Endogenous NUCB2/Nesfatin-1 Regulates Energy Homeostasis Under Physiological Conditions in Male Rats.

Nesfatin-1 is the proteolytic cleavage product of Nucleobindin 2, which is expressed both in a number of brain nuclei (e. g., the paraventricular nucleus of the hypothalamus) and peripheral tissues. While Nucleobindin 2 acts as a calcium binding protein, nesfatin-1 was shown to affect energy homeostasis upon central nervous administration by decreasing food intake and increasing thermogenesis. In turn, Nucleobindin 2 mRNA expression is downregulated in starvation and upregulated in the satiated state. Still, knowledge about the physiological role of endogenous Nucleobindin 2/nesfatin-1 in the control of energy homeostasis is limited and since its receptor has not yet been identified, rendering pharmacological blockade impossible. To overcome this obstacle, we tested and successfully established an antibody-based experimental model to antagonize the action of nesfatin-1. This model was then employed to investigate the physiological role of endogenous Nucleobindin 2/nesfatin-1. To this end, we applied nesfatin-1 antibody into the paraventricular nucleus of satiated rats to antagonize the presumably high endogenous Nucleobindin 2/nesfatin-1 levels in this feeding condition. In these animals, nesfatin-1 antibody administration led to a significant decrease in thermogenesis, demonstrating the important role of endogenous Nucleobindin 2/nesfatin-1in the regulation of energy expenditure. Additionally, food and water intake were significantly increased, confirming and complementing previous findings. Moreover, neuropeptide Y was identified as a major downstream target of endogenous Nucleobindin 2/nesfatin-1.

[Self-determination despite dementia! Participant protection instead of incapacitation in research : Report on the 5th Ethics Conference of the German Center for Neurodegenerative Diseases]. / Selbstbestimmung trotz Demenz! Probandenschutz statt Entmündigung in der Forschung : Tagungsbericht zur 5. Ethiktagung des Deutschen Zentrums für Neurodegenerative Erkrankungen (DZNE) Rostock/Greifswald.

In March 2019 the Ethics Conference of the German Center for Neurodegenerative Diseases (DZNE) was held for the fifth time. It was organized by the DZNE Rostock/Greifswald site and chaired by Prof. Wolfgang Hoffmann. The conference provided scientists, physicians, representatives of the German Alzheimer Society, (informal) caregivers of people with dementia (PwD), and other interested people with the opportunity to talk about the opportunities and limitations of research on and for PwD.Nationally and internationally recognized experts on healthcare services research, clinical research, nursing research, (geriatric) psychiatry, interdisciplinary ageing research, economic law, and psychotherapy discussed the pros and cons of a multitude of topics like self-determination, research participant decree, informed consent, and participation of PwD in research. The aim of the event was to reconcile the view of practice pleading for an ethically correct, human treatment of PwD and respecting their autonomy with participation in (clinical) studies. Experts controversially discussed and consolidated different points of view of practice and research.

Deficiency of the immunoproteasome subunit ß5i/LMP7 supports the anxiogenic effects of mild stress and facilitates cued fear memory in mice.

Proteolysis as mediated by one of the major cellular protein degradation pathways, the ubiquitin-proteasome system (UPS), plays an essential role in learning and memory formation. However, the functional relevance of immunoproteasomes in the healthy brain and especially their impact on normal brain function including processes of learning and memory has not been investigated so far. In the present study, we analyzed the phenotypic effects of an impaired immunoproteasome formation using a ß5i/LMP7-deficient mouse model in different behavioral paradigms focusing on locomotor activity, exploratory behavior, innate anxiety, startle response, prepulse inhibition, as well as fear and safety conditioning. Overall, our results demonstrate no strong effects of constitutive ß5i/LMP7-deficiency on gross locomotor abilities and anxiety-related behavior in general. However, ß5i/LMP7-deficient mice expressed more anxiety after mild stress and increased cued fear after fear conditioning. These findings indicate that the basal proper formation of immunoproteasomes and/or at least the expression of ß5i/LMP7 in healthy mice seem to be involved in the regulation of anxiety and cued fear levels.

