scholarly journals Clearing-factor lipase in adipose tissue. Distinction of different states of the enzyme and the possible role of the fat cell in the maintenance of tissue activity

1969 ◽  
Vol 112 (2) ◽  
pp. 203-209 ◽  
Author(s):  
V. J. Cunningham ◽  
D S Robinson

1. Incubation of intact epididymal adipose tissue from fed rats at 37° in an albumin solution at pH7·4 in vitro results in rapid loss of clearing-factor lipase activity until a low activity, stable to prolonged incubation, is attained. The clearing-factor lipase activity of intact tissue from starved rats, which is initially much less than that of tissue from fed rats, is mainly stable to incubation at 37°. 2. Much of the clearing-factor lipase activity of intact epididymal adipose tissue from fed rats is inactivated by collagenase. The enzyme activity of intact tissue from starved rats is not inactivated by collagenase. 3. The clearing-factor lipase activity of fat cells isolated from the epididymal adipose tissue of fed rats is stable to prolonged incubation at 37°. It represents only a small proportion of the total activity of the intact tissue. In starved rats, the isolated fat cells contain a much higher proportion of the activity of the intact tissue. Their activity is also stable at 37°. 4. Incubation of isolated fat cells in a serum-based medium leads to a progressive rise in clearing-factor lipase activity. Actinomycin increases the extent of this rise in activity. No rise in clearing-factor lipase activity occurs when stromal-vascular cells isolated from epididymal adipose tissue are incubated in the medium. 5. The findings indicate that less than 20% of the activity of intact adipose tissue from fed rats is retained when fat cells are isolated from the tissue by collagenase treatment. The activity that is lost could be that which normally functions in the uptake of triglyceride fatty acids by the tissue.

1978 ◽  
Vol 172 (2) ◽  
pp. 239-245 ◽  
Author(s):  
A Vanhove ◽  
C Wolf ◽  
M Breton ◽  
M C Glangeaud

This study supports the possibility for multiple subcellular forms of lipoprotein lipase. 1. The total activity of lipoprotein lipase per g of intact epididymal adipose tissue from fed rats is much higher than that from starved rats. 2. The isolated fat-cells of fed and of starved rats have lipoprotein lipase of almost the same activity per g of fat-pads. The isolated fat-cells of starved rats have a much higher proportion of total activity per g of the intact tissue than do those of fed rats. 3. Under the conditions of homogenization used, only a small proportion of the total activity per g of intact tissue from fed rats was associated with the fat layer which floated to the top of the homogenate during low-speed centrifugation. The different proportions of the specific enzyme activity found in each subcellular fraction are described. 4. Lipoprotein lipase from plasma membranes and microsomal fractions from starved and fed rats was purified by affinity chromatography. 5. The total activity of microsomal lipoprotein lipase per g of intact adipose tissue is enhanced by a normal diet. 6. In intact epididymal adipose tissue from fed rats, the activity per g of tissue of lipoprotein lipase of plasma membranes is much higher than that in the same fraction from starved rats. By contrast, the activities per g of tissue in plasma membranes obtained from starved or from fed rats by collagenase treatment were similar.


1975 ◽  
Vol 146 (2) ◽  
pp. 481-488 ◽  
Author(s):  
A Cryer ◽  
P Davies ◽  
E R Williams ◽  
D S Robinson

1. When fat-cells are isolated from the epididymal adipose tissue of 24h-starved rats and incubated at 25 degrees C in the presence of dialysed serum, glucose, insulin, amino acids and heparin, the total clearing-factor lipase acitivity of the incubation system increases progressively over a period of several hours. 2. All of the increase in activity is accounted for by the appearance of enzyme in the appearance of enzyme in the incubation medium and the fat-cell activity does not change significantly. Cycloheximids, at a concentration that prevents protein synthesis, does not affect the appearance of enzyme in the incubation medium, but the fat-cell enzyme activity is decreased in its presence. 3. The magnitude of the increase in total clearing factor lipase activity is unaffected by the omission of heparin from the medium. However, less enzyme is extracted in tis absence and the fat-cell activity increases. Cycloheximide again only affects the rise in cell activity and does not alter the activity in the incubation medium. 4. When serum in the incubation medium is replaced by casein, the distribution of enzyme between the cells and the medium is changed, but the magnitudes of the increases in total enzyme activity are similar. 5. These characteristics of the clearing-factor lipase response of isolated fat-cells differ in several respects from those observed earlier with intact adipose tissue from 24h-starved rats (Robinson & Wing, 1971; Cryer et al., 1973). The differences could be due, in part, to changes in the relative amounts of two different molecular forms of the enzyme that occur during the isolation of the fat-cells.


