Review: Animal model and the current understanding of molecule dynamics of adipogenesis.

Among several potential animal models that can be used for adipogenic studies, Wagyu cattle is the one that presents unique molecular mechanisms underlying the deposit of substantial amounts of intramuscular fat. As such, this review is focused on current knowledge of such mechanisms related to adipose tissue deposition using Wagyu cattle as model. So abundant is the lipid accumulation in the skeletal muscles of these animals that in many cases, the muscle cross-sectional area appears more white (adipose tissue) than red (muscle fibers). This enhanced marbling accumulation is morphologically similar to that seen in numerous skeletal muscle dysfunctions, disease states and myopathies; this might indicate cross-similar mechanisms between such dysfunctions and fat deposition in Wagyu breed. Animal models can be used not only for a better understanding of fat deposition in livestock, but also as models to an increased comprehension on molecular mechanisms behind human conditions. This revision underlies some of the complex molecular processes of fat deposition in animals.

INVITED REVIEW: Evolution of meat animal growth research during the past 50 years: Adipose and muscle stem cells.

If one were to compare today's animal growth research to research from a mere 50 yr ago, one would see programs with few similarities. The evolution of this research from whole-animal through cell-based and finally molecular and genomic studies has been enhanced by the identification, isolation, and in vitro evaluation of adipose- and muscle-derived stem cells. This paper will highlight the struggles and the milestones that make this evolving area of research what it is today. The contribution of adipose and muscle stem cell research to development and growth, tissue regeneration, and final carcass composition are reviewed.

Cell supermarket: adipose tissue as a source of stem cells.

Adipose tissue is derived from numerous sources, and in recent years this tissue has been shown to provide numerous cells from what seemingly was a population of homogeneous adipocytes. Considering the types of cells that adipose tissue-derived cells may form, these cells may be useful in a variety of clinical and scientific applications. The focus of this paper is to reflect on this area of research and to provide a list of potential (future) research areas.

Obesity, metabolic syndrome, and adipocytes.

Obesity and metabolic syndromes are examples whereby excess energy consumption and energy flux disruptions are causative agents of increased fatness. Because other, as yet elucidated, cellular factors may be involved and because potential treatments of these metabolic problems involve systemic agents that are not adipose depot-specific in their actions, should we be thinking of adipose depot-specific (cellular) treatments for these problems? For sure, whether treating obesity or metabolic syndrome, the characteristics of all adipose depot-specific adipocytes and stromal vascular cells should be considered. The focus of this paper is to begin to align metabolic dysfunctions with specific characteristics of adipocytes.

The effect of ractopamine hydrochloride on gene expression in adipose tissues of finishing pigs.

The long-term effect of feeding the catecholamine analog ractopamine (RAC; ractopamine hydrochloride, Elanco Animal Health, Indianapolis, IN) on the expression of genes involved in energy and lipid metabolism in subcutaneous adipose tissue was studied. Large White pigs (84 kg) were fed corn- and soybean meal-based diets supplemented with 0, 20, or 60 mg/kg of RAC for 14, 28, or 42 d. Expression (mRNA abundance) in adipose tissue of sterol regulatory binding protein-1 (SREBP-1), PPARα, PPARγ2, fatty acid synthase (FAS), glucose transporter 4 (GLUT4), and stearoyl-CoA desaturase was determined by Northern blotting. Feed intakes did not differ, and RAC (20 and 60 mg/kg) improved BW gain at d 14, 28, and 42 (P < 0.05) and increased loin eye area (measured on d 42 only; P < 0.05). Expression of SREBP-1 and PPARγ2 declined (P < 0.05) with RAC by d 28 and 42, whereas expression of PPARα was increased (P < 0.05) on d 14, 28, and 42. After 14 d, expression of FAS and GLUT4 was decreased (P < 0.05) with 60 mg/kg of RAC, whereas both RAC concentrations attenuated FAS expression on d 28 and 42. Overall, adipose tissue stearoyl-CoA desaturase expression was not affected by RAC but showed somewhat less expression (P < 0.15) on d 28 at 60 mg/kg of RAC. Although prolonged, chronic RAC feeding most likely downregulates adipose tissue membrane ß-adrenergic receptors, mRNA abundances of anabolic lipid metabolism transcription factors, glucose transporters, and enzymes (SREBP-1, PPARγ2, FAS, GLUT4) were still attenuated up to d 42. Conversely, a transcription factor related to oxidative metabolism expression (PPARα) was enhanced. We conclude that even after 42 d, RAC still decreased expression of lipogenic genes in adipose tissue by yet undefined cyclic adenosine monophosphate-directed mechanisms, but in contemporary lean pigs, this effect is likely of limited practical significance.

