scholarly journals The clearing-factor lipase activity of isolated fat-cells

1975 ◽  
Vol 146 (2) ◽  
pp. 481-488 ◽  
Author(s):  
A Cryer ◽  
P Davies ◽  
E R Williams ◽  
D S Robinson
Keyword(s):  
Enzyme Activity ◽  
Adipose Tissue ◽  
Lipase Activity ◽  
Incubation Medium ◽  
Cell Activity ◽  
Molecular Forms ◽  
Epididymal Adipose Tissue ◽  
Fat Cells ◽  
Isolated Fat Cells ◽  
Fat Cell

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.

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
Keyword(s):  
Adipose Tissue ◽  
Lipase Activity ◽  
Total Activity ◽  
Epididymal Adipose Tissue ◽  
Fat Cells ◽  
Isolated Fat Cells ◽  
Fat Cell ◽  
Prolonged Incubation ◽  
Intact Tissue

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.

scholarly journals Effect of nutrition on subcellular localization of rat fat-cell lipoprotein lipase

1978 ◽  
Vol 172 (2) ◽  
pp. 239-245 ◽  
Author(s):  
A Vanhove ◽  
C Wolf ◽  
M Breton ◽  
M C Glangeaud
Keyword(s):  
Adipose Tissue ◽  
Lipoprotein Lipase ◽  
Plasma Membranes ◽  
Total Activity ◽  
Normal Diet ◽  
Epididymal Adipose Tissue ◽  
Fat Cells ◽  
Isolated Fat Cells ◽  
Fat Cell ◽  
Intact Tissue

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.

scholarly journals Colchicine inhibition of the heparin-stimulated release of clearing-factor lipase from isolated fat-cells

1975 ◽  
Vol 152 (3) ◽  
pp. 717-720 ◽  
Author(s):  
A Cryer ◽  
A McDonald ◽  
E R Williams ◽  
D S Robinson
Keyword(s):  
Lipase Activity ◽  
Incubation Medium ◽  
Fat Cells ◽  
Isolated Fat Cells ◽  
Fat Cell

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.

scholarly journals Clearing-factor lipase in adipose tissue. Studies with puromycin and actinomycin

1968 ◽  
Vol 106 (3) ◽  
pp. 667-676 ◽  
Author(s):  
D. R. Wing ◽  
D S Robinson
Keyword(s):  
Enzyme Activity ◽  
Adipose Tissue ◽  
Lipase Activity ◽  
Half Life ◽  
Incubation Medium ◽  
Time Course

1. When adipose tissue from starved rats is incubated in a medium containing glucose, insulin, heparin and actinomycin (5μg./ml.) the total clearing-factor lipase activity of the system increases at least tenfold over a period of 9hr. In the absence of actinomycin, enzyme activity also increases, but to a lesser extent and for only about 3hr. Some enzyme activity appears in the incubation medium in both the presence and the absence of actinomycin. 2. When the glucose and insulin of the incubation medium are replaced by pyruvate and heparin is omitted, an increase in the total clearing-factor lipase activity in the presence of actinomycin still occurs, but only after a lag of several hours. When only heparin is omitted from the medium, the rise in enzyme activity begins immediately, but there is a shoulder in the time-course curve after a few hours. In the absence of heparin, little enzyme activity appears in the incubation medium. 3. The increases in enzyme activity in the presence of actinomycin are prevented if puromycin (0·5mg./ml.) is present in the incubation medium. 4. Catecholamines and corticotrophin inhibit the increase in enzyme activity caused by actinomycin. 5. The clearing-factor lipase activity of adipose tissue from fed animals declines with a half-life of between 1 and 1·5hr. when the tissue is incubated in the presence of puromycin. The clearing-factor lipase activity of adipose tissue from starved animals is stable under similar circumstances, as is the raised activity found after such tissue has been incubated in the presence of actinomycin. 6. Clearing-factor lipase extracted from adipose tissue of fed animals is less stable in solution than that extracted from the tissue of starved animals after this has been incubated in the presence of actinomycin.

