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Morphological changes in the testis induced by diethylcarbamazine.

Diethylcarbamazine (DEC) had been proved to be highly effective against lymphatic filariasis, however its effect on vertebrate cells remains uncertain. After 12 days treatment with DEC, most of the Leydig cells were hypertrophied with several lipid droplets, and others had no nucleus and presented characteristic steatosis features. Vacuolization of Sertoli cells was also noted. Ultrastructural analyses of DEC-treated testes revealed spermatogonies with morphological characteristics of apoptosis, as shrinkage of cytoplasm and increased chromosomal density. In addition, Leydig cells showed numerous lipid droplets scattered throughout the cytoplasm, multivesicular bodies and giant whorl-like smooth endoplasmic reticulum. Several spermatids presented vacuolated mitochondriae, which were disorganized in relation to the microtubular axis of the flagellae. These results indicate that DEC probably affects the microtubular function, however the present data does not exclude the possibility that DEC also can act directly on enzymatic hormonal pathways.

Administration, Oral↗

Effects of oral estrone on rat energy balance.

Oral doses of estrone from 10 nmol/(kg day) to 10 micromol/(kg day) were given to adult Wistar male rats for 10 days. Body composition, energy balance, total body estrone balance and plasma metabolites and hormones were measured at the end of the treatment. Body weight (as well as food intake, body energy, fat and water accrual) increased at doses in the 10--100 nmol/(kg day) range, but decreased at higher doses. Energy expenditure decreased with increasing doses of estrone. Plasma metabolite changes suggested the maintenance of energy homeostasis, and lipid parameters indicated that lipid mobilization increased with the increasing doses of estrone. Plasma estrone, acyl-estrone and estradiol levels decreased at low doses and increased at high doses of estrone. We conclude that: (a) repeated estrone gavages, even at very high doses, do not result in the accrual of estrone in the body; (b) low doses of estrone promote growth and high doses decrease body mass and fat accretion; (c) administration of estrone at low doses decreases its circulating levels and the levels of estradiol and acyl-estrone, a situation reverted at higher doses and (d) estrone administration induces a dose-dependent shift towards lower energy expenditure.

3-Hydroxybutyric Acid↗

Mechanistic toxicogenomic analysis of WAY-144122 administration in Sprague-Dawley rats.

Application of global gene expression analysis in the study of mechanisms of toxicity could provide a more comprehensive interpretation of the molecular basis of drug action. WAY-144122 has pharmacological activity against several targets improving insulin responsiveness and favorably altering lipid profiles. Normal rats treated with suprapharmacological doses of WAY-144122 for 28 days exhibited drug-related effects in the liver and ovary. To determine the molecular mechanism underlying these effects, we employed global gene expression profiling to measure RNA levels in these target organs obtained from WAY-144122-treated rats administered test article for 1, 3, 7, and 14 days. Genes altered in expression by WAY-144122 were functionally categorized and related to their biological activity. In the liver, WAY-144122 caused a widespread up-regulation of genes involved in lipid mobilization, peroxisomal proliferation, and fatty acid beta-oxidation. In the ovary, we observed reduced expression of genes encoding luteinizing hormone receptor, follistatin, and enzymes in the estradiol synthesis pathway. Transcriptional changes in both organs precede histopathological effects. Profiling analysis allowed us to formulate hypotheses for molecular mechanisms underlying the physiological observations. In the liver, transcriptional changes suggest that WAY-144122 induced increased metabolic activity and peroxisomal proliferation resulting in increased liver weight and hepatocellular hypertrophy. We propose decreased estradiol synthesis as the underlying mechanism for the observed follicular atrophy in the ovary. Importantly, in this study, we have identified potential molecular mechanisms of drug effect in expression profiles before observation of physiological changes.

Administration, Oral↗

Molecular cloning and functional expression of a Drosophila corazonin receptor.