Perspectives on the formation of an interdisciplinary research team.

As research funding becomes more competitive, it will be imperative for researchers to break the mentality of a single laboratory/single research focus and develop an interdisciplinary research team aimed at addressing real world challenges. Members of this team may be at the same institution, may be found regionally, or may be international. However, all must share the same passion for a topic that is bigger than any individual's research focus. Moreover, special consideration should be given to the professional development issues of junior faculty participating in interdisciplinary research teams. While participation may be "humbling" at times, the sheer volume of research progress that may be achieved through interdisciplinary collaboration, even in light of a short supply of grant dollars, is remarkable.

Effect of the degree and duration of early dietary amino acid restrictions on subsequent and overall pig performance and physical and sensory characteristics of pork.

The objective of this study was to investigate the effect of the degree and duration of early dietary AA restrictions on subsequent and overall pig performance and physical and sensory characteristics of pork. For the grower (G) and finisher-1 (F1) phases, 3 corn-soybean meal diets were formulated to contain 100, 80, or 60% of the 1998 NRC total Lys recommendations (100G, 80G, or 60G, and 100F1, 80F1, or 60F1, for the G and F1 phases, respectively). For the finisher-2 (F2) phase, a common corn-soybean meal diet was formulated to satisfy the 1998 NRC total Lys recommendation. Thirty gilts and 30 castrated males (2 gilts or 2 castrated males/pen) were randomly assigned to 5 dietary treatments (100G-100F1, 80G-100F1, 80G-80F1, 60G-100F1, and 60G-60F1) when BW was 22.7 +/- 0.3 kg. Pigs were switched to F1 and F2 diets at 50.7 +/- 0.4 and 79.9 +/- 0.5 kg of BW, respectively. Pigs had ad libitum access to feed and water. All pigs were slaughtered at 110.7 +/- 0.5 kg of BW, and LM samples were collected. Pigs fed the 60G diet had less (P < or = 0.05) ADG during the G phase and greater (P < or = 0.05) ultrasound backfat (UBF) at the end of the G phase than those fed the 100G diet. The ADG decreased linearly (R(2) = 0.70; P < 0.001) as the degree of AA restrictions became more severe. Although serum total protein (TP) and albumin concentrations in pigs fed the 60G-100F1 diets were less (P < or = 0.05) than those fed the 100G-100F1 diets at the end of the G phase, TP concentration was similar between the 2 groups at the end of the F1 phase. Likewise, ADG during the F1 phase and UBF at the end of the F1 phase in pigs fed the 60G-100F1 diets were similar to those fed the 100G-100F1 diets. Feeding the 80G diet resulted in numerically decreased ADG during the G phase, but there was no difference in ADG during the F1 and F2 phases or UBF at the end of F1 and F2 phases between pigs fed the 80G and 100G diets. Overall, pigs fed the 80G-80F1 diets had similar ADG, but less (P < or = 0.05) fat-free lean gain (LG) than those fed the 100G-100F1 diets. These pigs also had less (P < or = 0.05) serum TP and albumin concentrations than pigs fed the 100G-100F1 diets throughout the study. Pigs fed the 60G-60F1 diets had less (P < or = 0.05) overall ADG and G:F and less (P < or = 0.05) LM area and LG than those fed the 100G-100F1 diets. However, they had a greater (P < or = 0.05) subjective marbling score than those fed the 100G-100F1 diets. The results indicated that pigs fed the 80G-80F1 diets may have exhibited compensatory growth in BW gain, but not in terms of lean accretion. Growth performance and carcass traits of pigs fed the 60G-60F1 diets were reduced, indicating that the restriction may have been too severe or too long or both. Early dietary AA restrictions had no clear effect on physical and sensory characteristics of pork.

Board-invited review: the biology and regulation of preadipocytes and adipocytes in meat animals.

The quality and value of the carcass in domestic meat animals are reflected in its protein and fat content. Preadipocytes and adipocytes are important in establishing the overall fatness of a carcass, as well as being the main contributors to the marbling component needed for consumer preference of meat products. Although some fat accumulation is essential, any excess fat that is deposited into adipose depots other than the marbling fraction is energetically unfavorable and reduces efficiency of production. Hence, this review is focused on current knowledge about the biology and regulation of the important cells of adipose tissue: preadipocytes and adipocytes.

