Extrahypothalamic Brain Research Articles

Fasting-induced activity changes in MC3R neurons of the paraventricular nucleus of the thalamus.

The brain controls energy homeostasis by regulating food intake through signaling within the melanocortin system. Whilst we understand the role of the hypothalamus within this system, how extra-hypothalamic brain regions are involved in controlling energy balance remains unclear. Here we show that the melanocortin 3 receptor (MC3R) is expressed in the paraventricular nucleus of the thalamus (PVT). We tested whether fasting would change the activity of MC3R neurons in this region by assessing the levels of c-Fos and pCREB as neuronal activity markers. We determined that overnight fasting causes a significant reduction in pCREB levels within PVT-MC3R neurons. We then questioned whether perturbation of MC3R signaling, during fasting, would result in altered refeeding. Using chemogenetic approaches, we show that modulation of MC3R activity, during the fasting period, does not impact body weight regain or total food intake in the refeeding period. However, we did observe significant differences in the pattern of feeding-related behavior. These findings suggest that the PVT is a region where MC3R neurons respond to energy deprivation and modulate refeeding behavior.

A comprehensive chemotyping and gonadal regulation of seven kisspeptinergic neuronal populations in the mouse brain.

Herein, we present a systematic analysis, using dual and multiplex RNAscope methods, of seven kisspeptinergic neuronal populations, based on their chemotyping and distribution throughout the mouse brain. The co-expression of mRNAs coding for neuropeptides, for excitatory and inhibitory transmitter vesicular transporters, and for sex steroid receptors are described in four hypothalamic and three extra-hypothalamic nuclei. These include a newly characterized kisspeptin-expressing ventral premammillary nucleus cell group co-expressing vesicular glutamate transporter 2, pituitary adenylate cyclase-activating polypeptide and neurotensin mRNAs. Kisspeptin mRNA ( Kiss1) was observed within both somatic and dendritic compartments at a single-cell level in two hypothalamic sites, a prominent and previously undescribed feature of kisspeptin neurons in these two cell groups. Patterns of altered Kiss1 expression following gonadectomy among these seven KP populations suggest that androgen receptor signaling may also play a previously unremarked role in gonadal feedback regulation of kisspeptinergic neuronal function. Data from this study provide a chemoanatomical basis for hypothesis generation regarding the functional diversity of kisspeptinergic signaling in hypothalamic and extrahypothalamic brain.

Prenatal Hypoxia Triggers a Glucocorticoid-Associated Depressive-like Phenotype in Adult Rats, Accompanied by Reduced Anxiety in Response to Stress.

Fetal hypoxia and maternal stress frequently culminate in neuropsychiatric afflictions in life. To replicate this condition, we employed a model of prenatal severe hypoxia (PSH) during days 14-16 of rat gestation. Subsequently, both control and PSH rats at 3 months old were subjected to episodes of inescapable stress to induce learned helplessness (LH). The results of the open field test revealed an inclination towards depressive-like behavior in PSH rats. Following LH episodes, control (but not PSH) rats displayed significant anxiety. LH induced an increase in glucocorticoid receptor (GR) levels in extrahypothalamic brain structures, with enhanced nuclear translocation in the hippocampus (HPC) observed both in control and PSH rats. However, only control rats showed an increase in GR nuclear translocation in the amygdala (AMG). The decreased GR levels in the HPC of PSH rats correlated with elevated levels of hypothalamic corticotropin-releasing hormone (CRH) compared with the controls. However, LH resulted in a reduction of the CRH levels in PSH rats, aligning them with those of control rats, without affecting the latter. This study presents evidence that PSH leads to depressive-like behavior in rats, associated with alterations in the glucocorticoid system. Notably, these impairments also contribute to increased resistance to severe stressors.

Anorexia-Induced Hypoleptinemia Drives Adaptations in the JAK2/STAT3 Pathway in the Ventral and Dorsal Hippocampus of Female Rats.