The insect adipokinetic hormones (AKHs) constitute a large family of neuropeptides that mobilize lipids and sugar from the insect fat body during energy-requiring activities such as flight. We have previously identified the first insect AKH receptors from the fruitfly Drosophila melanogaster and the silkworm Bombyx mori (Staubli et al., PNAS 2002, 99: 3446-3451). Here, we have cloned the cDNA of a Drosophila G protein-coupled receptor that was closely related to the first Drosophila AKH receptor both with respect to amino-acid sequence and gene structure. We have subsequently expressed this orphan receptor in Chinese hamster ovary cells and identified Drosophila corazonin as the endogenous ligand for the receptor. Corazonin increases heart beat in some insects, but its function in Drosophila is unknown. These results are intriguing, because not only are the Drosophila AKH and corazonin receptors structurally and evolutionarily related, but also are their preprohormones, which suggests a co-evolution of ligands and receptors. The Drosophila corazonin receptor is expressed in embryos, larvae, pupae, and adult flies. Furthermore, a receptor that is structurally very similar to the Drosophila corazonin receptor can be found in the genomic database from the malaria mosquito Anopheles gambiae.

Amino Acid Sequence↗

Gln27Glu beta2-adrenergic receptor variant is associated with hypertriglyceridemia and the development of fatty liver.

BACKGROUND: Nonalcoholic steatohepatitis (NASH) is associated with the metabolism of lipid, glucose and energy. Beta-adrenergic receptors play an important role in the regulation of energy expenditure, in part, by stimulating lipid mobilization through lipolysis. METHODS: To assess whether it is common for the beta2-adrenergic receptor (B2AR) gene polymorphisms in codons 16 and 27 to play a role in the development of fatty liver, we investigated 251 unrelated healthy Japanese males who were drug-free and showed no signs of heavy drinking. RESULTS: The allelic frequency of B2AR gene mutation in codons 16 and 27 did not differ between obese subjects (BMI>25.0 kg/m(2), n=151) and non-obese subjects (BMI</=25.0 kg/m(2), n=100). The Gly16 homozygotes had a lower high-density lipoprotein cholesterol (HDL-C) level than the Arg16 homozygotes (1.50+/-0.4 vs. 1.32+/-0.3 mmol/l, p=0.014). However, no significant association with fatty liver was observed in the Gly16 allele frequency. The Gln27Glu27 heterozygotes showed higher concentrations of serum triglycerides (TG) than the Gln27Gln27 homozygotes (1.62+/-0.93 vs. 2.21+/-1.67 mmol/l, p=0.013). This correlation was also observed in all subjects regardless of weight classification. Univariate analysis indicated that subjects with the heterozygous Gln27Glu mutant alleles had a significantly higher prevalence of fatty liver vs. those without the mutation (Glu27 allele frequency, 0.07 vs. 0.12, p=0.047; odds ratio, 1.92; 95% confidence interval, 1.01-3.68). However, multivariate logistic regression models showed the prevalence of fatty liver to be significantly related to the homeostasis model assessment (HOMA) index, BMI, triglyceride and HDL-cholesterol. CONCLUSIONS: These results suggest that the amino-terminal polymorphisms of the beta2-adrenergic receptor gene in codon 27 were associated with hypertryglyceridemia and independent of obesity, and thereby could be involved in the molecular pathogenesis of fatty liver.

Adult↗

Identification of an Arabidopsis mitochondrial succinate-fumarate translocator.

Complementation of a yeast acr1 mutant carrying a deletion of the succinate/fumarate carrier gene enabled functional identification of a mitochondrial succinate translocator from Arabidopsis thaliana (AtmSFC1). Thus complementation of yeast mutants is applicable also for identification and characterization of organellar transporters. Reverse transcription polymerase chain reaction and promoter-GUS fusion showed expression of AtmSFC1 in 2 day old dark grown seedlings, which declined in cotyledons during further development, consistent with a role in export of fumarate for gluconeogenesis during lipid mobilization at early germination of Arabidopsis seeds. In mature plants, expression was found in developing and germinating pollen, suggesting a role in ethanolic fermentation.

Amino Acid Sequence↗

Relationships between obesity and metabolic hormones in the "cobalt" variant of rainbow trout.