The effect of dietary copper supplementation on fatty acid profile and oxidative stability of adipose depots in Boer x Spanish goats.

A feeding trial was designed to examine the effects of copper sulfate pentahydrate (CuSO(4).5H(2)O) on the fatty acid composition and oxidative stability in muscle and adipose tissues of Boer x Spanish goat kids. Fifteen (n = 5 per treatment) goats were fed 0, 100, or 200 mg of supplemental Cu per day as copper sulfate for 98 d. The animals were slaughtered, and LM, s.c. adipose from the sternal region, and mesenteric adipose tissues were collected. Total lipids were extracted with chloroform:methanol (2:1), methylated and isolated via GLC from all tissues. The subsequent peaks were then positively identified by mass spectrometry. Thiobarbituric acid-reactive substances were measured also. In s.c. adipose, dietary Cu significantly decreased C14:0 (P = 0.03) and C16:0 (P = 0.01). In muscle, C15:0 (P = 0.03) was linearly increased by Cu. Dietary Cu supplementation did not influence oxidative stability in goat muscle or s.c. adipose. Copper supplementation at 200 mg/d resulted in a significant increase in malondialdehyde in mesenteric adipose (P = 0.01) compared with the 0 or 100 mg/d groups. These results indicate that lipid composition may differ from depot to depot and that depending on the depot, dietary Cu seems to elicit a variable response on the fatty acid composition.

Measuring in vivo intracellular protein degradation rates in animal systems.

Continual synthesis and breakdown or remodeling of proteins (also called protein turnover) is a principal characteristic of protein metabolism. During animal production, the net differences between synthesis and breakdown represent the actual marketable muscle foods. Because protein synthesis is a highly end-ergonic and protein breakdown is metabolic energy dependent, efficiency of production can be markedly enhanced by lower muscle protein breakdown rates. Herein, various methodological approaches to studying protein breakdown, with particular emphasis toward food-producing animals, are presented. These include whole-animal tracer AA infusions in vivo, quantifying marker AA release from muscle proteins, and in vitro AA release-based methodologies. From such methods, protein synthesis rates and protein breakdown rates (mass units/time) may be obtained. The applications of such methods and innovations based on traditional methods are discussed. Whole-animal in vivo approaches are resource intensive and often not easily applied to high-throughput metabolic screening. Over the last 25 yr, biochemical mechanisms and molecular regulation of protein biosynthesis and protein breakdown have been extensively documented. Proteolysis is dependent in part on the extent of expression of genes for components of cellular proteolytic machinery during skeletal muscle atrophy. It is proposed that high-throughput methods, based on emerging understanding about protein breakdown, may be useful in enhancing production efficiency.

Use of bovine somatotropin for increased skeletal and lean tissue growth of Holstein steers.

One hundred-sixty Holstein growing-finishing steers (initial BW of 185 kg) were blocked by BW to determine the effectiveness of long-term bovine somatotropin (bST) administration on lean, skeletal, and carcass measurements. Steers were randomly allocated to 4 treatments (10 steers/treatment) within a block (n = 4 blocks). Treatments were control, no bST (C-C); bST from d 0 to 182 (bST-C); bST from d 183 to slaughter (C-bST); and bST from d 0 to slaughter (bST-bST). Steers received a s.c. injection of placebo or bST at 14-d intervals. Doses were 320 mg of bST/injection from d 0 to 112 and 640 mg of bST/injection from d 113 to slaughter. The last treatment was administered 31 d before slaughter. Steers received a 14% CP (DM basis) diet from d 0 to 182 and 11.5% CP from d 183 to slaughter that consisted of dry, whole-shelled corn and a pelleted protein-mineral supplement. Steers were slaughtered when BW per block averaged 615 kg (d 325, 353, 367, and 381 for the 4 blocks, respectively). Thirty steers were removed from the study because of poor performance with respect to their pen mates, illness, lameness, death, incomplete castration, and incorrect treatment. Serum IGF-I concentrations increased 151% (P < 0.01) from d 7 through 35 in bST-treated steers compared with control steers. During the first 182 d, bST-C and bST-bST steers were heavier (P < 0.01) and had greater (P < 0.01) ADG, G:F, hip height, and hip height gain compared with C-C and C-bST steers. From d 183 to slaughter, C-bST steers had reduced (P < 0.05) daily DMI and greater G:F than bST-C steers. At final slaughter, C-bST and bST-bST steers had greater (P < 0.05) hip height than C-C steers. Noncarcass weight was increased and dressing percent reduced (P < 0.05) in C-bST and bST-bST steers compared with C-C steers. Quality grade was least (P < 0.05) in bST-bST carcasses compared with C-C, whereas bST-C and C-bST carcasses were intermediate. At final slaughter, steers receiving bST had greater (P < 0.05) carcass protein and water composition and lower (P < 0.05) carcass lipid and lipid accretion than C-C steers. Bovine somatotropin was effective in reducing carcass fat and increasing edible lean. Administering bST to young, lightweight steers increased skeletal growth and noncarcass weight without an increase in total carcass weight, but decreased carcass quality.