Predator odour but not TMT induces 22-kHz ultrasonic vocalizations in rats that lead to defensive behaviours in conspecifics upon replay.

Predator odours induce defensive behaviour in prey animals such as rats. The present study investigated (1) whether laboratory rats exposed to predator odours emit 22-kHz calls which may have an alarming function and (2) whether playback of such calls induces behavioural changes in conspecifics. For this, Sprague-Dawley rats were exposed to samples of fox and lion urine, as well as to the synthetic predator odour TMT. Despite that all odours induced defensive behaviour, only predator urine samples but not TMT were able to induce 22-kHz calls in a few rats. In a second experiment, naive rats were exposed to playback presentations of the 22-kHz calls recorded in the first experiment, as well as to phase-scrambled and frequency-shifted control stimuli. Low intensity playback presentations led to a reduction of locomotor activity during the presentation of the 22-kHz calls but not of the control stimuli. This effect was less specific under high intensity conditions. Taken together the present findings show that natural predator odours are able to induce emission of 22-kHz calls in rats and support the hypothesis that these calls have an alarming function.

Predator odor induced defensive behavior in wild and laboratory rats: A comparative study.

Laboratory rats are frequently used as animal models in research. Since the 1920s rats are bred and reared in laboratories which affects anatomy, physiology, and behavior responses. In the present study we exposed laboratory and wild rats to predator odor and comparatively analyzed their behavioral and physiological responses. In detail, Warsaw Wild Captive Pisula Stryjek (WWCPS) rats and Lister Hooded (LH) rats were exposed to the predator odor 2,3,5-trimethyl-3-thiazoline (TMT), their behavior was videotaped and blood samples were collected for subsequent serum corticosterone analysis. In both rat stocks, exposure to TMT induced avoidance behavior and increased freezing behavior. Notably, the increase in freezing was based on an increase number of freezing events in LH rats whereas WWCPS rats prolonged the mean duration of the single freezing events. Interestingly, TMT exposure lead to a serum corticosterone increase in WWCPS rats but not in LH rats. Furthermore, WWCPS rats generally expressed decreased but faster locomotor activity, as well as more grooming behavior than LH rats. Taken together, these data indicate differences in behavioral and physiological defensive responses to predator odors in the two rat stocks.

Habenula and interpeduncular nucleus differentially modulate predator odor-induced innate fear behavior in rats.

Fear is an important behavioral system helping humans and animals to survive potentially dangerous situations. Fear can be innate or learned. Whereas the neural circuits underlying learned fear are already well investigated, the knowledge about the circuits mediating innate fear is still limited. We here used a novel, unbiased approach to image in vivo the spatial patterns of neural activity in odor-induced innate fear behavior in rats. We intravenously injected awake unrestrained rats with a 99m-technetium labeled blood flow tracer (99mTc-HMPAO) during ongoing exposure to fox urine or water as control, and mapped the brain distribution of the trapped tracer using single-photon emission computed tomography (SPECT). Upon fox urine exposure blood flow increased in a number of brain regions previously associated with odor-induced innate fear such as the amygdala, ventromedial hypothalamus and dorsolateral periaqueductal grey, but, unexpectedly, decreased at higher significance levels in the interpeduncular nucleus (IPN). Significant flow changes were found in regions monosynaptically connected to the IPN. Flow decreased in the dorsal tegmentum and entorhinal cortex. Flow increased in the habenula (Hb) and correlated with odor effects on behavioral defensive strategy. Hb lesions reduced avoidance of but increased approach to the fox urine while IPN lesions only reduced avoidance behavior without approach behavior. Our study identifies a new component, the IPN, of the neural circuit mediating odor-induced innate fear behavior in mammals and suggests that the evolutionarily conserved Hb-IPN system, which has recently been implicated in cued fear, also forms an integral part of the innate fear circuitry.

