bluuebunny/crossref_metadata_2025 huggingface dataset:
ANG II modifies cardiomyocyte function via extracardiac and intracardiac neurons: in situ and in vitro studies
Authors: Horackova M. , Armour J. A.
Published: December 2017

To determine whether angiotensin II (ANG II) affects cardiac performance via neurons in intrathoracic cardiac ganglia, studies were performed on anesthetized dogs. To exclude possible vascular regulatory effects of ANG II, experiments were also performed using long-term cultures of adult guinea pig ventricular cardiomyocytes with or without intrathoracic neurons. 1) In in situ experiments in 10 anesthetized dogs, cardiac augmentation occurred when ANG II (10 microl or 0.1 ml; 10-100 microM) was administered into limited loci within acutely decentralized stellate or middle cervical ganglia that were neurally connected to, but not those disconnected from, the heart. In another 18 dogs, ANG II increased intrinsic cardiac neuronal activity when administered adjacent to such neurons or into their local arterial blood supply. Ventricular ionotropic effects elicited by ANG II were eliminated by timolol, whereas increases in intrinsic cardiac neuronal activity were not affected. Effects elicited by ANG II were eliminated by administration of a selective AT1 receptor antagonist (losartan) but not by a selective AT2 receptor antagonist (PD-123319). 2) In in vitro experiments, ANG II (100 nM) induced positive chronotropic effects on cultured adult guinea pig cardiomyocytes innervated with adult extrinsic or intrinsic cardiac neurons, but not those cultured without neurons. The frequency of calcium inward current (Ca(i)) transients (recorded by fura 2 fluorescence) increased in innervated cocultures but not in the noninnervated cardiomyocyte cultures; however, the amplitude of Ca(i) transients was not affected by ANG II in cultures or in freshly isolated adult guinea pig cardiomyocytes. ANG II-induced effects in cocultures were blocked by losartan but not PD-123319 or timolol. Thus 1) ANG II-sensitive neurons exist in intrathoracic extracardiac and intrinsic cardiac ganglia; 2) these neurons possess AT1 receptors; and 3) these neurons appear to act directly and indirectly via adrenergic neurons to enhance cardiomyocyte function.
Intracellular regulation of protein degradation during sepsis is different in fast- and slow-twitch muscle
Authors: Tiao G. , Lieberman M. , Fischer J. E. , Hasselgren P. O.
Published: December 2017

We tested the hypothesis that the difference in the response to sepsis of protein breakdown between fast- and slow-twitch skeletal muscle reflects differential activation of the energy-ubiquitin-dependent proteolytic pathway. In addition, we defined the time course and the tissue specificity of sepsis-induced changes in the expression of the ubiquitin pathway. Sepsis was induced in rats by cecal ligation and puncture; control rats were sham operated. Energy-dependent protein breakdown was measured in incubated extensor digitorum longus (EDL) and soleus muscles. Ubiquitin mRNA levels were determined by Northern blot analysis. Sepsis resulted in increased energy-dependent protein breakdown and upregulated expression of ubiquitin mRNA in the fast-twitch EDL but not in the slow-twitch soleus muscle. The sepsis-induced increase in ubiquitin mRNA levels in the EDL muscle was noticeable before the increase in energy-dependent protein breakdown. Sepsis increased ubiquitin mRNA levels in the diaphragm (a mixed fiber-type muscle) but not in heart, liver, kidney, or intestine, consistent with a tissue-specific regulation of the ubiquitin system during sepsis. The results suggest that the difference in protein breakdown during sepsis between fast- and slow-twitch muscles reflects differential activation of the energy-ubiquitin-dependent proteolytic pathway. The data also suggest that the expression of the ubiquitin pathway is upregulated in a time-dependent fashion during sepsis and that this response is not a generalized phenomenon but is tissue specific.
Hyperpolarization of hepatocytes by 2,5-AM: implications for hepatic control of food intake
Authors: Scharrer E. , Rossi R. , Sutter D. A. , Seebacher M. C. , Boutellier S. , Lutz T. A.
Published: December 2017

Because 2,5-anhydro-D-mannitol (2,5-AM) seems to stimulate feeding by acting on the liver and because the hepatic membrane potential has been suggested to play an important role in control of feeding ("potentiostatic" hypothesis), we investigated the effect of 2,5-AM on the membrane potential of liver cells with microelectrodes using a superfused liver slice technique. 2,5-AM (2.5 mM), which reduces intracellular ATP in rat liver, hyperpolarized the liver cell membrane in mouse and rat liver slices by 4-7 mV. This hyperpolarization was reversed by quinine (1 mM), an unspecific blocker of Ca2+-dependent K+ channels, and abolished by apamin (20 nM), a blocker of Ca2+-activated K+ channels with low conductance. Amiloride at 10(-3) M, but not at 10(-6) M, or a low-Na medium (26 mM) also eliminated the hyperpolarization. The K+ channel blockers cetiedil (50 microM), glibenclamide (30 microM), and Ba2+ (5 mM); flufenamic acid (100 microM), a blocker of nonselective cation channels; and ouabain (1 mM), an inhibitor of the Na+-K+-adenosinetriphosphatase, did not significantly influence the 2,5-AM-induced hyperpolarization. It is concluded that 2,5-AM hyperpolarizes the liver cell membrane by activating Ca2+-dependent K+ channels. This activation seems to be impaired when the Na+/H+ exchanger is inhibited by amiloride or a low-Na+ medium. The findings also imply that the hyperphagic effect of 2,5-AM observed in rats is not associated with a decrease in the hepatic membrane potential, as postulated by the potentiostatic hypothesis.
Effects of pH on basolateral tetraethylammonium transport in snake renal proximal tubules
Authors: Kim Y. K. , Dantzler W. H.
Published: December 2017