Beta-adrenergic agonist hyperplastic effect is associated with increased fibronectin gene expression and not mitogen-activated protein kinase modulation in C2C12 cells.

Beta-adrenergic agonists (beta-AA) enhance protein accretion in skeletal muscles. This stimulation is characterized by increased protein synthesis, increased expression of myofibrillar protein genes and a depression in protein degradation in animals, and increased proliferation and DNA synthesis in muscle cells in vitro. The mechanism or signal path in muscle whereby beta-AA would elicit these physiological effects upon binding to the G protein-coupled beta-adrenergic receptor (beta-AR) is unclear. C2C12 myoblasts were used to determine beta-AR ligand binding characteristics, cyclic AMP synthesis in response to isoproterenol (ISO) stimulation, and effects of ISO on DNA synthesis, mitogen activated protein kinase (MAPK), and fibronectin (FN) gene expression. Results showed that C2C12 cells possess beta-AR which are specific, saturable, and of high affinity (Kd = 0.2 nM). Forskolin and ISO stimulated cAMP production by = 20-fold (P<0.001) and 17-fold (P<0.001), respectively. ISO and the cAMP analog, 8-bromo-cAMP (8-BC) stimulated DNA synthesis in proliferating cells by 150% (P<0.05) and 200% (P<0.01), respectively, without modulating MAPK activity, whereas addition of fetal bovine serum to culture resulted in a 500% increase (P<0.01) in DNA synthesis and MAPK activation. DNA synthesis in C2C12 cells treated with ISO, 8-BC, or FBS was abolished in the presence of 25 microM PD098059, an MAPK-kinase inhibitor, suggesting that an MAPK-dependent pathway is likely involved in C2C12 proliferation. During cAMP elevating agent stimulation, basal MAPK activity may be sufficient, in the presence of other putative signaling molecules, to support proliferation in these cells. ISO or 8-BC treatment increased FN mRNA by three- and seven-fold, respectively, in growing C2C12 cells implying a connection between increased DNA synthesis and FN gene expression.

Arachidonic acid inhibits lipogenic gene expression in 3T3-L1 adipocytes through a prostanoid pathway.

This report examines the effect of polyunsaturated fatty acids (PUFA) on lipogenic gene expression in cultured 3T3-L1 adipocytes. Arachidonic acid (20:4, n-6) and eicosapentaenoic acid (20:5, n-3) suppressed mRNAs encoding fatty acid synthase (FAS) and S14, but had no effect on beta-actin. Using a clonal adipocyte cell line containing a stably integrated S14CAT fusion gene, oleic acid (18:1, n-9), arachidonic acid (20:4, n-6) and eicosapentaenoic acid (20:5, n-3) inhibited chloramphenicol acetyltransferase (CAT) activity with an ED50 of 800, 50, and 400 microM, respectively. Given the high potency of 20:4, n-6, its effect on adipocyte gene expression was characterized. Arachidonic acid suppressed basal CAT activity, but did not affect glucocorticoid-mediated induction of S14CAT expression. The effect of 20:4, n-6 on S14CAT expression was blocked by an inhibitor of cyclooxygenase implicating involvement of prostanoids. Prostaglandins (PGE2 and PGF2alpha at 10 microM) inhibited CAT activity through a pertussis toxin-sensitive Gi/Go-coupled signalling cascade. Our results suggest that 20:4, n-6 inhibits lipogenic gene expression in 3T3-L1 adipocytes through a prostanoid pathway. This mechanism of control differs from the polyunsaturated fatty acid-mediated suppression of hepatic lipogenic gene expression.

