Unloading-Induced Skeletal Interoception Alters Hypothalamic Signaling to Promote Bone Loss and Fat Metabolism.

Microgravity is the primary factor that affects human physiology in spaceflight, particularly bone loss and disturbances of the central nervous system. However, little is known about the cellular and molecular mechanisms of these effects. Here, it is reported that in mice hindlimb unloading stimulates expression of neuropeptide Y (NPY) and tyrosine hydroxylase (TH) in the hypothalamus, resulting in bone loss and altered fat metabolism. Enhanced expression of TH and NPY in the hypothalamus occurs downstream of a reduced prostaglandin E2 (PGE2)-mediated ascending interoceptive signaling of the skeletal interoception. Sympathetic antagonist propranolol or deletion of Adrb2 in osteocytes rescue bone loss in the unloading model. Moreover, depletion of TH+ sympathetic nerves or inhibition of norepinephrine release ameliorated bone resorption. Stereotactic inhibition of NPY expression in the hypothalamic neurons reduces the food intake with altered energy expenditure with a limited effect on bone, indicating hypothalamic neuroendocrine factor NPY in the facilitation of bone formation by sympathetic TH activity. These findings suggest that reduced PGE2-mediated interoceptive signaling in response to microgravity or unloading has impacts on the skeletal and central nervous systems that are reciprocally regulated.

Zinc Supplementation Improved Neuropeptide Y, Nesfatin-1, Leptin, C-reactive protein, and HOMA-IR of Diet-Induced Obese Rats.

Obesity is a mild chronic inflammation that causes many metabolic diseases. It was aimed to investigate some parameters affective on the energy metabolism by adding zinc (Zn, ZnSO4) to drinking water of diet-induced obese rats. Five-week aged, male Sprague Dawley rats divided into as control group, consuming standard rat diet, and high-fat diet (HFD) group. After obesity induced by feeding HFD for 8 weeks, the obese rats were divided into Zn-supplemented obese group (HFD + obese + Zn; 150 mg Zn/L (for 6 weeks), 235 mg Zn/L (7th week), 250 mg Zn/L (8th week) in drinking water) and obese group (HFD + obese). Mean body weight, serum concentrations of C-reactive protein, neuropeptide-Y, leptin, insulin fasting blood glucose, and HOMA-IR were statistically decreased by given Zn in HFD + obese + Zn group compared to HFD + obese rats. It was observed that the total cholesterol, LDL, and HDL cholesterol levels of HFD + obese + Zn group became closer to the control group level, and Zn supplementation caused a statistically significant decrease in cholesterol profile than HFD + obese rats. Also, increased mean serum nesfatin-1 level, an effective protein for the formation of satiety, was analyzed in HFD + obese + Zn group when compared to HFD + obese ones. Serum triglyceride concentration tended to decrease with the effect of Zn in obese rats. In conclusion, it can be said that oral use of Zn could improve energy balance and prevent the occurrence of metabolic diseases related to obesity depending on the anti-inflammatory effect of Zn.

Glycyl-l-glutamine attenuates NPY-induced hyperphagia via the melanocortin system.

This study aimed to determine whether glycyl-l-glutamine (Gly-Gln; ß-endorphin (30-31)), a non-opioid peptide derived from ß-endorphin processing, modulates neuropeptide Y (NPY)-induced feeding and hypothalamic mRNA expression of peptide hormones in male broiler chicks. Intracerebroventricular injection of NPY (235 pmol) generated a hyperphagic response in ad libitum chicks within 30 min. Co-administration of Gly-Gln (100 nmol) attenuated this response, inducing a 30 % decrease. This was not attributable to Gly-Gln hydrolysis because co-administration of glycine (Gly) and glutamine (Gln) had no effect on NPY-induced hyperphagia. Gly-Gln injected alone also showed no effect. The hypothalamic pro-opiomelanocortin mRNA expression in the co-injection group was significantly higher than that in the NPY alone group. These data indicate that endogenous Gly-Gln may contribute to regulate feeding behavior via the central melanocortin system in chicks and acts as a counter regulator of the neural activity in energy metabolism.