1964 ◽  
Vol 207 (6) ◽  
pp. 1215-1220 ◽  
Author(s):  
Alisa Gutman ◽  
Eleazar Shafrir

Rat adipose tissue from different body sites was shown to contain uridine diphosphoglucose (UDPG)-transglucosylase activity, which on the basis of protein content was comparable to or higher than that reported for muscle or liver. In epididymal adipose tissue, the activity of UDPG-glycogen transglucosylase and phosphorylase, as well as the content of glycogen per wet weight, decreased with increasing age of the animals in parallel with the decrease of tissue protein content. On prolonged fast the activity of UDPG-glycogen transglucosylase and phosphorylase per milligram protein dropped by 25–50% of the control value. On refeeding, the extent of changes was variable but, in general, at 24 hr control or higher levels of activity were reached and at 48 hr the activities were elevated. The ratio of glucose 6-phosphate independent activity of UDPG-glycogen transglucosylase to total activity was not affected by fasting and refeeding or by the administration of glucose with insulin. In adrenalectomized rats, with high adipose tissue glycogen, no change in UDPG-glycogen transglucosylase was found, whereas the levels of phosphorylase were elevated. Epinephrine in vivo and in vitro did not affect the activity of UDPG-glycogen transglucosylase of adipose tissue.


1969 ◽  
Vol 43 (2) ◽  
pp. 285-294 ◽  
Author(s):  
D. R. LANGSLOW ◽  
C. N. HALES

SUMMARY The effects on lipolysis of various compounds have been studied in intact chicken adipose tissue and in isolated fat cells prepared from chicken adipose tissue. Glucagon stimulated lipolysis at concentrations down to 1 ng./ml. in intact pieces and 0·1 ng./ml. in isolated fat cells. The effect was enhanced by high concentrations of insulin. No anti-lipolytic effect of insulin was observed. Adrenaline, noradrenaline, porcine corticotrophin (ACTH) and long-acting ACTH were lipolytic but the effects were small and high concentrations were required. The adrenaline effect was blocked by propranolol hydrochloride. Dibutyryl 3′,5′-(cyclic)-AMP and theophylline stimulated lipolysis as did a combination of crude chicken growth hormone and hydrocortisone sodium succinate. It was concluded that the pattern of response of chicken adipose tissue was markedly different from that of the rat.


1973 ◽  
Vol 136 (2) ◽  
pp. 437-439 ◽  
Author(s):  
P. Davies ◽  
D. S. Robinson

The stability at 42°C of clearing-factor lipase in adipose tissue, and in intact fat-cells isolated from it, was investigated. That portion of the total activity of the tissue which is associated with the fat-cell is stable under such conditions. This stability is markedly diminished when the fat-cell is disrupted.


1975 ◽  
Vol 152 (3) ◽  
pp. 717-720 ◽  
Author(s):  
A Cryer ◽  
A McDonald ◽  
E R Williams ◽  
D S Robinson

When isolated fat-cells are incubated at 25 degrees C in serum-based media containing glucose, insulin and heparin, the rise that occurs in the clearing-factor lipase activity of the incubation medium is inhibited by colchicine. The rise in the fat-cell clearing-factor lipase activity that occurs during similar incubations in the absence of heparin is not affected by colchicine.


1984 ◽  
Vol 56 (5) ◽  
pp. 1157-1161 ◽  
Author(s):  
J. P. Despres ◽  
C. Bouchard ◽  
R. Savard ◽  
A. Tremblay ◽  
M. Marcotte ◽  
...  

