Obesity-resistant Rats Research Articles

Regulation of hypothalamic neuropeptides gene expression in diet induced obesity resistant rats: possible targets for obesity prediction?

Several factors play a role in obesity (i.e., behavior, environment, and genetics) and epigenetic regulation of gene expression has emerged as a potential contributor in the susceptibility and development of obesity. To investigate the individual sensitivity to weight gain/resistance, we here studied gene transcription regulation of several hypothalamic neuropeptides involved in the control of energy balance in rats developing obesity (diet-induced obesity, DIO) or not (diet resistant, DR), when fed with a high fat diet. Rats have been followed up to 21 weeks of high fat diet exposure. After 5 weeks high fat diet exposure, the obese phenotype was developed and we observed a selective down-regulation of the orexigenic neuropeptide Y (NPY) and peroxisome proliferator-activated receptor gamma (PPAR-γ) genes. No changes were observed in the expression of the agouti-related protein (AgRP), as well as for all the anorexigenic genes under study. After long-term high fat diet exposure (21 weeks), NPY and PPAR-γ, as well as most of the genes under study, resulted not be different between DIO and DR, whereas a lower expression of the anorexigenic pro-opio-melanocortin (POMC) gene was observed in DIO rats when compared to DR rats. Moreover we observed that changes in NPY and POMC mRNA were inversely correlated with gene promoters DNA methylation. Our findings suggest that selective alterations in hypothalamic peptide genes regulation could contribute to the development of overweight in rats and that environmental factor, as in this animal model, might be partially responsible of these changes via epigenetic mechanism.

Impact of a high energy diet on obesity-associated immune biomarkers in Sprague Dawley rats (MUC2P.929)

Abstract A trial was designed to examine effects of a high energy diet on obesity-associated biomarkers in Sprague-Dawley (SD) rats. Male rats (n = 176) were fed an AIN93G high-energy diet containing 16% dietary fat for 2 weeks, and segregated into obese-prone and obese-resistant phenotypes. Necropsies of animals displaying the highest (n = 10) and those displaying the lowest weight gain (n = 10) were conducted to examine potential differences in the concentration of immune biomarkers. Adipose, mucosal (ileum, ileal Peyer’s patches, cecum, proximal and distal colon), mesenteric lymph nodes (mln) and systemic (serum, liver, spleen) tissues were comparatively analyzed for cytokine or adipokine profiles. Obese rats had higher levels of leptin (P = 0.0001) and IL-22 (P = 0.059) in the distal colon and higher levels of IL-10 (P = 0.032) and TNF-α (P = 0.016) in the proximal colon compared to their obese-resistant counterparts. In obese-resistant rats, higher levels of CINC-2αβ (P = 0.048) occurred in mesenteric adipose tissue along with higher levels of soluble ICAM-1 (P = 0.018) in the mln. These findings suggest a high energy diet imparted differential effects on mucosal cytokine and adipokine production in colonic and mesenteric tissue between obese-prone and obese-resistant SD rats. Elevated levels of pro-inflammatory and regulatory cytokines are evident as little as 2 weeks after the initiation of a high-energy diet.

Impaired oxidative capacity due to decreased CPT1b levels as a contributing factor to fat accumulation in obesity.

To characterize mechanisms responsible for fat accumulation we used a selectively bred obesity-prone (OP) and obesity-resistant (OR) rat model where the rats were fed a Western diet for 76 days. Body composition was assessed by magnetic resonance imaging scans, and as expected, the OP rats developed a higher degree of fat accumulation compared with OR rats. Indirect calorimetry showed that the OP rats had higher respiratory exchange ratio (RER) compared with OR rats, indicating an impaired ability to oxidize fat. The OP rats had lower expression of carnitine palmitoyltransferase 1b in intra-abdominal fat, and higher expression of stearoyl-CoA desaturase 1 in subcutaneous fat compared with OR rats, which could explain the higher fat accumulation and RER values. Basal metabolic parameters were also examined in juvenile OP and OR rats before and during the introduction of the Western diet. Juvenile OP rats likewise had higher RER values, indicating that this trait may be a primary and contributing factor to their obese phenotype. When the adult obese rats were exposed to the orexigenic and adipogenic hormone ghrelin, we observed increased RER values in both OP and OR rats, while OR rats were more sensitive to the orexigenic effects of ghrelin as well as ghrelin-induced attenuation of activity and energy expenditure. Thus increased fat accumulation characterizing obesity may be caused by impaired oxidative capacity due to decreased carnitine palmitoyltransferase 1b levels in the white adipose tissue, whereas ghrelin sensitivity did not seem to be a contributing factor.

