Jonny St-Amand

Housekeeping and tissue-specific genes in mouse tissues

BackgroundThis study aims to characterize the housekeeping and tissue-specific genes in 15 mouse tissues by using the serial analysis of gene expression (SAGE) strategy which indicates the relative level of expression for each transcript matched to the tag.ResultsHere, we identified constantly expressed housekeeping genes, such as eukaryotic translation elongation factor 2, which is expressed in all tissues without significant difference in expression levels. Moreover, most of these genes were not regulated by experimental conditions such as steroid hormones, adrenalectomy and gonadectomy. In addition, we report previously postulated housekeeping genes such as peptidyl-prolyl cis-trans isomerase A, glyceraldehyde-3-phosphate dehydrogenase and beta-actin, which are expressed in all the tissues, but with significant difference in their expression levels. We have also identified genes uniquely detected in each of the 15 tissues and other tissues from public databases.ConclusionThese identified housekeeping genes could represent appropriate controls for RT-PCR and northern blot when comparing the expression levels of genes in several tissues. The results reveal several tissue-specific genes highly expressed in testis and pituitary gland. Furthermore, the main function of tissue-specific genes expressed in liver, lung and bone is the cell defence, whereas several keratins involved in cell structure function are exclusively detected in skin and vagina. The results from this study can be used for example to target a tissue for agent delivering by using the promoter of tissue-specific genes. Moreover, this study could be used as basis for further researches on physiology and pathology of these tissues.

Maximum tolerable dose of red pepper decreases fat intake independently of spicy sensation in the mouth

Dietary red pepper suppresses energy intake and modifies macronutrient intake. We have investigated whether a stimulus in the mouth and the sensation of spiciness are necessary for red pepper-induced changes in energy and macronutrient intake in human volunteers. In a preliminary test, sixteen Japanese male volunteers tasted samples of a soup with graded doses of red pepper in order to define a moderate and a maximum tolerable (strong) dose of red pepper. On the day of the experiment, a standardised breakfast was given to the volunteers. At lunchtime, the subjects ingested one of four experimental soups containing either a placebo, a moderate or a strong dose of red pepper plus placebo capsules, or a placebo soup plus capsules delivering a strong dose of red pepper. The rest of the meal was given ad libitum to all subjects. The amount of food, protein and carbohydrate ingested was similar for all conditions. Energy and fat intake were similar after the ingestion of the moderate soup compared with placebo. However, the strong soup significantly lowered fat intake compared with placebo (P=0.043), and ingestion of strong capsules also tended to suppress it (P=0.080). Moreover, energy intake after strong soup and capsules tended to be lower than placebo (P=0.089 and 0.076, respectively). The present results indicate that the maximum tolerable dose is necessary to have a suppressive effect of red pepper on fat intake. The main site of the action of red pepper is not in the mouth.

Serial analysis of gene expression in the skeletal muscle of endurance athletes compared to sedentary men

Physical exercise produces several adaptive changes in skeletal muscle. However, the molecular mechanisms of these effects are poorly understood. We performed serial analysis of gene expression (SAGE) to quantify the global gene expression profile in sedentary and endurance‐trained muscle. A total of 10869 SAGE tags was sequenced and represented 4727 genes. The genes most expressed in muscle are mainly involved in contraction and energy metabolism. Thirty‐three genes were differentially expressed between endurance athletes and sedentary individuals. Four genes such as myosin binding protein C fast‐type, glycogen phosphorylase, and pyruvate kinase were expressed less in endurance athletes, whereas eight genes coding for expressed sequence tag similar to (EST) crystallin alpha B, EST myosin light chain 2, EST surfactant pulmonary‐associated protein A1, EST thrombospondin, EST fructose‐bisphosphate aldolase A, EST cytochrome oxidase 1, NADH dehydrogenase 3, and G8 protein were up‐regulated. Most of the up‐regulated tags corresponded to novel genes. On the other hand, different isoforms of fructose‐bisphosphate aldolase A were also differentially expressed. The current study underlying the most highly expressed genes allows a better understanding of global muscle characteristics in normal and endurance‐trained individuals. Moreover, the current data suggest novel candidate genes that may be responsible for enhanced endurance performance.—Yoshioka, M., Tanaka, H., Shono, N., Snyder, E. E., Shindo, M., St‐Amand, J. Serial analysis of gene expression in the skeletal muscle of endurance athletes compared to sedentary men. FASEB J. 17, 1812–1819 (2003)

