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Relationship of adipokinetic hormone I and II to migratory propensity in the grasshopper, Melanoplus sanguinipes.

This report examines three aspects of adipokinetic hormone (AKH) involvement in migratory flight behavior in the grasshopper, Melanoplus sanguinipes. The titer of hemolymph AKH I during long-duration tethered flight was examined using radioimmunoassay (RIA) after narrow bore RP-HPLC. The hemolymph fraction containing AKH I was assayed using commercially available anti-Tyr1-AKH I serum. Titer determinations of hemolymph AKH were done at rest and after various periods of flight. The amount of AKH I released from the corpora cardiaca during flight was estimated. When resting levels of AKH I and II in corpora cardiaca (CC) of migrants and non-migrants were examined with HPLC, no significant differences in AKH levels were detected between non-migrants, animals that had flown for 1 h to identify them as migrants, and animals that had flown to exhaustion (i.e., voluntary cessation). CC levels of both AKH I and II were less in this species than in locusts. When the lipid mobilization in response to AKH I and II was compared in migrants (animals that had self-identified as migrants in a 1-h tethered flight test) and non-migrants (animals that would not perform a 1-h flight in a tethered flight test), the adipokinetic response to AKH I was greater in migrants than in non-migrants, possibly indicating differences in level of sensitivity or number of receptors in the target tissues. AKH II had little effect on hemolymph lipid levels in either flight group, and may not play a significant role in lipid mobilization in this species.

Analysis of Variance↗

Effects of exogenous intravenous glucose on plasma glucose and lipid homeostasis in anesthetized infants.

BACKGROUND: Whether intravenous glucose administration to infants during anesthesia is necessary remains to be resolved. The current study was designed to investigate the effect of exogenous glucose infusion on plasma glucose and lipid homeostasis in infants undergoing minor surgery. METHODS: Sixty infants (inpatients, ASA physical status 1) between 1 and 11 months of age were divided randomly into three groups as follows: LR group, lactated Ringer's solution (LR) alone; D2LR group, 2% glucose in LR; and D5LR group, 5% glucose in LR. Anesthesia was induced and maintained with halothane and nitrous oxide in oxygen. All fluids were infused at a rate of 6 ml.kg-1.h-1 until 1 h after surgery. Plasma concentrations of glucose, nonesterified fatty acids, ketone bodies, insulin, and cortisol were determined at induction of anesthesia, at the end of surgery, and 1 h after surgery. RESULTS: No infants in the three groups had hypoglycemia (< 50 mg.dl-1) throughout the study. In the LR group, plasma glucose concentration remained unchanged perioperatively compared with the basal values (at induction), whereas in the D2LR group, it increased during surgery but remained normoglycemic. In the D5LR group, plasma glucose concentration increased markedly both during and after surgery. In 6 of 20 infants, plasma glucose was greater than 200 mg.dl-1 at the end of surgery. In 8 of 20 infants receiving glucose-free infusion, plasma glucose concentrations decreased at the end of surgery. In contrast, the plasma glucose concentration increased in infants receiving glucose infusion. In the LR group, plasma concentrations of nonesterified fatty acids and ketone bodies increased at the end of and after surgery, suggesting lipid mobilization. The base excess decreased in the LR groups as concentration of the ketone bodies increased. Plasma insulin concentrations increased in the D2LR and D5LR groups and decreased after surgery in infants receiving a glucose-free solution. No intergroup differences in plasma cortisol concentrations existed at any sample point. CONCLUSIONS: These data indicate that, in otherwise healthy infants undergoing minor surgery, intravenous infusion of 2% glucose may be sufficient to maintain plasma glucose concentrations within physiologic ranges and to prevent a compensatory increase in lipid mobilization (lipolysis) when fluids are infused at a rate of 6 ml.kg-1.h-1. However, there are limitations in extrapolating the results to neonates.

Anesthesia↗

Melanocortin-4 receptor mRNA is expressed in sympathetic nervous system outflow neurons to white adipose tissue.

