Cpt1a silencing in AgRP neurons improves cognitive and physical capacity and promotes healthy aging in male mice.

Orexigenic neurons expressing agouti-related protein (AgRP) and neuropeptide Y in the arcuate nucleus (ARC) of the hypothalamus are activated in response to dynamic variations in the metabolic state, including exercise. We previously observed that carnitine palmitoyltransferase 1a (CPT1A), a rate-limiting enzyme of mitochondrial fatty acid oxidation, is a key factor in AgRP neurons, modulating whole-body energy balance and fluid homeostasis. However, the effect of CPT1A in AgRP neurons in aged mice and during exercise has not been explored yet. We have evaluated the physical and cognitive capacity of adult and aged mutant male mice lacking Cpt1a in AgRP neurons (Cpt1a KO). Adult Cpt1a KO male mice exhibited enhanced endurance performance, motor coordination, locomotion, and exploration compared with control mice. No changes were observed in anxiety-related behavior, cognition, and muscle strength. Adult Cpt1a KO mice showed a reduction in gastrocnemius and tibialis anterior muscle mass. The cross-sectional area (CSA) of these muscles were smaller than those of control mice displaying a myofiber remodeling from type II to type I fibers. In aged mice, changes in myofiber remodeling were maintained in Cpt1a KO mice, avoiding loss of physical capacity during aging progression. Additionally, aged Cpt1a KO mice revealed better cognitive skills, reduced inflammation, and oxidative stress in the hypothalamus and hippocampus. In conclusion, CPT1A in AgRP neurons appears to modulate health and protects against aging. Future studies are required to clarify whether CPT1A is a potential antiaging candidate for treating diseases affecting memory and physical activity.

Implantation of CPT1AM-expressing adipocytes reduces obesity and glucose intolerance in mice.

Obesity and its associated metabolic comorbidities are a rising global health and social issue, with novel therapeutic approaches urgently needed. Adipose tissue plays a key role in the regulation of energy balance and adipose tissue-derived mesenchymal stem cells (AT-MSCs) have gained great interest in cell therapy. Carnitine palmitoyltransferase 1A (CPT1A) is the gatekeeper enzyme for mitochondrial fatty acid oxidation. Here, we aimed to generate adipocytes expressing a constitutively active CPT1A form (CPT1AM) that can improve the obese phenotype in mice after their implantation. AT-MSCs were differentiated into mature adipocytes, subjected to lentivirus-mediated expression of CPT1AM or the GFP control, and subcutaneously implanted into mice fed a high-fat diet (HFD). CPT1AM-implanted mice showed lower body weight, hepatic steatosis and serum insulin and cholesterol levels alongside improved glucose tolerance. HFD-induced increases in adipose tissue hypertrophy, fibrosis, inflammation, endoplasmic reticulum stress and apoptosis were reduced in CPT1AM-implanted mice. In addition, the expression of mitochondrial respiratory chain complexes was enhanced in the adipose tissue of CPT1AM-implanted mice. Our results demonstrate that implantation of CPT1AM-expressing AT-MSC-derived adipocytes into HFD-fed mice improves the obese metabolic phenotype, supporting the future clinical use of this ex vivo gene therapy approach.

CPT1A in AgRP neurons is required for sex-dependent regulation of feeding and thirst.

BACKGROUND: Fatty acid metabolism in the hypothalamus has an important role in food intake, but its specific role in AgRP neurons is poorly understood. Here, we examined whether carnitinea palmitoyltransferase 1A (CPT1A), a key enzyme in mitochondrial fatty acid oxidation, affects energy balance. METHODS: To obtain Cpt1aKO mice and their control littermates, Cpt1a(flox/flox) mice were crossed with tamoxifen-inducible AgRPCreERT2 mice. Food intake and body weight were analyzed weekly in both males and females. At 12 weeks of age, metabolic flexibility was determined by ghrelin-induced food intake and fasting-refeeding satiety tests. Energy expenditure was analyzed by calorimetric system and thermogenic activity of brown adipose tissue. To study fluid balance the analysis of urine and water intake volumes; osmolality of urine and plasma; as well as serum levels of angiotensin and components of RAAS (renin-angiotensin-aldosterone system) were measured. At the central level, changes in AgRP neurons were determined by: (1) analyzing specific AgRP gene expression in RiboTag-Cpt1aKO mice obtained by crossing Cpt1aKO mice with RiboTag mice; (2) measuring presynaptic terminal formation in the AgRP neurons with the injection of the AAV1-EF1a-DIO-synaptophysin-GFP in the arcuate nucleus of the hypothalamus; (3) analyzing AgRP neuronal viability and spine formations by the injection AAV9-EF1a-DIO-mCherry in the arcuate nucleus of the hypothalamus; (4) analyzing in situ the specific AgRP mitochondria in the ZsGreen-Cpt1aKO obtained by breeding ZsGreen mice with Cpt1aKO mice. Two-way ANOVA analyses were performed to determine the contributions of the effect of lack of CPT1A in AgRP neurons in the sex. RESULTS: Changes in food intake were just seen in male Cpt1aKO mice while only female Cpt1aKO mice increased energy expenditure. The lack of Cpt1a in the AgRP neurons enhanced brown adipose tissue activity, mainly in females, and induced a substantial reduction in fat deposits and body weight. Strikingly, both male and female Cpt1aKO mice showed polydipsia and polyuria, with more reduced serum vasopressin levels in females and without osmolality alterations, indicating a direct involvement of Cpt1a in AgRP neurons in fluid balance. AgRP neurons from Cpt1aKO mice showed a sex-dependent gene expression pattern, reduced mitochondria and decreased presynaptic innervation to the paraventricular nucleus, without neuronal viability alterations. CONCLUSIONS: Our results highlight that fatty acid metabolism and CPT1A in AgRP neurons show marked sex differences and play a relevant role in the neuronal processes necessary for the maintenance of whole-body fluid and energy balance.

