Changes in lymphokine receptor expression and fatty acid composition of phospholipids and triacylglycerols in rat adipocytes associated with lymph nodes following a transient immune challenge.

Single-photon counting fluorimetry was used to record the time course of the expression of interleukin-10 receptors labelled with fluorescent antibodies on the surface of adipocytes over 24h, following an immune challenge to the rat popliteal lymph node. Homologous perinodal and remote-from-node samples from the stimulated and unstimulated popliteal depots were compared in rats fed on plain chow and chow supplemented with 10% w/w suet, fish or vegetable oils. Receptor expression was maximal 6 h after stimulation, and returned to baseline after 24 h, and was similar in the stimulated and unstimulated depots. Fewer receptors were elicited in tissues from rats fed lipid-supplemented diets compared with the control diet, with fewest of all following the fish oil diet. These data suggest that interleukin-10 is involved in local interactions between perinodal adipocytes and lymph node lymphoid cells. Both triacylglycerols and phospholipids contained more polyunsaturates and fewer saturates in perinodal adipose tissue than in samples from sites not associated with lymphoid tissue. These data are consistent with paracrine interactions between perinodal adipocytes and activated lymphoid cells.

Interactions of noradrenalin and tumour necrosis factor alpha, interleukin 4 and interleukin 6 in the control of lipolysis from adipocytes around lymph nodes.

The contributions of inflammatory and immunosuppressive cytokines and noradrenalin to the control of lipolysis in adipocytes surrounding and remote from lymph nodes were investigated in healthy adult guinea-pigs. A few hours after excision from fasting animals, spontaneous lipolysis in adipocytes from around the popliteal and mesenteric lymph nodes and omental "milky spots" was significantly lower than in those from elsewhere in the same depots, and much lower than in perirenal, epididymal or parametrial adipocytes. The perinodal adipocytes were consistently more sensitive to noradrenalin at 10(-8), 10(-7)and 10(-5) M, and their maximum rate of lipolysis was higher. They also responded more strongly to pre-incubation for 24 h with tumour necrosis factor alpha interleukin 6 and interleukin 4 than those elsewhere in the same depots. Tumour necrosis factor alpha and interleukin 6 applied alone stimulated lipolysis, but combined with interleukin 4, they suppressed glycerol release, especially in perinodal adipocytes, thereby creating large within-depot differences. These cytokines had minimal effects on lipolysis in perirenal or gonadal adipocytes. The authors conclude that adipocytes surrounding lymph nodes contribute little to whole-body energy supply during fasting, but are more sensitive than all others to cytokines and to noradrenalin, having higher maximum but lower minimum rates of lipolysis. These properties equip perinodal adipocytes for local interactions with lymphoid tissue.

In vivo evidence for the involvement of the adipose tissue surrounding lymph nodes in immune responses.

Spontaneous lipolysis in the adipocytes surrounding the popliteal lymph node rose within 1 h of its being activated with a subcutaneous injection of lipopolysaccharide (LPS), reached a peak after 6-9 h, then declined almost to basal levels after 24 h. The response of adipocytes from elsewhere in the same depot was delayed and smaller. Following the simulated immune challenge, perinodal adipocytes were consistently more sensitive to noradrenalin at 10(-8) and 10(-7) M than those elsewhere in the same depot, but the maximum lipolysis, in the presence of 10(-5) M noradrenalin, was similar in all popliteal samples. These effects were increased by incubating adipose tissue explants for 24 h in tissue culture medium, suggesting autocrine amplification of the initial stimuli. Incubation with interleukin-4 (IL-4, 10 ng/ml) abolished the increase in lipolysis in samples around the activated lymph node and depressed it to below control values in other adipocytes. In vivo stimulation of the popliteal node increased maximum lipolysis in the presence of 10(-5) M noradrenalin in samples from around mesenteric lymph nodes and after 24 h incubation, in omental perinodal adipocytes. No effects of any pre-treatments were detected in perirenal adipocytes. We conclude that the adipocytes surrounding lymph nodes are actively involved in local, transient immune responses. Their participation may explain why most major lymph nodes are embedded in adipose tissue.

