Insulin Controls Clock Gene Expression in the Liver of Goldfish Probably via Pi3k/Akt Pathway.

The liver circadian clock plays a pivotal role in driving metabolic rhythms, being primarily entrained by the feeding schedule, although the underlying mechanisms remain elusive. This study aimed to investigate the potential role of insulin as an intake signal mediating liver entrainment in fish. To achieve this, the expression of clock genes, which form the molecular basis of endogenous oscillators, was analyzed in goldfish liver explants treated with insulin. The presence of insulin directly increased the abundance of per1a and per2 transcripts in the liver. The dependency of protein translation for such insulin effects was evaluated using cycloheximide, which revealed that intermediate protein translation is seemingly unnecessary for the observed insulin actions. Furthermore, the putative interaction between insulin and glucocorticoid signaling in the liver was examined, with the results suggesting that both hormones exert their effects by independent mechanisms. Finally, to investigate the specific pathways involved in the insulin effects, inhibitors targeting PI3K/AKT and MEK/ERK were employed. Notably, inhibition of PI3K/AKT pathway prevented the induction of per genes by insulin, supporting its involvement in this process. Together, these findings suggest a role of insulin in fish as a key element of the multifactorial system that entrains the liver clock to the feeding schedule.

Resilience Among Direct Care Staff in Nursing Homes: Validation of the CD-RISC2.

OBJECTIVES: Caring for older adults is a demanding task. Some professionals may find themselves in complex situations for which they may not be prepared. However, the repercussions of these difficult experiences will depend on personal factors. Resilience acts as a buffer against difficulties and experiences. However, its study with first-line caregivers in nursing homes is limited. This study aimed to validate the Connor-Davidson Resilience Scale-2 (CD-RISC2), examine its psychometric properties, and analyze the variables associated with resilience (sociodemographic and professionals´ variables). METHODS: 312 nursing home professionals participated in the study. Besides the Connor-Davidson Resilience Scale-2, they answered questionnaires about good care, burnout, quality of life, person-centered care, and well-being. RESULTS: The CD-RISC2 showed adequate reliability and test-retest reliability. Good support was found for convergent validity with perceived health, person-directed care, personal growth, life purpose, quality of life and good care, and for discriminant validity with burnout. CONCLUSIONS: To our knowledge, this is the first study that validates the CD-RISC2 Spanish version with front-line workers in nursing homes showing adequate psychometric properties. CLINICAL IMPLICATIONS: An abbreviated, simpler, self-assessed version may be more useful than longer versions, especially with professionals with a heavy workload, such as direct care staff in nursing homes.

Periprandial response of central cannabinoid system to different feeding conditions in rainbow trout Oncorhynchus mykiss.

BACKGROUND: The mechanisms that regulate food intake are very complex since they comprise several neuroendocrine and metabolic signals responding to energetic or reward requirements. Previous studies in mammals indicate that cannabinoid system is implicated in homeostatic and hedonic regulation of food intake. In fish, several studies describe the components of this system, but only a little information is available regarding their role in food intake and energy balance regulation. OBJECTIVES: The objective of this study was to evaluate the main components of cannabinoid system related to feeding conditions in fish. METHODS: Samples of blood and different brain areas (telencephalon and hypothalamus) were taken from rainbow trout under different nutritional status (fasted, fed and refed) at different periprandial times (-30, 0, +30 and +180 min). RESULTS: Changes in AEA and 2-AG levels were observed in plasma related to the nutritional status and the sampling times assessed. At central levels, changes in endocannabinoids levels were observed in hypothalamus and in mRNA abundance of cnr1 and tprv1 in telencephalon and faah, gpr55 and fos in both brain areas. DISCUSSION: The results obtained suggest a role of endocannabinoid system in the regulation of food intake in fish at central level but further studies are required to fully elucidate the mechanisms involved.

Emotional distress among older adults during the COVID-19 outbreak: understanding the longitudinal psychological impact of the COVID-19 pandemic.

