Differential gene expression between the porcine morula and blastocyst.

The survival and development of pre-implantation embryos are determinant factors affecting the outcome of animal reproduction. It is essential to transfer the expression of the genetic material from maternal sources, that is the ovum to the zygote before implantation to ensure successful development. Differentiation and transformation of blastomeres initiated during the morula and blastocyst stages is an important step of the embryonic development prior to implantation. We collected morula and early blastocyst samples from pure-bred Landrace pigs in vivo to study the differential gene expression patterns at these two stages. Total RNA was extracted from individual embryos and two rounds of amplification were employed. Two micrograms of antisense RNA, targets, were prepared and hybridized with each of four custom made oligo microarrays representing 24,000 porcine genes. The analyses of replicate hybridizations showed that among the 24,000 genes, 162 genes were expressed fivefold or greater in the morula compared to early blastocysts and 2126 genes were expressed fivefold or greater in early blastocysts compared to the morula. Of these differentially expressed genes, 1429 genes were functionally annotated with related human Gene Ontology terms. In addition to basic metabolic processes, genes related to signal transduction, transportation and cell differentiation were found in both stages and were up-regulated as embryo development proceeded. Real time polymerase chain reaction was utilized to quantify 12 genes differentially expressed in the 2 embryonic stages and validated the reliability of major evidences shown in microarrays. In conclusion, we have obtained a preliminary landscape of genes differentially expressed during the transition from morula to early blastocysts in pigs and showed a generally increased transcriptional activity, perhaps in preparation for implantation. Our results provide an opportunity to study the functions of these genes in relation to the development and survival of pre-implantation porcine embryos.

Studies on the biochemistry of mitochondria and cell morphology in the neonatal swine hepatocyte.

Mitochondrial preparations isolated from neonatal swine hepatocytes show a marked increase in oxidative and concomitant phosphorylative capacity between birth and 2 days postpartum. There are no changes in the coupling parameters (respiratory control ratio and adenosine diphosphate/O ratio) with age. Changes in sedimentation properties in a sucrose gradient suggest qualitative changes in the mitochondria. Some of the lipid measurements (increased phospholipid) might be interpreted as supportive of this suggestion, although most could also be regarded as indicative of quantitative changes (increased number of mitochondria). Electron microscopy of isolated mitochondria and of the hepatocyte demonstrated an increased number of mitochondria but no change in shape, size, or structure as the pig developed. An increase in a number of cytoplasmic components (Golgi apparatus and endoplasmic reticulum) and a decrease in glycogen were also observed. The functional changes in mitochondria seem to occur within a short period of time (6-12 hr postpartum).

Nutritional deprivation reduces the transcripts for transcription factors and adipocyte-characteristic proteins in porcine adipocytes.

For an organism to survive during nutritional deprivation, it must be able to regulate the genes involved in energy metabolism. White adipose tissue is an energy source during fasting conditions. In adipose tissue, transcription factors regulate several adipocyte-characteristic proteins involved in differentiation and energy metabolism. We investigated the transcript concentrations of two key transcription factors, as well as the transcript concentrations of several adipocyte-characteristic proteins, and genes involved in adipocyte energy metabolism in the adipose tissue of pigs fasted for 72 hours. Nutritional deprivation resulted in decreased transcript concentrations of the transcription factors, peroxisome proliferator-activated receptor gamma, and CCAAT-enhancer-binding protein alpha. The transcript concentrations of several adipocyte-characteristic proteins, fatty acid synthase, glucose transporter 4, lipoprotein lipase, leptin, and adipocyte fatty acid binding protein were also significantly reduced. The insulin receptor transcript concentration did not change. We conclude that these transcript concentration changes are aimed collectively at adjusting energy partitioning to conserve energy during nutritional deprivation, thereby enabling survival.

Expression of porcine adipocyte transcripts: tissue distribution and differentiation in vitro and in vivo.

Transcription factor transcripts implicated in adipocyte differentiation (peroxisome proliferator-activated receptor gamma (PPAR gamma), retinoid x receptor alpha (RXR alpha), adipocyte determination and differentiation-dependent factor 1 (ADD1), and CCAAT/enhancer binding protein alpha (C/EBP alpha)) and adipocyte-characteristic protein transcripts (lipoprotein lipase (LPL) and adipocyte fatty acid binding protein (aP2)) were measured in pig tissues. Transcripts for PPAR gamma, ADD1, and aP2 were localized in porcine subcutaneous and perirenal adipose tissues; transcripts for C/EBP alpha and LPL were detected in other tissues, but the greatest concentrations were in the adipose tissues. In porcine stromal-vascular cells (S/V cells) differentiating in vitro, transcripts for PPAR gamma and aP2 increased gradually, transcripts for ADD1, and LPL increased early and transcripts for C/EBP alpha increased late. In pigs, adipose tissue transcripts for PPAR gamma, ADD1, and LPL were minimal at birth and increased to 28 days postpartum, transcripts for C/EBP alpha were low until 28 days and transcripts for aP2 were at high levels, regardless of age. Although transcript development was somewhat different in vitro and in vivo, the data suggest PPAR gamma (and ADD1 are involved in regulation of transcripts for LPL and that there may be more partially differentiated precursor cells in S/V cells at day 0 than in adipose tissue at birth.