Previous work on snake renal proximal tubules suggested that pH might influence tetraethylammonium (TEA) transport across the basolateral membrane. To examine this more directly, we determined the effects of altering either extracellular pH (pHo) or intracellular pH (pHi) on TEA uptake and efflux across the basolateral membrane of isolated snake renal proximal tubules. We found no evidence for trans-stimulation of either TEA uptake or efflux by H+. Therefore, there was no evidence for a TEA/H+ exchanger. However, we found evidence for trans-inhibition of both TEA uptake and efflux as well as for cis-inhibition of TEA uptake by increasing H+ concentration. H+ concentration appeared to have some type of direct effect on basolateral transport independent of any effect on membrane potential. Moreover, there appeared to be an optimal intracellular H+ concentration for entry of TEA into the cells that corresponded to the one found at the physiological pHi of 7.1. There also appeared to be an optimal extracellular H+ concentration for efflux of TEA from the cells that corresponded to the one found at the physiological pHo of 7.4. The mechanism involved in this relationship is unknown, but the data support a concept derived from previous studies that TEA transport across the basolateral membrane is asymmetric.
Regional blood flow in Dahl-Iwai salt-sensitive rats and the effects of dietary L-arginine supplementation
Authors: Tomohiro A. , Kimura S. , He H. , Fujisawa Y. , Nishiyama A. , Kiyomoto K. , Aki Y. , Tamaki T. , Abe Y.
Published: December 2017

The purpose of the present study was to determine 1) whether different organs undergo similar increase in vascular resistance in Dahl-Iwai salt-sensitive (S) rats, and 2) the effects of chronic oral L-arginine supplementation on the regional hemodynamics in S rats. Male 6-wk-old S rats and salt-resistant (R) rats were maintained on an 8% NaCl chow for 4 wk. One group (S or R rats) was maintained on tap water and the other group (S/Arg or R/Arg rats) received tap water containing L-arginine at a concentration of 1.5%. Organ blood flow and cardiac output were measured with microspheres in the conscious condition. Mean blood pressure in S, S/Arg, R, and R/Arg rats was 159 +/- 5, 138 +/- 3, 111 +/- 4, and 112 +/- 4 mmHg, respectively. Urinary excretion of protein and albumin in S/Arg rats was significantly suppressed compared with S rats. Concerning regional hemodynamics, the flow rate of the kidney was lower in S rats than in R rats, but there were no differences between S and R rats in the flow rates of the brain, heart, lung, liver, spleen, intestine, skeletal muscle, and skin. Thus the renal blood flow was solely reduced in S rats on a high-salt diet. The flow rate of the kidney in S/Arg rats was maintained at a higher level compared with that of S rats. L-Arginine treatment tended to produce a recovery in the urinary excretion of guanosine 3',5'-cyclic monophosphate in S rats, but had no effect in R rats. Thus the supplementation of L-arginine prevented the increase in blood pressure in S rats on a high-salt diet and normalized the abnormality of renal hemodynamics accompanying salt-induced hypertension.
Nitric oxide and renal effects of volume expansion in conscious monkeys
Authors: Peterson T. V. , Carter A. B. , Miller R. A.
Published: December 2017

Experiments were performed to determine the effects of nitric oxide (NO) synthase inhibition on the renal responses to volume expansion in conscious cynomolgus monkeys. All animals were volume expanded with 3% dextran in normal saline under three conditions: 1) during a control state, 2) during constant infusion of the NO synthase inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME, 30 microg x kg(-1) x min(-1)), and 3) during simultaneous infusion of L-NAME and excess NO substrate L-arginine (0.6 mg x kg(-1) x min(-1)). The control volume expansion increased urine flow from 0.27 +/- 0.05 to 0.94 +/- 0.28 ml/min and sodium excretion from 21 +/- 9 to 95 +/- 26 microeq/min. During L-NAME infusion, these responses were attenuated in that urine flow only increased from 0.13 +/- 0.03 to 0.28 +/- 0.09 ml/min and sodium excretion from 13 +/- 8 to 35 +/- 23 microeq/min. Addition of L-arginine to the L-NAME infusion abolished these renal excretory effects of L-NAME alone. With combined L-NAME/L-arginine, volume expansion increased urine flow from 0.37 +/- 0.23 to 1.09 +/- 0.23 ml/min and sodium excretion from 38 +/- 27 to 150 +/- 24 microeq/min, responses similar to control. L-Arginine also markedly attenuated the effect of L-NAME to increase mean arterial pressure and abolished the L-NAME decreases in creatinine and p-aminohippurate clearances. However, an L-NAME-induced bradycardia could only be partially reversed. These results demonstrate that a functioning NO system may be important in mediating normal renal responses to volume expansion in this primate species.
Manifestation of circadian rhythm under constant light depends on lighting conditions during lactation
Authors: Cambras T. , Canal M. M. , Torres A. , Vilaplana J. , Diez-Noguera A.
Published: December 2017