The "cobalt" variant of rainbow trout (Oncorhynchus mykiss) lacks most of the pars intermedia of the pituitary, and shows significant obesity with an enlarged liver and a fat accumulation in the abdominal cavity. Plasma levels of growth hormone, prolactin, and somatolactin were significantly lower in the cobalt variant than those in the normal trout. In contrast, plasma insulin level was four times higher than that in the normal. Plasma levels of total protein, free cholesterol, and triacylglycerol were higher in the cobalt, while those of glucose and fatty acids were not different from the normal levels. In the white muscle, red muscle, liver, and mesenteric fat, the cobalt showed higher contents of triacylglycerol than the normal fish. There was no significant difference in tissue contents of phosphatidylcholine between the two groups of the trout, except for that in the mesenteric fat, exhibiting significantly lower content than in the normal fish. Activity of triacylglycerol lipase in the liver in vivo was lower in the cobalt than that in the normal trout, while there was no significant difference between the two in the cultured liver slices. Desacetyl-alpha-MSH stimulated lipolysis of triacylglycerol similarly in the cultured liver slices from the normal trout and from the cobalt variant. Results from this study suggest that the lack of pars intermedia and the increased plasma level of insulin are involved in a depression of lipid mobilization and obesity in this variant of rainbow trout.

Animals↗

Chemical sympathectomy alters food intake and thermogenic responses to catecholamines in rats.

It has been suggested that the sympathetic nervous system contributes to the short-term control of feeding. The adrenergic innervation of some splanchnic organs seems to be especially involved in such processes, since catecholamines reduce feeding only when injected intraperitoneally or intraportally. In this work, the effects of neonatal sympathetic denervation with guanethidine (Gnt) upon food intake were assessed in adult rats. Gnt-treated male rats had lower body weight gain. The hypophagic response to intraperitoneal (ip) norepinephrine was 70% higher in Gnt-treated animals as compared to controls (P < 0.05); that of epinephrine (E) by 33% (P < 0.05) and that of isoproterenol was not significantly modified. As in normal rats, the hypophagic effect was much stronger after ip than after intramuscular (im) administration (P < 0.05). On the other hand, resting oxygen consumption (VO2) was consistently lower in denervated animals. Ip E administration did not modify VO2, while im E caused increased motor activity and VO2 (P < 0.05). In contrast to control rats, the respiratory exchange ratio in ad libitum fed Gnt rats did not decrease after Ip E administration, suggesting a lack of effect upon lipid mobilization. The lower rate of body weight gain induced by neonatal Gnt sympathectomy might be due to lower daily food intake possibly related, in part, to the sensitization of the alpha-adrenergic porto-hepatic response to endogenous catecholamines. Compared with controls, Gnt-treated rats also showed a limited thermogenic capacity not related to feeding, and a greater degree of carbohydrate oxidation, possibly due to a defect in E-induced lipolysis, which is beta-adrenergic.

Adrenergic Agonists↗

No difference in the lipolytic response to beta-adrenoceptor stimulation in situ but a delayed increase in adipose tissue blood flow in moderately obese compared with lean men in the postexercise period.

This study was undertaken to determine the effect of previous exercise on adipose tissue responsiveness to beta-adrenoceptor stimulation and on adipose tissue blood flow (ATBF). Eight lean and 8 obese men (body mass index [BMI], 23.6 +/- 2.1 [SD] v 29.0 +/- 1.9 kg x m(-2)) were investigated with abdominal subcutaneous microdialysis and 133Xe clearance. A stepwise isoprenaline infusion (10(-8), 10(-7), and 10(-6) mol x L(-1)) was administered in situ in the microdialysis catheter before and 2 hours after a submaximal exercise bout (90 minutes of cycling at 55% of maximal O2 uptake). No differences in the response (increase in interstitial glycerol v preinfusion level) to isoprenaline infusions were found between the 2 groups. In both groups, there was no difference in the response to postexercise versus preexercise infusion. When the vasodilating agent hydralazine (0.125 g x L(-1)) was infused into the microdialysis catheter to control for the vascular effects of isoprenaline, an interaction effect between exercise and isoprenaline dose was found. Analyses showed an attenuated response to the high isoprenaline dose after exercise (lean, 251 +/- 42 [SE] micromol x L(-1); obese, 288 +/- 77 micromol x L(-1)) versus before exercise (lean, 352 +/- 62 micromol x L(-1), P = .045 v after; obese, 380 +/- 94 micromol x L(-1), P = .021 v after), indicating a desensitization of lipolysis to beta-adrenoceptor stimulation. ATBF and arterial plasma glycerol increased after exercise in both groups, but the increase was delayed in obese subjects. Arterial plasma insulin was higher in the obese versus lean subjects at all times, and decreased during recovery in both groups. In conclusion, abdominal subcutaneous adipose tissue responsiveness to beta-stimulation is not enhanced postexercise in lean and obese men, whereas previous exercise increases ATBF. Furthermore, the data suggest slower lipid mobilization postexercise and resistance to the antilipolytic effect of insulin in the obese.