Leptin is an appetite-regulating adipokine that is reduced in patients with anorexia nervosa (AN), a psychiatric disorder characterized by self-imposed starvation, and has been linked to hyperactivity, a hallmark of AN. However, it remains unknown how leptin receptor (LepR) and its JAK2-STAT3 downstream pathway in extrahypothalamic brain areas, such as the dorsal (dHip) and ventral (vHip) hippocampus, crucial for spatial memory and emotion regulation, may contribute to the maintenance of AN behaviors. Taking advantage of the activity-based anorexia (ABA) model (i.e., the combination of food restriction and physical activity), we observed reduced leptin plasma levels in adolescent female ABA rats at the acute phase of the disorder [post-natal day (PND) 42], while the levels increased over control levels following a 7-day recovery period (PND49). The analysis of the intracellular leptin pathway revealed that ABA rats showed an overall decrease of the LepR/JAK2/STAT3 signaling in dHip at both time points, while in vHip we observed a transition from hypo- (PND42) to hyperactivation (PND49) of the pathway. These changes might add knowledge on starvation-induced fluctuations in leptin levels and in hippocampal leptin signaling as initial drivers of the transition from adaptative mechanisms to starvation toward the maintenance of aberrant behaviors typical of AN patients, such as perpetuating restraint over eating.

Novel insights into minipuberty and GnRH: Implications on neurodevelopment, cognition, and COVID-19 therapeutics.

In humans, the first 1000 days of life are pivotal for brain and organism development. Shortly after birth, gonadotropin-releasing hormone (GnRH) neurons in the hypothalamus are activated, a phenomenon known as minipuberty. This phenomenon, observed in all mammals studied, influences the postnatal development of the hypothalamic-pituitary-gonadal (HPG) axis and reproductive function. This review will put into perspective the results of recent studies showing that the impact of minipuberty extends beyond reproductive function, influencing sensory and cognitive maturation. Studies in mice have revealed the role of nitric oxide (NO) in regulating minipuberty amplitude, with NO deficiency linked to cognitive and olfactory deficits. Additionally, findings indicate that cognitive and sensory defects in adulthood in a mouse model of Down syndrome are associated with an age-dependent decline of GnRH production, whose origin can be traced back to minipuberty, and point to the potential therapeutic role of pulsatile GnRH administration in cognitive disorders. Furthermore, this review delves into the repercussions of COVID-19 on GnRH production, emphasizing potential consequences for neurodevelopment and cognitive function in infected individuals. Notably, GnRH neurons appear susceptible to SARS-CoV-2 infection, raising concerns about potential long-term effects on brain development and function. In conclusion, the intricate interplay between GnRH neurons, GnRH release, and the activity of various extrahypothalamic brain circuits reveals an unexpected role for these neuroendocrine neurons in the development and maintenance of sensory and cognitive functions, supplementing their established function in reproduction. Therapeutic interventions targeting the HPG axis, such as inhaled NO therapy in infancy and pulsatile GnRH administration in adults, emerge as promising approaches for addressing neurodevelopmental cognitive disorders and pathological aging.

Cholinergic Basal Forebrain Connectivity to the Basolateral Amygdala Modulates Food Intake.

Obesity results from excessive caloric input associated with overeating and presents a major public health challenge. The hypothalamus has received significant attention for its role in governing feeding behavior and body weight homeostasis. However, extrahypothalamic brain circuits also regulate appetite and consumption by altering sensory perception, motivation, and reward. We recently discovered a population of basal forebrain cholinergic (BFc) neurons that regulate appetite suppression. Through viral tracing methods in the mouse model, we found that BFc neurons densely innervate the basolateral amygdala (BLA), a limbic structure involved in motivated behaviors. Using channelrhodopsin-assisted circuit mapping, we identified cholinergic responses in BLA neurons following BFc circuit manipulations. Furthermore, in vivo acetylcholine sensor and genetically encoded calcium indicator imaging within the BLA (using GACh3 and GCaMP, respectively) revealed selective response patterns of activity during feeding. Finally, through optogenetic manipulations in vivo, we found that increased cholinergic signaling from the BFc to the BLA suppresses appetite and food intake. Together, these data support a model in which cholinergic signaling from the BFc to the BLA directly influences appetite and feeding behavior.