Adult rats transferred to continuous illumination (LL) show a disruption of circadian rhythms, although the mechanisms underlying this effect are not yet well known. In previous experiments, we found that when rats were born and raised under LL they showed an ultradian pattern during the first 10 days after weaning, but afterward they generated a circadian rhythm that was maintained until adulthood. It was not clear whether this evolution was attributable to the influence of the rhythm of the mother or to the effect of constant light. Here, we have studied the motor activity rhythm of young rats maintained under LL after weaning, taking into account the conditions to which they were exposed during lactation [LL or continuous darkness (DD)]. To check the possible effect of the rhythm of the dam, on the day of delivery some of the dams were blinded, others were subjected to a restricted feeding schedule of 3 h/day, and the others were used as controls. For each rat, the period of the circadian rhythm and the percentage of variance explained by this rhythm were calculated. Results show that all rats maintained under LL during lactation expressed a circadian rhythm in their motor activity. However, rats maintained under DD during lactation did not. This effect did not seem to be dependent on the type of dam. These results suggest that the rhythm of the dams does not affect the manifestation of the rhythm of the pups and that the expression of circadian rhythmicity under constant bright light depends on the lighting conditions under which the animals were maintained during lactation, which could affect the development of the circadian pacemaker or the retina.
Melatonin and S-20098 increase REM sleep and wake-up propensity without modifying NREM sleep homeostasis
Authors: Cajochen C. , Krauchi K. , Mori D. , Graw P. , Wirz-Justice A.
Published: December 2017

The pineal hormone melatonin has been implicated in the circadian regulation of sleep. In a crossover design, we investigated the effect of acute administration of 5 mg melatonin and a melatonin agonist (S-20098, 5 and 100 mg) in healthy young men when given 5 h before bedtime on sleep structure and electroencephalogram (EEG) power density. Each trial comprised a baseline, a treatment, and a posttreatment sleep episode. Relative to the placebo condition, all treatments phase advanced the core body temperature rhythm [Krauchi, K., C. Cajochen, D. Mori, C. Hetsch, and A. Wirz-Justice. Sleep Res. 24: 526, 1995; and Krauchi, K., C. Cajochen, D. Mori, and A. Wirz-Justice. Am. J. Physiol. 272 (Regulatory Integrative Comp. Physiol. 41): R1178-1188, 1997]. Rapid eye movement (REM) sleep was increased after both melatonin and S-20098. This increase in REM sleep was most pronounced in the first REM sleep episode. On the posttreatment night after melatonin and S-20098 administration, more wakefulness was present in the latter one-half of the sleep episode. EEG power density between 0.25 and 20 Hz during either non-REM (NREM) or REM sleep did not differ from placebo. Thus a single early evening dose of melatonin or the agonist S-20098 increases REM sleep propensity and advances sleep termination while, at the same time, the EEG in NREM sleep remains unaffected.
Leptin increases energy expenditure and selectively promotes fat metabolism in ob/ob mice
Authors: Hwa J. J. , Fawzi A. B. , Graziano M. P. , Ghibaudi L. , Williams P. , Van Heek M. , Davis H. , Rudinski M. , Sybertz E. , Strader C. D.
Published: December 2017

Obesity occurs whenever energy intake exceeds energy expenditure. The ob gene product leptin is a potent anorectic agent when administered to ob/ob mice, but its effects on energy expenditure have not been investigated in detail. The present study was designed to analyze the acute metabolic effects of leptin in vivo. Analysis of oxygen consumption in ob/ob mice demonstrated a reduction in energy expenditure compared with lean controls; this reduction showed a diurnal fluctuation and was most evident during the light cycle. A single intraperitoneal dose of leptin increased oxygen consumption during the light cycle in ob/ob mice, ablating the circadian fluctuation in this parameter. In addition, leptin had a profound effect on fuel selection: the respiratory quotient was markedly reduced, indicating a reduction in carbohydrate oxidation and an increase in fat oxidation. These acute effects of leptin on metabolic parameters are consistent with the selective loss of body fat observed on chronic leptin treatment and suggest that increased energy utilization plays an important role in the anti-obese actions of leptin.
Sucrose vs. maltose taste discrimination by rats depends on the input of the seventh cranial nerve
Authors: Spector A. C. , Markison S. , St John S. J. , Garcea M.
Published: December 2017

Although rats treat the taste of sucrose and maltose as perceptually similar, they nonetheless appear to be able to distinguish between the two sugars, as suggested from prior work examining the cross-generalization of conditioned taste aversions. This study explictly tested whether rats could behaviorally discriminate sucrose from maltose and examined the relative importance of the gustatory input of the seventh and ninth cranial nerves in maintaining such performance. Water-restricted rats were presurgically trained in a conditioned avoidance task to suppress licking to sucrose or maltose and to maintain licking to the other sugar. Concentration (0.05, 0.1, 0.2, and 0.4 M) was varied to make intensity an irrelevant cue. Stimuli were randomly presented in 5-s trials during 50-min sessions. Bilateral transection of the chorda tympani nerve (CT) or the glossopharyngeal nerve or sham surgery did not disrupt discrimination performance. In contrast, combined transection of the CT and greater superficial petrosal nerve, which collectively removes the taste input of the seventh cranial nerve, caused severe impairments in sugar discriminability. In these rats, performance was more disturbed at the lower concentrations. These findings confirm that rats can discriminate sucrose from maltose and that this capability relies heavily on the taste input of the seventh cranial nerve. Although the input of the ninth cranial nerve is unnecessary, it may help sustain partial competence in this task, especially at high concentrations, in the combined absence of the CT and greater superficial petrosal nerve.
Presynaptic sympathetic mechanism in the insulinostatic effect of epinephrine in mouse pancreatic islets
Authors: Karlsson S. , Myrsen U. , Nieuwenhuizen A. , Sundler F. , Ahren B.
Published: December 2017