Adipose Tissue↗

MTII administered peripherally reduces fat without invoking apoptosis in rats.

The melanocortin (MC) system in the brain is believed to be an important downstream effector of leptin signaling; interference with MC functioning results in severe obesity. Melanotan II (MTII), an MC3/4-receptor agonist, produces similar behavioral and metabolic outcomes to those observed after leptin treatments, which enhance apoptosis in specific fat depots. To determine whether MTII also mediates adipose apoptosis induced by leptin treatment, two groups of rats (n=8) received MTII (2 mg/kg, i.p.) or saline (2 ml/kg) once daily for 4 days and had free access to food and water, and a third group was injected with saline and pair-fed (PF) to MTII treated rats. Food intake, water intake, body temperature, and body weight were measured daily. MTII reduced food and water intake and body weight gain (P<.05) and decreased body temperature compared to PF and saline-treated control groups. Retroperitoneal white adipose tissue (WAT) mass and epididymal WAT mass were reduced 46.3% and 21.1%, respectively (P<.05), after MTII, but not after PF, compared with the saline control rats. Both MTII- (25.0%) and PF (33.3%)-treated rats had decreased brown fat weight (P<.05), whereas muscle mass remained unchanged. Free fatty acid concentrations in serum were not different between MTII and control groups, but increased by 56.4% in PF group. DNA fragmentation assay did not support a role for MTII as an apoptotic signal in any of the fat tissues tested. These results show that in addition to reducing food intake and inhibiting body weight gain, intraperitoneal administration of MTII reduces fat mass, most likely by accelerated lipid mobilization, but not by apoptosis.

Adipose Tissue↗

Effects of acetoacetate and D-beta-hydroxybutyrate on bovine in vitro embryo development in serum-free medium.

It is known that the ketone bodies acetoacetate and D-beta-hydroxybutyrate can be metabolized by the early bovine embryo for in vitro development. In the present work, we report experiments leading to the culture of bovine embryos in the absence of serum. In vitro-produced bovine zygotes were cultured in modified synthetic oviduct fluid medium supplemented with acetoacetate derivatives, acetoacetate and D-beta-hydroxybutyrate. Acetoacetate and its derivatives prevented blastocysts from forming in the absence of serum during the whole culture period. However, from Days 6 to 8 of culture in the absence of serum, acetoacetate did not affect development as compared to controls containing lactate and pyruvate or no substrate. Interestingly, D-beta-hydroxybutyrate stimulated blastocyst and expansion development, and allowed lipid mobilization. In feeder cells coculture, embryos produced with D-beta-hydroxybutyrate showed improved hatching. Embryos cultured in D-beta-hydroxybutyrate were viable upon transfer to recipients, although no pregnancies were confirmed later by ultrasonic scanning. The protective effect of serum upon embryos cultured in medium containing acetoacetate is apparently not required in the presence of D-beta-hydroxybutyrate.

3-Hydroxybutyric Acid↗

Identification of the cockroach neuropeptide Pea-CAH-II as a second adipokinetic hormone in the firebug Pyrrhocoris apterus.

A new member of the AKH/RPCH family was isolated from the corpora cardiaca of the firebug Pyrrhocoris apterus. It is the second adipokinetic peptide identified in this species. The peptide was characterized and its structure was deduced from the multiple MS(N) electrospray mass spectra as that of an octapeptide with the sequence pGlu-Leu-Thr-Phe-Thr-Pro-Asn-Trp-NH(2.) The peptide differs from the original P. apterus AKH (Pya-AKH) by one amino acid in position 3. Topical application and/or injection of the peptide induced lipid mobilization, but was inactive in mobilization of carbohydrates.