OR14-04 Musashi-1 Is A Cell-type Specific Regulator Of Type 2 Deiodinase-mediated Thyroid Hormone Activation

Abstract Disclosure: P. Mohacsik: None. E. Halmos: None. Y. Ruska: None. C. Fekete: None. B. Gereben: None. Thyroid hormone (TH) exerts a profound effect on the regulation of energy homeostasis, metabolism, brain development and the determination of cell fate in various tissues. Type 2 deiodinase enzyme (D2) is the crucial regulator of TH activation by converting the prohormone thyroxine to T3 that can bind the TH receptor. This represents a fundamental step of cell-type specific TH action. D2 regulation is tightly controlled at various molecular levels to allow quick adaptation of TH action to specific challenges. Within the brain, D2 is expressed in hypothalamic tanycytes that play a crucial role in the regulation of the hypothalamo-pituitary-thyroid (HPT) axis and in astrocytes of extrahypothalamic brain regions. Under changing TH economy, D2 activity undergoes homeostatic alteration in astrocytes while remains largely unaffected in hypothalamic tanycytes. Our aim was to identify regulators that could underlie this phenomenon as a cell-type specific regulator of D2-dependent TH action. Musashi (Msi)-1 is a highly conserved RNA-binding protein that regulates cell cycle and play a crucial role during brain development. We found, that Msi-1 was co-expressed with D2 both in hypothalamic tanycytes and cortical astrocytes in mice and T3 treatment (24h T3 1µg/bwg, i.p.) increased Msi-1 protein level in both cell types indicating its TH-dependent regulation. Then we identified multiple potential Msi-1 binding sites in the extremely long 3’UTR both of the human, mouse, rat and chicken D2 mRNA and used RNA electromobility shift assay that revealed direct binding of Msi-1 to the 3’UTR of the D2 mRNA. Studies on the impact of Msi-1 level on D2 activity showed, that Msi-1 overexpression decreased D2 activity in HEK293 cells while this effect was abolished by the deletion of a 3 kb region of the 3’UTR of D2 containing the Msi-1 binding sites. Furthermore, Msi-1 knock down by shRNA in H4 glioma cells increased endogenous D2 activity. A mouse model, lacking Msi-1 protein exclusively in the brain and pituitary, showed increased D2 activity in the cortex accompanied with a slightly decreased serum fT4 and decreased liver dio1 expression. Increased cortical D2 activity was not a consequence of the mildly hypothyroid periphery since cortical expression of dio3, the T3-regulated T3-degrading enzyme remained unchanged along with enpp2, another T3-regulated gene. In contrast, D2 activity remained unchanged in hypothalamic tanycytes. Our results identify Msi-1 as a TH-dependent cell-type specific regulator of D2 activity in the mouse brain that acts via direct binding to the 3’UTR of the D2 mRNA. In summary, Msi-1 regulates D2 activity cell type specifically and may contribute to the homeostatic regulation of cortical D2 while at the same time allows TH-resistant D2 function in tanycytes that is required by proper HPT function. Presentation: Friday, June 16, 2023

Disturbed circadian rhythm and retinal degeneration in a mouse model of Alzheimer’s disease

The circadian clock is synchronized to the 24 h day by environmental light which is transmitted from the retina to the suprachiasmatic nucleus (SCN) primarily via the retinohypothalamic tract (RHT). Circadian rhythm abnormalities have been reported in neurodegenerative disorders such as Alzheimer's disease (AD). Whether these AD-related changes are a result of the altered clock gene expression, retina degeneration, including the dysfunction in RHT transmission, loss of retinal ganglion cells and its electrophysiological capabilities, or a combination of all of these pathological mechanisms, is not known. Here, we evaluated transgenic APP/PS1 mouse model of AD and wild-type mice at 6- and 12-month-old, as early and late pathological stage, respectively. We noticed the alteration of circadian clock gene expression not only in the hypothalamus but also in two extra-hypothalamic brain regions, cerebral cortex and hippocampus, in APP/PS1 mice. These alterations were observed in 6-month-old transgenic mice and were exacerbated at 12 months of age. This could be explained by the reduced RHT projections in the SCN of APP/PS1 mice, correlating with downregulation of hypothalamic GABAergic response in APP/PS1 mice in advanced stage of pathology. Importantly, we also report retinal degeneration in APP/PS1 mice, including Aβ deposits and reduced choline acetyltransferase levels, loss of melanopsin retinal ganglion cells and functional integrity mainly of inner retina layers. Our findings support the theory that retinal degeneration constitutes an early pathological event that directly affects the control of circadian rhythm in AD.