[Effects on cervical spondylosis of vertebral artery type and the concentrations of plasma NPY and UII in the patients treated with the modified acupuncture at unilateral/bilateral Renying (ST 9)].

OBJECTIVE: To observe the differences in the clinical therapeutic effects on cervical spondylosis of vertebral artery type (CSA) between the modified acupuncture and the routine acupuncture at unilateral/bilateral Renying (ST 9) as well as the impacts on the concentrations of plasma neuropeptide Y (NPY) and urotensinⅡ(UⅡ) in the patients. METHODS: A total of 160 patients were divided into a modified bilateral acupuncture group, a modified unilateral acupuncture group, a routine bilateral acupuncture group and a routine unilateral acupuncture group, 40 cases in each one according to the random number table. In the modified bilateral acupuncture group, the modified acupuncture was applied bilaterally to Renying (ST 9). In the modified unilateral acupuncture group, the modified acupuncture was applied unilaterally to Renying (ST 9). In the routine bilateral acupuncture group, the routine acupuncture was applied bilaterally to Renying (ST 9). In the routine unilateral acupuncture group, the routine acupuncture was applied unilaterally to Renying (ST 9). The treatment was given once every day, continuously for 6 days as one course. Two courses of treatment were required at the interval of 1 day. In each group, before and after treatment, we observed the peak systolic blood flow velocity (Vs) of the vertebral artery (VA) and the basilar artery (BA), cervical vertigo symptoms and functional assessment scales (ESCV) and the concentration of plasma NPY and UⅡ. The clinical therapeutic effects were compared among the groups. RESULTS: After treatment, the clinical therapeutic effect in the modified bilateral acupuncture group was 90.0% (36/40), which was better than 80.0% (32/40) in the modified unilateral acupuncture group, 77.5% (35/40) in the routine bilateral acupuncture group and 65.0% (26/40) in the routine unilateral acupuncture group (all P<0.05). After treatment, Vs of VA and BA was improved remarkably in every group (all P<0.01), and the result in the modified bilateral acupuncture group was higher than those in the other groups (all P<0.01). After treatment, ESCV scores were all increased remarkably in every group (all P<0.01). ESCV score and improvement index in the modified bilateral acupuncture group were all higher than those in the other groups (P<0.05, P<0.01). After treatment, the concentrations of plasma NPY and UⅡ were all reduced remarkably in every group (all P<0.01) and the differences were significant among the groups (all P<0.01). CONCLUSION: The modified bilateral acupuncture at Renying (ST 9) effectively regulates the blood supply of the vertebral basilar artery and improves the cerebral circulation. The effects are superior to those of the unilateral acupuncture at Renying (ST 9).

[Early acupuncture for traumatic intracerebral hematoma: a randomized controlled trial].

