Subcellular localization of vitellogenin in crustacean adipocytes by the unlabelled antibody enzyme method

The subepidermal fat body of the Amphipod Crustacean Orchestia gammarellus shows ultrastructural modifications related to vitellogenin synthesis. In the adipocytes of vitellogenic females, the rough endoplasmic reticulum is well developed whereas in those of males and non-vitellogenic females it is almost entirely absent; lipids and glycogen are, on the contrary, less abundant. The unlabelled antibody enzyme method shows the presence of vitellogenin in the dense bodies of the adipocytes of vitellogenic females. Adipocytes of males and non-vitellogenic females are not immunoreactive.     

Differential long-term effects of tannic acid on adenyl cyclase activity and lipolysis in rat adipocytes

Plants elaborate a variety of secondary metabolites such as hydrolysable tannins which are relatively abundant in fruits, vegetables and beverages in the human diet. We have studied the in vivo long-term effect consumption of tannic acid-supplemented drinking water (0.05%, w/v) on the rat adipocyte adenyl cyclase system and on lipolysis. We found that 14-day tannic acid supplementation did not significantly affect either body growth or food consumption, while fat pads weight was higher than that of the control, although the difference was not significant. On the other hand, tannic acid supplementation decreased both basal and isoproterenol-stimulated lipolysis significantly whereas cyclic AMP production as well as adenyl cyclase activity increased significantly. These results are at a first glance contradictory as cyclic AMP accumulation and lipolysis are positively correlated in rat adipocytes. They suggest at least that the tannic acid diet led to an inhibition of cyclic AMP-dependent protein kinase activity followed by a decrease in lipolysis in rat adipocytes, and to an increased activity of the type VI adenyl cyclase subunit of rat fat cells. This subunit is known to be negatively regulated under phosphorylation by cyclic AMP-dependent protein kinase. More in-depth studies are required to examine whether tannic acid could at least modify the expression of the catalytic subunit of adenyl cyclase, G-proteins and cyclic AMP-dependent protein kinase and/or alter their activities.     

Shedding new light on lipid biology with coherent anti-Stokes Raman scattering microscopy

Despite the ubiquitous roles of lipids in biology, the detection of lipids has relied on invasive techniques, population measurements, or nonspecific labeling. Such difficulties can be circumvented by a label-free imaging technique known as coherent anti-Stokes Raman (CARS) microscopy, which is capable of chemically selective, highly sensitive, and high-speed imaging of lipid-rich structures with submicron three-dimensional spatial resolution. We review the broad applications of CARS microscopy to studies of lipid biology in cell cultures, tissue biopsies, and model organisms. Recent technical advances, limitations of the technique, and perspectives are discussed.     

Proteomic profiling of metformin effects in 3T3-L1 adipocytes by SILAC-based quantification

Metformin is a common and generally the first medication prescribed for treatment of type 2 diabetes. Its mechanism involves affecting pathways that regulate glucose and lipid metabolism in metabolic cells such as that of muscle and liver cells. In spite of various studies exploring its effects, the proteome changes in adipocytes in response to metformin remains poorly understood. In this study, we performed stable isotope labeling by amino acids in cell culture (SILAC)-based quantitative proteomic profiling to study the effects of metformin specifically on 3T3-L1 adipocytes. We define proteins that exhibited altered levels with metformin treatment, 400 of them showing statistically significant changes in our study. Our results suggest that metformin affects not only the PPAR signaling pathway, as well as glucose and lipid metabolism, but also protein folding, endoplasmic reticulum stress, negative regulation of appetite, and one-carbon folate metabolism in adipocytes. This proteomic investigation provides important insight into effects of metformin in adipocytes.     

