In vivo study of metabolic dynamics and heterogeneity in brown and beige fat by label‐free multiphoton redox and fluorescence lifetime microscopy

In this work, the metabolic characteristics of adipose tissues in live mouse model were investigated using a multiphoton redox ratio and fluorescence lifetime imaging technology. By analyzing the intrinsic fluorescence of metabolic coenzymes, we measured the optical redox ratios of adipocytes in vivo and studied their responses to thermogenesis. The fluorescence lifetime imaging further revealed changes in protein bindings of metabolic coenzymes in the adipocytes during thermogenesis. Our study uncovered significant heterogeneity in the cellular structures and metabolic characteristics of thermogenic adipocytes in brown and beige fat. Subgroups of brown and beige adipocytes were identified based on the distinct lipid size distributions, redox ratios, fluorescence lifetimes and thermogenic capacities. The results of our study show that this label‐free imaging technique can shed new light on in vivo study of metabolic dynamics and heterogeneity of adipose tissues in live organisms.     

Brown adipose tissue: from thermal physiology to bioenergetics

Brown adipose tissue is an organ in mammals specialized for the generation of heat. The tissue plays an important role in thermoregulatory heat production (nonshivering thermogenesis), and in nutritional energetics (through the process of diet-induced thermogenesis). Much of the current interest in brown adipose tissue has been catalysed by the postulate (1970’s) that a reduced capacity for thermogenesis underlies the development of obesity. Heat is generated in brown fat by a controlled uncoupling of oxidative phosphorylation, a process regulated by a tissue-specific mitochondrial uncoupling protein,M _r 32–33,000. The immunological identification of uncoupling protein is now used as a biochemical criterion for distinguishing brown fat from white adipose tissue. The gene coding for uncoupling protein has been cloned in several species, and a number of factors regulating the expression of the gene, as well as the amount and activity of the protein itself, have been documented. In addition to its direct role in heat production, brown adipose tissue has some notable general metabolic properties, such as in the conversion of thyroxine to triiodothyronine. An overview of the biology of brown adipose tissue is presented in this article, with an emphasis on some recent developments.     

Brown adipose tissue dynamics in wild-type and UCP1-knockout mice: in vivo insights with magnetic resonance

We used noninvasive magnetic resonance imaging (MRI) and magnetic resonance spectroscopy to compare interscapular brown adipose tissue (iBAT) of wild-type (WT) and uncoupling protein 1 (UCP1)-knockout mice lacking UCP1-mediated nonshivering thermogenesis (NST). Mice were sequentially acclimated to an ambient temperature of 30°C, 18°C, and 5°C. We detected a remodeling of iBAT and a decrease in its lipid content in all mice during cold exposure. Ratios of energy-rich phosphates (ATP/ADP, phosphocreatine/ATP) in iBAT were maintained stable during noradrenergic stimulation of thermogenesis in cold- and warm-adapted mice and no difference between the genotypes was observed. As free fatty acids (FFAs) serve as fuel for thermogenesis and activate UCP1 for uncoupling of oxidative phosphorylation, brown adipose tissue is considered to be a main acceptor and consumer of FFAs. We measured a major loss of FFAs from iBAT during noradrenergic stimulation of thermogenesis. This mobilization of FFAs was observed in iBAT of WT mice as well as in mice lacking UCP1. The high turnover and the release of FFAs from iBAT suggests an enhancement of lipid metabolism, which in itself contributes to the sympathetically activated NST and which is independent from uncoupled respiration mediated by UCP1. Our study demonstrates that MRI, besides its potential for visualizing and quantification of fat tissue, is a valuable tool for monitoring functional in vivo processes like lipid and phosphate metabolism during NST.     

Metabolism and thermoregulation in Maximowiczi's voles (Microtus maximowiczii) and Djungarian hamsters (Phodopus campbelli)

1 Metabolic rates (Vo₂), body temperature (T_b), and thermal conductance (C) were first determined in newly captured Maximowiczi's voles (Microtus maximowiczii) and Djungarian hamsters (Phodopus campbelli) from the Inner Mongolian grasslands at a temperature range from 5 to 35 °C.

2 The thermal neutral zone (TNZ) was between 25 and 32.5 °C for Maximowiczi's voles and between 25 and 30 °C for Djungarian hamsters. Mean T_b was 37.0±0.1 °C for voles and 36.2±0.1 °C for hamsters. Minimum thermal conductance was 0.172±0.004 ml O₂/g h °C for voles and 0.148±0.003 ml O₂/g h °C for hamsters.