Animals↗

Peptides in the locusts, Locusta migratoria and Schistocerca gregaria.

The first peptide identified in locusts was adipokinetic hormone I (AKH-I), a neurohormone mobilizing lipids from the fat body. No other locusts peptides were isolated until 1985. From then on peptide identification started to boom at such a tremendously fast rate that even specialists in the field could hardly keep track. At this moment the total number of different insect neuropeptide sequences exceeds 100. Currently, the locusts Locusta migratoria and Schistocerca gregaria are the species from which the largest number of neuropeptides has been isolated and sequenced, namely 56. Myotropic bioassays have played a major role in the isolation and subsequent structural characterization of locust neuropeptides. They have been responsible for the discovery of locustamyotropins, locustapyrokinins, locustatachykinins, locustakinin, locusta accessory gland myotropins, locustasulfakinin, cardioactive peptide, and locustamyoinhibiting peptides. Members of the myotropin peptide families have been associated with a variety of physiological activities such as myotropic activities, pheromonotropic activities, diapause induction, stimulation of cuticular melanization, diuresis, pupariation, and allatostatic activities. Recently, we have identified in Schistocerca 10 peptides belonging to the allatostatin peptide family, which inhibit peristaltic movements of the oviduct. Some of the myotropins appear to be important neurotransmitters or modulators innervating the locust oviduct, the salivary glands, the male accessory glands, and the heart, whereas others are stored in neurohemal organs until release in the hemolymph. Some myotropic peptides have been found to be releasing factors of neurohormones from the corpora cardiaca. Several peptides isolated in locusts appear to be unique to insects or arthropods; others seem to be members of peptides families spanning across phyla: two vasopressin-like peptides, FMRFamide-related peptides, Locusta diuretic hormone (CRF-like), Locusta insulin-related peptide, locustatachykinins, locustasulfakinin (gastrin/CCK-like). In a systematic structural study of neuropeptides in Locusta, several novel peptides have been isolated from the corpora cardiaca and the pars intercerebralis. They include the neuroparsins, two 6-kDa dimeric peptides, and three proteinase inhibitors. Ovary maturating parsin is the first gonadotropin identified in insects. The isolation of a peptide from an ovary extract that inhibits ovary maturation in Schistocerca gregaria is currently underway in our lab. The proteinase inhibitors, recently found to be mainly transcribed in the fat body, are believed to play a role in defense reactions of insects. Finally, a locust ion transport peptide and a peptide stimulating salivation recently can be added to this extensive list of locust peptides.

Amino Acid Sequence↗

[Development of nutritional obesity in transgenic mice with an adrenergic receptivity in adipose tissue comparable with that of humans].

Obesity is characterized by an excessive development of fat mass which is a consequence of increased fat cell size and/or fat cell number. Several hormones and neurotransmitters are regulators of adipose tissue development and metabolism. Among them, catecholamines play a major role by acting through alpha 2- and beta-adrenergic receptors. Stimulation of alpha 2-adrenergic receptors induce inhibition of lipolysis in mature adipocytes as well as preadipocyte proliferation. The antilipolytic effect mediated by alpha 2-adrenergic receptors is in part responsible for the weak lipid mobilization of some fat deposits in humans (subcutaneous fat in particular). Changes in beta- and alpha 2-adrenergic receptors ratio and function have been proposed to explain the lipolytic disturbances described in some obese subjects. Human and rodent adipocytes differ considerably with respect to the balance between beta- and alpha 2-adrenergic receptors. Human adipocytes express mainly alpha 2- but very few beta 3-adrenergic receptors while the reverse is true for rodent adipocytes. Since no suitable animal model was available to study the contribution of alpha 2/beta-adrenergic balance in adipocytes in vivo, we combined gene targeting and transgenic approaches to create a mice with increased alpha 2/beta-adrenergic ratio in adipose tissue. Specifically, we have generated transgenic mice strains on a beta 3-adrenergic receptor knock-out background which express human alpha 2-adrenergic receptors. No particular phenotype was observed in mice maintained in normal diet whereas when fed a high fat diet, transgenic mice increased significantly body weight and fat mass. These results underline the physiologic relevance of the interaction of the presence of alpha 2-adrenergic receptors with a high fat diet in the control of adipose tissue development.