The effects of feeding suet-enriched chow on site-specific differences in the composition of triacylglycerol fatty acids in adipose tissue and its interactions in vitro with lymphoid cells.

The effects of diet on the composition and properties of adipose tissue in relation to lymph nodes were studied in adult guinea-pigs. The proportions of monoenoic triacylglycerol fatty acids were constant in all sites in adipose tissue of similarly fed guinea-pigs, but were substantially greater in samples from guinea-pigs fed on suet-enriched chow. Triacylglycerols in adipose tissue from near nodes contained significantly fewer saturated fatty acids, and significantly more 18:2n-6 and 18:3n-3 than those in samples from sites remote from nodes within the same depot. Depots that interact most strongly with lymphoid cells in vitro had the largest and most consistent within-depot differences. The gradients of triacylglycerol fatty acid composition with distance from lymph nodes in two small intermuscular depots were similar in guinea-pigs fed on plain or suet-enriched chow. These findings are consistent with the hypothesis that adipose tissue around lymph nodes is specialized for local interactions with the lymphoid cells therein, and help to explain the variability of serial or duplicate measurements of adipose tissue composition. When cultured alone, lipopolysaccharide-stimulated lymph node lymphoid cells from suet-fed guinea-pigs incorporated as much labelled thymidine as the controls. Adipose tissue explants from suet-fed guinea-pigs inhibited lymphocyte proliferation much less than those of the controls, although the site-specific differences were similar. The pattern of site-specific differences in glycerol released from explants incubated alone was generally similar for both dietary groups, but except in the popliteal depot, the increases following co-culturing with lymphoid cells were smaller for samples from suet-fed guinea-pigs. These experiments show that minor changes in the fatty acid composition of the diet can substantially alter the interactions between adipose tissue and lymphoid cells.

Interactions between adipose tissue around lymph nodes and lymphoid cells in vitro.

The functional relationships between lymphoid cells and the adipose tissue that surrounds lymph nodes were investigated in healthy adult guinea pigs. Lymphoid cells extracted from healthy adult guinea pigs were co-cultured for 48 h with adipose tissue explants from 18 sites defined by their anatomical relations to lymph nodes. Such explants from near a node suppressed lymphocyte proliferation stimulated with concanavalin A or lipopolysaccharide more than those from sites 5-10 mm from nodes. Inhibition was almost completely abolished by 500 microU insulin. The presence of lymphoid cells increased lipolysis (measured as glycerol release) in adipose tissue from all depots containing lymph nodes (i.e., except perirenal), especially in the presence of mitogens and with near-node samples from intermuscular and mesenteric depots. Inhibition of lymphocyte proliferation by adipose tissue was proportional to the additional lipolysis stimulated by the presence of lymphoid cells. For all depots except the mesenteric, glycerol release stimulated by lymphoid cells was inversely proportional to spontaneous lipolysis in adipose tissue cultured alone. These experiments demonstrate reciprocal interactions between lymphoid cells and adipose tissue, especially that around lymph nodes. The mediators of the action of adipose tissue on lymphoid cells probably include lipolytic products; mediators of the inverse effects are unknown.

Site-specific properties of human adipose depots homologous to those of other mammals.

1. Mean adipocyte volume, collagen content and the maximum activities of hexokinase and phosphofructokinase were measure in 15 depots of 8 men who were in good health until their sudden death. The data were used to establish homologies between depots in humans and other mammals. 2. The basic organization of adipose tissue in humans is similar to that of other mammals, although additional depots that are minimal or absent in rodents may be massive in humans. 3. Known site-specific properties relevant to manipulation of adipose tissue are reviewed.

The effects of exercise and feeding on the activity of lipoprotein lipase in nine different adipose depots of guinea pigs.