Energy balance results from the coordination of multiple pathways affecting energy expenditure and food intake. Candidate neuropeptides involved in energy balance are the melanocortins. Several species, including Siberian hamsters studied here, decrease and increase food intake in response to stimulation and blockade of the melanocortin 4-receptor (MC4-R). In addition, central application of the MC3/4-R agonist melanotan-II decreases body fat (increases lipolysis) beyond that accounted for by its ability to decrease food intake. Because an increase in the sympathetic nervous system drive to white adipose tissue (WAT) is the principal initiator of lipolysis, we tested whether the sympathetic outflow circuitry from brain to WAT contained MC4-R mRNA expressing cells. This was accomplished by labeling the sympathetic outflow to inguinal WAT using the pseudorabies virus (PRV), a transneuronal retrograde viral tract tracer, and then processing the brain for colocalization of PRV immunoreactivity with MC4-R mRNA, the latter assessed by in situ hybridization. MC4-R mRNA was impressively colocalized in PRV-labeled cells (approximately greater than 60%) in many brain areas across the neuroaxis, including those typically implicated in lipid mobilization (e.g., hypothalamic paraventricular, suprachiasmatic, arcuate and dorsomedial nuclei, lateral hypothalamic area), as well as those not traditionally identified with lipolysis (e.g., preoptic area, subzona incerta of the lateral hypothalamus, periaqueductal gray, solitary nucleus). These data provide compelling neuroanatomical evidence that could underlie a direct central modulation of the sympathetic outflow to WAT by the melanocortins through the MC4-Rs resulting in changes in lipid mobilization and adiposity.

Adipose Tissue↗

1H NMR visible lipids are induced by phosphonium salts and 5-fluorouracil in human breast cancer cells.

Cationic lipophilic phosphonium salts (CLPS) selectively accumulate in the mitochondria of neoplastic cells and inhibit mitochondrial function. The effects of the CLPS p-(triphenylphosphoniummethyl) benzaldehyde chloride (drug A), and [4-(hydrazinocarboxy)-1-butyl] tris-(4-dimethylaminophenyl) phosphonium chloride (drug B), on human breast cells of differing biological properties were assessed using growth inhibition assays and 1H NMR. Drug A and, to a lesser extent, drug B demonstrated selective growth inhibition of the highly tumorigenic DU4475 breast carcinoma cell line compared to the transformed HBL-100 human breast cell line. However, in contrast to previous studies using other cell lines, no synergistic activity was found when the drugs were used in combination. 1H NMR demonstrated significant increases in mobile lipid acyl chain resonances in both cell lines treated with cytotoxic doses (IC50, 48 h) of the drugs used either alone or in combination. Two-dimensional NMR revealed accompanying decreases in phosphocholine/Lys levels in HBL-100 cells treated with A, B, or a 1:1 combination A+B at the IC50, and in DU4475 cells treated with drug A (IC50). This was accompanied by significant increases in cho/Lys ratios with IC50 A or combination A+B treatment. Similar spectra were observed in cells treated with 5-fluorouracil but not methotrexate, indicating that mobile lipid accumulation is a general but not universal response to cytotoxic insult.

Antineoplastic Agents↗

Regulation of lipolysis in severely burned children.

In this study, the rates of lipid mobilization and of lipolysis have been quantified in severely burned children. In all 12 patients studied, the basal rates were determined. In seven patients, the lipolytic responsiveness to an infusion of epinephrine (0.015 micrograms/kg/min) was tested, and in the other five patients, the response to beta-adrenergic blockade (propranolol, 1 mg/kg) was tested. The rate of appearance (Ra) of free fatty acids (FFA) was quantified by means of the infusion of 1-13C-palmitate to determine the rate of lipid mobilization, and Ra glycerol was determined using d5-glycerol to assess the rate of lipolysis more directly. In five patients, body composition was determined after recovery by means of H2(18)O dilution. The basal rate of lipolysis was higher than normal in the burned children. In four of the seven patients infused with epinephrine, there was a pronounced increase in Ra glycerol. In all patients given beta-blockade, Ra glycerol decreased greatly. Changes in Ra FFA corresponded with the changes in Ra glycerol in each case. Total body fat was very low (approximately 2% body weight), reflecting the surgical removal of fat in the process of burn wound excision. From these data it is concluded that lipolytic responsiveness to catecholamines in severely burned children is variable, but not absent, despite chronically elevated levels of catecholamines. The total extent of lipolysis may be limited by the available fat mass in children treated with fascial excision. In such patients, the limitation in the ability to mobilize an adequate amount of FFA to fully meet energy requirements provides an important rationale for the clinical practice of providing nutritional support in hourly boluses, as opposed to infrequent meals, since any period of even a few hours in which nutrients are not being absorbed will result in an energy substrate deficiency and consequent increase in amino acid oxidation.