Level of physical fitness and adipocyte lipolysis in humans

1984 ◽  
Vol 56 (5) ◽  
pp. 1157-1161 ◽  
Author(s):  
J. P. Despres ◽  
C. Bouchard ◽  
R. Savard ◽  
A. Tremblay ◽  
M. Marcotte ◽  
...  
Keyword(s):  
Adipose Tissue ◽  
Exercise Training ◽  
Lipolytic Activity ◽  
Fat Cells ◽  
Isolated Fat Cells ◽  
Fat Cell ◽  
Marathon Runners ◽  
Trained Subjects ◽  
Male Subjects ◽  
Sedentary Subjects

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.

scholarly journals The early development of white adipose tissue. Effects of litter size on the lipoprotein lipase activity of four adipose-tissue depots, serum immunoreactive insulin and tissue cellularity during the first four weeks of life in the rat

1979 ◽  
Vol 178 (3) ◽  
pp. 711-724 ◽  
Author(s):  
Anthony Cryer ◽  
Heather M. Jones
Keyword(s):  
Adipose Tissue ◽  
Lipoprotein Lipase ◽  
White Adipose Tissue ◽  
Lipase Activity ◽  
Nonesterified Fatty Acid ◽  
Immunoreactive Insulin ◽  
Postnatal Life ◽  
Fat Cells ◽  
Lipoprotein Lipase Activity ◽  
Fat Cell

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.

Water content of rat adipose tissue and isolated adipocytes in relation to cell size

1976 ◽  
Vol 231 (5) ◽  
pp. 1568-1572 ◽  
Author(s):  
M DiGirolamo ◽  
JL Owens
Keyword(s):  
Adipose Tissue ◽  
Water Content ◽  
Cell Volume ◽  
Cell Size ◽  
Intracellular Water ◽  
Male Rats ◽  
Fat Cells ◽  
Fat Cell ◽  
Tissue Water ◽  
Adipose Mass

Epididymal adipose tissue composition and adipocyte water content were studied in male rats during growth and development of spontaneous obesity. The data show that a highly significant positive correlation exists between fat-cell volume and intracellular water space (IWS) (r=.967, P less than .001). Intracellular water, expressed as picoliters per fat cell, varied from 1.5-2 in small fat cells (mean vol, 30-50 pl) to 9-10 in large cells (800-1,000 pl). When expressed as percent of fat-cell volume, IWS varied from 5-7% in the small fat cells to 1-1.3% in the large ones. Total adipose tissue water continued to increase with increasing adipose mass. Similarly, total adipocyte water increased with enlarging cell size and tissue mass. The contribution of total adipocyte water (as contrasted to that of nonadipocyte water) to total tissue water, however, was found to be limited (less than 23%) and to decline progressively with adipose mass expansion.

scholarly journals Factors affecting the permeability of isolated fat-cells from the rat to [42K]potassium and [36Cl]chloride ions

1970 ◽  
Vol 117 (3) ◽  
pp. 615-621 ◽  
Author(s):  
M. C. Perry ◽  
C. N. Hales
Keyword(s):  
Initial Phase ◽  
Chloride Ions ◽  
Fat Cells ◽  
Adrenocorticotrophic Hormone ◽  
Isolated Fat Cells ◽  
Free Medium ◽  
Fat Cell ◽  
Factors Affecting ◽  
Exchange Diffusion ◽  
High K

1. The effluxes of 42K+ and 36Cl− from isolated fat-cells from the rat were studied under a variety of conditions known to affect the metabolism of the cells. 2. 42K+ efflux from isolated fat cells was increased in a Na+-free–high-K+ medium and decreased in a K+-free medium. The existence of K+ exchange diffusion across the fat-cell membrane is suggested. 3. 36Cl− efflux from isolated fat-cells was decreased when the Cl− component of the wash medium was replaced by acetate. The basal 36Cl− efflux is suggested to be partly by Cl− exchange diffusion and partly in company with a univalent cation. 4. A variety of lipolytic stimuli, adrenaline, adrenocorticotrophic hormone, N-6,O-2′-dibutyryladenosine cyclic 3′:5′-monophosphate and theophylline, increased 42K+ efflux from isolated fat-cells. The adrenaline stimulation was biphasic; an initial, rapid and transient increase in 42K+ loss from the fat-cells was followed by a slower, more prolonged, increase in 42K+ efflux. The initial phase was inhibited by phentolamine but not by propranolol. 5. Insulin increased 42K+ efflux only after preincubation with the cells.

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