MP8-04 UNDERACTIVE BLADDER IN OBESE-PRONE RATS FED A HIGH FAT DIET

You have accessJournal of UrologyUrodynamics/Incontinence/Female Urology: Basic Research I1 Apr 2015MP8-04 UNDERACTIVE BLADDER IN OBESE-PRONE RATS FED A HIGH FAT DIET Nazema Siddiqui, Alexis Dieter, Cindy Amundsen, Jillene Brooks, Danielle Degoski, and Matthew Fraser Nazema SiddiquiNazema Siddiqui More articles by this author , Alexis DieterAlexis Dieter More articles by this author , Cindy AmundsenCindy Amundsen More articles by this author , Jillene BrooksJillene Brooks More articles by this author , Danielle DegoskiDanielle Degoski More articles by this author , and Matthew FraserMatthew Fraser More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2015.02.271AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Changes in systemic metabolism lead to alterations in lower urinary tract (LUT) physiology. High fat diets (HFD) are implicated in the development of insulin resistance. Obese Prone (OP) and Obese Resistant (OR) rats (Charles River) are used in obesity research, as OP rats become obese on HFD while OR remain lean. To assess the effect of obesity on LUT function, we characterized LUT function in both rat strains during chronic HFD feeding. METHODS Four OP and 4 OR female rats were placed on the same HFD at 9 weeks (wks) of age (30% fat for 12 wk, followed by 60% fat for 4 wk, Research Diets). Conscious restrained cystometry was performed at 7, 11 and 15 wk HFD. Body mass index (BMI) was calculated, and serum glucose was measured. For all cystometric evaluations, single fill cystometrograms were performed to determine true bladder capacity (TBC), followed by 60 min of continuous cystometry to determine functional bladder capacity (FBC). Voiding efficiency (%VE) was calculated as follows: %VE = [average FBC/TBC]∗100. For animals showing overflow incontinence (OI), TBC was a priori considered to be 5 milliliters. RESULTS Mean BMIs were higher in OP compared to OR rats (0.64 vs. 0.46 at 7wk, 0.72 vs. 0.52 at 11 wk, 0.74 vs. 0.52 at 15 wk, respectively; p = 0.002). At 15 wk HFD, there were no significant differences in mean blood glucose between OP and OR groups (102 +/-14 vs. 97 +/- 11, respectively; p=0.58). TBC and %VE were compared across all time points and results are shown in the figure. At 7 wk HFD, all rats exhibited normal voiding patterns. At 11 wk HFD, 2/4 (50%) of OP rats exhibited OI while the remaining 2/4 OP and all 4 OR rats exhibited normal voiding patterns. At 15 wk HFD, one of the OP rats with OI at 11 wk HFD could no longer be assessed due to catheter issues. Of the remaining OP rats, 2/3 (66%) exhibited overflow incontinence while all OR rats showed normal voiding patterns. Mean %VE decreased in the OP group. CONCLUSIONS Obese non-diabetic animals exhibited elevated bladder capacities, to the point of OI in the majority, and decreased voiding efficiency after 15 wk HFD. OP rats given a chronic HFD may serve as a promising new model to study underactive bladder. © 2015 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 193Issue 4SApril 2015Page: e74 Advertisement Copyright & Permissions© 2015 by American Urological Association Education and Research, Inc.MetricsAuthor Information Nazema Siddiqui More articles by this author Alexis Dieter More articles by this author Cindy Amundsen More articles by this author Jillene Brooks More articles by this author Danielle Degoski More articles by this author Matthew Fraser More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Cardiovascular, metabolic, and coronary dysfunction in high-fat-fed obesity-resistant/prone rats.