The catecholamines inhibit insulin release. It is not established whether presynaptic mechanisms contribute to this effect. We therefore examined the relative contribution of presynaptic and postsynaptic mechanisms to the insulinostatic effects of epinephrine and norepinephrine. Mice were injected with 6-hydroxydopamine (6-OHDA; 0.19 mmol/kg) or its vehicle. Islets were isolated after 48 h. Islets from vehicle-injected control animals contained numerous tyrosine hydroxylase (TH)-immunoreactive nerve terminals (marker for sympathetic nerves). In contrast, TH-immunoreactive nerves were not detected in islets from 6-OHDA-treated animals, indicating sympathetic denervation. Basal (5.6 mmol/l glucose) or glucose-stimulated (16.7 mmol/l) insulin secretion did not differ between incubated islets from vehicle-injected control animals and islets from 6-OHDA-treated animals. The insulinostatic effect of epinephrine, but not that of norepinephrine, was markedly impaired in islets from 6-OHDA-treated animals: the lowest effective insulinostatic concentration of epinephrine was 0.01 nmol/l in islets from vehicle-injected animals and 1 nmol/l in islets from 6-OHDA-treated animals. We conclude that in isolated mouse islets the insulinostatic effect of epinephrine, but not that of norepinephrine, partially depends on sympathetic nerve terminals, suggesting an important role for presynaptic mechanisms in epinephrine-induced inhibition of insulin secretion.
Regulated hypothermia in the hypothyroid rat induced by administration of propylthiouracil
Authors: Yang Y. , Gordon C. J.
Published: December 2017

Propylthiouracil (PTU), an antithyroidal drug that reduces serum L-thyroxine (T4) and 3,5,3'-triiodothyronine (T3), is presumed to lower core temperature (T0) by impairing metabolic thermogenesis. However, it is not understood why PTU-treated animals cannot use behavioral and other thermoeffectors to maintain normal Tc. Male rats were administered PTU in drinking water (0.05 mg/ml) while the following parameters were measured: 1) Tc and motor activity (MA) recorded by radiotelemetry for 24 h at ambient temperatures (Ta) of 10-30 degrees C; 2) selected Ta, MA, and Tc in a temperature gradient; and 3) Tc, MA, and grooming behavior during exposure to heat stress (TH = 34.5 degrees C) for 2 h. PTU reduced serum levels of T4, and T3 by 95 and 60%, respectively. Tc decreased after 3 days of PTU treatment; a 0.5 degree C decrease in Tc persisted throughout the PTU treatment. PTU rats exposed to Ta of 10-30 degrees C maintained a consistent hypothermic Tc during the light phase; however, a deficit in the stability of Tc at night was noted during exposure to 10 degrees C. In the temperature gradient, PTU rats selected warmer Ta, but their Tc was maintained at the same hypothermic levels as observed at fixed Ta values of 15-30 degrees C. Heat stress caused Tc of control rats to increase to 39 degrees C, whereas Tc of the PTU rats was maintained below 38 degrees C. The regulation of Tc at hypothermic levels over a wide range of Ta values and when rats were housed in a temperature gradient indicates that chronic PTU induces a state of regulated hypothermia.
Role of Na(+)-K+ pump and Na+ channel concentrations in the contractility of rat soleus muscle
Authors: Harrison A. P. , Nielsen O. B. , Clausen T.
Published: December 2017

The dependence of contractile performance on the leak-to-pump ratio for Na+ has been examined. In isolated rat soleus muscle the concentration of Na(+)-K+ pumps was shown to decrease with age (-57%) or K+ deficiency (-69%), whereas Na+ channel concentration remained constant. This relative increase in the ratio between Na+ channels and Na(+)-K+ pumps was associated with a markedly faster rate of force decline (58 and 97%, respectively; both P < 0.001) during stimulation at 90 Hz and reduced subsequent force recovery (-34 and -38%, respectively; both P < 0.001). Similar effects were elicited by acute inhibition of Na(+)-K+ pump activity with ouabain. Preincubation with aconitine and veratridine, resulting in a 91 and 118% increase in Na+ influx per contraction, respectively (both P < 0.05), significantly hastened the initial rate of force decline (19%; P < 0.05 for aconitine and 69%; P < 0.001 for veratridine) and slowed recovery (-59 and -86%, respectively, both P < 0.001). It is concluded that the ratio between excitation-induced Na+ influx and Na(+)-K+ pump capacity is an important determinant for endurance and rate of recovery of force in skeletal muscle.
The kidneys stimulate vasopressin release during hemorrhage in rats with chronic NTS lesions
Authors: Schreihofer A. M. , Hoffman G. E. , Sved A. F.
Published: December 2017

Elimination of baroreceptor afferent input to the brain produced by chronic lesion of nucleus of the solitary tract (NTS) does not alter vasopressin (VP) release during hypotensive hemorrhage in conscious rats. To investigate whether the kidneys play a critical role in stimulating VP release during hemorrhage in chronic NTS-lesioned rats, we examined the effects of removing potential signals arising from the kidneys. In NTS-lesioned rats, nephrectomy or renal denervation, but not captopril injection, markedly attenuated (but did not abolish) hemorrhage-induced VP release. In contrast, none of these manipulations attenuated the VP response in NTS-intact rats. Hemorrhage increased plasma renin activity in control and NTS-lesioned rats, and this response was not altered by renal denervation. In rats with NTS lesions and renal denervation, hemorrhage induced the expression of Fos in hypothalamic magnocellular VP neurons in a pattern similar to that of hemorrhage in intact rats. Collectively, these results indicate that in chronic NTS-lesioned rats an afferent signal arising from the kidneys stimulates VP release during hemorrhage, possibly through renal nerves. However, with the NTS intact or after the selective removal of arterial baroreceptor inputs, such a role for the kidneys is not apparent. Furthermore, in the absence of the NTS and renal nerves, another signal generated by hypotensive hemorrhage continues to stimulate VP neurons.
Hepatic fatty acid metabolism in pigs and rats: major differences in endproducts, O2 uptake, and beta-oxidation
Authors: Adams S. H. , Lin X. , Yu X. X. , Odle J. , Drackley J. K.
Published: December 2017

