Matthew S. Gill

Serum leptin through childhood and adolescence

Leptin is the protein product of the recently cloned ob gene, that has been implicated in the control of body weight and thermogenesis, but also independently stimulates the reproductive axis. As major changes in body composition and gonadal function occur during human adolescence, we have assessed serum leptin concentration through childhood.

An automated high-throughput assay for survival of the nematode Caenorhabditis elegans

Many genetic or environmental manipulations that extend life span in the nematode Caenorhabditis elegans (C. elegans) also enhance survival following acute stresses such as oxidative damage and thermal stress. This coupling of stress response and aging mechanisms has proved a useful tool in identifying new genes that affect the aging process without the need for performing lengthy life span analyses. Therefore, it is likely that this approach may also be applied to the identification of pharmacological agents that extend life span through enhanced resistance to oxygen radicals or other stressors. To facilitate high-throughput drug screens in the nematode, we have developed a microtitre plate survival assay that uses uptake of the fluorescent dye SYTOX green as a marker of nematode death. An increase in throughput compared with the conventional survival assay was achieved by combining automated worm-handling technology with automated real-time fluorescence detection. We have validated this assay by examining survival during acute heat stress and protection against oxidative stress with the superoxide dismutase/catalase mimetic Euk-134. We propose that this novel method of survival analysis will accelerate the discovery of new pharmacological interventions in aging and oxidative stress.

Intramyocellular Fatty-Acid Metabolism Plays a Critical Role in Mediating Responses to Dietary Restriction in Drosophila melanogaster

Changes in fat content have been associated with dietary restriction (DR), but whether they play a causal role in mediating various responses to DR remains unknown. We demonstrate that upon DR, Drosophila melanogaster shift their metabolism toward increasing fatty-acid synthesis and breakdown, which is required for various responses to DR. Inhibition of fatty-acid synthesis or oxidation genes specifically in the muscle tissue inhibited life-span extension upon DR. Furthermore, DR enhances spontaneous activity of flies, which was found to be dependent on the enhanced fatty-acid metabolism. This increase in activity was found to be at least partially required for the life-span extension upon DR. Overexpression of adipokinetic hormone (dAKH), the functional ortholog of glucagon, enhances fat metabolism, spontaneous activity, and life span. Together, these results suggest that enhanced fat metabolism in the muscle and physical activity play a key role in the protective effects of DR.

N-acylethanolamine signalling mediates the effect of diet on lifespan in Caenorhabditis elegans.

Dietary restriction is a robust means of extending adult lifespan and postponing age-related disease in many species, including yeast, nematode worms, flies and rodents. Studies of the genetic requirements for lifespan extension by dietary restriction in the nematode Caenorhabditis elegans have implicated a number of key molecules in this process, including the nutrient-sensing target of rapamycin (TOR) pathway and the Foxa transcription factor PHA-4 (ref. 7). However, little is known about the metabolic signals that coordinate the organismal response to dietary restriction and maintain homeostasis when nutrients are limited. The endocannabinoid system is an excellent candidate for such a role given its involvement in regulating nutrient intake and energy balance. Despite this, a direct role for endocannabinoid signalling in dietary restriction or lifespan determination has yet to be demonstrated, in part due to the apparent absence of endocannabinoid signalling pathways in model organisms that are amenable to lifespan analysis. N-acylethanolamines (NAEs) are lipid-derived signalling molecules, which include the mammalian endocannabinoid arachidonoyl ethanolamide. Here we identify NAEs in C. elegans, show that NAE abundance is reduced under dietary restriction and that NAE deficiency is sufficient to extend lifespan through a dietary restriction mechanism requiring PHA-4. Conversely, dietary supplementation with the nematode NAE eicosapentaenoyl ethanolamide not only inhibits dietary-restriction-induced lifespan extension in wild-type worms, but also suppresses lifespan extension in a TOR pathway mutant. This demonstrates a role for NAE signalling in ageing and indicates that NAEs represent a signal that coordinates nutrient status with metabolic changes that ultimately determine lifespan.

Compounds that Confer Thermal Stress Resistance and Extended Lifespan

The observation that long-lived and relatively healthy animals can be obtained by simple genetic manipulation prompts the search for chemical compounds that have similar effects. Since aging is the most important risk factor for many socially and economically important diseases, the discovery of a wide range of chemical modulators of aging in model organisms could prompt new strategies for attacking age-related disease such as diabetes, cancer and neurodegenerative disorders [Collins, J.J., Evason, K., Kornfeld, K., 2006. Pharmacology of delayed aging and extended lifespan of Caenorhabditis elegans. Exp. Gerontol.; Floyd, R.A., 2006. Nitrones as therapeutics in age-related diseases. Aging Cell 5, 51-57; Gill, M.S., 2006. Endocrine targets for pharmacological intervention in aging in Caenorhabditis elegans. Aging Cell 5, 23-30; Hefti, F.F., Bales, R., 2006. Regulatory issues in aging pharmacology. Aging Cell 5, 3-8]. Resistance to multiple types of stress is a common trait in long-lived genetic variants of a number of species; therefore, we have tested compounds that act as stress response mimetics. We have focused on compounds with antioxidant properties and identified those that confer thermal stress resistance in the nematode Caenorhabditis elegans. Some of these compounds (lipoic acid, propyl gallate, trolox and taxifolin) also extend the normal lifespan of this simple invertebrate, consistent with the general model that enhanced stress resistance slows aging.

