Daily profile in ghrelin and hypothalamic ghrelin receptors in obese and lean mice Neotomodon alstoni

Mice Neotomodon alstoni develops, in some individuals, overweight and obesity when kept in vivarium conditions and fed on standard rodent diet. In the present study, we explored the possible differences between lean and obese mice, on the daily expression of the hormone ghrelin, its total and active form, as well as the relative expression of ghrelin receptors (GSHR) in hypothalamic tissue. Plasma hormones were detected and quantified by immunological techniques by ELISA; the relative presence of GSHR was assessed by western blot. Mice were sacrificed at different times throughout the day. The results obtained indicate that there is a rhythm of presence of total and acylated ghrelin in lean mice, which dampers and changes its peak’s phase in obese mice. The GHSR expression tend to be more abundant in obese mice than in lean mice. The results indicate that, in the obese mice there is a possible resistance to ghrelin.     

Day-night variations in pro-oxidant reactions of hypothalamic,hepatic and pancreatic tissue in mice with spontaneous obesity (Neotomodon alstoni)

Objectives: The present work compares the pro-oxidant properties in hypothalamic, hepatic, and pancreatic tissue of spontaneously obese and lean Neotomodon alstoni during day and night. Methods: Lipid peroxidation from hypothalamus, liver, and pancreas from lean and obese mice were quantified by the two-thiobarbituric acid method. Lipid peroxidation in vivo was also determined by means of detection of conjugated dienes in lipid extracts. Results: Hypothalamic tissue from obese Neotomodon showed a notorious increase (nearly 700%) in the production of thiobarbituric acid-reactive substances (TBARS) at day, either in basal as well as in an assay supplemented with Fe²⁺; the presence of conjugated dienes was also notably greater (76%) at day. Hypothalamus of lean mice presented an increase (170%) in assay supplemented with Fe²⁺. Hepatic tissue in obese mice showed diurnal increasing in TBARS + Fe²⁺ (34%) and in conjugated dienes (38%), while lean mice showed only a diurnal increase (45%) in TBARS + Fe²⁺. Pancreatic tissue from obese mice presented a diurnal increasing in basal TBARS (130%) but a decrease (72%) in TBARS + Fe²⁺. Presence of conjugated dienes was also decreased during the day in lean (60%) and in obese (40%) mice. Conclusions: In the obese Neotomodon, there is a larger day–night change in pro-oxidant status in the hypothalamus and in pancreas than the one observed in the liver, suggesting a differential equilibrium between oxidative reactions and antioxidant defenses in the different tissues during the day–night cycle.     

Effects of obesity on circadian photic entrainment of locomotor activity in wild mice Neotomodon alstoni

Obesity is increasing in industrialized countries at an alarming rate. Recent studies have linked this condition with changes in the circadian regulation, and circadian clock dysfunctions have also been linked to metabolic disorders. When in captivity and fed a regular rodent chow diet ad libitum some volcano mice, Neotomodon alstoni (endemic species of Mexico) become overweight and display symptoms equivalent to metabolic syndrome. The aim of this work was to observe whether there are significant changes in the functional properties of the circadian system, namely in the entraining circadian locomotor activity rhythm, between mice that became obese and normal adult mice. Freely moving circadian rhythms of locomotor activity were tested under constant conditions as well as under conditions of discrete and continuous entrainment. Our results show that volcano lean mice present a phase response curve with larger delays than advances, indicating that re-entrainment is more easily achieved by delays. Volcano obese mice are less efficient at re-entraining because they show smaller phase shifts in delays than control mice. These results indicate that obesity in N. alstoni has a negative effect on the circadian mechanisms that integrate the photic entrainment.     