Protein and fat metabolism in cows given somavubove before parturition.

Forty-one Holstein cows were injected with 0, 5, or 14 mg/d of bST for the last 46 +/- 6 d before parturition. Compared with data for controls, the 5- and 14-mg doses of bST increased apparent protein synthesis about 16% before parturition. Exogenous bST before parturition increased apparent protein degradation 30% during wk 1 after parturition. During wk 1 of lactation, 14 mg of bST also increased milk protein yield 33%. No treatment differences were present in concentration of serum NEFA, body condition score, or thickness of subcutaneous fat. Therefore, administration of bST before parturition did not alter metabolism of subcutaneous fat. Prepartum treatment with 5 and 14 mg of bST increased and maintained serum somatotropin at 6.5 and 22.7 ng/ml, respectively, compared with 1.6 ng/ml in controls. Concentrations of serum IGF-I were initially increased but were not maintained as parturition approached. On d -23, IGF binding protein 3 was increased 65% but was not different among groups by d -7. For groups administered the 5 and 14 mg/d of bST, IGF binding protein 2 was decreased 40%. Administration of bST before parturition increased protein reserves and stimulated milk protein yield for 1 wk but did not alter metabolism of subcutaneous fat. Furthermore, energy balance appeared to be a major regulator of concentrations of IGF binding protein 3 and responsiveness of IGF-I to exogenous somatotropin before parturition.

RNA transcription in porcine skeletal muscle nuclei during postnatal development.

Postnatal developmental pretranslational regulation of skeletal muscle alpha-actin gene expression was investigated. Northern blot analysis of skeletal muscle alpha-actin and beta-tubulin mRNA from 1- and 28-day-old pigs indicated that there are developmental increases in alpha-actin mRNA abundance (P < 0.03) and no significant changes in beta-tubulin mRNA (P > 0.1). A system for isolation of nuclei from porcine skeletal muscle and for transcriptional "run-on" analysis was established in order to investigate the regulatory mechanism of developmental changes in porcine skeletal muscle protein. Skeletal muscle nuclei were isolated from longissimus dorsi (LD) muscle of 1- and 28-day-old pigs by adapting a method to isolate nuclei from cardiac muscle. Results from a [3H]-UTP incorporation assay indicate that these nuclei preparations have the capacity to synthesize RNA and attain maximum incorporation after 40-45 min at 26 degrees C. Messenger RNA syntheses from skeletal muscle nuclei from 1- and 28-day-old pigs were not significantly different (P > 0.25). All nascent tRNA, rRNA, and mRNA in the nuclei were elongated since [3H]-UTP incorporation was reduced after addition of 0.05 micrograms/ml alpha-amanitin to the transcription mixture. Transcription "run-on" assay results indicated that more (P < 0.02) skeletal muscle alpha-actin pre-mRNA was synthesized in the 28-day-old pig skeletal muscle nuclei than in the 1-day-old pig skeletal muscle nuclei. These results indicate that the relative increase in skeletal muscle alpha-actin mRNA observed in the older animals was due, at least in part, to an increase in the transcriptional activity of the skeletal muscle alpha-actin gene.

Skeletal muscle growth and protein turnover in neonatal boars and barrows.

The objective of this study was to determine the effect of castration, within 24 h after birth, on skeletal muscle growth and protein metabolism in neonatal pigs at 1, 2, and 4 wk of age. Four additional pigs were slaughtered at birth to obtain initial body composition. All other pigs were infused with [14C]tyrosine for 6 h before slaughter to determine in vivo fractional protein synthesis rates (FSR). At slaughter, muscle bundles were removed from the semitendinosus and incubated with [3H]tyrosine to determine in vitro protein synthesis rates. Nucleic acids and protein were determined on the semitendinosus muscle. Testosterone concentrations, determined at weekly intervals, peaked in boars at 3 wk of age. Castration at birth did not affect combined weights of the semitendinosus, longissimus, triceps brachii, and brachialis muscles. Likewise, neither in vitro protein synthesis rates nor in vivo FSR were affected by castration. However, a developmental decline in in vivo FSR and in vitro protein synthesis rates occurred from 1 to 4 wk. Neither concentrations nor total quantity of protein, RNA, or DNA in the semitendinosus muscle differed between neonatal boars and barrows at any age. Concentrations of DNA and RNA at 4 wk were two- and threefold lower, respectively, than at birth. Protein:DNA and protein:RNA ratios increased three- and sixfold, respectively, from birth to 4 wk. Testosterone concentrations had little effect on skeletal muscle growth and protein turnover rates during this neonatal period.