OBJECTIVE: To observe the effect of early acupuncture intervention on brain edema in patients with traumatic intracerebral hematoma and explore its mechanism on the basis of conventional western medicine. METHODS: With stratified block randomization, sixty-four patients with glasgow coma scale (GCS) of 6 to 12 were divided into an acupuncture combined with medicine group (a combination group) and a western medication group, 32 cases in each one. In the western medication group, dehydration to reduce intracranial pressure and nutritional nerves were given as the basic treatment. In the combination group, on the basis of the treatment as the western medication group, acupuncture was applied at Xuehai (SP 10), Taixi (KI 3), Fenglong (ST 40), Yinlingquan (SP 9), Zusanli (ST 36), etc. The treatment was given once every day, for 6 times as one course; there was an interval of 1 day between two courses; a total of 4 courses were required. GCS score and recovery time were recored before treatment and on the 7 th, 14 th and 28 th days. 90 days follow-up after treatment, the GOS was observed, and the mortality and effective survival rate were calculated. The Barthel index (BI) score was evaluated before treatment and on the 14th, 21st, 28th days and 90 days follow-up after treatment. Before treatment and 3rd, 7th, 14th, 21st, 28th days, cranial CT or MR scan was performed to calculate the brain edema index (BEI); Plasma interleukin-6 (6IL-6), neuropeptide Y (NPY) and nitric oxide (NO) were measured before treatment and on the 3rd, 7th and 14th days after treatment. RESULTS: (1) The GCS scores increased gradually in the two groups during treatment, and there was significant difference between the 28th days and before treatment (both P<0.05). There were no significant difference between the two groups about GCS score and average recovery time on the 28th days treatment (all P>0.05). (2) The mortality rate of the combination group was 6.3% (2/32) on 90 days follow-up, 9.4% (3/32) in the western medication group (P>0.05). The effective survival rate was 81.3% (26/32) in the combination group, which was higher than 59.4% (19/32) in the western medication group (P<0.05). (3) The BI score was significantly higher than that before treatment on the 28th days and 90 days follow-up in the two groups (all P<0.05), and the result in the combination group was superior to that in the western medication group (both P <0.05). (4) The BEI decreased on the 14th, 21st and 28th days in the two groups (all P<0.05), and on the 14th day, the BEI decreased more significantly in the combination group than that in the western medication group (P<0.05). (5) The levels of IL-6, NPY and NO decreased on the 7th and 14th days in the two groups (all P<0.05), and decreased more significantly in the combination group than that in the western medication group on the 7th day (P<0.05). CONCLUSION: On the basis of conventional western medicine, early acupuncture can reduce cerebral edema and improve the prognosis of patients, and acupuncture combined with medicine are superior to western medicine alone. Acupuncture mechanism may be related to reducing the expression of inflammatory response.

Neuropeptide Y (NPY) or cocaine- and amphetamine-regulated transcript (CART) fiber innervation on central and medial amygdaloid neurons that project to the locus coeruleus and dorsal raphe in the rat.

The amygdaloid nuclear complex has been linked to the regulation of emotional behavior and energy regulation in that emotional stress might cause either reduction or enhancement of eating. We examined hypothalamic neuronal origin of feeding/arousal-related peptidergic fibers containing cocaine- and amphetamine-regulated transcript (CART) and neuropeptide Y (NPY) located in the rat amygdala along with its efferent projections to the brainstem monoaminergic nuclei. First, central (CeA) as well as medial (MeA) amygdala, among several amygdaloid subdivisions, exhibited the most prominent NPY or CART immunostaining which consisted of a substantial number of somata as well as labeled fibers. When we examined hypothalamic neuronal origin of NPY or CART fibers projecting to the CeA and MeA, medial and lateral arcuate nuclei were neuronal origins of NPY and CART fibers, respectively. However, the majority (>70%) of amygdala-projecting CART neurons which co-contained melanin-concentrating hormone (MCH) originated from the lateral hypothalamus (LH), zona incerta (ZI), and dorsal hypothalamic area (DA). This observation implied that the CeA as well as the MeA might receive potent second-order (and downstream) feeding-related CART input from the lateral hypothalamic regions in addition to first-order CART or NPY input from the Arc. Second, a large number of CeA neurons projected to the locus coeruleus (LC), whereas only a small number of MeA cells projected to the dorsal raphe (DR); none of the CeA or MeA cells provided dual projections to the LC and DR. Finally, a portion of MCH cells in the LH, ZI, and DA sent divergent axon collaterals to the CeA and LC. Considering that the CeA sends substantial GABAergic input to the LC, the present observation might serve as an anatomical substrate to support the potent hypnogenic role of MCH neurons in the LH regions during cataplexy and REM sleep.

Targeted Orexin and Hypothalamic Neuropeptides for Migraine.