Obesity is a global epidemic leading to several comorbidities including diabetes and cardiovascular disease. The hypothesis that the genetic background of the obesity-prone rat (OP) predisposes to physiologic, metabolic, and microvascular dysfunction which is exacerbated by a diet high in saturated fats was tested. Male OP and obesity-resistant (OR) rats were fed either a diet containing 10% (chow) or 45% kcal fat (HF) for 42 weeks. Weight of OP rats was greater than OR rats by 8 weeks on both diets. Blood pressure was increased in OP rats on chow and further augmented by HF diet compared to OR rats on similar diets. In contrast to weight and blood pressure, glucose clearance was similar in OR and OP rats on chow and impaired in both models on HF diet. Relaxation to acetylcholine was attenuated in OP rats compared to OR rats by 8 weeks and remained reduced throughout the study. A longer period of time was required to observe vascular dysfunction in HF-fed OR rats. When compared to OR rats, OP rats are prone to develop not only greater obesity but also hypertension and vascular dysfunction on a normal diet which is further augmented with HF diet.

Obesity Resistance Promotes Mild Contractile Dysfunction Associated with Intracellular Ca2+ Handling.

BackgroundDiet-induced obesity is frequently used to demonstrate cardiac dysfunction. However, some rats, like humans, are susceptible to developing an obesity phenotype, whereas others are resistant to that.ObjectiveTo evaluate the association between obesity resistance and cardiac function, and the impact of obesity resistance on calcium handling.MethodsThirty-day-old male Wistar rats were distributed into two groups, each with 54 animals: control (C; standard diet) and obese (four palatable high-fat diets) for 15 weeks. After the experimental protocol, rats consuming the high-fat diets were classified according to the adiposity index and subdivided into obesity-prone (OP) and obesity-resistant (OR). Nutritional profile, comorbidities, and cardiac remodeling were evaluated. Cardiac function was assessed by papillary muscle evaluation at baseline and after inotropic maneuvers.ResultsThe high-fat diets promoted increase in body fat and adiposity index in OP rats compared with C and OR rats. Glucose, lipid, and blood pressure profiles remained unchanged in OR rats. In addition, the total heart weight and the weight of the left and right ventricles in OR rats were lower than those in OP rats, but similar to those in C rats. Baseline cardiac muscle data were similar in all rats, but myocardial responsiveness to a post-rest contraction stimulus was compromised in OP and OR rats compared with C rats.ConclusionObesity resistance promoted specific changes in the contraction phase without changes in the relaxation phase. This mild abnormality may be related to intracellular Ca2+ handling.

High-fat diet induces endothelial dysfunction through a down-regulation of the endothelial AMPK-PI3K-Akt-eNOS pathway.

Activation of endothelial adenosine monophosphate-activated protein kinase (AMPK) contributes to increase nitric oxide (NO) availability. The aim of this study was to assess if high-fat diet (HFD)-induced endothelial dysfunction is linked to AMPK deregulation. Twelve-week-old Sprague Dawley male rats were assigned either to control (10 kcal % from fat) or to HFD (45 kcal % from fat) for 8 wk. HFD rats segregated in obesity-prone (OP) or obesity-resistant (OR) rats according to body weight. HFD triggered an impaired glucose management together with impaired endothelium-dependent relaxation, reduced endothelial AMPK activity and lower NO availability in aortic rings of OP and OR cohorts. Relaxation evoked by AMPK activator, 5-aminoimidazole-4-carboxamide-1-β-d-ribofuranoside (AICAR) was reduced in both OP and OR rings, which exhibited lower p-AMPKα-Thr(172) /AMPKα ratios that negatively correlated with plasma non-esterified fatty acids (NEFA) and triglycerides (TG). Inhibition of PI3K (wortmannin, 10(-7) M) or Akt (triciribine, 10(-5) M) reduced relaxation to AICAR only in the control group (p < 0.001). Akt (p-Akt-Ser(473) ) and eNOS phosphorylation (p-eNOS-Ser(1177) ) were significantly reduced in OP and OR (p < 0.01). Endothelial dysfunction caused by HFD is related to a dysfunctional endothelial AMPK-PI3K-Akt-eNOS pathway correlating with the increase of plasma NEFA, TG, and an impaired glucose management.

