal hypothalamus for any of the genes studied. Overall, our data suggest that pituitary-derived adipokines may play an unexpected role in the neuroendocrine regulation of energy homeostasis.

Adipose Tissue↗

PubMed2003

Leptin receptors are developmentally regulated in rat pituitary and hypothalamus.

We have previously reported that leptin is expressed in adult rat brain and pituitary gland, though the role of leptin in these sites has not been determined. Leptin mRNA is developmentally regulated in the brain and pituitary of male and female rats during early postnatal development, suggesting a role in the maturation of the brain-pituitary system. Here, we sought to extend our previous studies by evaluating (1) the ontogeny of leptin receptor mRNA levels in rat brain and pituitary and (2) pituitary leptin protein levels in neonatal and pre-pubertal rats. Pituitary leptin concentration was highest shortly after birth (postnatal day (PD) 4, 25 ng/mg protein) and fell significantly throughout postnatal development and into adulthood (PD 60, 3.5 ng/mg protein; P<0.005) coincident with a decline in pituitary leptin mRNA levels. Significant age-related effects on leptin receptor mRNA levels were also observed in the pituitary and the hypothalamus of male and female rats using semi-quantitative RT-PCR analysis. In the pituitary, the short form (OBRa) mRNA levels were highest in neonatal rats (PD 4) but declined throughout postnatal development (PD 4-22) paralleling the fall in pituitary leptin mRNA and protein levels. The long form (OBRb) mRNA levels were unaffected by age between PD 4 and 22. In contrast, hypothalamic, levels of OBRb mRNA were very low to undetectable shortly after birth (PD 4) and rose significantly between PD 4 and 14/22 while levels of OBRa mRNA were not significantly different between PD 4 and 22. Immunohistochemical detection of leptin receptor immunoreactivity (all forms) revealed the presence of OBR-like protein in pituitary and hypothalamus as early as PD 4. Cortical leptin receptor mRNA levels were similar throughout early postnatal development. No gender-related differences in leptin receptor mRNA levels were noted in brain or pituitary. In conclusion, these data, together with our previous work, indicate that the neonatal pituitary gland expresses leptin and leptin receptors at levels far in excess of those observed in mature rats. The pituitary is thus quite different from adipose tissue, hypothalamus and cerebral cortex, in which neonatal leptin expression is lowest at birth. Since neonatal pituitary leptin receptor expression is also elevated, it is possible that pituitary-derived leptin plays some role in the development of the hypothalamic-pituitary system.

Animals↗

PubMed2003

Daily methylphenidate administration attenuates c-fos expression in the striatum of prepubertal rats.

Methylphenidate (Ritalin) is a psychostimulant drug used to treat children with attention deficit hyperactivity disorder. Despite its widespread and increasing clinical use, little is known about the long-term consequences of drug treatment. We compared the effects of a single injection of methylphenidate with that of long-term methylphenidate injections (one/day; 14 days) on immediate-early gene expression (c-fos) in the striatum of prepubertal male rats. Rats (25 days old) were injected once daily for 14 days with either saline or methylphenidate (1, 2 or 10 mg/kg), or for 13 days with saline followed by one injection of methylphenidate (1, 2 or 10 mg/kg) on day 14, and were sacrificed 2 h post-injection. Methylphenidate dose-dependently increased FOS immunoreactivity in the striatum. A single injection of methylphenidate (2 or 10 mg/kg) on day 14, following saline treatment for 13 days, caused a dramatic elevation in c-fos expression. This effect was significantly attenuated in animals treated chronically with methylphenidate (2 or 10 mg/kg) for the entire 14 days. Our data suggest that repeated methylphenidate treatment, at a clinically relevant dose (2 mg/kg), markedly inhibits immediate-early gene expression in the brain. This is the first demonstration of methylphenidate-induced modification of gene expression in developing rat striatum and may have implications for chronic methylphenidate use in children.