Models of mammalian hepatic lipid metabolism are based largely on observations made in adult rats, emphasizing ketogenesis as a primary adjunct to mitochondrial beta-oxidation. Studies using piglets have illustrated the divergent nature of intermediary metabolism in this model, wherein ketogenesis and beta-oxidation are small and acetogenesis is an important route of fuel carbon flux. To clarify potential species differences in hepatic lipid metabolism and its control, we compared O2 consumption and metabolic end products in fasted pig and rat liver homogenates treated with 1-[14C]C16:0. Carboxyl carbon accumulation in acid-soluble products (ASP) plus CO2 was threefold greater and O2 consumption was twofold greater in rats (P < 0.05). Unlike rats, pigs showed negligible carboxyl carbon accumulation in ketone bodies (3-7% of ASP), whereas acetate was a carboxyl carbon reservoir in both animals (17-31% of ASP in pigs). Malonate increased (approximately 2-fold) and antimycin/rotenone decreased (40-60%) radiolabel accumulation in acetate. These data concur with the hypotheses that comparatively low hepatic beta-oxidative flux in piglets is partially related to a smaller metabolic rate and that substantial acetogenesis occurs intramitochondrially in both pigs and rats.
Reflex pathways controlling urethral striated and smooth muscle function in the male rat
Authors: Kakizaki H. , Fraser M. O. , De Groat W. C.
Published: December 2017

The organization of vesicourethral reflex mechanisms in the male rat was studied by monitoring intraurethral pressure and the external urethral sphincter (EUS) electromyogram. EUS striated and urethral smooth muscle activities were elicited by reflex isovolumetric bladder contractions evoked by bladder filling or electrical stimulation of nerves in the bladder wall. Evoked EUS bursting activity in normal rats was eliminated in chronic spinal rats and replaced by tonic activity. Reflex urethral smooth muscle activity mediated by an increase in urethral pressure after paralysis of the EUS with alpha-bungarotoxin occurred in normal and chronic spinal rats. The response was significantly larger in chronic spinal (21.3 +/- 3.0 cmH2O) than in normal rats (4.2 +/- 0.7 cmH2O). NG-nitro-L-arginine methyl ester (a nitric oxide synthase inhibitor, 20 mg/kg i.v.) increased the smooth muscle response in normal (5.9 +/- 1.3 cmH2O) and chronic spinal rats (6.9 +/- 1.8 cmH2O). This increase in urethral pressure was not changed by sympathetic nerve transection or prazosin (0.2-0.3 mg/kg i.v.) but was abolished by hexamethonium and reduced 74-89% by atropine. These results indicate that coordinated EUS function (bursting activity) in the male rat is dependent on supraspinal pathways and that the urethral smooth muscle response during voiding is composed of a predominant cholinergic, atropine-sensitive contraction as well as a nitric oxide-mediated relaxation. Both are mediated by activation of parasympathetic pathways and are maintained or significantly larger after spinal cord injury, indicating that they are dependent on spinal reflex pathways.
Relationships between gastric motility and gastric vagal afferent responses to CCK and GRP in rats differ
Authors: Schwartz G. J. , Moran T. H. , White W. O. , Ladenheim E. E.
Published: December 2017

The brain-gut peptides cholecystokinin (CCK) and the mammalian bombesin-like peptide gastrin-releasing peptide (GRP) suppress food intake. Vagotomy blocks CCK- but not bombesin (BN)-induced feeding suppression, demonstrating differential vagal contributions. We examined the relationship between the ability of CCK and the active fragment of GRP, GRP-(18-27), to stimulate gastric vagal afferent activity and their ability to elicit changes in gastric motility. We also examined ligated cervical vagal segments and revealed specific 125I-CCK vagal binding without evidence of radiolabeled BN binding sites. Both close arterial and intraperitoneal CCK and GRP-(18-27) produced dose-dependent increases in activity in gastric vagal mechanoreceptive afferents. CCK dose dependently decreased gastric pressure without altering antral wall tension, whereas GRP-(18-27) dose dependently increased both gastric pressure and peak antral wall muscle tension. These results suggest that GRP-(18-27) activates gastric vagal afferents secondary to its stimulation of gastric motor effects. CCK activates this same population of vagal afferents independent of changes in gastric tension, suggesting a direct action of CCK at functional vagal CCK receptors.
Recombinant leptin exchanges between parabiosed mice but does not reach equilibrium
Authors: Harris R. B. , Zhou J. , Weigle D. S. , Kuijper J. L.
Published: December 2017

Parabiosis experiments suggest that ob/ob mice are deficient in a circulating "lipostatic" signal but respond to such a signal when it is delivered in the cross circulation from their parabiotic partner. Identification of leptin as the mutation in ob/ob mice leads to the assumption that leptin is the lipostatic signal. The objective of these experiments was to determine the circulating half-life of leptin and to demonstrate whether it exchanged between parabiosed mice. Measurement of disappearance of recombinant leptin from serum in SWRJ mice indicated a circulating half-life of approximately 36 min. Single ob/ob mice or one member of a parabiosed pair of ob/ob mice received 50 micrograms recombinant murine leptin in two intraperitoneal injections a day for 10 days, starting 40 days after parabiosis surgery. Control mice and pairs received equivalent injections of vehicle. In single mice, leptin significantly reduced food intake, body weight, serum insulin, and pancreatic and liver weight. Leptin treatment of one member of a parabiosed pair of ob/ob mice reduced serum insulin, gut content (an index of food intake), and body fat in both partners. The injected parabiont lost more fat than its partner, and body temperature was increased only in the injected mouse, indicating that leptin did not reach equilibrium in the two animals. This was confirmed by Western blot analysis of serum leptin measured 2 h after injection. Therefore, although leptin can exchange between parabionts, its half-life is inadequate to allow equilibrium when a large concentration gradient exists between partners.
Identification and characterization of the zeta-opioid receptor in developing rat heart
Authors: McLaughlin P. J. , Wu Y.
Published: December 2017