Repeated allopregnanolone exposure induces weight gain in schedule fed rats on high fat diet

Ingestion of energy rich high fat diets is one of the determining factors associated with the obesity epidemic. Therefore, much can be learned from studies of obesity-related substances given to animals fed a high fat diet. The progesterone metabolite allopregnanolone is a potent positive modulator of the gamma-aminobutyric acid (GABA)A-receptor, and both allopregnanolone and GABA have been implicated in evoking hyperphagia. In this study, food intake and body weight gain were investigated during repeated allopregnanolone exposure. Male Wistar rats were studied when fed chow ad libitum, with chow access for 4h per day or with 45% high fat pellets for 4h per day. Rats on the high fat diet were separated into obesity prone and obesity resistant individuals. Subcutaneous injections of allopregnanolone were given once daily over five consecutive days. Repeated exposure to allopregnanolone lead to increased weight gain, significantly so in schedule fed rats on a high fat diet. The increased weight gain was correlated to an increased energy intake. Both obesity resistant and obesity prone rats responded to allopregnanolone with increased weight gain. Obesity resistant rats treated with allopregnanolone increased their energy intake and ate as much as vehicle treated obesity prone rats. Their weight gain was also increased to the level of obesity prone rats injected with just the vehicle carrier oil. Thus, it appears that allopregnanolone may be one of the endogenous factors involved in weight gain, especially when the diet is rich in fat.

Dietary restriction in moderately obese rats improves body size and glucose handling without the renal and hepatic alterations observed with a high-protein diet.

Obesity is increasing worldwide, and high-protein (HP) diets are widely used for weight loss. However, the overall safety of HP diets is not well established in obese individuals, who make up a significant proportion of the population. To evaluate the health effects of an HP diet in obesity, obesity-prone (OP) Sprague-Dawley rats were given high-fat diets for 12 weeks to induce obesity. Following this, for 8 more weeks, these rats were given either a normal-protein (NP) (15% of energy) or an HP (35% of energy) diet ad libitum, or the NP diet at a restricted level to achieve body weights similar to those of the HP group (pair-weighted (PW) group). Obesity-resistant (OR) control rats were also given the NP diet throughout the feeding period. The HP-OP group had higher food intake but lower body weight, improved glucose handling, and lowered serum haptoglobin compared with the NP-OP group. These benefits were also observed in PW-OP rats. In addition, PW-OP rats had less fat accumulation when compared with NP-OP rats, and an improved Lee index, lower liver size, and lower serum alanine aminotransferase when compared with HP-OP rats. On the other hand, kidney size, proteinuria, and serum homocysteine were increased in HP-OP rats compared with NP-OP rats, whereas PW-OP rats did not experience these effects. These results indicate that in obese rats, more benefits are obtained via dietary restriction with an NP diet and without some of the potentially detrimental effects of an HP diet.