1. The activity of lipoprotein lipase (LPL) was measured in whole adipose tissue from 9 identified adipose depots of sedentary, fasting adult guinea pigs and following 30 min of exercise or voluntary ingestion of chow, and in adipocyte and stromal-vascular fractions from exercised specimens. 2. In sedentary, fasting specimens, LPL activity was up to 4 times higher in the small intermuscular depots than in the perirenal and epididymal depot (Table 1). 3. LPL activity increased significantly after feeding only in the large superficial depot, groin, and in the perirenal depot. LPL activity decreased after exercise only in the 2 intermuscular depots and in small anterior superficial depots. These effects of exercise were consistently greater in males than in females (Table 3). 4. Following exercise, there was up to twice as much LPL in the adipocytes as in the stromal-vascular fraction of the intermuscular depots, about 50% more in adipocytes from the minor superficial depots and about equal quantities in the 2 fractions of the intra-abdominal and groin depots (Table 2). 5. The data demonstrate the physiological inhomogeneity of both superficial and internal adipose depots, and are consistent with the hypothesis that LPL originating from adipose tissue may enter the circulation.

The effects of noradrenaline and insulin on lipolysis in adipocytes isolated from nine different adipose depots of guinea-pigs.

The rates of glycerol release in adipocytes isolated from nine identified adipose depots of sedentary or exercised guinea-pigs were measured in the presence of adenosine deaminase and 10(-9) to 10(-5) M noradrenaline and/or 1-1000 muunit/ml of bovine insulin. Twenty minutes of exercise increased the basal noradrenaline-stimulated rates of lipolysis in all depots, but these effects, and their interactions with in vitro application of the neurotransmitter differed between depots, showing that the long-lasting effects of exercise and the response to acute application of NA involve different mechanisms that may occur separately or together in different adipose depots. In general, large depots had the highest resting rates of lipolysis and the lowest responses to both noradrenaline and insulin, and lipolysis was only slightly different from the basal rate in adipocytes incubated with mixtures of the two agents. The two small intermuscular depots had the lowest unstimulated rates of lipolysis, but the fastest change and greatest maximum response to both agents. Noradrenaline-stimulated lipolysis was most effectively inhibited by small quantities of insulin in these depots. Different combinations of these properties were demonstrated in two smaller superficial depots, the mesenteric and omental depot, and in the cardiac depots. The data demonstrate the physiological inhomogeneity of both 'subcutaneous' and 'intra-abdominal' depots, and are consistent with the hypothesis that intermuscular adipose tissue interacts locally with adjacent muscle. Noradrenaline-stimulated lipolysis was more effectively inhibited by 100 muunit/ml insulin in adipocytes from the mesenteric and omental depot in those from any other site. A possible role for this property in the enlargement of this depot in hyperinsulinaemia in humans is proposed.

Dermatofibromas and arthropod bites: is there any evidence to link the two?

30 patients with dermatofibromas were questioned closely about contacts with stinging or blood-sucking arthropods before the development of their lesions. Their responses were not significantly different from those of a matched control group. 100 dermatofibromas were examined by dissection, histology, and after acid digestion of the collagenous material; no traces of arthropod skeletal tissues were found. These data are not consistent with the theory that the mechanical presence of arthropod tissues is necessarily, or even usually, a cause of dermatofibromas.

Adipose tissue in the mammalian heart and pericardium: structure, foetal development and biochemical properties.

1. The occurrence and relative abundance of adipose tissue around the heart and in the pericardium of wild and domesticated mammals are reviewed and some new data reported. 2. For macaque monkeys and a wide range of other adult mammals, the mean volume of epicardial adipocytes is constant at about half the average of that of other depots, although the relative mass of this depot is unrelated to the abundance of adipose tissue in the rest of the body. 3. In young adult guinea-pigs, the maximum rate of fatty acid synthesis is significantly higher in epicardial adipose tissue than that in the pericardial, perirenal and popliteal depots. 4. The rate of fatty acid release by epicardial adipose tissue is approximately twice that of the pericardial and perirenal depots. 5. The protein contents of guinea-pig epicardial and pericardial adipose tissue are similar, and are significantly higher than those of the perirenal and popliteal adipose tissue and there are no site-specific differences in the abundance of mitochondria. 6. In adult Macaca monkeys, the capacity of the epicardial adipose tissue for glucose utilization is about half that of the intra-abdominal depots. 7. The principal difference between epicardial adipose tissue and that elsewhere in the body is its greater capacity for fatty acid release. 8. It is suggested that cardiac adipose tissue may act as a local energy supply for adjacent myocardium and/or as a buffer against toxic levels of free fatty acids.