Relationship between growth hormone (GH) status, serum leptin and body composition in healthy and GH deficient elderly subjects

Growth hormone (GH) secretion declines with age and is affected by body composition. The signal that mediates the latter relationship remains ill‐defined. Leptin, the protein product of the adipocyte specific ob gene, is thought to accurately reflect fat mass and could therefore be a candidate to influence GH secretion. We have therefore investigated the relationship between GH status, leptin and body composition in normal and GH‐deficient elderly subjects.

DAF-12-dependent rescue of dauer formation in Caenorhabditis elegans by (25S)-cholestenoic acid

Population density, temperature and food availability all regulate the formation of the Caenorhabditis elegans dauer larva by modulating endocrine signaling pathways. The orphan nuclear receptor DAF‐12 is pivotal for the decision to form a dauer or to undergo normal reproductive development. The DAF‐12 ligand has been predicted to be a sterol that is metabolized by DAF‐9, a cytochrome P450. Here we chemically characterize purified lipophilic nematode extracts and show that the ligand for DAF‐12 contains a carboxyl moiety and is likely to be derived from a sterol. Using a candidate ligand approach we find that the C27 bile acid cholestenoic acid (5‐cholesten‐3β‐ol‐(25S)‐carboxylic acid) promotes reproductive growth in dauer‐constitutive mutants in a daf‐9‐ and daf‐12‐dependent manner. Furthermore, we find that cholestenoic acid can act as a DAF‐12 ligand by activating DAF‐12 in a cell‐based transcription assay. Analysis of dauer‐rescuing lipophilic extracts from nematodes by gas chromatography–mass spectrometry indicates the presence of several regioisomers of cholestenoic acid that are distinct from Δ5‐cholestenoic acid and are not present in extracts from daf‐9 mutants. These data suggest that carboxylated sterols may be key determinants of life history.

Constitutional delay in growth and puberty (CDGP) is associated with hypoleptinaemia

Serum leptin concentrations are higher in early adolescence compared with childhood and may play a facilitatory role in pubertal development. Constitutional delay in growth and puberty (CDGP) is a disorder of the tempo of physical maturation and may be associated with relative hypoleptinaemia. We have therefore compared serum leptin concentrations in normal boys with those in boys exhibiting constitutional delay of growth, controlling for pubertal status, age and body mass index (BMI).

The relationship between stature, growth, and short-term changes in height and weight in normal prepubertal children

Human growth is a nonlinear process with marked variation in growth rate during the short-term. It is not known how long-term height gain or stature is influenced by short-term changes in height and weight. This study has addressed these issues by using thrice weekly height and weight measurements during 1 year in 43 normal prepubertal children (aged 5.7-7.7 y) to construct individual height and weight velocity curves by regression analysis. The former were comprised of 3 to 6 growth spurts separated by stasis, whereas the latter were characterized by 2 to 5 periods of weight gain separated by periods of weight loss. Stepwise regression analysis to determine characteristics of these curves that influence stature and growth showed that height SD score was correlated to the mean absolute weight velocity amplitude (+), the mean lenght of height velocity peaks (-), and the number of periods of weight gain (-) (r2 = 38%). In contrast, change in height SD score (Δheight SD score) was correlated to the number (+) and mean amplitude (+) of the periods of weight gain and the mean height velocity peak amplitude (+) (r2 = 44%). Examination of changes in height relative to weight during the year in the whole group revealed that height increased relative to weight in autumn and spring, whereas the reverse occurred during the winter months. We conclude that 1) both height and weight velocities during 1 year show a biphasic pattern, 2) there is seasonal variation in the short-term change in height relative to weight, and 3) prepubertal stature and the amount grown through the year are related to short-term changes in height and weight. Our data indicate that large but infrequent changes in weight with growth spurts of short duration are found in tall children. Good growth during the year was related to large but frequent gains in weight and large individual spurts in height.

Urinary IGF and IGF binding protein‐3 in children with disordered growth

OBJECTIVE Both IGF‐l and IGFBP‐3 reflect spontaneous GH secretion in healthy individuals. We have evaluated the clinical usefulness of urinary IGF‐I and IGFBP‐3 measurements in the diagnosis of children with disordered growth.