3 The mean resting metabolic rate within TNZ was 2.21±0.05 ml O₂/g h in voles and 2.01±0.07 ml O₂/g h in hamsters. Nonshivering thermogenesis was 5.36±0.30 ml O₂/g h for voles and 6.30±0.18 ml O₂/g h for hamsters.

4 All these thermal physiological properties are adaptive for each species and are shaped by both macroenvironmental and microenvironmental conditions, food habits, phylogeny and other factors.

Chronic dietary vitamin A supplementation regulates obesity in an obese mutant WNIN/Ob rat model

Objective: To understand the possible role of chronic dietary high vitamin A supplementation in body weight regulation and obesity using a novel WNIN/Ob obese rat model developed at the National Centre for Laboratory Animal Sciences of National Institute of Nutrition, India. Research Methods and Procedures: Thirty‐six 7‐month‐old male rats of lean, carrier, and obese phenotypes were broadly divided into two groups; each group was subdivided into three subgroups consisting of six lean, six carrier, and six obese rats and received diets containing either 2.6 or 129 mg vitamin A/kg of diet for 2 months. Body weight gain, food intake, and weights of various organs were recorded. Adiposity index and BMI were calculated. Serum and liver retinol and brown adipose tissue (BAT)‐uncoupling protein1 (UCP1) mRNA expression levels were quantified. Results: Chronic feeding of high but non‐toxic doses of vitamin A through diet significantly reduced (P ≤ 0.05) body weight gain, adiposity index, and retroperitoneal white adipose tissue mass (without affecting food intake) in obese rats compared with their lean and carrier counterparts. In general, vitamin A treatment significantly improved hepatic retinol stores (P ≤ 0.05) in all phenotypes without affecting serum free retinol levels. However, augmented BAT‐UCP1 expression was observed only in carrier and obese rats (whose basal expression was low). Discussion: Our data suggest that chronic dietary vitamin A supplementation at high doses effectively regulates obesity in obese phenotype of the WNIN/Ob strain, possibly through up‐regulation of the BAT‐UCP1 gene and associated adipose tissue loss. However, in vitamin A‐supplemented lean and carrier rats, changes in adiposity could not be related to BAT‐UCP1 expression levels.     

Blocking mitochondrial pyruvate import in brown adipocytes induces energy wasting via lipid cycling

Combined fatty acid esterification and lipolysis, termed lipid cycling, is an ATP‐consuming process that contributes to energy expenditure. Therefore, interventions that stimulate energy expenditure through lipid cycling are of great interest. Here we find that pharmacological and genetic inhibition of the mitochondrial pyruvate carrier (MPC) in brown adipocytes activates lipid cycling and energy expenditure, even in the absence of adrenergic stimulation. We show that the resulting increase in ATP demand elevates mitochondrial respiration coupled to ATP synthesis and fueled by lipid oxidation. We identify that glutamine consumption and the Malate‐Aspartate Shuttle are required for the increase in Energy Expenditure induced by MPC inhibition in Brown Adipocytes (MAShEEBA). We thus demonstrate that energy expenditure through enhanced lipid cycling can be activated in brown adipocytes by decreasing mitochondrial pyruvate availability. We present a new mechanism to increase energy expenditure and fat oxidation in brown adipocytes, which does not require adrenergic stimulation of mitochondrial uncoupling.     

笼养普通朱雀适应北方冬季气候的 体温调节特征

内温动物的能量代谢和体温调节表现出普遍的季节性适应,这对于动物的能量分配、存活和繁殖等具有积极意义。本研究中,利用陆生动物呼吸代谢测量系统,测定了在北方适应了3年(2018至2020年)的普通朱雀(Carpodacus erythrinus)冬季代谢产热特征,以期理解该物种对北方冬季气候的响应。结果发现,普通朱雀冬季基础代谢率(以单位时间耗氧量表示)为(6.5±0.3)ml/(g·h);热中性区为20～35℃,环境温度低于下临界温度时,代谢率随环境温度降低显著升高,环境温度高于上临界温度时,代谢率随环境温度升高显著增加。体温在环境温度为10～25℃时,保持相对稳定,为(42.9±0.15)℃;环境温度为5℃时体温降低,高于30℃时,体温升高。最小热传导(0.25±0.02)ml/(g·h·℃),热传导随环境温度上升显著增加。与已有关于普通朱雀的研究报道相比较,本结果表明,普通朱雀在冬季代谢产热增加,以维持较高体温;热中性区变宽,下临界温度下调,以节省体温调节的能量支出;体温维持较高水平以抵抗寒冷,当气温过低时,适当降低体温,以减少直接能量消耗。     