Gender difference of androgen actions on skeletal muscle transcriptome

Sarcopenia is related to metabolic syndrome in postmenopausal women. Hormone replacement therapies with androgens improve muscle functions by molecular mechanisms that are still unknown, at least partly because the skeletal muscle transcriptome has been less characterized in females. We performed the serial analysis of gene expression method in six experimental groups, intact (male and female), ovariectomy (OVX), OVX+dihydrotestosterone (DHT) injection 1, 3, or 24 h before kill in mice. The 438 transcript species differentially expressed between gender showed that females had higher expression levels of mRNA related to cytoskeleton/contractile apparatus and mitochondrial processes as well as protein, lipid, and amino acid metabolisms. In females, OVX and DHT modulated 109 and 128 transcript species respectively. OVX repressed transcripts of fast/glycolytic fiber, glycolysis, and glucose transport, whereas all these effects were reversed 3 h after the DHT injection. Moreover, DHT treatment induced transcripts which reduce intracellular Ca(2+) level at early time points. These results may suggest that DHT treatment in OVX mice increases muscle contractility by affecting fiber distribution and intracellular Ca(2+) concentration as well as improving glucose metabolism. On the other hand, transcripts of fast/oxidative fiber, oxidative phosphorylation, and ATP production were repressed 24 h after DHT administration. In our previous study using male mice, transcripts in oxidative phosphorylation and ATP production were induced 24 h after DHT injection (Yoshioka M, Boivin A, Ye P, Labrie F & St-Amand J 2006 Effects of dihydrotestosterone on skeletal muscle transcriptome in mice measured by serial analysis of gene expression. Journal of Molecular Endocrinology 36 247-259 ). These results demonstrate gender differences in DHT actions on skeletal muscle, and contribute to a precise understanding of the molecular mechanisms of androgen actions in the female skeletal muscle.

Regulation of skeletal muscle transcriptome in elderly men after 6 weeks of endurance training at lactate threshold intensity.

A compromised muscle function due to aging, sarcopenia and reduced level of physical activity can lead to metabolic complications and chronic diseases. Endurance exercise counters these diseases by inducing beneficial adaptations whose molecular mechanisms remain unclear. We have investigated the transcriptomic changes following mild-intensity endurance training in skeletal muscle of elderly men. Seven healthy subjects followed an exercise program of cycle ergometer training at lactate threshold (LT) level for 60 min/day, five times/week during six weeks. Physiological and transcriptomic changes were analyzed before and after training. LT training decreased percentage body fat and fasting levels of plasma glucose, while increasing high-density lipoprotein cholesterol and lecithin-cholesterol acyltransferase levels. Transcriptomic analysis revealed fast-to-slow fiber type transition, increased amount of mtDNA encoded transcripts and modulation of 12 transcripts notably related to extracellular matrix (ECM), oxidative phosphorylation (OXPHOS), as well as partially characterized and novel transcripts. The training simultaneously induced the expression of genes related to slow fiber type transition, OXPHOS and ECM, which might contribute to the improvement of glucose and lipid metabolisms and whole body aerobic capacity.

Transcriptomic characterization of the long-term dihydrotestosterone effects in adipose tissue

Objective: To study the long‐term transcriptomic effects of dihydrotestosterone (DHT) in adipose tissue. Fat distribution is regulated by sexual hormones. It is still unclear if androgens are promoting or reducing intra‐abdominal fat accumulation.

High-fat meal-induced changes in the duodenum mucosa transcriptome.