Prenatal Hypoxia-Induced Adverse Reaction to Mild Stress is Associated with Depressive-Like Changes in the Glucocorticoid System of Rats.

The effects of prenatal hypoxia on neurodevelopment are predominantly associated with impaired maternal glucocorticoid stimulation of the fetus, which is "imprinted" in altered sensitivity of glucocorticoid reception in brain structures of offspring and can affect brain plasticity during lifespan. This study aimed to investigate response of the brain glucocorticoid system to mild stress (MS) in adult rats that survived prenatal severe hypoxia (PSH) on embryonic days 14-16. In response to MS the control (but not PSH) rats demonstrate increased corticosterone levels, a decrease in exploratory activity and increased anxiety. In the raphe nuclei of adult PSH rats the expression of glucocorticoid receptors (GR) is increased without changes in serotonin levels in comparison with the control. MS induces a decrease in GR expression accompanied by up-regulation of tryptophan hydroxylase 2 (tph2) and down-regulation of monoamine oxidase A (maoa) transcription in the raphe nuclei of both control and PSH groups. PSH also causes significant deviations in GR expression and GR-dependent transcription in the hippocampus, the medial prefrontal cortex, but not in the amygdala of rats. However, in response to MS, PSH rats demonstrate mild changes in their activity, while in control animals the MS-induced activity of the glucocorticoid system in these brain structures is similar to intact PSH animals. Impaired activity of the glucocorticoid system in the extrahypothalamic brain structures of PSH rats is accompanied by increase in the hypothalamic corticotropin-releasing hormone (CRH) levels in comparison with the control regardless of MS. Synthesis of proopiomelanocortin (POMC) and release of adrenocorticotropic hormone (ACTH) into the blood are decreased in response to MS in the pituitary of control rats, which demonstrates a negative glucocorticoid feedback mechanism. Meanwhile, in the pituitary of PSH rats reduced POMC levels were found regardless of MS. Thus, prenatal hypoxia causes depression-like patterns in the brain glucocorticoid system with adverse reaction to mild stressors.

Mapping key neuropeptides involved in the melanocortin system in Atlantic salmon (Salmo salar) brain.

The melanocortin system is a key regulator of appetite and food intake in vertebrates. This system includes the neuropeptides neuropeptide y (NPY), agouti-related peptide (AGRP), cocaine- and amphetamine-regulated transcript (CART), and pro-opiomelanocortin (POMC). An important center for appetite control in mammals is the hypothalamic arcuate nucleus, with neurons that coexpress either the orexigenic NPY/AGRP or the anorexigenic CART/POMC neuropeptides. In ray-finned fishes, such a center is less characterized. The Atlantic salmon (Salmo salar) has multiple genes of these neuropeptides due to whole-genome duplication events. To better understand the potential involvement of the melanocortin system in appetite and food intake control, we have mapped the mRNA expression of npy, agrp, cart, and pomc in the brain of Atlantic salmon parr using in situ hybridization. After identifying hypothalamic mRNA expression, we investigated the possible intracellular coexpression of npy/agrp and cart/pomc in the tuberal hypothalamus by fluorescent in situ hybridization. The results showed that the neuropeptides were widely distributed, especially in sensory and neuroendocrine brain regions. In the hypothalamic lateral tuberal nucleus, the putative homolog to the mammalian arcuate nucleus, npya, agrp1, cart2b, and pomca were predominantly localized in distinct neurons; however, some neurons coexpressed cart2b/pomca. This is the first demonstration of coexpression of cart2b/pomca in the tuberal hypothalamus of a teleost. Collectively, our data suggest that the lateral tuberal nucleus is the center for appetite control in salmon, similar to that of mammals. Extrahypothalamic brain regions might also be involved in regulating food intake, including the olfactory bulb, telencephalon, midbrain, and hindbrain.