The hypothalamus is involved in the regulation of homeostatic mechanisms and migraine-related trigeminal nociception and as such has been hypothesized to play a central role in the migraine syndrome from the earliest stages of the attack. The hypothalamus hosts many key neuropeptide systems that have been postulated to play a role in this pathophysiology. Such neuropeptides include but are not exclusive too orexins, oxytocin, neuropeptide Y, and pituitary adenylate cyclase activating protein, which will be the focus of this review. Each of these peptides has its own unique physiological role and as such many preclinical studies have been conducted targeting these peptide systems with evidence supporting their role in migraine pathophysiology. Preclinical studies have also begun to explore potential therapeutic compounds targeting these systems with some success in all cases. Clinical efficacy of dual orexin receptor antagonists and intranasal oxytocin have been tested; however, both have yet to demonstrate clinical effect. Despite this, there were limitations in these cases and strong arguments can be made for the further development of intranasal oxytocin for migraine prophylaxis. Regarding neuropeptide Y, work has yet to begun in a clinical setting, and clinical trials for pituitary adenylate cyclase activating protein are just beginning to be established with much optimism. Regardless, it is becoming increasingly clear the prominent role that the hypothalamus and its peptide systems have in migraine pathophysiology. Much work is required to better understand this system and the early stages of the attack to develop more targeted and effective therapies aimed at reducing attack susceptibility with the potential to prevent the attack all together.

Early acupuncture for traumatic intracerebral hematoma: a randomized controlled trial / 中国针灸

<p><b>OBJECTIVE</b>To observe the effect of early acupuncture intervention on brain edema in patients with traumatic intracerebral hematoma and explore its mechanism on the basis of conventional western medicine.</p><p><b>METHODS</b>With stratified block randomization, sixty-four patients with glasgow coma scale (GCS) of 6 to 12 were divided into an acupuncture combined with medicine group (a combination group) and a western medication group, 32 cases in each one. In the western medication group, dehydration to reduce intracranial pressure and nutritional nerves were given as the basic treatment. In the combination group, on the basis of the treatment as the western medication group, acupuncture was applied at Xuehai (SP 10), Taixi (KI 3), Fenglong (ST 40), Yinlingquan (SP 9), Zusanli (ST 36), etc. The treatment was given once every day, for 6 times as one course; there was an interval of 1 day between two courses; a total of 4 courses were required. GCS score and recovery time were recored before treatment and on the 7 th, 14 th and 28 th days. 90 days follow-up after treatment, the GOS was observed, and the mortality and effective survival rate were calculated. The Barthel index (BI) score was evaluated before treatment and on the 14th, 21st, 28th days and 90 days follow-up after treatment. Before treatment and 3rd, 7th, 14th, 21st, 28th days, cranial CT or MR scan was performed to calculate the brain edema index (BEI); Plasma interleukin-6 (6IL-6), neuropeptide Y (NPY) and nitric oxide (NO) were measured before treatment and on the 3rd, 7th and 14th days after treatment.</p><p><b>RESULTS</b>(1) The GCS scores increased gradually in the two groups during treatment, and there was significant difference between the 28th days and before treatment (both <0.05). There were no significant difference between the two groups about GCS score and average recovery time on the 28th days treatment (all >0.05). (2) The mortality rate of the combination group was 6.3% (2/32) on 90 days follow-up, 9.4% (3/32) in the western medication group (>0.05). The effective survival rate was 81.3% (26/32) in the combination group, which was higher than 59.4% (19/32) in the western medication group (<0.05). (3) The BI score was significantly higher than that before treatment on the 28th days and 90 days follow-up in the two groups (all <0.05), and the result in the combination group was superior to that in the western medication group (both <0.05). (4) The BEI decreased on the 14th, 21st and 28th days in the two groups (all <0.05), and on the 14th day, the BEI decreased more significantly in the combination group than that in the western medication group (<0.05). (5) The levels of IL-6, NPY and NO decreased on the 7th and 14th days in the two groups (all <0.05), and decreased more significantly in the combination group than that in the western medication group on the 7th day (<0.05).</p><p><b>CONCLUSION</b>On the basis of conventional western medicine, early acupuncture can reduce cerebral edema and improve the prognosis of patients, and acupuncture combined with medicine are superior to western medicine alone. Acupuncture mechanism may be related to reducing the expression of inflammatory response.</p>