Adipocyte cellularity in different adipose depots in bulls of seven Spanish breeds slaughtered at two body weights

The influence of body weight (BW) at slaughter and genotype on adipocyte size and number in the omental (OM), perirenal (PR), subcutaneous (SC) and intermuscular (IM) adipose tissues was studied in 168 bulls of Spain's local Asturiana, Avileña, Morucha, Parda Alpina, Pirenaica, Retinta, and Rubia Gallega cattle breeds. The young bulls were slaughtered at two BWs, 320 and 540 kg. The results obtained showed the higher amounts of lipids that accumulated between 320 and 540 kg BW (P < 0.001) to be ascribable primarily to adipose cell hypertrophy, i.e. larger adipocyte size, in the OM and PR depots (P < 0.001). In addition to hypertrophy, there was also an increase (P < 0.001) in the number of adipose cells, i.e. hyperplasia, in the SC and IM adipose depots. Significant differences were observed when comparing the different genotypes, with the Morucha, Retinta and Avileña breeds having the highest amount of adipose tissue and the largest adipocytes. The Asturiana and Rubia Gallega breeds had the lowest amount of adipose tissue and the smallest adipocytes. The Pirenaica and Parda Alpina breeds had intermediate values in between the two groups identified above. In short, the results were indicative of different lipid deposition patterns in the different breeds depending on the individual growth and maturation rates in each. Similar findings were made when comparing the different adipose tissue depots, with adipocyte hypertrophy being the main factor responsible for lipid accumulation in the OM and PR depots, as opposed to adipocyte hyperplasia in the SC and IM depots.     

PI3K/AKT抑制剂渥曼青霉素对猪前体脂肪细胞增殖和凋亡的影响

为探寻PI3K/AKT抑制剂渥曼青霉素(Wortmannin,WM)对猪前体脂肪细胞增殖和凋亡均无影响的适宜浓度,文章首先分离并验证了猪原代前体脂肪细胞的分化潜能,然后对不同浓度渥曼青霉素处理11 d的细胞采用Annexin V-FITC/PI双标法检测细胞凋亡,并通过凋亡相关基因的表达以及DNA损伤程度进行验证,同时利用甲烷硫代磺酸盐(Methanethiosulfonate,MTS)检测了细胞的增殖活性。结果表明,100 nmol/L渥曼青霉素对猪前体脂肪细胞的增殖和凋亡均无显著影响,而200 nmol/L的渥曼青霉素对猪原代脂肪细胞的增殖活性虽没有显著影响,但对细胞凋亡有显著促进作用。研究发现,处理后促凋亡因子caspase8和TNFR1表达显著上调,非caspase依赖促凋亡因子GZMA表达无显著性差异,而GZMB表达则显著上调,抗凋亡因子Bcl-x1表达显著上调,cFLIP表达则无显著性差异。100 nmol/L的渥曼青霉素对细胞DNA的损伤不显著。因此,100 nmol/L的渥曼青霉素对猪前体脂肪细胞的增殖和凋亡均无显著影响,是在不影响细胞生长的情况下研究PI3K通路对脂肪细胞分化的较为理想的浓度。     

Increased PC-1 phosphodiesterase activity and inhibition of glucose uptake in adipocytes of type 2 diabetic rats

This study was designed to understand the cellular mechanisms responsible for defects in the insulin-stimulated signal transduction pathway in a type 2 diabetic animal model. We examined the in vitro PC-1 phosphodiesterase activity and glucose uptake in adipose tissue of streptozotocin (STZ)-induced type 2 diabetic rats. The PC-1 activity was significantly increased in adipose tissue of diabetic rats (0.54 ± 0.08 nmol PNTP hydrolyzed/mg protein/min) compared with controls (0.29 ± 0.05 nmol PNTP hydrolyzed/mg protein/min, p < 0.05). Upon insulin stimulation (100 nM), glucose uptake in the adipose tissue of the controls (4.17 ± 1.28×10⁻⁸ μmol/mg/min) was significantly higher than that in the diabetic rats (1.26 ± 0.35×10⁻⁸; p < 0.05). These results suggest that elevated PC-1 phosphodiesterase activity and decreased glucose uptake in adipose tissues may be acquired characteristics contributing to the development of type 2 diabetes mellitus.