In order to identify the potential peripheral signals of appetite and satiety from duodenum, we have performed a transcriptomic study in the mucosa after high‐fat (HF) and low‐fat (LF) meal ingestion. After fasting, one group of mice was killed and the others were fed ad libitum with HF or LF diet, and killed 30 min, 1 h, and 3 h after the beginning of the meal. The duodenum mucosa was sampled, and the serial analysis of gene expression (SAGE) method was performed. The mRNA regulations were confirmed by real‐time PCR. Energy, protein, and fat intakes were higher in the HF than in the LF group. Gene expression profile revealed 118 characterized or partially characterized differentially expressed transcripts. The HF meal delayed the expressions of peptidases compared to the LF groups. Most of mRNAs related to fat absorption, including apolipoprotein A‐IV (Apoa4), were decreased in HF1h group, whereas plasma triglyceride (TG) levels were comparable between HF and LF groups. Noteworthy, these downregulations were concomitant to a break in fat intake 1 h after HF meal. At the same time, the HF meal induced transcripts related to cell growth and organization, whereas transcripts involved in cell defense were repressed. Moreover, we have identified fat‐responsive transcripts. This study has characterized the molecular responses of duodenum mucosa after HF or LF meal ingestion. Characterization of novel fat‐specific candidates whose relations with feeding behavior have never been reported may contribute to the development of new therapeutic targets for appetite and satiety controls.

Regulation of Muscle Genes by Moderate Exercise

Moderate-intensity exercise at the lactate threshold (LT) is considered to be a safe and effective training regimen for improving metabolic syndrome. The aim of the current study was to investigate the effects of moderate exercise performed at the LT on skeletal muscle gene expression. 6 healthy men participated in cycle ergometer training at LT, 60 min/d, 5 d/wk for 12 wks. Muscle samples were collected after 5 d of training, and then 2 d after training at wks 6 and 12. Quantitative real-time PCR analysis revealed that the expression of peroxisome proliferator activated receptor co-activated 1alpha was significantly increased at 1 h after the training session on day 5. Moreover, using serial analysis gene expression, we found that moderate training for 6 and 12 wks simultaneously induced the expression of a number of metabolic genes involved in the TCA cycle, beta-oxidation, and electron transport. Furthermore, several genes encoding antioxidant enzymes and contractile apparatus were induced. The expression levels of 233 novel transcripts were also altered in response to moderate exercise. Thus, moderate training at the LT is a sufficient stimulus to induce the expression of numerous genes implicated in the development of metabolic syndrome, transcripts involved in the contractile apparatus, and novel transcripts.

Atlas of dihydrotestosterone actions on the transcriptome of prostate in vivo.

Using serial analysis of gene expression (SAGE), we studied the transcriptomic changes in vivo by dihydrotestosterone (DHT) treatment in mice to better understand androgen effects in the prostate.

Feeding induced changes in the hypothalamic transcriptome.

BACKGROUND Obesity is a complex multifactorial disorder which needs a comprehensive approach for prevention and treatment. We investigated the modifications in the hypothalamic gene expression induced by high-fat (HF) and low-fat (LF) meal ingestion in mice, in order to identify the signals rapidly mediating the hypothalamic control on energy intake. METHODS After fasting, 1 group of mice was sacrificed and the others were fed ad libitum with HF or LF diet, and sacrificed 3 h after the beginning of the meal. The hypothalamus was sampled and the serial analysis of gene expression method was performed. RESULTS Approximately 254,588 tags, which correspond to 65,548 tag species were isolated from the 3 groups. The data showed twelve transcripts regulated by food intake. Among these, 2 transcripts have mitochondrial functions (MtCo1, Ppid), 3 are involved in protein transport and regulation (Ube2q2, Mup1, Sec13), 1 in cellular pH control (Slc4a3) and another 1 has a role in the epigenetic control of gene expression (Setd3). In addition, 5 potentially novel transcripts were differentially modulated. CONCLUSION We identified genes that may regulate hypothalamic circuits governing the early response to food intake. 3 genes were specifically modulated by high-fat intake.