Abstract 155: Cardiac Function And Calcium Handling in Obesity-Resistant rats

Experimental studies have demonstrated that obesity induced by different types of high-fat and/or high-energy diets also lead to myocardial dysfunction. Nevertheless, few studies have evaluated the myocardial function in obesity-resistant rats. Moreover, the mechanisms underlying the participation of calcium (Ca 2+ ) handling on cardiac function in this model remain unknown. The aim of this study was to investigate in rodent model of obesity-resistant. In addition, as obesity-resistant group is fed a high-fat diet but did not become obese, this study tested whether obesity-resistant model develops cardiac abnormalities and impairment of calcium handling as obesity-prone. Male 30-day-old Wistar rats were fed standard (C) and alternately four palatable unsaturated high-fat diets (Ob) for 15 weeks. After experimental protocol, Ob rats consuming the unsaturated high-fat diets were ranked based on adiposity index. Rats on the unsaturated high-fat diets exhibiting the greatest adiposity index were referred to as OP, whereas those exhibiting the lowest adiposity index were referred to as OR. Obesity was determined by adiposity index and comorbidities were evaluated. Myocardial function was evaluated in isolated left ventricle papillary muscles under basal conditions and after inotropic and lusitropic maneuvers. After 15 weeks, final body weight, total body fat, adiposity index, triglycerides levels and heart weight were significantly greater in OP rats than C and OR rats, however, there was no change in systolic blood pressure between groups. The C, OP and OR muscles developed similar baseline data, but myocardial responsiveness to post-rest contraction stimulus was compromised in OP rats. In conclusion, obesity-resistant model by unsaturated high-fat diet, after 15 weeks, does not display nutritional and metabolic characteristics of obesity-prone. Furthermore, obesity-resistant does not promote cardiac abnormalities and impairment of calcium handling visualized in obesity-prone.

Glutamatergic signaling and low prodynorphin expression are associated with intact memory and reduced anxiety in rat models of healthy aging.

The LOU/C/Jall (LOU) rat strain is considered a model of healthy aging due to its increased longevity, maintenance of stable body weight (BW) throughout life and low incidence of age-related diseases. However, aging LOU rat cognitive and anxiety status has yet to be investigated. In the present study, male and female LOU rat cognitive performances (6–42 months) were assessed using novel object recognition and Morris Water Maze tasks. Recognition memory remained intact in all LOU rats up to 42 months of age. As for spatial memory, old LOU rat performed similarly as young animals for learning acquisition, reversal learning, and retention. While LOU rat BW remained stable despite aging, 20-month-old ad-libitum-fed (OAL) male Sprague Dawley rats become obese. We determined if long-term caloric restriction (LTCR) prevents age-related BW increase and cognitive deficits in this rat strain, as observed in the obesity-resistant LOU rats. Compared to young animals, recognition memory was impaired in OAL but intact in 20-month-old calorie-restricted (OCR) rats. Similarly, OAL spatial learning acquisition was impaired but LTCR prevented the deficits. Exacerbated stress responses may favor age-related cognitive decline. In the elevated plus maze and open field tasks, LOU and OCR rats exhibited high levels of exploratory activity whereas OAL rats displayed anxious behaviors. Expression of prodynorphin (Pdyn), an endogenous peptide involved in stress-related memory impairments, was increased in the hippocampus of OAL rats. Group 1 metabotropic glutamate receptor 5 and immediate early genes Homer 1a and Arc expression, both associated with successful cognitive aging, were unaltered in aging LOU rats but lower in OAL than OCR rats. Altogether, our results, supported by principal component analysis and correlation matrix, suggest that intact memory and low anxiety are associated with glutamatergic signaling and low Pdyn expression in the hippocampus of non-obese aging rats.

Replication of Obesity and Associated Signaling Pathways Through Transfer of Microbiota From Obese-Prone Rats

Aberrations in gut microbiota are associated with metabolic disorders, including obesity. However, whether shifts in the microbiota profile during obesity are a characteristic of the phenotype or a consequence of obesogenic feeding remains elusive. Therefore, we aimed to determine differences in the gut microbiota of obese-prone (OP) and obese-resistant (OR) rats and examined the contribution of this microbiota to the behavioral and metabolic characteristics during obesity. We found that OP rats display a gut microbiota distinct from OR rats fed the same high-fat diet, with a higher Firmicutes-to-Bacteroidetes ratio and significant genera differences. Transfer of OP but not OR microbiota to germ-free (GF) mice replicated the characteristics of the OP phenotype, including reduced intestinal and hypothalamic satiation signaling, hyperphagia, increased weight gain and adiposity, and enhanced lipogenesis and adipogenesis. Furthermore, increased gut permeability through conventionalization resulted in inflammation by proinflammatory nuclear factor (NF)-κB/inhibitor of NF-κB kinase subunit signaling in adipose tissue, liver, and hypothalamus. OP donor and GF recipient animals harbored specific species from Oscillibacter and Clostridium clusters XIVa and IV that were completely absent from OR animals. In conclusion, susceptibility to obesity is characterized by an unfavorable microbiome predisposing the host to peripheral and central inflammation and promoting weight gain and adiposity during obesogenic feeding.