BACKGROUND: Older adults have proven their ability to overcome adversities throughout their life. This study aims to assess the impact of the COVID-19 pandemic on older adults' psychological distress (anxiety and depression) over time. METHODS: A community-dwelling Spaniard population (N = 192) completed a survey and reported on their sociodemographic characteristics, appraisal and personal resources. Older adults took part in a longitudinal online survey collected in April 2020 (during the lockdown restrictions) and at two subsequent time points 3 and 9 months after baseline (without lockdown restrictions and during the third wave of the pandemic respectively). RESULTS: Older adults did not evidence higher emotional distress than during the initial lockdown. Furthermore, depression remained stable and anxiety significantly decreased. Results also suggest that some sociodemographic characteristics, appraisals and personal resources are relevant. Older participants showed less anxiety than younger ones. Furthermore, being a male, resilience, and acceptance were related with the decrease of anxiety. Otherwise, fear of the COVID-19 outbreak and depression were related with the increase of anxiety. CONCLUSION: Older adults may adapt to the adverse pandemic impact by using more adaptive resources that reduce their distress. Efforts to ameliorate older adults' anxiety by focusing on older adults' personal resources should be considered.

Central administration of endocannabinoids exerts bimodal effects in food intake of rainbow trout.

The endocannabinoid system (ECs) is known to participate in several processes in mammals related to synaptic signaling including regulation of food intake, appetite and energy balance. In fish, the relationship of ECs with food intake regulation is poorly understood. In the present study, we assessed in rainbow trout Oncorhynchus mykiss the effect of intracerebroventricular administration (ICV) of low and high doses of the endocannabinoids anandamide (AEA) and 2-arachidonoylglycerol (2-AG) on food intake. We assessed endocannabinoid levels in hypothalamus, telencephalon and plasma as well as the effect of AEA and 2-AG administration at central level on gene expression of receptors involved in ECs (cnr1, gpr55 and trpv1) and markers of neural activity (fos, ntrk2 and GABA-related genes). The results obtained indicate that whereas high doses of endocannabinoids did not elicit changes in food intake levels, low doses of the endocannabinoids produce an orexigenic effect that could be due to a possible inhibition of gabaergic neurotransmission and the modulation of neural plasticity in brain areas related to appetite control, such as hypothalamus and telencephalon.

Role of the G protein-coupled receptors GPR84 and GPR119 in the central regulation of food intake in rainbow trout.

We evaluated the role of the G protein-coupled receptors GPR84 and GPR119 in food intake regulation in fish using rainbow trout (Oncorhynchus mykiss) as a model. In the first experiment, we assessed the effects on food intake of intracerebroventricular treatment with agonists of these receptors. In the second experiment, we assessed the impact of the same treatments on mRNA abundance in the hypothalamus and hindbrain of neuropeptides involved in the metabolic control of food intake (npy, agrp1, pomca1 and cartpt) as well as in changes in parameters related to signalling pathways and transcription factors involved in the integrative response leading to neuropeptide production. Treatment with both agonists elicited an anorectic response in rainbow trout attributable to changes observed in the mRNA abundance of the four neuropeptides. Changes in neuropeptides relate to changes observed in mRNA abundance and phosphorylation status of the transcription factor FOXO1. These changes occurred in parallel with changes in the phosphorylation status of AMPKα and Akt, the mRNA abundance of mTOR as well as signalling pathways related to PLCß and IP3. These results allow us to suggest that (1) at least part of the capacity of fish brain to sense medium-chain fatty acids such as octanoate depends on the function of GPR84, and (2) the capacity of fish brain to sense N-acylethanolamides or triglyceride-derived molecules occurs through the binding of these ligands to GPR119.

Validation of the professional good care scale in nursing homes (GCS-NH).