Current Perspectives on Kisspeptins Role in Behaviour.

The neuropeptide kisspeptin is now well-established as the master regulator of the mammalian reproductive axis. Beyond the hypothalamus, kisspeptin and its cognate receptor are also extensively distributed in extra-hypothalamic brain regions. An expanding pool of animal and human data demonstrates that kisspeptin sits within an extensive neuroanatomical and functional framework through which it can integrate a range of internal and external cues with appropriate neuroendocrine and behavioural responses. In keeping with this, recent studies reveal wide-reaching effects of kisspeptin on key behaviours such as olfactory-mediated partner preference, sexual motivation, copulatory behaviour, bonding, mood, and emotions. In this review, we provide a comprehensive update on the current animal and human literature highlighting the far-reaching behaviour and mood-altering roles of kisspeptin. A comprehensive understanding of this important area in kisspeptin biology is key to the escalating development of kisspeptin-based therapies for common reproductive and related psychological and psychosexual disorders.

The Role of Glial Cells in Regulating Feeding Behavior: Potential Relevance to Anorexia Nervosa.

Eating behavior is controlled by hypothalamic circuits in which agouti-related peptide-expressing neurons when activated in the arcuate nucleus, promote food intake while pro-opiomelanocortin-producing neurons promote satiety. The respective neurotransmitters signal to other parts of the hypothalamus such as the paraventricular nucleus as well as several extra-hypothalamic brain regions to orchestrate eating behavior. This complex process of food intake may be influenced by glia cells, in particular astrocytes and microglia. Recent studies showed that GFAP+ astrocyte cell density is reduced in the central nervous system of an experimental anorexia nervosa model. Anorexia nervosa is an eating disorder that causes, among the well-known somatic symptoms, brain volume loss which was associated with neuropsychological deficits while the underlying pathophysiology is unknown. In this review article, we summarize the findings of glia cells in anorexia nervosa animal models and try to deduce which role glia cells might play in the pathophysiology of eating disorders, including anorexia nervosa. A better understanding of glia cell function in the regulation of food intake and eating behavior might lead to the identification of new drug targets.

Erectile Function and Sexual Behavior: A Review of the Role of Nitric Oxide in the Central Nervous System.

Nitric oxide (NO), the neuromodulator/neurotransmitter formed from l-arginine by neuronal, endothelial and inducible NO synthases, is involved in numerous functions across the body, from the control of arterial blood pressure to penile erection, and at central level from energy homeostasis regulation to memory, learning and sexual behavior. The aim of this work is to review earlier studies showing that NO plays a role in erectile function and sexual behavior in the hypothalamus and its paraventricular nucleus and the medial preoptic area, and integrate these findings with those of recent studies on this matter. This revisitation shows that NO influences erectile function and sexual behavior in males and females by acting not only in the paraventricular nucleus and medial preoptic area but also in extrahypothalamic brain areas, often with different mechanisms. Most importantly, since these areas are strictly interconnected with the paraventricular nucleus and medial preoptic area, send to and receive neural projections from the spinal cord, in which sexual communication between brain and genital apparatus takes place, this review reveals that central NO participates in concert with neurotransmitters/neuropeptides to a neural circuit controlling both the consummatory (penile erection, copulation, lordosis) and appetitive components (sexual motivation, arousal, reward) of sexual behavior.

Norepinephrine and Glucocorticoids Modulate Chronic Unpredictable Stress-Induced Increase in the Type 2 CRF and Glucocorticoid Receptors in Brain Structures Related to the HPA Axis Activation.