Attenuated insulin induced dilation in coronary arteries from obesity resistant and obesity prone high fat fed rats (1071.6)

The purpose of this study was to compare physiologic, metabolic and microvascular complications in obesity resistant (OR) and obesity prone (OP) rats fed a high fat (HF) diet. 8 week old male rats were fed a diet containing 45% kcal saturated fat for 42 weeks. Blood pressure was measured by telemetry. Reactivity of small (75 ‐ 150 μm intraluminal diameter) coronary arteries was evaluated. Body weights were 504±7, 669±25*, 685±8*, and 924±39* grams for OR chow, OR HF, OP chow OP HF rats, respectively. Blood pressure was increased in OP rats on control diet (121.5 ± 4.3mmHg*) and further increased with HF diet (129.7 ± 2.4mmHg*), compared to OR rats on control (98.3 ± 4.3mmHg) or HF diet (99.5 ± 3.1mmHg). Glucose clearance was impaired in high fat fed OR and OP rats. Insulin mediated relaxation of coronary arteries from HF fed OR and OP rats was impaired (% relaxation to 100 ng/ml: OR chow = 37.1 ± 15, OR HF = 4.6 ± 5.3, OP chow = 29.3 ± 6.6 and OP HF = 7.6 ± 2.3). Acetylcholine (ACh)‐mediated relaxation of coronary arteries from HF fed rats was impaired (%relaxation to 10 μM Ach: OR chow=67 ± 10, OR HF = 36±12*, OP chow = 41±8*, and OP HF = 22±6*). Sodium nitroprusside (SNP) responses were not altered. *p&lt;0.05 vs OR chow diet. These studies indicate that HF diet augments metabolic and microvascular complications in obesity prone rats.Grant Funding Source: Supported by VA Merit to Dr. Oltman

Hyperinsulinemia potentiates airway responsiveness to parasympathetic nerve stimulation in obese rats.

Obesity is a substantial risk factor for developing asthma, but the molecular mechanisms underlying this relationship are unclear. We tested the role of insulin in airway responsiveness to nerve stimulation using rats genetically prone or resistant to diet-induced obesity. Airway response to vagus nerve stimulation and airway M2 and M3 muscarinic receptor function were measured in obese-prone and -resistant rats with high or low circulating insulin. The effects of insulin on nerve-mediated human airway smooth muscle contraction and human M2 muscarinic receptor function were tested in vitro. Our data show that increased vagally mediated bronchoconstriction in obesity is associated with hyperinsulinemia and loss of inhibitory M2 muscarinic receptor function on parasympathetic nerves. Obesity did not induce airway inflammation or increase airway wall thickness. Smooth muscle contraction to acetylcholine was not increased, indicating that hyperresponsiveness is mediated at the level of airway nerves. Reducing serum insulin with streptozotocin protected neuronal M2 receptor function and prevented airway hyperresponsiveness to vagus nerve stimulation in obese rats. Replacing insulin restored dysfunction of neuronal M2 receptors and airway hyperresponsiveness to vagus nerve stimulation in streptozotocin-treated obese rats. Treatment with insulin caused loss of M2 receptor function, resulting in airway hyperresponsiveness to vagus nerve stimulation in obese-resistant rats, and inhibited human neuronal M2 receptor function in vitro. This study shows that it is not obesity per se but hyperinsulinemia accompanying obesity that potentiates vagally induced bronchoconstriction by inhibiting neuronal M2 muscarinic receptors and increasing acetylcholine release from airway parasympathetic nerves.

High-Fat Diet Alters the Orosensory Sensitivity to Fatty Acids in Obesity-Resistant but not Obesity-Prone Rats.