Estradiol regulates leptin sensitivity to control feeding via hypothalamic Cited1

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Endurance training prevents inflammation and apoptosis in hypothalamic neurons of obese mice

This study investigated the effects of exercise training in regulating inflammatory processes, endoplasmic reticulum stress, and apoptosis in hypothalamic neurons of obese mice. Swiss mice were distributed into three groups: Lean mice (Lean), sedentary animals fed a standard diet; obese mice (Obese), sedentary animals fed a high-fat diet (HFD); trained obese mice (T. Obese), animals fed with HFD and concurrently subjected to an endurance training protocol for 8 weeks. In the endurance training protocol, mice ran on a treadmill at 60% of peak workload for 1 hr, 5 days/week for 8 weeks. Twenty-four hours after the last exercise session, the euthanasia was performed. Western blot, quantitative real-time polymerase chain reaction, and terminal deoxynucleotide transferase biotin-dUTP nick end-labeling (TUNEL) techniques were used for the analysis of interest. The results show exercise training increased phosphorylation of leptin signaling pathway proteins (pJAK2/pSTAT3) and reduced the content of tumor necrosis factor α, toll-like receptor 4, suppressor of cytokine signaling 3, protein–tyrosine phosphatase 1B as well as the phosphorylation of IkB kinase in the hypothalamus of T. Obese animals. A reduction of macrophage activation and phosphorylation of eukaryotic initiation factor 2α, and protein kinase RNA-like endoplasmic reticulum kinase (PERK) were also observed in exercised animals. Furthermore, exercise decreased the expression of the proapoptotic protein (PARP1) and increased anti-inflammatory (IL-10) and antiapoptotic (Bcl2) proteins. Using the TUNEL technique, we observed that the exercised animals had lower DNA fragmentation. Finally, physical exercise preserved pro-opiomelanocortin messenger RNA content. In conclusion, exercise training was able to reorganize the control of the energy balance through anti-inflammatory and antiapoptotic responses in hypothalamic tissue of obese mice.     

Effects of intracerebroventricular leptin administration on feeding and sexual behaviors in lean and obese female Zucker rats

Obese Zucker rats (fa/fa) are characterized by inadequate leptin signaling caused by a mutation in the leptin receptor gene. Obese Zucker females are infertile and hyporesponsive to the inductive effects of ovarian hormones on sexual behaviors. Leptin treatment reverses aspects of reproductive dysfunction due to perturbations in energy balance in other animal models. Our first experiment tested the hypothesis that intracerebroventricular (icv) leptin administration would enhance the display of sexual behaviors in obese Zucker females. A second experiment compared lean and obese Zucker females' responses to leptin, during fed and fasted conditions. Ovariectomized (OVX) Zucker rats were implanted with lateral ventricular cannulae. In Experiment 1, fasted, obese females received estradiol benzoate, progesterone, and icv injections of 3, 18, or 36 microg murine leptin or vehicle. Leptin administration reduced food intake, but did not enhance sexual behaviors. In Experiment 2, steroid-replaced, OVX lean and obese females (from a different source than those in Experiment 1) received icv injections of vehicle or 3 or 36 microg leptin under fed and fasted conditions. Leptin treatment reduced food intake and weight gain in the fed, but not the fasted, condition in both genotypes. Sexual receptivity and locomotion were not affected, but icv leptin injections reduced proceptive behaviors in ad libitum-fed rats. These data confirm previous reports that centrally administered leptin decreases food intake and weight gain in obese Zucker rats; results from Experiment 2 suggest that lean and obese females are similarly responsive to these actions of leptin. Contrary to our hypothesis, leptin treatment did not stimulate sexual behaviors; rather, the hormone appears to inhibit the display of sexual proceptivity in ad libitum-fed lean and obese Zucker female rats.     

The rheumatoid arthritis drug auranofin lowers leptin levels and exerts antidiabetic effects in obese mice

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A truncating mutation of Alms1 reduces the number of hypothalamic neuronal cilia in obese mice