The cowl does not make the monk: scarab beetle pollination of the Neotropical aroid Taccarum ulei (Araceae: Spathicarpeae)

Taccarum ulei (Araceae, Spathicarpeae) is a seasonal geophytic aroid, native to north‐eastern Brazil, that flowers during two months of the rainy season. Patterns of floral thermogenesis, pollination biology, and floral traits associated with pollination syndromes were studied and compared with those of other Araceae. Two species of cyclocephaline scarabs (Scarabaeidae, Cyclocephalini) were recognized as effective pollinators: Cyclocephala celata and Cyclocephala cearae. Larvae of an unidentified species of fruit fly (Melanoloma spp., Richardiidae, Diptera) were also frequently observed in inflorescences at various maturation stages, feeding on the connectives of male florets and fruits, and thus lowering the reproductive success of individual plants. Beetles were attracted by odoriferous inflorescences in the early evening of the first day of anthesis, during the female phase. The emission of attractive volatiles was coupled with intense thermogenic activity in the entire spadix, unlike other aroids in which only certain zones of the spadix heat up. Pollen release, which marks the beginning of the male phase on the subsequent evening, was not related to floral thermogenesis. Comparative multivariate analysis of the floral traits of T. ulei points to a beetle‐pollinated aroid, although some of the observed traits of the species are not common to other taxa sharing this pollination strategy. Such incongruence might be explained by the evolutionary history of the tribe Spathicarpeae and potential pollinator shifts. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, ??, ??–??.     

Etude de la respiration, à l’état isolé, des haplotypes de l’abeille tunisienne Apis mellifera intermissa (Buttel Reepen 1906) (Hymenoptera : Apidae)

L’étude de la respiration de l’abeille domestique, Apis mellifera intermissa, à l’état isolé a été réalisée sur les quatre haplotypes existant en Tunisie : A1, A4, A8 et A9 et sous différentes températures : 0°, 10°, 15°, 20°, 25°, 30° et 35°C. Des mesures effectuées sur 1000 abeilles isolées (250 de chaque haplotype) ont montré que la consommation d’oxygène de l’abeille varie en fonction de la température. L’intensité respiratoire des haplotypes augmente progressivement à partir de 0°C pour atteindre son maximum à 15°C. Au-delà de cette température, la consommation d’oxygène de l’abeille diminue progressivement. Pour les basses températures (0° et 10°C), la respiration de l’abeille dépend aussi du facteur génétique et la consommation d’oxygène est faible mais ne s’annule pas. Cette étude a montré qu’il existe une différence de consommation d’oxygène entre les haplotypes A1 et A8 et les haplotypes A4 et A9 pour les basses températures. Le déclenchement de la thermogenèse chez les haplotypes A1 et A8 se produit à 15°C, alors que chez les haplotypes A4 et A9, il a lieu à une température plus basse et qui est de 10°C.     

光周期和高脂食物对雌性高山姬鼠能量代谢和产热的影响

为了研究光周期和高脂食物对小型哺乳动物能量代谢和产热的影响,将成年雌性高山姬鼠分别驯化于长光照低脂、高脂食物和短光照低脂、高脂食物条件下,7周后测定动物的体重、能量摄入、产热、身体组成、血清瘦素浓度以及体脂含量等参数。结果发现:1)短光照抑制体重增长,降低血清瘦素浓度,增加非颤抖性产热;2)高脂食物使摄入能减少,消化率和体脂含量提高,但未显著影响体重、基础代谢率、非颤抖性产热和血清瘦素浓度;3)血清瘦素浓度与摄入能不相关,但与体脂重量正相关。结果暗示:短光照下瘦素作用敏感性增加和产热能力增强,可能介导了抵抗高脂食物诱导的肥胖。在野外条件下,高山姬鼠能通过能量代谢和产热的适应性调节避免体重的过度增长,有利于降低捕食风险,增强生存能力。