BACKGROUND: There is extensive concern about older people's care in institutions, especially recently in the past years. One of the reasons is linked to the cases of elder abuse, not only shown by academic and scientific sources, but also by social and mass media and their impact on public perception of the institutional setting. What is more, current COVID-19 pandemic consequences on older people have provoked alarm and worry especially about what is happening in institutions. METHODS: The sample for this study consists of 286 staff working in nursing homes in Spain. This study aimed to assess the psychometric properties of the Professional Good Care Scale in Nursing Homes (GCS-NH). RESULTS: Results of parallel analyses and exploratory factor analyses (EFAs) showed a four-factor model for the 32-item scale: humanization (9 items), non-infantilization (10 items), respect (7 items) and empowerment (6 items). Then, psychometric properties were tested analysing internal consistency (reliability) and convergent, divergent and criterion validity. High internal consistency (reliability) and different validity evidence were obtained for the total scores of the GCS-NH and its subscales. GCS-NH scores were also capable of detecting risk of probable institutional elder abuse. CONCLUSIONS: Results show that this scale is an appropriate, valid, and reliable multidimensional instrument to evaluate good care in older institutionalized people by staff. Good care is an outcome of a complex construct in which a wide range of factors converge (staff, older people, and environmental characteristics). The GCS-NH has potential to be used as a multidimensional tool to assess good care.

The role of acceptance in parents whose child suffers from cancer.

OBJECTIVE: Parents are the main caregivers when there is a child diagnosed with cancer. Receiving a diagnosis of childhood cancer is a stressful experience. The objective of this study is to explore the mediating role of emotional avoidance and acceptance in the emotional adjustment of parents throughout the disease. METHODS: Using the qualitative methodology Interpretative Phenomenological Analysis (IPA), parent's experiences of having a child with a diagnosis of cancer were explicated. Participants (n = 10) took part in one-on-one semi-structured interviews. RESULTS: Acceptance serves to cope with paediatric cancer because parents live day by day, actively taking care of their children. Acceptance and emotional avoidance with oneself, with the sick child and with the others play a key role in emotional adjustment. Differences between acceptance and avoidance were considered. CONCLUSION: Three main themes emerged from the analyses. Parents' ability to overcome their emotional struggle provides a non-pathological view of the experience of a child's illness. The results also showed the possibility for parents to access new ways to support their loved ones and to maintain the quality of their relationships. Acceptance has a positive impact on parents' adjustment while avoidance showed a negative effect on parents of children with cancer.

Hypothalamic AMPKα2 regulates liver energy metabolism in rainbow trout through vagal innervation.

Hypothalamic AMPK plays a major role in the regulation of whole body metabolism and energy balance. Present evidence has demonstrated that this canonical mechanism is evolutionarily conserved. Thus, recent data demonstrated that inhibition of AMPKα2 in fish hypothalamus led to decreased food intake and liver capacity to use and synthesize glucose, lipids, and amino acids. We hypothesize that a signal of abundance of nutrients from the hypothalamus controls hepatic metabolism. The vagus nerve is the most important link between the brain and the liver. We therefore examined in the present study whether surgical transection of the vagus nerve in rainbow trout is sufficient to alter the effect in liver of central inhibition of AMPKα2. Thus, we vagotomized (VGX) or not (Sham) rainbow trout and then intracerebroventricularly administered adenoviral vectors tagged with green fluorescent protein alone or linked to a dominant negative isoform of AMPKα2. The inhibition of AMPKα2 led to reduced food intake in parallel with changes in the mRNA abundance of hypothalamic neuropeptides [neuropeptide Y (npy), agouti-related protein 1 (agrp1), and cocaine- and amphetamine-related transcript (cartpt)] involved in food intake regulation. Central inhibition of AMPKα2 resulted in the liver having decreased capacity to use and synthesize glucose, lipids, and amino acids. Notably, these effects mostly disappeared in VGX fish. These results support the idea that autonomic nervous system actions mediate the actions of hypothalamic AMPKα2 on liver metabolism. Importantly, this evidence indicates that the well-established role of hypothalamic AMPK in energy balance is a canonical evolutionarily preserved mechanism that is also present in the fish lineage.

The endocannabinoid system is affected by a high-fat-diet in rainbow trout.