Effects on cervical spondylosis of vertebral artery type and the concentrations of plasma NPY and UII in the patients treated with the modified acupuncture at unilateral/bilateral Renying (ST 9) / 中国针灸

<p><b>OBJECTIVE</b>To observe the differences in the clinical therapeutic effects on cervical spondylosis of vertebral artery type (CSA) between the modified acupuncture and the routine acupuncture at unilateral/bilateral Renying (ST 9) as well as the impacts on the concentrations of plasma neuropeptide Y (NPY) and urotensinⅡ(UⅡ) in the patients.</p><p><b>METHODS</b>A total of 160 patients were divided into a modified bilateral acupuncture group, a modified unilateral acupuncture group, a routine bilateral acupuncture group and a routine unilateral acupuncture group, 40 cases in each one according to the random number table. In the modified bilateral acupuncture group, the modified acupuncture was applied bilaterally to Renying (ST 9). In the modified unilateral acupuncture group, the modified acupuncture was applied unilaterally to Renying (ST 9). In the routine bilateral acupuncture group, the routine acupuncture was applied bilaterally to Renying (ST 9). In the routine unilateral acupuncture group, the routine acupuncture was applied unilaterally to Renying (ST 9). The treatment was given once every day, continuously for 6 days as one course. Two courses of treatment were required at the interval of 1 day. In each group, before and after treatment, we observed the peak systolic blood flow velocity (Vs) of the vertebral artery (VA) and the basilar artery (BA), cervical vertigo symptoms and functional assessment scales (ESCV) and the concentration of plasma NPY and UⅡ. The clinical therapeutic effects were compared among the groups.</p><p><b>RESULTS</b>After treatment, the clinical therapeutic effect in the modified bilateral acupuncture group was 90.0% (36/40), which was better than 80.0% (32/40) in the modified unilateral acupuncture group, 77.5% (35/40) in the routine bilateral acupuncture group and 65.0% (26/40) in the routine unilateral acupuncture group (all <0.05). After treatment, Vs of VA and BA was improved remarkably in every group (all <0.01), and the result in the modified bilateral acupuncture group was higher than those in the other groups (all <0.01). After treatment, ESCV scores were all increased remarkably in every group (all <0.01). ESCV score and improvement index in the modified bilateral acupuncture group were all higher than those in the other groups (<0.05, <0.01). After treatment, the concentrations of plasma NPY and UⅡ were all reduced remarkably in every group (all <0.01) and the differences were significant among the groups (all <0.01).</p><p><b>CONCLUSION</b>The modified bilateral acupuncture at Renying (ST 9) effectively regulates the blood supply of the vertebral basilar artery and improves the cerebral circulation. The effects are superior to those of the unilateral acupuncture at Renying (ST 9).</p>

Hypothalamic neuronal origin of neuropeptide Y (NPY) or cocaine- and amphetamine-regulated transcript (CART) fibers projecting to the tuberomammillary nucleus of the rat.

Based on the importance of tuberomammillary nucleus (TMN) as a target for feeding/arousal-related functions, we aimed in the present study to investigate hypothalamic neuronal origin of neuropeptide Y (NPY) and cocaine- and amphetamine-regulated transcript (CART) fibers projecting to the histaminergic nucleus. In the first series of experiments, we examined NPY (or CART) fiber distribution within the boundary of adenosine deaminase (ADA)-immunoreactive (ir) TMN regions; extensive NPY (or CART)-ir axon terminals were observed in E4 (TMMd), E3 (TMMv), and E2 (TMVr) subdivisions. NPY varicosities co-contained vesicular GABA transporters (vGAT). CART boutons, however, contained either vGAT or vesicular glutamate transporters (vGLU), which suggested dual (or multiple) origins of CART fibers. Based on the previous observation on melanin-concentrating hormone (MCH)-ir neuronal elements in the TMN, their coexistence with CART peptide was examined in detail. In E4 subdivision, approximately 40.8% of MCH-ir somata co-contained CART, but the proportion was reduced to 24.1% in E3 region. In E2 and E1 (TMVc) regions, only MCH-ir axon terminals existed without any MCH-ir somata. In the second series of experiments, we investigated hypothalamic neuronal origin of NPY (or CART) fibers projecting to the TMN. The arcuate nucleus (Arc) was the sole source of hypothalamic NPY fibers projecting to the nucleus. In contrast, CART fibers in the TMN originated from the Arc as well as the other hypothalamic nuclei including the retrochiasmatic nucleus, paraventricular nucleus, lateral hypothalamus (LH), zona incerta (ZI), and dorsal hypothalamic area. Quantitative analysis showed that arcuate CART projection to the TMN occupied approximately 23.5% of the total hypothalamic CART input to the nucleus, while the rest originated mainly from the LH and ZI. The present observations suggested that the TMN might play a key role in energy balance and arousal, by receiving periphery-derived, first-order NPY (or CART) inputs from the Arc as well as second-order (and downstream) CART inputs from the other hypothalamic nuclei.