Animals↗

Delayed myelination in a rhizomelic chondrodysplasia punctata case: MR spectroscopy findings.

Rhizomelic chondrodysplasia punctata is a member of genetic peroxisomal disorders. Delayed myelination, which is probably related to the inadequacy of plasmalogens biosynthesis, is an important feature of this disorder. Direct assessment of neuropathologic aspects of RCDP syndrome such as neuronal degeneration and delayed myelination is possible with MR spectroscopy. In this report, MR spectroscopy findings (decreased Cho/Cr and increased Ins-Gly/Cr ratios and increased levels of mobile lipids) of a rhizomelic chondrodysplasia punctata case supporting delayed myelination are presented. This is the second report of MR spectroscopy examination of the specific brain metabolic changes associated with rhizomelic chondrodysplasia punctata.

Aspartic Acid↗

Paradoxical increases of circulating nonesterified fatty acids in somatotropin treated cattle undergoing mild disturbances.

Effects of various doses of bovine somatotropin (bST) on plasma concentrations of nonesterified fatty acids (NEFA) were studied in a 14-d Latin square with six Holstein heifers. Animals were given daily injections of excipient or bST at 12:00 p.m. and fed twice daily at 7:00 a.m. and 7:00 p.m. On Day 14, plasma NEFA remained low through the day except around the 7:00 p.m. feeding when they were substantially elevated. The elevation was significantly greater in bST-treated animals and corresponded to the excitement of the animals in anticipation of the evening feeding. To further investigate this phenomenon, a second experiment was conducted in which nine growing Holstein steers were fed hourly and received either daily intramuscular (i.m.) injection of excipient or bST (120 mg/kg BW) for 15 d in a crossover design. Daily profiles of NEFA were obtained under undisturbed conditions or concurrently with intensive handling. Although no elevations could be detected in any case in control animals, bST caused a substantial rise in NEFA concentration only when animals were subjected to intensive handling. This suggested that NEFA peaks noted in bST-treated heifers in the first experiment resulted from increased ability of adipose tissue to respond to adrenergic stimulation associated with the anticipation of feeding. Consistent with this hypothesis, plasma NEFA concentrations in bST-treated steers were increased to a greater extent during a challenge involving i.v. injection of epinephrine. This amplification of adipose tissue response by bST must be considered when conducting intensive studies. Even the minimal excitement associated with blood sampling can confound the results regarding lipid mobilization, and this may have contributed to the notion that ST is a lipolytic hormone.

Adipose Tissue↗

The role of circulating catecholamines in the regulation of fish metabolism: an overview.

The physiological role of the catecholamines (CA), adrenaline and noradrenaline in fish has been frequently reviewed, but the metabolic consequences of these hormones have received less attention. The purpose of this review is to examine the recent literature dealing with CA actions on whole fish and tissue metabolism. The CA increase glucose production both in vivo and in vitro, at least in isolated hepatocytes. Although the data are less clear, lipid mobilization is also a consequence of elevated circulating CA. The difficulty with using the whole fish for such studies is that CA may alter other circulating hormone levels, CA turnover in the circulation quickly, and it is difficult to define precisely the tissue being affected. Much of our understanding is derived, therefore, from the study of isolated tissues, and especially the hepatocyte. Catecholamines stimulate both glycogenolysis and gluconeogenesis in hepatocytes isolated from a large number of fish species. This review examines the steps involved in the signal transduction system, from the binding of CA to alpha- and beta-adrenoceptors to the ultimate effects of specific enzyme phosphorylation. Recent literature demonstrates that the complexity of the adrenoceptor system noted for mammals, also is expressed in fish. Adrenoceptor subtypes are specific to species, to tissues and to function of the tissues, and these issues are discussed especially as they are related to external and to internal stressors. Future research will pursue better definitions of the adrenoceptor systems, molecular biology of the components of these receptor systems and development of alternative cell models. There still remains a poor explanation of the reason for the diversity of adrenoceptor systems, and there are a number of fish systems that may provide unique opportunities to understand this question.

Animals↗