Predator odor exposure increases food-carrying behavior in rats.

To cover their energy demands, prey animals are forced to search for food. However, during foraging they also expose themselves to the risk of becoming the prey of predators. Consequently, in order to increase their fitness foraging animals have to trade-off efficiency of foraging against the avoidance of predation risk. For example, the decision on whether a found food piece should be eaten at the food source or whether it should be carried to a protective site such as the nest (food-carrying behavior), is strongly dependent on different incentive factors (e.g., hunger level, food size, distance to the nest). It has been shown that food-carrying behavior increases the more risky the foraging situation becomes. Since predator odors are clearly fear-inducing in rats, we ask here whether the detection of predator odors in close proximity to the food source modulates food-carrying behavior. In the present study, the food-carrying behavior of rats for six different food pellet sizes was measured in a "low risk" and a "high risk" testing condition by presenting water or a fox urine sample, respectively, next to the food source. For both testing conditions, food-carrying behavior of rats increased with increasing food pellet weight. Importantly, the proportion of food-carrying rats was significantly higher during exposure to fox urine ("high risk") than when rats were tested with the water control ("low risk"). Taken together, these results demonstrate that food-carrying behavior of rats is increased by the detection of a predator odor. Our data also support the idea that such food-carrying behavior can be considered as a pre-encounter defensive response.

The olfactory hole-board test in rats: a new paradigm to study aversion and preferences to odors.

Odors of biological relevance (e.g., predator odors, sex odors) are known to effectively influence basic survival needs of rodents such as anti-predatory defensiveness and mating behaviors. Research focused on the effects of these odors on rats' behavior mostly includes multi-trial paradigms where animals experience single odor exposures in subsequent, separated experimental sessions. In the present study, we introduce a modification of the olfactory hole-board test that allows studying the effects of different odors on rats' behavior within single trials. First, we demonstrated that the corner holes of the hole-board were preferentially visited by rats. The placement of different odors under the corner holes changed this hole preference. We showed that holes with carnivore urine samples were avoided, while corner holes with female rat urine samples were preferred. Furthermore, corner holes with urine samples from a carnivore, herbivore, and omnivore were differentially visited indicating that rats can discriminate these odors. To test whether anxiolytic treatment specifically modulates the avoidance of carnivore urine holes, we treated rats with buspirone. Buspirone treatment completely abolished the avoidance of carnivore urine holes. Taken together, our findings indicate that the olfactory hole-board test is a valuable tool for measuring avoidance and preference responses to biologically relevant odors.

Nesfatin-1 increases energy expenditure and reduces food intake in rats.

OBJECTIVE: Energy homeostasis results from a balance of food intake and energy expenditure, accomplished by the interaction of peripheral and central nervous signals. The recently discovered adipokine nesfatin-1 is involved in the central control of food intake, but whether it also participates in the regulation of thermogenesis is unknown. METHODS: Nesfatin-1 was administered intracerebroventricularly to freely moving, male Wistar rats and direct calorimetry was performed to assess its effects on thermogenesis. Furthermore, food intake was measured and hypothalamic and N. tractus solitarius (NTS) neuropeptide expression was determined by quantitative real-time polymerace chain reaction. Leptin, which is involved in both the regulation of food intake and thermogenesis, was used as positive control. RESULTS: For the first time it was shown that central nervous administration of nesfatin-1 profoundly increases thermogenesis in rats to a similar extent as leptin and the role of both peptides in the control of food intake was confirmed. Nesfatin-1 significantly downregulated neuropeptide Y (NPY) mRNA expression in both hypothalamus and NTS. CONCLUSIONS: The results strongly support the prominent role of nesfatin-1 for both energy expenditure and food intake and NPY neurons appear to be involved in this effect.