The endocannabinoid system (ECs) is a well known contributor to the hedonic regulation of food intake (FI) in mammals whereas in fish, the knowledge regarding hedonic mechanisms that control FI is limited. Previous studies reported the involvement of ECs in FI regulation in fish since anandamide (AEA) treatment induced enhanced FI and changes of mRNA abundance of appetite-related neuropeptides through cannabinoid receptor 1 (cnr1). However, no previous studies in fish evaluated the impact of palatable food like high-fat diets (HFD) on mechanisms involved in hedonic regulation of FI including the possible involvement of ECs. Therefore, we aimed to evaluate the effect of feeding a HFD on the response of ECs in rainbow trout (Oncorhynchus mykiss). First, we demonstrated a higher intake over 4 days of HFD compared with a control diet (CD). Then, we evaluated the postprandial response (1, 3 and 6 h) of components of the ECs in plasma, hypothalamus, and telencephalon after feeding fish with CD and HFD. The results obtained indicate that the increased FI of HFD occurred along with increased levels of 2-arachidonoylglycerol (2-AG) and AEA in plasma and in brain areas like hypothalamus and telencephalon putatively involved in hedonic regulation of FI in fish. Decreased mRNA abundance of EC receptors like cnr1, gpr55 and trpv1 suggest a feed-back counter-regulatory mechanism in response to the increased levels of EC. Furthermore, the results also suggest that neural activity players associated to FI regulation in mammals as cFOS, γ-Amino butyric acid (GABA) and brain derived neurotrophic factor (BDNF)/neurotrophic receptor tyrosine kinase (NTRK) systems could be involved in the hedonic eating response to a palatable diet in fish.

Oral and pre-absorptive sensing of amino acids relates to hypothalamic control of food intake in rainbow trout.

To assess the putative role of taste and pre-absorptive sensing of amino acids in food intake control in fish, we carried out an oral administration with l-leucine, l-valine, l-proline or l-glutamic acid in rainbow trout (Oncorhynchus mykiss). Treatment with proline significantly reduced voluntary food intake at 2 h and 3 h after oral administration, while glutamic acid showed a less pronounced satiating effect at 3 h. The mRNA expression of taste receptor subunits tas1r1, tas1r2a, tas1r2b and tas1r3 was measured in the epithelium overlying the bony basihyal of the fish (analogous to the tetrapod tongue) at 10, 20 or 30 min following treatment. No significant changes were observed, except for a tas1r down-regulation by valine at 30 min. Of the downstream taste signalling genes that were analysed in parallel, plcb2 and possibly trpm5 (non-significant trend) were down-regulated 20 min after proline and glutamic acid treatment. The signal originated in the oropharyngeal and/or gastric cavity presumably relays to the brain as changes in genes involved in the regulation of food intake occurred in hypothalamus 10-30 min after oral treatment with amino acids. In particular, proline induced changes consistent with an increased anorexigenic potential in the hypothalamus. We have therefore demonstrated, for the first time in fish, that the peripheral (pre-absorptive) detection of an amino acid (l-proline), presumably by taste-related mechanisms, elicits a satiety signal that in hypothalamus is translated into changes in cellular signalling and neuropeptides regulating food intake, ultimately resulting in decreased food intake.

The long-chain fatty acid receptors FFA1 and FFA4 are involved in food intake regulation in fish brain.

We hypothesized that the free fatty acid receptors FFA1 and FFA4 might be involved in the anorectic response observed in fish after rising levels of long-chain fatty acids (LCFAs) such as oleate. In one experiment we demonstrated that intracerebroventricular (i.c.v.) treatment of rainbow trout with FFA1 and FFA4 agonists elicited an anorectic response 2, 6 and 24 h after treatment. In a second experiment, the same i.c.v. treatment resulted after 2 h in an enhancement in the mRNA abundance of anorexigenic neuropeptides pomca1 and cartpt and a decrease in the values of orexigenic peptides npy and agrp1 These changes occurred in parallel with those observed in the mRNA abundance and/or protein levels of the transcription factors Creb, Bsx and FoxO1, protein levels and phosphorylation status of Ampkα and Akt, and mRNA abundance of plcb1 and itrp3 Finally, we assessed in a third experiment the response of all these parameters after 2 h of i.c.v. treatment with oleate (the endogenous ligand of both free fatty acid receptors) alone or in the presence of FFA1 and FFA4 antagonists. Most effects of oleate disappeared in the presence of FFA1 and FFA4 antagonists. The evidence obtained supports the involvement of FFA1 and FFA4 in fatty acid sensing in fish brain, and thus involvement in food intake regulation through mechanisms not exactly comparable (differential response of neuropeptides and cellular signalling) to those known in mammals.