The present experiment was conducted to study the influence of exercise training on adipose tissue lipolytic activity and to identify the amount of training required to induce maximal adaptation in humans. Fifty-one male subjects were divided into three groups according to their training regimen: 1) sedentary subjects (SS) (n = 21); 2) trained subjects (TS) (n = 15) who had exercised during a period of 20 wk, 5 days/wk, 45 min/session; and 3) experienced marathon runners (MR) (n = 15) who ran an average of 120 km/wk for many years. Biopsies of fat were performed in the suprailiac region after an overnight fast. Adipocyte diameter (AD) and epinephrine-stimulated lipolysis ( ESL ) were assessed on collagenase-isolated fat cells. A lower AD was noted in the MR group compared with the two other groups. Basal lipolysis (BL) and ESL were significantly higher in TS and MR than in controls. Moreover, BL values were comparable in the two trained groups, whereas ESL in the TS group was higher than in the MR group. These results indicate that training increases suprailiac fat cell lipolysis, which seems to adapt maximally within about 4 mo.


1979 ◽  
Vol 178 (3) ◽  
pp. 711-724 ◽  
Author(s):  
Anthony Cryer ◽  
Heather M. Jones

1. Newborn rats were reared in litters of either four or sixteen individuals. The animals from the small litters gained body weight more rapidly than those from large litters during the first 29 days of postnatal life studied. 2. The relative weights of the perigenital, perirenal, subcutaneous and intramuscular white-adipose-tissue sites in the animals from small litters indicated their relative obesity compared with controls. 3. The adipose depots from animals reared in small litters had a greater proportion of lipid present, by weight, and had a greater number of larger fat-cells present in them compared with the depots of animals reared in large litters. 4. Compared with both normal-sized litter controls and animals reared in sixteens, during the period of study the animals from small litters were hypertriacylglycerolaemic but normocholesterolaemic. 5. During suckling the blood glucose concentrations of animals reared in fours were increased, as were the concentrations of circulating immunoreactive insulin. 6. During the 29 days of life studied, in general, the lipoprotein lipase activity of adipose depots from animals reared in fours was greater than for animals in large litters when expressed as μmol of nonesterified fatty acid released from the substrate/h per g fresh weight of tissue, per depot, or per million fat-cells, but were similar per cm2 of fat-cell surface area. 7. The previously noted [Cryer & Jones (1978) Biochem. J.172, 319–325] pattern of mid-suckling elevation, late-suckling decline and post-weaning increase in the lipoprotein lipase activity of the four white-adipose depots studied was not obliterated by the nutritional manipulations employed. 8. The relation of the enzyme-activity changes and their hormonal stimuli to triacylglycerol accumulation in fat-cells of animals from large and small litters is discussed in relation to the possible significance they may have to our understanding of neonatally induced obesity.


Endocrinology ◽  
2021 ◽  
Author(s):  
Christel Björk ◽  
Narmadha Subramanian ◽  
Jianping Liu ◽  
Juan Ramon Acosta ◽  
Beatriz Tavira ◽  
...  

Abstract Objective Healthy hyperplasic (many but smaller fat cells) white adipose tissue (WAT) expansion is mediated by recruitment, proliferation and/or differentiation of new fat cells. This process (adipogenesis) is controlled by transcriptional programs mostly identified in rodents. A systemic investigation of adipogenic human transcription factors (TFs) that are relevant for metabolic conditions has not been revealed previously. Methods TFs regulated in WAT by obesity, adipose morphology, cancer cachexia and insulin resistance were selected from microarrays. Their role in differentiation of human adipose tissue-derived stem cells (hASC) was investigated by RNA interference (RNAi) screen. Lipid accumulation, cell number and lipolysis were measured for all screened factors (148 TFs). RNA (RNAseq), protein (western blot) expression, insulin and catecholamine responsiveness were examined in hASC following siRNA treatment of selected target TFs. Results Analysis of TFs regulated by metabolic conditions in human WAT revealed that many of them belong to adipogenesis-regulating pathways. The RNAi screen identified 39 genes that affected fat cell differentiation in vitro, where 11 genes were novel. Of the latter JARID2 stood out as being necessary for formation of healthy fat cell metabolic phenotype by regulating expression of multiple fat-cell phenotype-specific genes. Conclusions This comprehensive RNAi screening in hASC suggests that a large proportion of WAT TFs that are impacted by metabolic conditions might be important for hyperplastic adipose tissue expansion. The screen also identified JARID2 as a novel TF essential for the development of functional adipocytes.


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