Attenuated Weight Gain with the Novel Analog of Olanzapine Linked to Sarcosinyl Moiety (PGW5) Compared to Olanzapine.

Olanzapine-induced weight gain is associated with atherosclerosis, hypertension, dyslipidemia, and diabetes. We synthesized a novel antipsychotic drug (PGW5) possessing an olanzapine moiety linked to sarcosine, a glycine transporter 1 inhibitor. In this study, we compared the metabolic effects of PGW5 and olanzapine in a female rat model of weight gain. Female rats were treated daily with oral olanzapine (4 mg/kg), PGW5 (25 mg/kg), or vehicle for 16 days. Behavioral tests were conducted on days 12-14. Biochemical analyses were performed at the end of the treatment. A significant increase in body weight was observed in the olanzapine-treated group, while the PGW5 group did not differ from the controls. The open field test showed hypo-locomotion in the olanzapine-treated animals as compared to PGW5 and control groups. A significant increase in hypothalamic protein expression of the neuropeptide Y5 receptor and a decrease in pro-opiomelanocortin messenger ribonucleic acid (mRNA) levels were detected following PGW5 treatment, but not after olanzapine administration. PGW5 appears to possess minor metabolic effects compared with the parent compound olanzapine. The differential modulation of brain peptides associated with appetite regulation is possibly involved in the attenuation of metabolic effects by PGW5.

The role of pancreatic polypeptide in the regulation of energy homeostasis.

Imbalances in normal regulation of food intake can cause obesity and related disorders. Inadequate therapies for such disorders necessitate better understanding of mechanisms that regulate energy homeostasis. Pancreatic polypeptide (PP), a robust anorexigenic hormone, effectively modulates food intake and energy homeostasis, thus potentially aiding anti-obesity therapeutics. Intra-gastric and intra-intestinal infusion of nutrients stimulate PP secretion from the gastrointestinal tract, leading to vagal stimulation that mediates complex actions via the neuropeptide Y4 receptor in arcuate nucleus of the hypothalamus, subsequently activating key hypothalamic nuclei and dorsal vagal complex of the brainstem to influence energy homeostasis and body composition. Novel studies indicate affinity of PP for the relatively underexplored neuropeptide y6 receptor, mediating actions via the suprachiasmatic nucleus and pathways involving vasoactive intestinal polypeptide and insulin like growth factor 1. This review highlights detailed mechanisms by which PP mediates its actions on energy balance through various areas in the brain.

Dynamics of appetite-mediated gene expression in daidzein-fed female rats in the meal-feeding method.

We previously found that daidzein decreased food intake in female rats. The present study aimed to elucidate the relationship between dynamics of appetite-mediated neuropeptides and the anorectic effect of daidzein. We examined appetite-mediated gene expression in the hypothalamus and small intestine during the 3 meals per day feeding method. Daidzein had an anorectic effect specifically at the second feeding. Neuropeptide-Y (NPY) and galanin mRNA levels in the hypothalamus were significantly higher after feeding in the control but not in the daidzein group, suggesting that daidzein attenuated the postprandial increase in NPY and galanin expression. The daidzein group had higher corticotrophin-releasing hormone (CRH) mRNA levels in the hypothalamus after feeding, and increased cholelcystokinin (CCK) mRNA levels in the small intestine, suggesting that CCK is involved in the hypothalamic regulation of this　anorectic effect. Therefore, daidzein may induce anorexia by suppressing expression of NPY and galanin and increasing expression of CRH in the hypothalamus.