The stress response is multifactorial and enrolls circuitries to build a coordinated reaction, leading to behavioral, endocrine, and autonomic changes. These changes are mainly related to the hypothalamus-pituitary-adrenal (HPA) axis activation and the organism's integrity. However, when self-regulation is ineffective, stress becomes harmful and predisposes the organism to pathologies. The chronic unpredictable stress (CUS) is a widely used experimental model since it induces physiological and behavioral changes and better mimics the stressors variability encountered in daily life. Corticotropin-releasing factor (CRF) and glucocorticoids (GCs) are deeply implicated in the CUS-induced physiological and behavioral changes. Nonetheless, the CUS modulation of CRF receptors and GR and the norepinephrine role in extra-hypothalamic brain areas were not well explored. Here, we show that 14days of CUS induced a long-lasting HPA axis hyperactivity evidenced by plasmatic corticosterone increase and adrenal gland hypertrophy, which was dependent on both GCs and NE release induced by each stress session. CUS also increased CRF2 mRNA expression and GR protein levels in fundamental brain structures related to HPA regulation and behavior, such as the lateral septal nucleus intermedia part (LSI), ventromedial hypothalamic nucleus (VMH), and central nucleus of the amygdala (CeA). We also showed that NE participates in the CUS-induced increase in CRF2 and GR levels in the LSI, reinforcing the locus coeruleus (LC) involvement in the HPA axis modulation. Despite the CUS-induced molecular changes in essential areas related to anxiety-like behavior, this phenotype was not observed in CUS animals 24h after the last stress session.

The role of the corticotropin-releasing hormone and its receptors in the regulation of stress response.

Stress is an essential part of everyday life. The neuropeptide corticotropin-releasing hormone (CRH, also called CRF and corticoliberin) plays a key role in the integration of neuroendocrine, autonomic and behavioral responses to stress. The activation of the hypothalamic-pituitary-adrenal axis (HPA axis) by neurons of the paraventricular hypothalamic nucleus (PVN), the primary site of synthesis CRH, triggers stress reactions. In addition to the hypothalamus, CRH is widespread in extrahypothalamic brain structures, where it functions as a neuromodulator for coordination and interaction between the humoral and behavioral aspects of a stress response. The axons of neurons expressing CRH are directed to various structures of the brain, where the neuropeptide interacts with specific receptors (CRHR1, CRHR2) and can affect various mediator systems that work together to transmit signals to different brain regions to cause many reactions to stress. Moreover, the effect of stress on brain functions varies from behavioral adaptation to increased survival and increased risk of developing mental disorders. Disturbances of the CRH system regulation are directly related to such disorders: mental pathologies (depression, anxiety, addictions), deviations of neuroendocrinological functions, inflammation, as well as the onset and development of neurodegenerative diseases such as Alzheimer’s disease. In addition, the role of CRH as a regulator of the neurons structure in the areas of the developing and mature brain has been established. To date, studies have been conducted in which CRHR1 is a target for antidepressants, which are, in fact, antagonists of this receptor. In this regard, the study of the participation of the CRH system and its receptors in negative effects on hormone-dependent systems, as well as the possibility of preventing them, is a promising task of modern physiological genetics. In this review, attention will be paid to the role of CRH in the regulation of response to stress, as well as to the involvement of extrahypothalamic CRH in pathophysiology and the correction of mental disorders.

Prenatal Introduction of Dexamethasone Causes Disruption of Glucocorticoid Feedback Associated with a Change in the Amount of Corticosteroid Receptors in Extrahypothalamic Brain Structures of Adult Rats