Gene-environment interactions play a role in the development of obesity but specific effects of diet on the orosensory detection of fatty acids have yet to be clarified. The objective of this study is to characterize the effect of prolonged (5-week) exposure to a high-fat (60%) diet on the behavioral sensitivity to the fatty acid linoleate following a conditioned taste aversion in obesity-prone and obesity-resistant rats. Exposure to the high-fat diet significantly enhanced the sensitivity of obesity-resistant (S5B/Pl) rats to linoleate while producing no effect on the fatty acid sensitivity for obesity-prone rats. Specifically, high-fat diet fed S5B/Pl rats showed stronger initial avoidance of linoleate and slower extinction rates than their normal diet cohorts. Our study suggests that prolonged dietary fat consumption may alter the behavioral sensitivity to fatty acids particularly in obesity-resistant animals.

Preference for linoleic acid in obesity-prone and obesity-resistant rats is attenuated by the reduction of CD36 on the tongue.

Differential sensing of dietary fat and fatty acids by the oral cavity is proposed to regulate the susceptibility to obesity. In the current experiments, animals that differ in their susceptibility to obesity were used to investigate the influence of the oral cavity on the preference for the polyunsaturated fatty acid, linoleic acid. In experiment 1, the preference for differing concentrations of linoleic acid was determined in obesity-prone Osborne-Mendel (OM) and obesity-resistant S5B/Pl (S5B) rats. The preference threshold for linoleic acid was lower in S5B rats, compared with OM rats. To determine whether differences in linoleic acid preference threshold were related to innate strain differences in the fatty acid receptors on the tongue, the expression of GPR120, GPR40, and CD36 on the circumvallate papillae were assessed in OM and S5B rats. Results indicated that the expression of CD36, GPR40, and GPR120 did not differ between these two strains. Numerous studies have examined the role of CD36 on fat intake; therefore, in experiment 3, RNA interference was used to decrease the expression of CD36 on the tongues of OM and S5B rats, and the effect of decreased CD36 expression on linoleic acid preference was determined. CD36 siRNA attenuated linoleic acid preference for the most preferred concentration in both OM and S5B rats. Overall, these data indicate that there are innate differences in the preference threshold for linoleic acid in obesity-prone and obesity-resistant rats. Experimentally reducing the expression of CD36 on the circumvallate papillae attenuated the preference for linoleic acid in both strains.

Mechanisms underlying obesity resistance associated with high spontaneous physical activity

Obesity resistance due to elevated orexin signaling is accompanied by high levels of spontaneous physical activity (SPA). The behavioral and neural mechanisms underlying this observation have not been fully worked out. We determined the contribution of hypothalamic orexin receptors (OXRs) to SPA stimulated by orexin A (OXA), whether OXA-stimulated SPA was secondary to arousal and whether voluntary wheel running led to compensations in 24-h SPA. We further tested whether orexin action on dopamine one receptors (DA1R) in the substantia nigra (SN) plays an important role in the generation of SPA. To test this, SPA response was determined in lean and obese rats with cannulae targeted toward the rostral lateral hypothalamus (rLH) or SN. Sleep/wake states were also measured in rats with rLH cannula and electroencephalogram/electromyogram radiotelemetry transmitters. SPA in lean rats was more sensitive to antagonism of the OX1R and in the early response to the orexin 2 agonist. OXA increased arousal equally in lean and obese rodents, which is discordant from the greater SPA response in lean rats. Obesity-resistant rats ran more and wheel running was directly related to 24-h SPA levels. The OX1R antagonist, SB-334867-A, and the DA1R antagonist, SCH3390, in SN more effectively reduced SPA stimulated by OXA in obesity-resistant rats. These data suggest OXA-stimulated SPA is not secondary to enhanced arousal, propensity for SPA parallels inclination to run and that orexin action on dopaminergic neurons in SN may participate in the mediation of SPA and running wheel activity.