Regulation of energy homeostasis by the NPY system.

Obesity develops when energy intake exceeds energy expenditure over time. Numerous neurotransmitters, hormones, and factors have been implicated to coordinately control energy homeostasis, centrally and peripherally. However, the neuropeptide Y (NPY) system has emerged as the one with the most critical functions in this process. While NPY centrally promotes feeding and reduces energy expenditure, peptide YY (PYY) and pancreatic polypeptide (PP), the other family members, mediate satiety. Importantly, recent research has uncovered additional functions for these peptides that go beyond the simple feeding/satiety circuits and indicate a more extensive function in controlling energy homeostasis. In this review, we will discuss the actions of the NPY system in the regulation of energy balance, with a particular focus on energy expenditure.

Hypothalamic, feeding/arousal-related peptidergic projections to the paraventricular thalamic nucleus in the rat.

Based on the importance of paraventricular thalamic nucleus (PVT) as a relay station of energy balance, arousal, and food reward, we aimed in the present study to determine projection patterns of neuropeptide Y (NPY), cocaine- and amphetamine-regulated transcript (CART), melanin-concentrating hormone (MCH), and orexin (ORX)-ergic fibers to the PVT. First, the distribution of peptidergic axon terminals within the PVT was examined. NPY and CART terminals were confined within the boundary of the thalamic nucleus, exhibiting almost identical distribution. MCH terminals were rarely observed. In contrast, ORX terminals were as extensive as NPY/CART terminals, but spread into the peri-PVT region. Second, neuronal origin of feeding/arousal-related peptides projecting to the PVT was investigated. NPY neurons were observed in the medial subdivision of the arcuate nucleus (Arc), whereas CART cells were in the lateral Arc as well as other hypothalamic regions including the paraventricular hypothalamic nucleus, lateral hypothalamus (LH), dorsal hypothalamic area, and zona incerta. Both NPY- and CART-fiber projections to the PVT were bilateral; ipsilateral proportion was 54.0% ± 3.6% (n = 6) for NPY and 57.1% ± 2.5% (n = 6) for CART. The total number of CART neurons projecting to the PVT exceeded that of NPY cells; the ratio of labeled CART neurons to NPY cells was 2.4 ± 0.2 (n = 6). In contrast, ORX-ergic fiber projection to the PVT exhibited a slight ipsilateral dominance (62.7% ± 1.6%, n = 6), with majority of labeled cells located in the LH medial to the fornix (72.2% ± 2.3%, n = 6). Third, based on heavy projection from the PVT to the nucleus accumbens shell (NAcSh), the convergence of NPY and CART terminals on a single PVT neuron was identified; the proportion of labeled PVT neurons that received converging NPY/CART terminals compared with the total PVT neurons projecting to the NAcSh was 2.7% ± 0.6% (n = 3). Finally, PVT cells receiving NPY, CART, or ORX terminals provided divergent axon collaterals to NAcSh and medial prefrontal cortex. The present observations provided the anatomical evidence that the PVT might play an essential role in the integration of antagonistically-acting, feeding/arousal-related peptidergic inputs on their way to the cortical reward circuit.

Delineating the regulation of energy homeostasis using hypothalamic cell models.