Opioid growth factor (OGF; [Met5]enkephalin) inhibits DNA synthesis of epicardial and myocardial cells in the neonatal rat heart in a receptor-mediated fashion. Ligand binding assays using newborn rat heart and [3H][Met5]enkephalin were performed to characterize the receptor responsible for the cell replicative effects of OGF in developing heart. Specific and saturable binding was detected, and Scatchard analysis revealed that the data were consistent for a single binding site with a binding affinity of 6.8 +/- 0.3 nM and a binding capacity of 21.8 +/- 1.9 fmol/mg protein. Subcellular fractionation studies revealed that binding was restricted to the nuclear fraction. Competition experiments showed that cold [Met5]enkephalin was the most effective ligand at displacing [3H][Met5]enkephalin. Binding was recorded on gestation days 18 and 20, reached its highest level at birth, and steadily decreased from postnatal days 1 to 15; binding at days 21 and 35 and in adults was negligible. The function, pharmacological and biochemical characteristics, distribution, and subcellular location of this OGF receptor in developing mammalian heart are consistent with the zeta-opioid receptor.
Convection as one of the limiting factors of human respiration during normoxic exercise
Authors: Heller H. , Schuster K. D.
Published: December 2017

Each of the pathways within respiration has been suspected of limiting maximal performance, suggesting that O2 transport may be affected by each single pathway. The use of the stable, isotopic O2 molecules 16O2 and 16O18O is presented as a novel method for assessing respiration. Because of their different molecular weights, 16O2 diffuses 3% more rapidly than 16O18O, whereas 16O2 is convectively transported as rapidly as 16O18O. This can be quantified by using the overall fractionation factor alpha O. The more diffusion becomes limiting, the more 16O2 is transported in preference to 16O18O and alpha O is increased to 1.03. By contrast, the more respiration is limited by convection, the closer alpha O comes to 1.0 during the entire O2 transport. Six untrained subjects underwent normoxic exercise on a cycle ergometer. Isotopic analysis was performed at rest and during exercise loads of 50, 100, 150, 200, and 250 W using respiratory mass spectrometry. With increasing workload, a decrease in alpha O from 1.0072 at rest to 1.0033 at 250 W was determined in all subjects. On the basis of a serial resistance model of respiration, we concluded that, in our subjects, O2 transport was increasingly affected by convection but decreasingly limited by diffusion. The relative contribution of convection to the entire resistance to O2 flow ranged from > or = 44.6% at rest to > or = 74.6% at the most strenuous level of exercise, whereas the diffusive pathways decreasingly contributed to resistance to O2 flow by < or = 24% at rest and < or = 11% at 250 W.
Central lipopolysaccharide elevates plasma IL-6 concentration by an alpha-adrenoreceptor-mediated mechanism
Authors: Finck B. N. , Dantzer R. , Kelley K. W. , Woods J. A. , Johnson R. W.
Published: December 2017

High circulating levels of interleukin-6 (IL-6) are evident after intracerebroventricular injection of lipopolysaccharide (LPS). To investigate the pathway of centrally induced IL-6 production, in the present study we evaluated the effects of specific alpha-adrenergic receptor antagonists administered peripherally on IL-6 production and hypertriglyceridemia induced by LPS administered centrally. In the first study, adult male Wistar-Furth rats were injected intracerebroventricularly with LPS. Centrally injected LPS increased plasma IL-6 and triglycerides (TG) in a dose-dependent fashion. To determine if LPS increased plasma IL-6 and TG through an alpha-adrenoreceptor mechanism, rats were pretreated intraperitoneally with either vehicle, phentolamine (alpha 1- and alpha 2-receptor antagonist), prazosin (alpha 1-receptor antagonist), or yohimbine (alpha 2-receptor antagonist). Thirty minutes later, rats were injected intracerebroventricularly with LPS. Whereas prazosin and yohimbine attenuated the increases in plasma IL-6 caused by LPS, phentolamine completely blocked the peripheral effects of central LPS. These data suggest that increased sympathetic activity and subsequent activation of alpha 1- and alpha 2-adrenergic receptors are important for controlling peripheral metabolic and endocrine systems when inflammatory stimuli are present in the brain.
Dysregulated proteolytic balance as the basis of excess extracellular matrix in fibrotic disease
Authors: Peters C. A. , Freeman M. R. , Fernandez C. A. , Shepard J. , Wiederschain D. G. , Moses M. A.
Published: December 2017

To investigate mechanisms of tissue fibrosis, we developed a model of ovine fetal bladder fibrosis due to surgically induced obstruction. Tissues were analyzed for matrix metalloproteinases (MMPs) and their inhibitors, the tissue inhibitors of metalloproteinases (TIMPs). Active MMP-2 was not detected in obstructed bladders, while latent and active forms were detected in normal bladders. MMP-1 (interstitial collagenase) activity was lower in obstructed bladders. MMP inhibitory activity was increased with obstruction, as were levels of TIMP mRNA and protein. These results indicate that the proteins responsible for collagen degradation are present in the developing bladder, and a shift in the proteolytic balance favoring inhibition of degradation occurs in a model of obstruction-induced fibrosis. This altered proteolytic balance favors accumulation of extracellular matrix and decreased tissue compliance characteristic of this and perhaps other fibrotic conditions.
Differential role of glucocorticoids in mediating endotoxin-induced changes in IGF-I and IGFBP-1
Authors: Li Y. H. , Fan J. , Lang C. H.
Published: December 2017