Evidence for the presence in rainbow trout brain of amino acid-sensing systems involved in the control of food intake.

To assess the hypothesis of central amino acid-sensing systems involved in the control of food intake in fish, we carried out two experiments in rainbow trout. In the first one, we injected intracerebroventricularly two different branched-chain amino acids (BCAAs), leucine and valine, and assessed food intake up to 48 h later. Leucine decreased and valine increased food intake. In a second experiment, 6 h after similar intracerebroventricular treatment we determined changes in parameters related to putative amino acid-sensing systems. Different areas of rainbow trout brain present amino acid-sensing systems responding to leucine (hypothalamus and telencephalon) and valine (telencephalon), while other areas (midbrain and hindbrain) do not respond to these treatments. The decreased food intake observed in fish treated intracerebroventricularly with leucine could relate to changes in mRNA abundance of hypothalamic neuropeptides [proopiomelanocortin (POMC), cocaine- and amphetamine-related transcript (CART), neuropeptide Y (NPY), and agouti-related peptide (AgRP)]. These in turn could relate to amino acid-sensing systems present in the same area, related to BCAA and glutamine metabolism, as well as mechanistic target of rapamycin (mTOR), taste receptors, and general control nonderepressible 2 (GCN2) kinase signaling. The treatment with valine did not affect amino acid-sensing parameters in the hypothalamus. These responses are comparable to those characterized in mammals. However, clear differences arise when comparing rainbow trout and mammals, in particular with respect to the clear orexigenic effect of valine, which could relate to the finding that valine partially stimulated two amino acid-sensing systems in the telencephalon. Another novel result is the clear effect of leucine on telencephalon, in which amino acid-sensing systems, but not neuropeptides, were activated as in the hypothalamus.

Response of rainbow trout's (Oncorhynchus mykiss) hypothalamus to glucose and oleate assessed through transcription factors BSX, ChREBP, CREB, and FoxO1.

We aimed to obtain information regarding mechanisms that link glucose- and fatty acid-sensing systems to expression of neuropeptides that regulate food intake in the fish brain. We assessed the relative expression and protein levels of the transcription factors BSX, ChREBP, FoxO1, and CREB in the hypothalamus of rainbow trout (Oncorhynchus mykiss) treated for 6 h with either glucose or oleate in vivo (intra-cerebroventricular treatment with 1 µl 100 g- 1 body weight of 40 µg glucose or 1 µmol oleate) or in vitro (incubation with 4-8 mM glucose or 100-500 µM oleate). BSX levels decreased after oleate treatment for mRNA (10% in vitro and 47% in vivo) and protein (25%), while minor changes occurred after glucose treatment. CREB values generally decreased after glucose or oleate treatment for mRNA (50% in vivo) as well as the phosphorylation status of protein (80%). Foxo1 mRNA levels increased in vivo with glucose (129%) and decreased in vivo with oleate (60%), and protein phosphorylation status increased with glucose (in vivo) and oleate. mRNA values of chrebpα decreased in response to glucose and oleate, while protein levels decreased with oleate and increased with glucose. The results support the association of several transcription factors with metabolic control of food intake in fish.

The anorectic effect of central PYY1-36 treatment in rainbow trout (Oncorhynchus mykiss) is associated with changes in mRNAs encoding neuropeptides and parameters related to fatty acid sensing and metabolism.