Adolescent↗

Recent developments on lipolysis regulation in humans and discovery of a new lipolytic pathway.

In man, the major hormones controlling the lipolytic function are insulin (inhibition of lipolysis) and catecholamines (stimulation of lipolysis). Catecholamines are of major importance for the regulation of lipid mobilization in human adipose tissue and for the increase of non-esterified fatty acid supply to the working muscle. In vitro studies have shown that there are differences in the catecholaminergic control of fat cells from various fat deposits and a number of physiological and pathological alterations of catecholamine-induced lipolysis have been reported. Lipolytic resistance to catecholamines has been reported in subcutaneous adipose tissue, the major fat depot in obese subjects. Multiple alterations in catecholamine signal transduction pathways have been reported. In situ microdialysis allows a physiological exploration of adipose tissue biology. Recent data obtained on the catecholaminergic regulation of lipolysis and lipid mobilization, using microdialysis in humans, will be analysed. A potent lipolytic and lipomobilizing effect of atrial natriuretic peptide has recently been discovered; the mechanisms of action and physiological relevance will also be discussed.

Adipose Tissue↗

Hepatic lipidosis in pregnant captive American bison (Bison bison).

Hepatic lipidosis, a hallmark lesion of lipid mobilization disorders in ruminants, was noted in four 3-year-old, pregnant bison (Bison bison) after periods of anorexia that progressed to recumbency and death. The affected bison were part of a herd at the National Animal Disease Center (NADC) that was used for brucellosis vaccine research. Microscopically, the liver contained swollen hepatocytes with numerous, variably sized, round, smoothly contoured vacuoles that displaced cytoplasmic structures. Hepatocytes in all zones of the lobule were affected equally. Hypoglycemia, decreased total carbon dioxide, elevated gamma-glutamyltransferase, elevated alkaline phosphatase, and increased nonesterified fatty acid levels were noted. As in the case of cattle, altered nutritional demands of late gestation combined with management factors such as obesity, nutrition, stress, and concomitant disease may be critical in the pathophysiology of lipid mobilization disorders in bison. Additionally, stressors unique to this research herd likely contributed to fatal hepatic lipidosis.

Animals↗

Lateral mobility of lipid analogues and GPI-anchored proteins in supported bilayers determined by fluorescent bead tracking.

Lipid analogues and glycosylphosphatidylinositol (GPI)-anchored proteins incorporated in glass-supported phospholipid bilayers (SBL) were coupled to small (30 nm diameter) fluorescent beads whose motion in the liquid phase was tracked by intensified fluorescence video microscopy. Streptavidin (St), covalently attached to the carboxyl modified surface of the polystyrene bead, bound either the biotinylated membrane component, or a biotinylated monoclonal antibody (mAb) directed against a specific membrane constituent. The positions of the beads tethered to randomly diffusing membrane molecules were recorded at 0.2 sec intervals for about 1 min. The mean square displacement (rho) of the beads was found to be a linear function of diffusion time t, and the diffusion coefficient, D, was derived from the relation, rho(t) = 4Dt. The values of D for biotinylated phosphatidylethanolamine (Bi-PE) dispersed in an egg lecithin:cholesterol (80:20%) bilayer obtained by this methodology range from 0.05 to 0.6 micron 2/sec with an average of mean value of D = 0.26 micron 2/sec, similar to the value of mean value of D = 0.24 micron 2/sec for fluorescein-conjugated phosphatidylethanolamine (Fl-PE) linked to St-coupled beads by the anti-fluorescein mAb 4-4-20 or its Fab fragment. These values of D are comparable to those reported for Fl-PE linked to 30 nm gold particles but are several times lower than that of Fl-PE in the same planar bilayer as measured by fluorescence photobleaching recovery, D = 1.3 microns 2/sec. The mobilities of two GPI-anchored proteins in similar SBL were also determined by use of the appropriate biotinylated mAb and were found to be mean value of D = 0.25 and 0.56 micron 2/sec for the decay accelerating factor (DAF, CD55) and the human Fc gamma RIIIB (CD16) receptors, respectively. The methodology described here is suitable for tracking any accessible membrane component.