As are other artificial glucocorticoids, dexamethasone is widely used in everyday obstetric practice. The threat of termination of pregnancy is an indication for the use of glucocorticoids. However, there is evidence that the introduction of glucocorticoids during pregnancy can lead to impaired brain development and behavior of offspring that can be caused by a change in the activity of the hypothalamic–pituitary–adrenocortical system (HPAS) due to past effects. The aim of the present work was to study the effect of the introduction of synthetic hormone dexamethasone at the 14th–16th (DM14–16) and 17th–19th (DM17–19) days of prenatal ontogenesis on the stress reactivity of the HPAS in adult 3-month-old rats, as well as on the level of gluco- (GR) and mineralocorticoid (MR) receptors in the most vulnerable areas of the brain (hippocampus and neocortex). Significant intergroup differences in the dynamics of a change in the level of corticosterone in the blood of adult rats in response to a weak stress effect (rapid stress reactivity test) were detected. Using the immunohistochemical method, changes in the expression (the amount of immunopositive cells) of GR and MR were studied in CA1 field and dentate gyrus (DG) of the hippocampus, as well as in second and fifth neocortex layers in adult male rats. In animals of the DM14–16 group, a significant decrease in the number of cells intensively stained using antibodies to GR in CA1 field, hippocampus DG, and neocortex layer V (to 24.5, 32.4, and 5.5%, respectively) as compared with the control were observed. The introduction of dexamethasone at the 17th–19th day of gestation also led to a decrease in the amount of intensively stained GR-immunopositive cells in the hippocampus CA1 field and neocortex layer V (to 31.9 and 35.7%, respectively), but to a lesser degree than in animals from the group DM14–16. A trend toward a decrease in the number of GR-positive cells is accompanied by an increase in immunoreactivity of the cells to MR. Thus, the introduction of dexamethasone at different periods of prenatal ontogenesis modifies the work of gluco- and mineralocorticoid receptors; however, the degree, localization, and direction of these changes differ depending on the time of the effect.

Chapter 19 - The infundibular peptidergic neurons and glia cells in overeating, obesity, and diabetes

Dysfunctional regulation of energy homeostasis results in increased bodyweight and obesity, eventually leading to type 2 diabetes mellitus. The infundibular nucleus (IFN) of the hypothalamus is the main regulator of energy homeostasis. The peptidergic neurons and glia cells of the IFN receive metabolic cues concerning energy state of the body from the circulation. The IFN can monitor hormones like insulin and leptin and nutrients like glucose and fatty acids. All these metabolic cues are integrated into an output signal regulating energy homeostasis through the release of neuropeptides. These neuropeptides are released in several inter- and extrahypothalamic brain regions involved in regulation of energy homeostasis. This review will give an overview of the peripheral signals involved in the regulation of energy homeostasis, the peptidergic neurons and glial cells of the IFN, and will highlight the main intra-hypothalamic projection sites of the IFN.

Oxytocin Influences Male Sexual Activity via Non-synaptic Axonal Release in the Spinal Cord

Oxytocinergic neurons in the paraventricular nucleus of the hypothalamus that project to extrahypothalamic brain areas and the lumbar spinal cord play an important role in the control of erectile function and male sexual behavior in mammals. The gastrin-releasing peptide (GRP) system in the lumbosacral spinal cord is an important component of the neural circuits that control penile reflexes in rats, circuits that are commonly referred to as the "spinal ejaculation generator (SEG)." We have examined the functional interaction between the SEG neurons and the hypothalamo-spinal oxytocin system in rats. Here, we show that SEG/GRP neurons express oxytocin receptors and are activated by oxytocin during male sexual behavior. Intrathecal injection of oxytocin receptor antagonist not only attenuates ejaculation but also affects pre-ejaculatory behavior during normal sexual activity. Electron microscopy of potassium-stimulated acute slices of the lumbar cord showed that oxytocin-neurophysin-immunoreactivity was detected in large numbers of neurosecretory dense-cored vesicles, many of which are located close to the plasmalemma of axonal varicosities in which no electron-lucent microvesicles or synaptic membrane thickenings were visible. These results suggested that, in rats, release of oxytocin in the lumbar spinal cord is not limited to conventional synapses but occurs by exocytosis of the dense-cored vesicles from axonal varicosities and acts by diffusion-a localized volume transmission-to reach oxytocin receptors on GRP neurons and facilitate male sexual function.