The purpose of the present study was to determine whether endogenous elevations in glucocorticoids mediate the changes in insulin-like growth factor (IGF) 1 and IGF binding protein (IGFBP) 1 levels in plasma and tissues observed after in vivo administration of lipopolysaccharide (LPS). In overnight-fasted male rats LPS injected via the tail vein decreased the IGF-I concentration in plasma, liver, and skeletal muscle (30-45%) and increased IGF-I content in kidney (approximately 3-fold). LPS also decreased IGF-I mRNA abundance in liver and muscle and increased gene expression in kidney. Concomitantly, IGFBP-1 levels in plasma, liver, and muscle were markedly elevated by LPS. All these changes were associated with a greater than fourfold elevation in plasma corticosterone. Pretreatment of rats with the glucocorticoid receptor antagonist RU-486 completely prevented or blunted the LPS-induced changes in IGF-I content in plasma, liver, muscle, and kidney. In liver and muscle RU-486 significantly attenuated the reduction in IGF-I mRNA abundance produced by LPS, but in kidney the LPS-induced increase in IGF-I mRNA was still evident. In contrast, pretreatment with RU-486 did not prevent or attenuate the LPS-induced increase in IGFBP-1 levels in plasma, liver, or muscle. These data suggest that glucocorticoids play a major role in regulating IGF-I mRNA and peptide content in tissues in response to LPS, but the increased IGFBP-1 in blood and tissues induced by LPS appears largely glucocorticoid independent.
Cloning, expression, and tissue localization of the calcium-sensing receptor in chicken (Gallus domesticus)
Authors: Diaz R. , Hurwitz S. , Chattopadhyay N. , Pines M. , Yang Y. , Kifor O. , Einat M. S. , Butters R. , Hebert S. C. , Brown E. M.
Published: December 2017

In previous studies, we characterized an extracellular Ca2+ (Cao(2+))-sensing receptor (CaR) that plays a central role in regulating parathyroid hormone secretion in mammals by sensing Cao2+. In the present study, we have cloned and characterized the chicken (Gallus domesticus) homolog of the CaR. The chicken parathyroid CaR shares a high degree of homology (84% amino acid identity) with the human CaR and displays a similar topology. Moreover, amino acid residues where mutations cause disorders of Cao(2+)-sensing in the human CaR share the wild-type human sequence in the chicken CaR. However, a single region in the extracellular domain of the chicken CaR differs substantially from its mammalian homologs. Xenopus laevis oocytes injected with chicken CaR cRNA respond to elevated ambient levels of Cao2+, extracellular Mg2+, or extracellular Gd3+ with the characteristic activation of inositol trisphosphate-dependent, intracellular Ca(2+)-induced Cl- currents elicited by mammalian CaRs as well as by G protein-linked receptors coupled to activation of phospholipase C. By in situ hybridization, clusters of cells in chicken parathyroid glands were shown to express CaR messenger RNA. Northern analysis and immunohistochemistry demonstrated expression of receptor transcripts and/or protein in kidney tubules and intestine as well as in brain. The close conservation of the amino acid sequence of the chicken CaR with its mammalian homologs as well as its similar tissue distribution suggest that the receptor may also play an important role in avian calcium homeostasis.
Spontaneous skin temperature oscillations in normal human subjects
Authors: Shusterman V. , Anderson K. P. , Barnea O.
Published: December 2017

A noninvasive method based on high-resolution measurements and bandpass filtering of spontaneous skin temperature oscillations (approximately 4.0 x 10(-2) degrees C) in the low-frequency range (0.01-0.04 Hz) was investigated in normal human subjects. We hypothesized that the oscillations (temperature variability) originate from vasomotor activity of small arteries and arterioles in subcutaneous tissues. To test this hypothesis, continuous blood pressure waveforms were obtained with the use of an external piezoelectric sensor. The peak-to-peak envelope of the pressure signal (pressure variability) was used as an indicator of vasomotor activity. The variabilities of temperature and pressure were compared using cross-spectral and coherence analysis. The correlation between the peak frequency of the signals was 0.92, and the coherence was greater than 0.9. The signals demonstrated similar changes in spectral energy and peak frequency in response to mental stress. Reproducibility of the temperature variability in individual subjects was verified by repeating measurements 1-12 wk later. The differences in peak frequency were small (0.0155 +/- 0.001 Hz), and in each subject the signals exhibited similar patterns in response to stress. Correlation between spectral characteristics of the signals suggests that temperature variability can be attributed to changes in blood flow resulting from oscillations in vasomotor smooth muscle tone.
Nitric oxide within the paraventricular nucleus mediates changes in renal sympathetic nerve activity
Authors: Zhang K. , Mayhan W. G. , Patel K. P.
Published: December 2017