Animals↗

Regional-dependent increase of sympathetic innervation in rat white adipose tissue during prolonged fasting.

White adipose tissue (WAT) is innervated by the sympathetic nervous system. A role for WAT sympathetic noradrenergic nerves in lipid mobilization has been suggested. To gain insight into the involvement of nerve activity in the delipidation process, WAT nerves were investigated in rat retroperitoneal and epididymal depots after prolonged fasting. A significant increase in tyrosine hydroxylase (TH) content was found in epididymal and, especially, retroperitoneal WAT by Western blotting. Accordingly, an increased immunoreactivity for TH was detected by immunohistochemistry in epididymal and, especially, retroperitoneal vascular and parenchymal noradrenergic nerves. Neuropeptide Y (NPY)-containing nerves were found around arteries and in the parenchyma. Double-staining experiments and confocal microscopy showed that most perivascular and some parenchymal noradrenergic nerves also contained NPY. Detection of protein gene product (PGP) 9.5, a general marker of peripheral nerves, by Western blotting and PGP 9.5-TH by double-staining experiments showed significantly increased noradrenergic nerve density in fasted retroperitoneal, but not epididymal depots, suggesting that formation of new nerves takes place in retroperitoneal WAT in fasting conditions. On the whole, these data confirm the important role of sympathetic noradrenergic nerves in WAT lipid mobilization during fasting but also raise questions about the physiological role of regional-dependent nerve adjustments and their functional significance in relation to white adipocyte secretory products.

Adipose Tissue↗

[Effect of estradiol on lipolytic processes in the blood and adipose tissue of female rats].

The effect of estradiol on lipolysis in blood and in adipose tissue of female rat has been investigated. The hormone was administered to the studied animals subcutaneously during 20 days. The lipid mobilizing activity was determined in blood serum according to the four different experimental protocols: test serum--test tissue, test serum--control tissue, control serum--test tissue, and control serum--control tissue. Blood serum concentrations of triglycerides, free fatty acids and glucose, as well as lipolytic activity against exogenous substrate, were determined in addition. Estradiol administration was found to enhance the activity of factors potentiating lipolysis both in the blood and in adipose tissue of female rats, inasmuch as an increase in free fatty acids and triglyceride concentration, and a decrease in glucose concentration were observed during the period of administration of the hormone.

Adipose Tissue↗

Lipomobilizing effects of procaterol and yohimbine in the conscious dog: comparison of endocrinological, metabolic and cardiovascular effects.

1. Lipid mobilization during a hypocaloric diet may be enhanced by a pharmacological approach using beta 2-adrenoceptor agonists or alpha 2-adrenoceptor antagonists. Studies were undertaken in the dog, an animal model presenting fat cell antilipolytic alpha 2- and lipolytic beta-adrenoceptors, in order, firstly, to demonstrate the presence of beta 2 subtype adrenoceptors on adipocytes and, secondly, to compare the effects of procaterol (beta 2-adrenoceptor agonist) and of yohimbine (alpha 2-adrenoceptor antagonist) on metabolic, endocrinological and cardiovascular parameters. 2. Procaterol strongly stimulates lipolysis in dog adipocytes in vitro. The utilisation of selective beta 1- and beta 2-adrenoceptor antagonists (bisoprolol and ICI 118,551) in both lipolysis and binding studies (displacement of [3H]-dihydroalprenolol binding) demonstrated the presence of the two beta-adrenoceptor subtypes in dog fat cells. 3. Infusion of either yohimbine or procaterol (10 and 0.4 nmol min-1 kg-1, respectively for 30 min), provoked an equivalent increase in plasma non-esterified fatty acids (+100%). Procaterol, but not yohimbine, induced hyperglycaemia (+120%). Plasma insulin was weakly enhanced by yohimbine (+120%) as compared to the increase given by procaterol (+500%). 4. Both drugs stimulated sympathetic nervous system activity, as indicated by the increased plasma noradrenaline concentration, but only yohimbine increased the plasma adrenaline level. 5. Cardiovascular measurements indicated that procaterol strongly enhances heart rate and transiently decreases mean blood pressure. Yohimbine exhibits a weaker effect on heart rate and slightly increases mean blood pressure. 6. The present work clearly indicates that lipid mobilization is enhanced during fasting in the dog by selective beta 2-adrenoceptor stimulation or by alpha 2-adrenoceptor blockade. This enhanced lipolytic effect may result either from a direct action of the drugs on the adrenoceptors of fat cells or from an activation of the sympathetic nervous system. Procaterol suffers major limitations since it strongly increases heart rate, immunoreactive insulin and glycaemia. On the other hand, yohimbine induces only minor modifications both in cardiovascular and endocrinological parameters.