Biochemical correlates of the structural allometry and site-specific properties of mammalian adipose tissue.

1. The maximum activities of the glycolytic enzymes hexokinase (HK) and phosphofructokinase (PFK) were measured in defatted homogenates of adipose tissue from nine homologous depots of 57 wild and captive mammals belonging to 17 species and eight orders and differing in body mass by six orders of magnitude. 2. Site-specific differences in the enzyme activities were similar in all terrestrial species and were not consistently related to adipocyte volume. 3. The specimen-mean maximum activities of HK and PFK did not correlate with body mass, body composition or natural diet. 4. When specimens of different body composition and body mass were compared, glycolytic enzyme activity per adipocyte was directly proportional to adipocyte volume. 5. Site-specific differences in collagen content of adipose tissue did not correspond to those adipocyte volume. When homologous depots of different specimens were compared, the collagen content of adipose tissue was directly proportional to body mass. 6. Adipose tissue of large cetaceans contains more collagen than predicted from the allometric equations fitted to the data from terrestrial mammals. 7. Neither the scaling of the collagen content with body mass nor the site-specific differences in its abundance are consistent with a role as protection or support for adjacent tissues. 8. There are consistent site-specific differences in the extracellular components of adipose tissue as well as in the structure and metabolism of the adipocytes. 9. Adipose tissue differs from most other tissues in that its maximum metabolic capacities do not scale to body mass. 10. Adjustment of the biochemical activity of adipose tissue to changes in body mass and body composition must depend upon neural and endocrine controls, not upon intrinsic differences in its metabolic capabilities.

Site-specific and sex differences in the rates of fatty acid/triacylglycerol substrate cycling in adipose, tissue and muscle of sedentary and exercised dwarf hamsters (Phodopus sungorus).

Site-specific and sex differences in fatty acid/triacylglycerol substrate cycling in adipose tissue and muscle were found in vivo in adult hamsters (Phodopus sungorus) fed ad libitum, both at rest and immediately after moderate exercise. In adipose tissue, rates of fatty acid/triacylglycerol cycling at rest were highest in two small intermuscular depots and lowest in the groin, behind forelimb, epididymal and kidney sites. Cycling rates were significantly higher in the groin and in the two depots around the forelimb in females. Following an hour of exercise in a hamster ball, the rates of fatty acid/triacylglycerol cycling rose significantly in the intermuscular and certain superficial adipose fatty depots of both sexes, but the increases were more uniform in males. In females only, cycling rates in the skeletal muscles also increased significantly. The rates of fatty acid/triacylglycerol cycling in adipose tissue correlate closely with the site-specific activities of hexokinase and phosphofructokinase, but not with mean adipocyte volume per se. The data are consistent with the hypotheses that adipose depots are depleted selectively during exercise and that there are sex differences in the pattern of lipid mobilization.

Comparative aspects of hexokinase and phosphofructokinase activity in intermuscular adipose tissue.

1. The gross mass, mean adipocyte volume and activities of hexokinase (HK) and phosphofructokinase (PFK) were measured in adipose tissue from precisely identified intermuscular, superficial and intra-abdominal depots of 56 randomly collected wild and captive mammals and one bird. 2. In all intermuscular depots studied except that medial to the trapezius muscle, the activities of HK and PFK per adipocyte in adipose tissue in the centre of the depot were greater than in superficial and intra-abdominal depots of the same specimen. 3. These data are consistent with the suggestion that intermuscular adipose tissue may act as a local energy supply for adjacent muscles.

The effects of exercise and dietary restriction on the activities of hexokinase and phosphofructokinase in superficial, intra-abdominal and intermuscular adipose tissue of guinea-pigs.