Role of the locus coeruleus in enhanced orexin A-induced spontaneous physical activity in obesity-resistant rats

Orexin/hypocretin terminals innervate noradrenergic locus coeruleus (LC) neurons that project to the prefrontral cortex, which may influence spontaneous physical activity (SPA) and energy balance. Obesity-resistant (OR) rats have higher orexin receptors (OXR) mRNA in the LC and other brain regions, as well as lower adiposity compared with obese rats. These findings led us to hypothesize that orexin activity in the LC is relevant for the OR phenotype. We compared OR rats to Sprague-Dawley rats. We predicted that: 1) brain OXR expression pattern is sufficient to differentiate OR from non-bred Sprague-Dawley rats; 2) nonresting energy expenditure (NREE) and orexin A (OXA)-stimulated SPA after injection in LC would be greater in OR rats; and 3) the effect of OXA on SPA would be greater than its effect on feeding. OXR mRNA from 11 brain sites and the SPA and feeding responses to OXA in the LC were determined. Body composition, basal SPA, and EE were determined. Principal component analysis of the OXR expression pattern differentiates OR and Sprague-Dawley rats and suggests the OXR mRNA in the LC is important in defining the OR phenotype. Compared with Sprague-Dawley rats, OR rats had greater SPA and NREE and lower resting EE and adiposity. SPA responsivity to OXA in the LC was greater in OR rats compared with Sprague-Dawley rats. OXA in the LC did not stimulate feeding in OR or Sprague-Dawley rats. These data suggest that the LC is a prominent site modulating OXA-stimulated SPA, which promotes lower adiposity and higher nonresting EE.

Sympathetic cardiac hyperinnervation and atrial autonomic imbalance in diet-induced obesity promote cardiac arrhythmias

Obesity increases the risk of arrhythmias and sudden cardiac death, but the mechanisms are unknown. This study tested the hypothesis that obesity-induced cardiac sympathetic outgrowth and hyperinnervation promotes the development of arrhythmic events. Male Sprague-Dawley rats (250-275 g), fed a high-fat diet (33% kcal/fat), diverged into obesity-resistant (OR) and obesity-prone (OP) groups and were compared with rats fed normal chow (13% kcal/fat; CON). In vitro experiments showed that both OR and OP rats exhibited hyperinnervation of the heart and high sympathetic outgrowth compared with CON rats, even though OR rats are not obese. Despite the hyperinnervation and outgrowth, we showed that, in vivo, OR rats were less susceptible to arrhythmic events after an intravenous epinephrine challenge compared with OP rats. On examining total and stimulus-evoked neurotransmitter levels in an ex vivo system, we demonstrate that atrial acetylcholine content and release were attenuated in OP compared with OR and CON groups. OP rats also expressed elevated atrial norepinephrine content, while norepinephrine release was suppressed. These findings suggest that the consumption of a high-fat diet, even in the absence of overt obesity, stimulates sympathetic outgrowth and hyperinnervation of the heart. However, normalized cardiac parasympathetic nervous system control may protect the heart from arrhythmic events.

Combination of Obesity and High-Fat Feeding Diminishes Sensitivity to GLP-1R Agonist Exendin-4

Gastrointestinal mechanisms involved in the suppression of appetite are compromised in obesity. Glucagon-like peptide-1 (GLP-1) is released in response to nutrients, suppresses food intake, and has been shown to play a role in regulation of energy balance. It is not known whether obese-prone (OP) rats exhibit dysfunctional GLP-1 signaling that could contribute to decreased nutrient-induced satiation and hyperphagia. Therefore, we examined the effects of exogenous intraperitoneal administration of the GLP-1R agonist, exendin-4 (Ex-4), on food intake in OP and obese-resistant (OR) rats during chow or high-energy/high-fat (HE/HF) feeding. All doses of Ex-4 effectively suppressed intake in OP and OR rats fed chow; however, during HE/HF-feeding, OP rats suppressed intake significantly less than OR rats at all Ex-4 doses tested. This was associated with downregulation of GLP-1R mRNA expression in the vagal nodose ganglia of OP rats. Furthermore, HE/HF-fed OP rats had significantly lower plasma GLP-1 levels, decreased protein levels of GLP-1 in the intestinal epithelium, and reduced number of L cells in the distal ileum. These results demonstrate that HE/HF-feeding, coupled with OP phenotype, results in reduced endogenous GLP-1 and GLP-1R activation, indicating that impaired GLP-1 signaling during obesity may exacerbate hyperphagia and weight gain.