We hypothesized that peptide YY (PYY) is involved in the metabolic regulation of food intake in fish. Therefore, we assessed in rainbow trout (Oncorhynchus mykiss) the effects of intracerebroventricular treatment with 10 ng/g PYY1-36 on food intake, expression of neuropeptides involved in food intake control, and the activity of fatty acid-sensing systems. The administration of PYY1-36 caused a significant reduction in food intake up to 24 h post-treatment. This anorectic action was associated with changes 2 h after treatment in mRNA abundance of neuropeptides involved in metabolic regulation of food intake in hypothalamus (decreased NPY and raised CART values) and hindbrain (increased POMCa1 values). We also observed that PYY1-36 treatment induced changes in mRNA abundance of parameters related to fatty acid sensing and metabolism in hypothalamus (decreased values of ACLY, PPARγ, and SREBP1c) and hindbrain (increased values of LPL, FAT/CD36, PPARα, PPARγ, and SREBP1c and decreased values of UCP2a). PYY1-36 treatment also increased mRNA abundance of mTOR. In general, it seems that mRNAs encoding some components of the machinery required for fatty acid sensing and metabolism are activated by PYY1-36. The response observed was higher in the hindbrain than in the hypothalamus, supporting the greater importance of this brain area in mediating the modulatory effects of gastrointestinal hormones on feeding regulation.

Obesity modulates diaphragm curvature in subjects with and without COPD.

Obesity is a common comorbidity of chronic obstructive pulmonary disease (COPD) and has been associated with worse outcomes. However, it is unknown whether the interaction between obesity and COPD modulates diaphragm shape and consequently its function. The body mass index (BMI) has been used as a correlate of obesity. We tested the hypothesis that the shape of the diaphragm muscle and size of the ring of its insertion in non-COPD and COPD subjects are modulated by BMI. We recruited 48 COPD patients with postbronchiodilator forced expiratory volume in 1 s (FEV1)-to-forced vital capacity (FVC) < 0.7 and 29 age-matched smoker/exsmoker control (non-COPD) subjects, who underwent chest computed tomography (CT) at lung volumes ranging from functional residual capacity (FRC) to total lung capacity (TLC). We then computed maximum principal diaphragm curvature in the midcostal region of the left hemidiaphragm at the end of inspiration during quiet breathing (EI) and at TLC. The radius of maximum curvature of diaphragm muscle increased with BMI in both COPD and non-COPD subjects. The size of diaphragm ring of insertion on the chest wall also increased significantly with increasing BMI. Surprisingly, COPD severity did not appear to cause significant alteration in diaphragm shape except in normal-weight subjects at TLC. Our data uncovered important factors such as BMI, the size of the diaphragm ring of insertion, and disease severity that modulate the structure of the ventilatory pump in non-COPD and COPD subjects.

Orally administered fatty acids enhance anorectic potential but do not activate central fatty acid sensing in Senegalese sole post-larvae.

Studies in fish have reported the presence and function of fatty acid (FA)-sensing systems comparable in many aspects to those known in mammals. Such studies were carried out in juvenile and adult fish, but the presence of FA-sensing systems and control of food intake have never been evaluated in early life stages, despite the importance of establishing when appetite regulation becomes functional in larval fish. In this study, we aimed to elucidate the possible effects of different specific FAs on neural FA-sensing systems and neuropeptides involved in the control of food intake in Senegalese sole post-larvae. To achieve this, we orally administered post-larvae with different solutions containing pure FA - oleate (OA), linoleate (LA), α-linolenate (ALA) or eicosapentaenoate (EPA) - and evaluated changes in mRNA abundance of neuropeptides involved in the control of food intake and of transcripts related to putative FA-sensing systems, 3 and 6 h post-administration. The changes in neuropeptide gene expression were relatively consistent with the activation of anorectic pathways (enhanced cart4 and pomcb) and a decrease in orexigenic factors (npy) following intake of FA. Even though there were a few differences depending on the nature of the FA, the observed changes appear to suggest the existence of a putative anorectic response in post-larvae fish to the ingestion of all four tested FAs. However, changes in neuropeptides cannot be explained by the integration of metabolic information regarding FAs in circulation through FA-sensing mechanisms in the brain. Only the reduction in mRNA levels of the FA metabolism gene acc in OA-treated (6 h), ALA-treated (3 h) and EPA-treated (3 and 6 h) post-larvae could be indicative of the presence of a FA-sensing system, but most genes either were not significantly regulated (fat/cd36-lmp2, acly, kir6.x, srebp1c) or were affected in a way that was inconsistent with FA-sensing mechanisms (fat/cd36-pg4l, fas, cpt1.1, cpt1.2, cpt1.3, sur, pparα and lxrα).