Dietary Options for Rodents in the Study of Obesity.

Obesity and its associated metabolic diseases are currently a priority research area. The increase in global prevalence at different ages is having an enormous economic and health impact. Genetic and environmental factors play a crucial role in the development of obesity, and diet is one of the main factors that contributes directly to the obesogenic phenotype. Scientific evidence has shown that increased fat intake is associated with the increase in body weight that triggers obesity. Rodent animal models have been extremely useful in the study of obesity since weight gain can easily be induced with a high-fat diet. Here, we review the dietary patterns and physiological mechanisms involved in the dynamics of energy balance. We report the main dietary options for the study of obesity and the variables to consider in the use of a high-fat diet, and assess the progression of obesity and diet-induced thermogenesis.

Impact of Adaptive Thermogenesis in Mice on the Treatment of Obesity.

Obesity and associated metabolic diseases have become a priority area of study due to the exponential increase in their prevalence and the corresponding health and economic impact. In the last decade, brown adipose tissue has become an attractive target to treat obesity. However, environmental variables such as temperature and the dynamics of energy expenditure could influence brown adipose tissue activity. Currently, most metabolic studies are carried out at a room temperature of 21 °C, which is considered a thermoneutral zone for adult humans. However, in mice this chronic cold temperature triggers an increase in their adaptive thermogenesis. In this review, we aim to cover important aspects related to the adaptation of animals to room temperature, the influence of housing and temperature on the development of metabolic phenotypes in experimental mice and their translation to human physiology. Mice studies performed in chronic cold or thermoneutral conditions allow us to better understand underlying physiological mechanisms for successful, reproducible translation into humans in the fight against obesity and metabolic diseases.

Efecto de ingesta de calcio del desayuno en la termogénesis alimentaria y oxidación de grasas postprandial en mujeres con sobrepeso / Effect of Breakfast Calcium Intake on Food Thermogenesis and Posprandial Fat Oxidation in Overweight Women

Resumen Antecedentes: una alta ingesta de calcio se relaciona con mayor termogénesis alimentaria y oxidación de grasa posprandial. Objetivo: evaluar el efecto de la ingesta de calcio del desayuno con termogénesis alimentaria y oxidación de grasas posprandial, en mujeres con sobrepeso. Materiales y métodos: estudio experimental, aleatorizado, conformado por 16 mujeres (ocho en el grupo experimental y ocho en el grupo control) entre 20-25 años. Se evaluó IMC, composición corporal mediante bioimpedanciometría, tasa metabólica en reposo en ayuno y posprandial mediante calorimetría indirecta, oxidación de grasa mediante cociente respiratorio y vitamina D sérica por radioinmunoensayo. Se administró al azar un desayuno isocalórico (377 kcal), alto en calcio (625 mg) o habitual en calcio (306 mg). Se describió con mediana y percentiles, y se comparó con pruebas Mann-Whitney y Wilcoxon para muestras pareadas. Resultados: la mediana de masa grasa y masa libre de grasa fue 30,9 % (27,5-33,9); 69,1 % (66,2-72,5) en el grupo experimental y 32,2 % (30,1-34,7); 67,8 % (65,3-69,9) en el grupo control (p=0,372). El grupo experimental mostró un aumento estadísticamente significativo en la termogénesis posprandial después del desayuno (p=0,035). Ambos grupos mostraron una mediana en cociente respiratorio posprandial aproximado a 1, (p=0,207), oxidando preferentemente carbohidratos. Conclusiones: las mujeres con desayuno alto en calcio presentan posterior al desayuno mayor termogénesis alimentaria, pero no mayor oxidación de grasa posprandial.

Abstract Background: High calcium intake is related to higher food thermogenesis and posprandial fat oxidation. Objective: Evaluate the calcium intake level at breakfast and both food thermogenesis and posprandial fat oxidation in overweight women. Materials and Methods: Experimental study with a random sample of 16 women, experimental group (8) and control group (8) aged 20-25 years. BMI, body composition by bioimpedance, resting metabolic rate at fasting, and posprandial were evaluated by indirect calorimetry; fat oxidation by respiratory quotient; and serum vitamin D by radioimmunoassay. Two types of isocaloric (377 kcal) breakfasts that were high (625 mg) or habitual (306 mg) in calcium were randomly administered. Results were described by medians and percentiles, which were compared by the Mann-Whitney test and Wilcoxon matched-paired test. Results: Median fat mass and fat-free mass was 30.9 % (27.5-33.9) and 69.1 % (66.2-72.5), and 32.2 % (30.1-34.7) and 67.8% (65.3-69.9) in the experimental and control group, respectively (p=0.372). The experimental group exhibited a statistically significant increase in posprandial breakfast thermogenesis (p=0.035). Both groups showed an approximate posprandial RQ median of 1 (p=0.207); they tended to oxidize carbohydrates. Conclusions: Women who consumed a high calcium breakfast exhibited higher post-breakfast food thermogenesis, but posprandial fat oxidation was not higher.