The paraventricular nucleus (PVN) of the hypothalamus is known to be involved in the control of sympathetic outflow. The goal of the present study was to examine the role of nitric oxide within the PVN in the regulation of renal sympathetic nerve activity. Renal sympathetic nerve discharge (RSND), arterial blood pressure, and heart rate in response to the microinjection of nitric oxide synthase inhibitor NG-monomethyl-L-arginine (L-NMMA; 50, 100, and 200 pmol) into the PVN were measured in male Sprague-Dawley rats. Microinjection of L-NMMA elicited an increase in RSND, arterial blood pressure, and heart rate. Administration of NG-monomethyl-D-arginine (D-NMMA, 50-200 pmol) into the PVN did not change RSND, arterial pressure, or heart rate. Similarly, microinjection of another nitric oxide inhibitor NG-nitro-L-arginine methyl ester (L-NAME; 100 nmol) also elicited an increase in RSND, arterial blood pressure, and heart rate. L-Arginine (100 nmol) reversed the effects of L-NAME in the PVN. Furthermore, microinjection of sodium nitroprusside (SNP; 50, 100, and 200 nmol) into the PVN elicited a significant decrease in RSND, arterial blood pressure, and heart rate. These effects of L-NMMA, L-NAME, and SNP on RSND and arterial blood pressure were not mediated by their vasoactive action because microinjection of phenylephrine and hydralazine did not elicit similar respective changes. In conclusion, our data indicate that endogenous nitric oxide within the PVN regulates sympathetic outflow via some inhibitory mechanisms. Altered nitric oxide mechanisms within the PVN may contribute to elevated sympathetic nerve activity observed during various diseases states such as heart failure and hypertension.
Catechol changes in the rat rostral ventrolateral medulla following changes in systemic CO2
Authors: Rentero N. , Bruandet N. , Pequignot J. M. , Quintin L.
Published: December 2017

A catechol signal recorded with in vivo voltammetry within the rat rostral ventrolateral medulla (RVLM) was taken as an index of the activity of RVLM adrenergic neurons and related to the level of arterial PCO2, under halothane anesthesia. Reversible increases in catechol signal were observed during reversible increases in arterial partial CO2 pressure (PaCO2) from 20 to 60 mmHg after alteration of tidal volume (n = 5 intact rats, n = 5 after carotid sinus deafferentation). A reversible increase in inspiratory CO2 combined with constant tidal volume led to changes in PaCO2 from 40 mmHg to 50 or 60 or 70 mmHg for 60 min (n = 5 in each group) and to a reversible increase in catechol signal (r = 0.76). These changes were also observed after carotid sinus deafferentation (PaCO2 = 40 to 60 to 40 mmHg, n = 5). Lowering the PaCO2 from 40 to 20 mmHg led to a minor, nonsignificant reduction in catechol signal (n = 5). Changes in arterial pressure were minimal, although they reached statistical significance in some groups of experiments. The level of catechol metabolism in the RVLM 1) is continuously related to the level of arterial CO2, 2) functions close to its resting level under baseline nonstimulated condition with respect to CO2, and 3) is reversibly modified on changes in capnia. Sensitivity of the catechol signal, recorded in the rostral ventrolateral medulla, to CO2 appears primarily to be centrally mediated. Thus adrenergic RVLM neurons may relay inputs from the central respiratory generator to the sympathetic chemoreflex or act as chemosensors for CO2, next to the ventrolateral medulla surface.
Role of the nitric oxide pathway in hypoxia-induced hypothermia of rats
Authors: Branco L. G. , Carnio E. C. , Barros R. C.
Published: December 2017

Hypothermia is a response to hypoxia that occurs in organisms ranging from protozoans to mammals, but very little is known about the mechanisms involved. Recently, the NO pathway has been suggested to be involved in thermoregulation. In the present study, we assessed the participation of nitric oxide in hypoxia-induced hypothermia by means of NO synthase inhibition using NG-nitro-L-arginine methyl ester (L-NAME). The rectal temperature of awake, unrestrained rats was measured before and after hypoxia or L-NAME injection or both treatments together. Control animals received saline injections of the same volume. We observed a significant (P < 0.05) reduction in body temperature of 1.32 +/- 0.36 degrees C after hypoxia (7% inspired O2) and of 0.96 +/- 0.42 degree C after L-NAME (30 mg/kg body wt) injected intravenously. When the two treatments were combined, no significant difference in body temperature was observed. To assess the role of central thermo-regulatory mechanisms, a smaller dose of L-NAME (1 mg/kg) was injected into the third cerebral ventricle or intravenously. Intracerebroventricular injection of L-NAME caused an increase in body temperature, but when L-NAME was combined with hypoxia (7% inspired O2) no change in body temperature was observed. Intravenous injection of 1 mg/kg L-NAME had no effect. The data indicate that NO plays a major role in hypoxia-induced hypothermia at central rather than peripheral sites.
Carbohydrate utilization in rat soleus muscle is influenced by carbonic anhydrase III activity
Authors: Côté Claude H. , Perreault Guylaine , Frenette Jérôme
Published: December 2017

Inhibition of carbonic anhydrase III (CA III; EC 4.2.1.1 ) activity in type I muscle can influence resistance to fatigue and glycogen utilization. Our aim was to determine if CA III inhibition could influence muscle pH and glycolytic rate. Muscle pH, hexosemonophosphates (HMP), glycolytic intermediates, ATP, and creatine phosphate (CP) were measured at rest and during a fatigue protocol in rat soleus muscles in vitro with or without CA inhibitors (CAI). In resting muscles, CAI resulted in a significant drop in pH (7.11 vs. 7.06, P < 0.05) and in a two- to threefold increase in HMP content compared with control muscles. Measurements of HMP and glycolytic intermediates during the fatigue protocol suggested, however, that the glycolytic flux was not influenced. Globally, muscles incubated with CAI showed larger perturbations of their CP and ATP content than control muscles. The accumulation of HMP induced by the CAI was found to be totally dependent on the combined presence of external glucose and contractile activity, suggesting that inhibiting CA III may augment the responsitivity of the contraction-induced glucose uptake process.