Primary cilia are ubiquitous cellular antennae whose dysfunction collectively causes various disorders, including vision and hearing impairment, as well as renal, skeletal, and central nervous system anomalies. One ciliopathy, Alström syndrome, is closely related to Bardet–Biedl syndrome (BBS), sharing amongst other phenotypic features morbid obesity. As the cellular and molecular links between weight regulation and cilia are poorly understood, we used the obese mouse strain foz/foz, bearing a truncating mutation in the Alström syndrome protein (Alms1), to help elucidate why it develops hyperphagia, leading to early onset obesity and metabolic anomalies. Our in vivo studies reveal that Alms1 localizes at the base of cilia in hypothalamic neurons, which are implicated in the control of satiety. Alms1 is lost from this location in foz/foz mice, coinciding with a strong postnatal reduction (～70%) in neurons displaying cilia marked with adenylyl cyclase 3 (AC3), a signaling protein implicated in obesity. Notably, the reduction in AC3‐bearing cilia parallels the decrease in cilia containing two appetite‐regulating proteins, Mchr1 and Sstr3, as well as another established Arl13b ciliary marker, consistent with progressive loss of cilia during development. Together, our results suggest that Alms1 maintains the function of neuronal cilia implicated in weight regulation by influencing the maintenance and/or stability of the organelle. Given that Mchr1 and Sstr3 localization to remaining cilia is maintained in foz/foz animals but known to be lost from BBS knockout mice, our findings suggest different molecular etiologies for the satiety defects associated with the Alström syndrome and BBS ciliopathies. © 2012 Wiley Periodicals, Inc. Develop Neurobiol, 2013     

Neuronal nitric oxide synthase and calbindin delineate sex differences in the developing hypothalamus and preoptic area

Throughout the hypothalamus there are several regions known to contain sex differences in specific cellular, neurochemical, or cell grouping characteristics. The current study examined the potential origin of sex differences in calbindin expression in the preoptic area and hypothalamus as related to sources of nitric oxide. Specific cell populations were defined by immunoreactive (ir) calbindin and neuronal nitric oxide synthase (nNOS) in the preoptic area/anterior hypothalamus (POA/AH), anteroventral periventricular nucleus (AVPv), and ventromedial nucleus of the hypothalamus (VMN). The POA/AH of adult mice was characterized by a striking sex difference in the distribution of cells with ir-calbindin. Examination of the POA/AH of androgen receptor deficient Tfm mice suggests that this pattern was in part androgen receptor dependent, since Tfm males had reduced ir-calbindin compared with wild-type males and more similar to wild-type females. At P0 ir-calbindin was more prevalent than in adulthood, with males having significantly more ir-calbindin and nNOS than have females. Cells that contained either ir-calbindin or ir-nNOS in the POA/AH were in adjacent cell groups, suggesting that NO derived from the enzymatic activity of nNOS may influence the development of ir-calbindin cells. In the region of AVPv, at P0, there was a sex difference with males having more ir-nNOS fibers than have females while ir-calbindin was not detected. In the VMN, at P0, ir-nNOS was greater in females than in males, with no significant difference in ir-calbindin. We suggest that NO as an effector molecule and calbindin as a molecular biomarker illuminate key aspects of sexual differentiation in the developing mouse brain.     

Multifaceted origins of sex differences in the brain

Studies of sex differences in the brain range from reductionistic cell and molecular analyses in animal models to functional imaging in awake human subjects, with many other levels in between. Interpretations and conclusions about the importance of particular differences often vary with differing levels of analyses and can lead to discord and dissent. In the past two decades, the range of neurobiological, psychological and psychiatric endpoints found to differ between males and females has expanded beyond reproduction into every aspect of the healthy and diseased brain, and thereby demands our attention. A greater understanding of all aspects of neural functioning will only be achieved by incorporating sex as a biological variable. The goal of this review is to highlight the current state of the art of the discipline of sex differences research with an emphasis on the brain and to contextualize the articles appearing in the accompanying special issue.     

Oxidative status and serum leptin levels in obese prepubertal children

The prevalence of obesity in children has increased dramatically over the last 20-30 years in developed countries. The aim of this study was to evaluate the oxidative and antioxidant status and any correlation with leptin in obese prepubertal children. A cross-sectional study was made of healthy children from ten elementary schools in the province of Elazig, Eastern Turkey. Blood samples were drawn from children comprising obese and control groups, on a visit to their school in the morning after an overnight fast. The mean body mass index (BMI) was 24.03 +/- 4.09 kg/m(2) in the obese group and was 17.51 +/- 2.33 kg/m(2) in the control group. Mean plasma leptin concentration was significantly higher in the obese children. Homocysteine and malondialdehyde (MDA) levels were also significantly higher in the obese group. In contrast superoxide dismutase (SOD) and glutathione peroxidase activities were significantly decreased in the obese group (p < 0.001). In conclusion, in prepubertal obese children oxidative stress was increased and MDA and homocysteine levels were well correlated with serum leptin level and BMI. In contrast with the increase in oxidative stress, antioxidant activities of SOD and glutathione peroxidase were decreased in obese prepubertal children.     