Skeletal muscle growth and expression of skeletal muscle alpha-actin mRNA and insulin-like growth factor I mRNA in pigs during feeding and withdrawal of ractopamine.

Sixty crossbred barrows were used to study the effect of ractopamine (a phenethanolamine/beta-adrenergic agonist) treatment and its withdrawal on muscle growth and on the relative abundance of skeletal muscle alpha-actin (sk-alpha-actin) mRNA and of liver and longissimus muscle IGF-I mRNA at 4 wk. Ractopamine was fed (20 ppm) for periods of 2, 4, and 6 wk (six pigs per group). Additional pigs (four per group) were fed ractopamine (20 ppm) for 6 wk and then slaughtered 1, 3, and 7 d after withdrawal of ractopamine. Ractopamine increased (P < .05) longisimus muscle weight and protein content, although protein concentrations were not different. The increased muscle weight and protein content attained by feeding ractopamine for 6 wk was retained when ractopamine was withdrawn. The RNA and DNA concentrations did not change, whereas total DNA and RNA content per muscle was 18 and 26.7% greater, respectively, in ractopamine-treated pigs at 4 wk, but there were no differences at 2 or 6 wk or among the withdrawal groups. The relative abundance of sk-alpha-actin mRNA in the longissimus muscle was 41 and 62% greater (P < .05) in treated animals at 2 and 4 wk but was similar to that in controls at 6 wk and during the withdrawal period. The relative abundance of IGF-I mRNA in liver and longissimus muscle was not altered with ractopamine treatment for 4 wk. These results indicate that the ractopamine-enhanced muscle growth may result from increased myofibrillar gene expression at the pretranslational level, which is maximal with short-term treatment of ractopamine.(ABSTRACT TRUNCATED AT 250 WORDS)

Fatty acid profiles, lipogenesis, and lipolysis in lipid depots in finishing pigs treated with recombinant porcine somatotropin.

Thirty-two pigs (average weight, 74 kg) were used to determine the effects of recombinant porcine somatotropin (rpST) and dietary CP concentration on carcass and noncarcass tissues. Pigs were injected intramuscularly with either rpST (50 micrograms.kg BW-1.d-1; n = 16) or vehicle (n = 16) at 0900 for 24 d. Half the treated and control pigs were given ad libitum access to either a 14 or 20% CP corn-soybean meal diet. The left side of the carcass was physically dissected into separable skin, soft tissues, and bone. Tissue samples were obtained for enzyme assays, proximate analysis, and fatty acid profiles. Proximate analysis (fat, water, protein) of adipose tissue (AT) samples indicated that rpST decreased the percentage of ether extractable lipid (P < .01) and increased the percentage of protein and water (P < .05, P < .01) in intermuscular (IM), subcutaneous (SC), and intrafascicular (IF) AT depots and slightly altered fatty acid profiles of longissimus muscle IF and SC AT. Fatty acid synthesis and malic enzyme activity were decreased by rpST (P < .01), suggesting that lipogenesis was decreased; however, lipolysis was unaffected. Pigs fed the high-CP diet (20%) had decreased AT malic enzyme activity (P < .05) and fatty acid synthesis (P < .05). Additionally, pigs fed the high-CP diet had greater kidney weights (P < .01). Heart, liver, and kidney weights were heavier (P < .01) in pigs treated with rpST, whereas skin and total bone weight were unaffected. Neither weights nor lengths of four individual long bones were affected by dietary protein concentration or administration of rpST.

Alpha-2 adrenergic receptor activity in porcine adipocytes is androgen and age/cell size dependent.

1. Using epinephrine plus propranolol we demonstrated alpha-2 adrenergic receptor (alpha 2AR) activity (49% inhibition of theophylline stimulated lipolysis) in adipocytes of adult intact male pigs (61 kg body wt and 97 microns adipocyte diameter). 2. Dose titration with an alpha 2AR antagonist (yohimbine) confirmed alpha 2AR associated activity. 3. No alpha 2AR activity was observed in younger male or castrated male pigs. 4. The inhibitory action on lipolysis via the alpha 2AR in pigs is dependent on androgen status and adipocyte size or age.