Adipose Tissue↗

Central nervous system origins of the sympathetic nervous system outflow to white adipose tissue.

White adipose tissue (WAT) is innervated by postganglionic sympathetic nervous system (SNS) neurons, suggesting that lipid mobilization could be regulated by the SNS [T. G. Youngstrom and T. J. Bartness. Am. J. Physiol. 268 (Regulatory Integrative Comp. Physiol. 37): R744-R751, 1995]. A viral transsynaptic retrograde tract tracer, the pseudorabies virus (PRV), was used to identify the origins of the SNS outflow from the brain to WAT neuroanatomically. PRV was injected into epididymal or inguinal WAT (EWAT and IWAT, respectively) of Siberian hamsters and IWAT of rats. PRV-infected neurons were visualized by immunocytochemistry and found in the spinal cord, brain stem (medulla, nucleus of the solitary tract, caudal raphe nucleus, C1 and A5 regions), midbrain (central gray), and several areas within the forebrain. The general pattern of infection of WAT in both species was more similar than different and resembled that seen after PRV injections into the adrenal medulla in rats (A. M. Strack, W. B. Sawyer, J. H. Hughes, K. B. Platt, and A. D. Loewy. Brain Res. 491: 156-162, 1989). EWAT versus IWAT injected hamsters had relatively less labeling in the suprachiasmatic, dorsomedial, and arcuate nuclei. Overall, it appeared that the SNS innervation of WAT originates from the general SNS outflow of the central nervous system and therefore may play a significant role in lipid mobilization.

Adipose Tissue↗

Structure and mobility of lipid membranes on a thermoplastic substrate.

Supported lipid membranes constitute one of the most important model systems for cell membranes. The properties of lipid membranes supported by the hydrophobic solid polymer cyclic olefin copolymer (COC) were investigated. Lipid layers consisting of varying amounts of 1,2-dioleoyl-3-trimethylammonium propane (DOTAP, cationic) and 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC, neutral) prepared by vesicle fusion and solvent exchange were compared. All lipid mixtures coated the COC surface homogeneously forming a fluid membrane as verified by fluorescence microscopy and fluorescence recovery after photobleaching (FRAP). The exact structure of the supported membranes was determined by synchrotron reflectivity experiments using a microfluidic chamber. The X-ray data are in agreement with a compressed (head-to-head distance = 29 angstroms) and less densely packed bilayer.

Cations↗

Lipid binding to the amphipathic membrane protein cytochrome b5.

The lipid binding properties of the membrane protein cytochrome b(5) (detergent-extracted from calf liver microsomal preparations) were characterized by studying the interaction of spin-labeled lipids (5-, 12-, and 16-doxylstearic acid and 5- and 16-doxylphosphatidyl-choline, where doxyl refers to the nitroxide moiety) with cytochrome b(5), using electron spin resonance spectroscopy. The intact cytochrome b(5) molecule immobilizes all of the lipid spin labels, while the segment of cytochrome b(5) released by trypsin does not affect lipid mobility. The immobilization of lipid spin labels on the hydrophobic surface of intact cytochrome b(5) is not appreciably altered by associating the protein with liposomes. Differences in polarity of the lipid binding sites between cytochrome b(5) and phospholipid vesicles were also observed. The lipid binding sites on cytochrome b(5) are hydrophobic by conventional criteria, but are more polar than the interior of fluid phospholipid bilayers.

Animals↗