1. The activities per gram of adipose tissue of hexokinase and phosphofructokinase were higher in the centre two intermuscular adipose depots of guinea-pigs than in any of the seven superficial and intra-abdominal depots studied. 2. The capacity for glucose utilization was reduced following about 3 months of regular exercise in all adipose depots studied except in the popliteal depot. The proportion of lipid decreased, and that of protein increased, in this intermuscular depot following regular, strenuous exercise, but it did not enlarge disproportionately. 3. We suggest that certain intermuscular adipose depots may act as a local energy source for adjacent muscles. The anatomical relations of adipose tissue to muscle may be an integral part of its physiological function.

The anatomy of adipose tissue in captive Macaca monkeys and its implications for human biology.

In a sample of 31 sedentary, ad libitum-fed monkeys, most specimens had less than 5% adipose tissue by weight. Total fatness correlated closely with the number of adipocytes per kilogram lean body mass, but not at all with mean adipocyte volume, except in specimens below 5% fat. The total number of adipocytes per kilogram of lean body mass increased more than tenfold in the most obese specimens. These data suggest that, like humans but in contrast to laboratory rodents, adipocyte proliferation, not adipocyte enlargement, is the chief mechanism of adipose tissue expansion except in very lean monkeys. Adipose tissue was found in all the typical mammalian depots and in the superficial abdominal paunch, which enlarged disproportionately in obese specimens, forming an almost continuous layer over most of the body. Site-specific differences in the activities of some glycolytic enzymes were similar to those of other mammals. Adipocytes in the paunch depot showed biochemical properties in common with those in the groin depots. The distribution and cellularity of adipose tissue in normal humans were similar to those of exceptionally obese monkeys. Many of the interspecific and sex differences can be attributed to the much greater abundance of adipose tissue in humans, and may not be associated with hair reduction or aquatic habits. Some minor changes in the size or shape of certain adipose depots may have arisen recently under sexual selection. The relevance of laboratory rodents as animal models of human obesity is assessed from comparison of the cellular structure, anatomical distribution and enzyme profiles of adipose tissue in monkeys with those of human and other mammals.

The effects of age, dietary restriction, exercise and maternity on the abundance and volume of adipocytes in twelve adipose depots of adult guinea-pigs.

1. The mean adipocyte volume and cellularity of adipose tissue in twelve anatomically defined depots were measured in 190 virgin and reproductive guinea-pigs aged 6 months to 2 years, maintained on five different regimens of diet and exercise. 2. The total adipocyte complement was constant up to the age of about 10 months and increased by 65-70% during the 2nd year of life. At constant percentage body-weight as fat, age-related accumulation of adipocytes was accompanied by reduction in average adipocyte volume. A period of high percentage body-weight as fat was not a necessary antecedent to age-related accumulation of adipocytes, and the effect was not observed in specimens that exercised regularly. 3. In guinea-pigs over 13 months old, changes in adipose tissue cellularity made a major contribution to total fatness. The mean volume of samples of adipocytes was a satisfactory indicator of fatness only in younger specimens in which adipose tissue cellularity was not changing. 4. The lean body mass and the total adipocyte complement were lower in guinea-pigs on the sedentary, restricted-diet regimen, whether the regimen began at the age of 19 weeks or 31 weeks, but the depots studied were not equally affected. Both moderate and strenuous exercise on an ad lib. diet caused a reduction in the percentage body-weight as fat in males but not in females. The total adipocyte complement and lean body mass were unchanged, although the males became almost as thin following exercise as those on the restricted-diet regimen. 5. Among guinea-pigs under 400 d old, there were no significant differences between males and virgin females in the site-specific volume of adipocytes relative to the size of those in other depots of the same specimen; there was only one significant difference in this index between the older and the younger specimens. The diet and exercise regimens modified the site-specific adipocyte volume relative to those in other depots; adipocytes under the trapezius muscle of the neck (UMN), at the interscapular depot (HUMP) and those in the depots anterior to the forelimb became relatively smaller following exercise, while those in the popliteal fat mass were relatively larger. Adipocytes in the UMN and HUMP also became more numerous relative to those in the other depots following both moderate and strenuous exercise. Those in the groin site, the intra-abdominal depots and the intermuscular depots became relatively smaller following dietary restriction without exercise.(ABSTRACT TRUNCATED AT 400 WORDS)

The effects of dietary restriction and exercise on the volume of adipocytes in two intra-orbital depots in the guinea-pig.