Ceramide counteracts the effects of ghrelin on the metabolic control of food intake in rainbow trout.

In mammals, ceramides are involved in the modulation of the orexigenic effects of ghrelin (GHRL). We previously demonstrated in rainbow trout that intracerebroventricular (ICV) treatment with ceramide (2.5 µg/100 g fish) resulted in an anorexigenic response, i.e. a response opposed to that described in mammals, where ceramide treatment is orexigenic. Therefore, we hypothesized that the putative interaction between GHRL and ceramide must be different in fish. Accordingly, in a first experiment, we observed that ceramide levels in the hypothalamus of rainbow trout did not change after ICV treatment with GHRL. In a second experiment, we assessed whether the effects of GHRL treatment on the regulation of food intake in rainbow trout changed in the presence of ceramide. Thus, we injected ICV GHRL and ceramide alone or in combination to evaluate in hypothalamus and hindbrain changes in parameters related to the metabolic control of food intake. The presence of ceramide generally counteracted the effects elicited by GHRL on fatty acid-sensing systems, the capacity of integrative sensors (AMPK, mTOR and SIRT-1), proteins involved in cellular signalling pathways (Akt and FoxO1) and neuropeptides involved in the regulation of food intake (AgRP, NPY, POMC and CART). The results are discussed in the context of regulation of food intake by metabolic and endocrine inputs.

Changes in the levels and phosphorylation status of Akt, AMPK, CREB and FoxO1 in hypothalamus of rainbow trout under conditions of enhanced glucosensing activity.

There is no available information about mechanisms linking glucosensing activation in fish and changes in the expression of brain neuropeptides controlling food intake. Therefore, we assessed in rainbow trout hypothalamus the effects of raised levels of glucose on the levels and phosphorylation status of two transcription factors, FoxO1 and CREB, possibly involved in linking these processes. We also aimed to assess the changes in the levels and phosphorylation status of two proteins possibly involved in the modulation of these transcription factors: Akt and AMPK. Therefore, in pooled preparations of hypothalamus incubated for 3 and 6 h in the presence of 2, 4 or 8 mmol l-1 d-glucose, we evaluated the response of parameters related to glucosensing mechanisms, neuropeptide expression and levels and phosphorylation status of the proteins of interest. The activation of hypothalamic glucosensing systems and the concomitant enhanced anorectic potential occurred in parallel with activation of Akt and inhibition of AMPK. The changes in these proteins relate to neuropeptide expression through changes in the level and phosphorylation status of transcription factors under their control, such as CREB and FoxO1, which displayed inhibitory (CREB) or activatory (FoxO1) responses to increased glucose.

Ceramides are involved in the regulation of food intake in rainbow trout (Oncorhynchus mykiss).

We hypothesize that ceramides are involved in the regulation of food intake in fish. Therefore, we assessed in rainbow trout (Oncorhynchus mykiss) the effects of intracerebroventricular treatment with C6:0 ceramide on food intake. In a second experiment, we assessed the effects in brain areas of ceramide treatment on neuropeptide expression, fatty acid-sensing systems, and cellular signaling pathways. Ceramide treatment induced a decrease in food intake, a response opposed to the orexigenic effect described in mammals, which can be related to enhanced mRNA abundance of cocaine and amphetamine-related transcript and proopiomelanocortin and decreased mRNA abundance of Agouti-related protein and neuropeptide Y. Fatty acid-sensing systems appear to be inactivated by ceramide treatment. The mRNA abundance of integrative sensors AMPK and sirtuin 1, and the phosphorylation status of cellular signaling pathways dependent on protein kinase B, AMPK, mammalian target of rapamycin (mTOR), and forkhead box protein O1 (FoxO1) are generally activated by ceramide treatment. However, there are differences between hypothalamus and hindbrain in the phosphorylation status of AMPK (decreased in hypothalamus and increased in hindbrain), mTOR (decreased in hypothalamus and increased in hindbrain), and FoxO1 (increased in hypothalamus and decreased in hindbrain) to ceramide treatment. The results suggest that ceramides are involved in the regulation of food intake in rainbow trout through mechanisms comparable to those characterized previously in mammals in some cases.