Animals↗

PubMed2002

FOS and FOSB expression in the medial preoptic nucleus pars compacta of maternally active C57BL/6J and DBA/2J mice.

C57BL/6J and DBA/2J inbred mice differ in aspects of maternal behavior and in the morphology of the medial preoptic nucleus (MPO), suggesting a possible association. DBA/2J mice have a compact subnucleus in the MPO, the MPOpc, that is sexually dimorphic and absent in C57BL/6J mice. To determine whether MPOpc cells are activated by maternal behavior, FOS and FOSB immunohistochemistry was performed on brain sections of C57BL/6J and DBA/2J mothers following the return of their pups after a separation of 2 days. In both light and dark phases of the daily cycle, stimulation of DBA/2J mothers evoked an increase in FOS- and FOSB-immunoreactivity in the MPOpc. Stimulated C57BL/6J mice, which lack the MPOpc, did not show an increase in cellular activity in the corresponding MPO region. Cells immediately lateral to the MPOpc were activated by pup stimulation, in both strains. These results suggest that MPOpc cells are active during maternal behavior, and that strain differences in maternal behavior are related to anatomical differences in the MPO.

Animals↗

PubMed2002

Resistin expression and regulation in mouse pituitary.

Resistin, a new adipocytokine, is expressed in human, rat and mouse adipose tissue. Its putative role as a mediator of insulin resistance is controversial. We hypothesized that resistin, in common with leptin, has multiple roles in non-adipose tissues. Using reverse transcription polymerase chain reaction (RT-PCR) we show that the resistin gene (Retn) is expressed in mouse brain (hypothalamus and cortex) and pituitary gland. Immunohistochemistry revealed resistin protein in the arcuate nucleus and pituitary gland. Semi-quantitative RT-PCR analysis indicated that Retn mRNA is developmentally regulated in the pituitary. Expression was lowest at birth, increased abruptly between postnatal days 14 and 25 (four-fold; P<0.001), and declined thereafter. This peak in pituitary Retn mRNA was unaffected by early weaning but was abolished by neonatal treatment with monosodium glutamate, suggesting that the basal hypothalamus regulates pituitary Retn. Although the role(s) of endogenous resistin in mouse brain and pituitary remains to be determined, it may be distinct from its controversial involvement in insulin resistance. Our data suggest that local resistin expression could have functional implications during prepubertal maturation of the hypothalamic-pituitary system.

Aging↗

PubMed2002

Leptin immunoreactivity is localized to neurons in rat brain.

Leptin is secreted from adipocytes and is thought to enter the brain to regulate and coordinate metabolism, feeding behaviour, energy balance and reproduction. It is now clear that there are many additional sites of leptin production, including human placenta, ovary, stomach, skeletal muscle, mammary gland, pituitary gland and brain. In the present work, we employed double-label immunofluorescent histochemistry to establish the neuronal localization of leptin immunoreactivity (IR). To accomplish this, we used the neuron-specific marker NeuN to label cells in the arcuate nucleus (ARC), piriform cortex and hippocampus. In the supraoptic nucleus (SON) and paraventricular nucleus (PVN), we used antisera to oxytocin and vasopressin as neuronal markers. Double labelling revealed leptin IR in neurons of the ARC and piriform cortex. Leptin IR was confined to the nucleus and to distinct perinuclear sites. In contrast, neurons in the CA 2/CA 3 region of the hippocampus showed little nuclear staining. Leptin IR was clustered around the nucleus in these cells. Neurons of the dentate gyrus exhibited both nuclear and perinuclear localization of leptin IR. In the SON/PVN, most oxytocin- and vasopressin-IR neurons also contained leptin IR, often in perinuclear sites. In conclusion, the neuronal, perinuclear localization of leptin IR in rat brain corresponds closely to that of leptin receptor (OB-R) IR, which has also been detected intracellularly. Our observation of leptin IR associated with cell nuclei suggests the existence of an OB-R distinct from the well-described membrane forms.

Animals↗