Cachexia in MAC16 adenocarcinoma: suppression of hunger despite normal regulation of leptin, insulin and hypothalamic neuropeptide Y

Weight loss normally stimulates hunger, through mechanisms that include falls in circulating leptin and insulin, leading to stimulation of hypothalamic neuropeptide Y (NPY). Here, we investigated the leptin, insulin and NPY to clarify why hunger is suppressed in mice with severe cachexia due to the MAC16 adenocarcinoma. MAC16-bearing mice progressively lost weight (19% below controls) and fat (- 61%) over 16 days after tumour transplantation, while total food intake fell by 10%. Pair-fed mice showed less wasting, with final weight being 9% and fat mass 25% below controls. Plasma leptin fell by 85% in MAC16 and 51% in pair-fed mice, in proportion to loss of fat. Plasma insulin was also reduced by 49% in MAC16 and 53% in pair-fed groups. Hypothalamic leptin receptor (OB-Rb) mRNA was significantly increased in both MAC16 (+ 223%) and pair-fed (+192%) mice. Hypothalamic NPY mRNA was also significantly raised in MAC16 (+152%) and pair-fed (+ 99%) groups, showing negative correlations with plasma leptin and insulin, and a positive association with OB-Rb mRNA. In MAC16-induced cachexia, leptin production and hypothalamic OB-Rb and NPY expression are regulated appropriately in response to fat depletion. Therefore, suppression of hunger is probably due to tumour products that inhibit NPY transport or release, or that interfere with neuronal targets downstream of NPY.     

Cardiac proteomics reveals sex chromosome-dependent differences between males and females that arise prior to gonad formation

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Prepubertal castration eliminates sex differences in lifespan and growth trajectories in genetically heterogeneous mice

Sex differences in aging and longevity have been widely observed, with females consistently outliving males across human populations. However, the mechanisms driving these disparities remain poorly understood. In this study, we explored the influence of post-pubertal testicular effects on sex differences in aging by prepubertally castrating genetically heterogeneous (UM-HET3) mice, a unique mouse model that emulates human sex differences in age-related mortality. Prepubertal castration eliminated the longevity disparity between sexes by reducing the elevated early- to mid-life mortality rate observed in males and extending their median lifespan to match that of females. Additionally, castration extended the duration of body weight growth and attenuated the inverse correlation between early-age body weight and lifespan in males, aligning their growth trajectories with those of females. Our findings suggest that post-pubertal testicular actions in genetically diverse mice are primarily responsible for sex differences in longevity as well as growth trajectories. These findings offer a foundation for further investigation into the fundamental mechanisms driving sex-specific aging patterns and the development of potential pro-longevity interventions.     

Spatial learning in deer mice: sex differences and the effects of endogenous opioids and 60 Hz magnetic fields

We examined the effects of brief exposure to weak 60 Hz extremely low frequency (ELF) magnetic fields and opioid systems on spatial behavior and learning in reproductive adult male and female deer mice, Peromyscus maniculatus. Sex differences were evident in spatial performance, with male deer mice displaying significantly better performance than female mice in the Morris water maze, whereby animals had to acquire and retain the location of a submerged hidden platform. Brief (maximum 5 min) exposure to weak (100 T) 60 Hz magnetic fields during task acquisition significantly improved female performance, eliminating the sex differences in acquisition. The opiate antagonist, naltrexone, also improved female acquisition, though significantly less than the magnetic fields. These facilitatory effects involved alterations of non-spatial (task familiarization and reduction of related anxiety/aversive related behaviors) and possibly spatial aspects of the task. Enhancement of enkephalin activity with the enkephalinase inhibitor, SCH 34826, significantly reduced task performance by male deer mice. Both naltrexone and the 60 Hz magnetic fields attenuated the enkephalin mediated reductions of spatial performance. These findings indicate that brief exposure to 60 Hz magnetic fields can enhance water maze task acquisition by deer mice and suggest that these facilitatory effects on spatial performance involve alterations in opioid activity.Abbreviations ELF extremely low frequency - Hz hertz