Acute Elevations in Cortisol Increase the In Vivo Binding of [11C]NOP-1A to Nociceptin Receptors: A Novel Imaging Paradigm to Study the Interaction Between Stress- and Antistress-Regulating Neuropeptides

BackgroundAn imbalance between neuropeptides that promote stress and resilience, such as corticotropin-releasing factor and nociceptin, has been postulated to underlie relapse in addiction. The objective of this study was to develop a paradigm to image the in vivo interaction between stress-promoting neuropeptides and nociceptin (NOP) receptors in humans. Methods[11C]NOP-1A positron emission tomography was used to measure the binding to NOP receptors at baseline (BASE) and following an intravenous hydrocortisone challenge (CORT) in 19 healthy control subjects. Hydrocortisone was used as a challenge because in microdialysis studies it has been shown to increase corticotropin-releasing factor release in extrahypothalamic brain regions such as the amygdala. [11C]NOP-1A total distribution volume (VT) in 11 regions of interest were measured using a 2-tissue compartment kinetic analysis. The primary outcome measure was hydrocortisone-induced ΔVT calculated as (VT CORT − VT BASE)/VT BASE. ResultsHydrocortisone led to an acute increase in plasma cortisol levels. Regional [11C]NOP-1A VT was on average 11% to 16% higher in the post-hydrocortisone condition compared with the baseline condition (linear mixed model, condition, p = .005; region, p < .001; condition × region, p < .001). Independent Student's t tests in all regions of interest were statistically significant and survived multiple comparison correction. Hydrocortisone-induced ΔVT was significantly negatively correlated with baseline VT in several regions of interest. ConclusionsHydrocortisone administration increases NOP receptor availability. Increased NOP in response to elevated cortisol might suggest a compensatory mechanism in the brain to counteract corticotropin-releasing factor and/or stress. The [11C]NOP-1A and hydrocortisone imaging paradigm should allow for the examination of interactions between stress-promoting neuropeptides and NOP in addictive disorders.

174-OR: Dorsal Vagal Complex Is a Site for Brain Lipid Sensing to Regulate Lipid Homeostasis

Obesity and diabetes are characterized by dyslipidemia due partly to an overproduction of hepatic triglyceride-rich very-low-density lipoproteins (VLDL-TG). The hypothalamus regulates whole-body lipid homeostasis in rodents but whether extra-hypothalamic brain regions can impact lipid homeostasis remain unclear. We here assessed whether the dorsal vagal complex (DVC), a region in the brainstem which contains a circumventricular organ termed the area postrema, as well as the nucleus of the solitary tract and the dorsal motor nucleus of the vagus, senses oleic acids to regulate whole-body lipid homeostasis. We first surgically implanted a guide cannula into the DVC of male 280-300g rats for vehicle (3% cyclodextrin in saline) or oleic acid (1 mM dissolved in vehicle) infusion, followed by catheter placements (7 days after) in the left carotid artery and right jugular vein for sampling and infusion. After an additional 5 days of recovery, 10h-fasted rats received either vehicle or oleic acid pre-infusion to the DVC for 10 min, and DVC infusion was maintained for an additional 150 min after an i.v. injection of poloxamer (600 mg/kg; a lipoprotein lipase inhibitor), while plasma samples were collected every 15 min for TG measurement. The rate of plasma TG appearance as a function of time (slope) reflects the rate of VLDL-TG secretion in the current experimental context. DVC oleic acid vs. vehicle infusion (0.33 ul/h) lowered VLDL-TG secretion (mM/min) in healthy rats (0.032±0.003, n=7 vs. 0.049±0.006, n=5; p&amp;lt;0.001), but interestingly failed to do so in high-fat fed rats. Taken together, our findings indicate that: (i) the DVC is a novel site of fatty acid sensing that impacts whole-body lipid homeostasis by regulating hepatic VLDL-TG secretion, and (ii) a disruption in DVC fatty acid sensing may contribute to the overproduction of hepatic VLDL-TG and a disruption in lipid homeostasis in obesity and diabetes. Disclosure B. Batchuluun: None. T. Waise: None. J.T. Yue: None. T.K. Lam: None. Funding Canadian Institutes of Health Research