Attesting to its intimate peripheral connections, hypothalamic neurons integrate nutritional and hormonal cues to effectively manage energy homeostasis according to the overall status of the system. Extensive progress in the identification of essential transcriptional and post-translational mechanisms regulating the controlled expression and actions of hypothalamic neuropeptides has been identified through the use of animal and cell models. This review will introduce the basic techniques of hypothalamic investigation both in vivo and in vitro and will briefly highlight the key advantages and challenges of their use. Further emphasis will be place on the use of immortalized models of hypothalamic neurons for in vitro study of feeding regulation, with a particular focus on cell lines proving themselves most fruitful in deciphering fundamental basics of NPY/AgRP, Proglucagon, and POMC neuropeptide function.

An investigation of appetite-related peptide transcript expression in Atlantic cod (Gadus morhua) brain following a Camelina sativa meal-supplemented feeding trial.

Camelina sativa is a hardy oilseed crop with seeds that contain high levels of ω3 polyunsaturated fatty acids and protein, which are critical components of fish feed. Camelina might thus be used as a cheaper and more sustainable supplement to fish-based products in aquaculture. Atlantic cod, Gadus morhua, is a species of interest in the aquaculture industry due to a decrease in wild populations and subsequent collapse of some cod fisheries. As cod are carnivorous fish, it is necessary to determine how this species physiologically tolerates plant-based diets. In this study, juvenile Atlantic cod were subjected to 13 weeks of either 15 or 30% camelina meal (CM)-supplemented diets or a control fish meal feed. Growth and food intake were evaluated and the mRNA expression of appetite-related hormones [pro-melanin-concentrating hormone (pmch), hypocretin (synonym: orexin, hcrt), neuropeptide Y (npy) and cocaine- and amphetamine-regulated transcript (cart)] was assessed using quantitative real-time PCR in brain regions related to food intake regulation (telencephalon/preoptic area, optic tectum/thalamus and hypothalamus). CM inclusion diets caused decreases in both growth and food intake in Atlantic cod. Optic tectum pmch transcript expression was significantly higher in fish fed the 30% CM diet compared to fish fed the 15% CM diet. In the hypothalamus, compared to fish fed the control diet, hcrt expression was significantly higher in fish fed the 30% CM diet, while npy transcript expression was significantly higher in fish fed the 15% CM diet. cart mRNA expression was not affected by diet in any brain region. Further studies are needed to determine which factors (e.g. anti-nutritional factors, palatability and nutritional deficits) contribute to reduced feed intake and growth, as well as the maximum CM inclusion level that does not negatively influence feed intake, growth rate and the transcript expression of appetite-related factors in Atlantic cod.

Neuroendocrine phenotypes in a boy with 5q14 deletion syndrome implicate the regulatory roles of myocyte-specific enhancer factor 2C in the postnatal hypothalamus.

The 5q14.3 deletion syndrome is a rare chromosomal disorder characterized by moderate to severe intellectual disability, seizures and dysmorphic features. We report a 14-year-old boy with 5q14.3 deletion syndrome who carried a heterozygous deletion of the myocyte-specific enhancer factor 2c (MEF2C) gene. In addition to the typical neurodevelopmental features of 5q14.3 deletion syndrome, he showed recurrent hypoglycemia, appetite loss and hypothermia. Hormonal loading tests using insulin, arginine and growth hormone-releasing factor revealed that growth hormone was insufficiently released into serum in response to these stimuli, thus disclosing the hypothalamic dysfunction in the present case. To uncover the biological roles of MEF2C in the hypothalamus, we studied its expression in the postnatal mouse brain. Notably, neuropeptide Y (NPY)-positive interneurons in the hypothalamic arcuate nuclei highly expressed MEF2C. In contrast, the Rett syndrome-associated protein, Methyl-CpG binding Protein 2 (MECP2) was barely expressed in these neurons. MEF2C knockdown or overexpression experiments using Neuro2a cells revealed that MEF2C activated the endogenous transcription of NPY. Conversely, siRNA-mediated knockdown of MECP2 led to derepression of the Npy gene. These data support the concept that MEF2C and MECP2 share common molecular pathways regulating the homeostatic expression of NPY in the adult hypothalamus. We propose that individuals with 5q14.3 deletion syndrome may exhibit neuroendocrine phenotypes through the functional loss of MEF2C in the postnatal hypothalamus.