The volume of adipocytes in two intra-orbital sites and fourteen superficial and intra-abdominal sites, and the total adipocyte complement have been measured in virgin and reproductive guinea-pigs maintained on several different regimens of diet and exercise. The adipocytes around the ocular muscles at the back of the orbit (peripheral fat) are always larger than those just behind the eyeball (orbital fat). The adipocytes in both the intra-orbital sites are significantly larger in guinea-pigs whose total adipocyte complement is smaller than one standard deviation from the mean, than in those which have a normal-size or large adipocyte complement. The volume of intra-orbital adipocytes correlates very significantly with the volume of adipocytes in superficial and intra-abdominal sites in guinea-pigs which have large adipocyte complements, correlates weakly in those with normal adipocyte complements and not at all in those with small adipocyte complements. It is suggested that there may be fewer intra-orbital adipocytes in animals which have small adipocyte complements, and that, because the intra-orbital adipose tissue occupies a constant volume, the adipocytes in these sites become larger when they are less numerous.

The effects of food restriction and exercise on site-specific differences in adipocyte volume and adipose tissue cellularity in the guinea-pig. 2. Intermuscular sites.

Two intermuscular adipose depots, the popliteal mass (POP) and the fat under the gluteus superficialis muscle ( GLS ) were studied in guinea-pigs maintained under three different regimes of diet and exercise. Eleven different sites in POP and four sites in GLS were defined using associated muscle and fascia as 'landmarks'. Samples of twenty to twenty-five adipocytes from each site were removed and their mean volume measured; a consistent pattern of distribution of larger and smaller adipocytes within POP and GLS was found in all animals. The correlation between the volume of adipocytes from sites between the same pairs of muscles was substantially greater than the correlation between the volume of adipocytes from comparable sites between different muscles, whether the sites were within the same fat mass (POP) or in different fat masses (POP and GLS ). The volume of adipocytes from the edge of the fat mass correlated more closely with those from the edge of the same or a synergistic muscle than with the volume of cells from the centre sites or edge sites from an interface with another muscle. These effects were most pronounced in exercised animals and weak or insignificant in sedentary ad lib.-fed animals. The volume of adipocytes from edge sites correlated with the gross weight of the adjacent muscles in exercised animals only. It is concluded that muscle activity has a local effect on the volume of adjacent adipocytes. It is suggested that the mechanism involved is mechanical rather than biochemical.

The effects of food restriction and exercise on site-specific differences in adipocyte volume and adipose tissue cellularity in the guinea-pig. 1. Superficial and intra-abdominal sites.

The volume and number of adipocytes were measured in fourteen anatomical sites of adult guinea-pigs kept in small cages and fed ad lib., kept in small cages and on restricted diet or fed ad lib. and exercised. In the sedentary ad lib.-fed animals, there was no significant correlation between percentage body-weight as adipose tissue, as determined by direct dissection, and mean adipocyte volume based on samples from many different sites. The correlation was significant, though not high, for sedentary, restricted-diet animals and for exercised specimens. The correlations between the volume of adipocytes from left-right pairs of sites, and from sites around the same limb, were highly significant under all conditions studied. The correlation between the volume of adipocytes from sites other than left-right pairs of sites was weaker and in some cases statistically insignificant in sedentary ad lib.-fed guinea-pigs. The volume of adipocytes from sites in the groin region, the mesenteries and medial to the trapezius muscle failed to correlate in many cases with the volume of adipocytes from other sites sampled. The number of adipocytes at each site was similar in the exercised and sedentary ad lib.-fed animals. The restricted-diet, sedentary group had fewer adipocytes at all sites studied except the omental and mesenteric fat mass and the groin sites. It is suggested that moderate regular exercise or fasting gives rise to closer coordination between adipocytes at different sites because central factors regulating adipocyte volume become more prominent than local factors.