Expression of porcine adipocyte transcripts during differentiation in vitro and in vivo.

Transcript concentrations for the transcription factors, CCAAT enhancer binding protein beta and alpha (C/EBPbeta and C/EBPalpha), plus the adipocyte-characteristic proteins, fatty acid synthase (FAS), glucose transporter 4 (Glut 4), hormone-sensitive lipase (HSL), insulin receptor (InsR), lipoprotein lipase (LPL), and leptin were measured during differentiation of porcine stromal-vascular (S/V) cells in vitro. These same transcripts, excluding FAS and InsR, were measured in porcine adipose tissue from birth to 7 weeks of age. In S/V cells, C/EBPbeta and InsR were continuously elevated. At day 0, C/EBPalpha was approximately 20% of the day 9 value. The LPL increased gradually from day 0 to 9, whereas most other transcripts had a lag period of several days. In tissue, C/EBPbeta was substantial at birth and increased gradually. The C/EBPalpha was relatively low at birth and increased at day 17. The LPL and leptin increased continuously. The Glut 4 was low at birth and increased at day 28. The HSL was relatively low at birth, increased at day 10, and plateaued at day 28. Transcripts in porcine S/V cells develop somewhat differently from adipocyte differentiation models established in clonal cells, but the porcine cells represent a model that should be more applicable to pigs.

Effect of bovine somatotropin and food deprivation on beta-adrenergic and A1 adenosine receptor binding in adipose tissue of lactating cows.

Lactating Holstein cows were used to assess the effect of bovine somatotropin (bST; n = 8) and fasting (FAST; n = 4) on ligand binding to beta-adrenergic (BAR) and Type-1 adenosine (A1R) receptors in adipose tissue. Cows received exogenous bST (sometribove; 40 mg/d) or no hormone (control) for 4 d in a single-reversal design with a 7-d interval between treatment periods. Subcutaneous adipose tissue biopsies were taken on day 4 of each treatment. Eight d after the bST regimen, 4 cows were fasted for 3 d and adipose biopsies were taken. Ligand binding was quantified with a postnuclear, total adipose tissue membrane preparation (100,000 x g pellet). Binding to BAR and A1R was assessed with the antagonists [125I]iodocyanopindolol (ICP) and [3H]8-cyclopentyl-1,3-dipropylxanthine (DCPCX), respectively. The binding affinity (Kd) of BAR for ICP was not affected by bST but was enhanced by FAST; maximal binding (Bmax) was increased with bST treatment (P < 0.06) and reduced by FAST (61%, P < 0.01). Kd values for DCPCX binding to A1R were not changed by bST or FAST. bST did not affect Bmax for A1R; however, FAST reduced the Bmax by 38%. Data highlight the differential regulation of BAR and A1R by bST and FAST.

Comparison of plasma free-fatty-acid and blood-glycerol concentrations with measurement of lipolysis in porcine adipose tissue in vitro.

Rates of adipose tissue lipid metabolism in vitro are often measured to evaluate the function in vivo of metabolic pathways and thus appraise the accretion or loss of depot fat. This study directly addressed the comparison of degradative metabolism in vitro and in vivo. The concentrations of plasma free-fatty-acids and blood-glycerol as putative representatives of lipolysis in vivo and the lipolytic rate in adipose tissue in vitro obtained at the time of blood sampling were both measured in the same pig. Concentrations of plasma free-fatty-acids and blood-glycerol were increased or decreased by infusion of the norepinephrine analog, isoproterenol or by infusion of the adrenergic antagonist, propranolol, respectively. Although lipolytic rates and sensitivity to isoproterenol in vitro changed during some acute hormonal manipulations of the pig, the modulation in vitro was usually small relative to the large changes observed in plasma free-fatty-acid and blood-glycerol concentrations. Some of the subtle changes in vitro may reflect biological responses to hormone infusion, e.g., desensitization of the response to adrenergic agonists, but the magnitude of rate changes in vitro negates prediction of the rates in vivo from rates in vitro. Extrapolation of lipolytic rates in vitro and several adipose tissue anabolic rates obtained from the literature indicate the improbability for prediction of rates and degree of fat accretion in pigs from metabolic rates in vitro.

Postnatal expression of adipose tissue metabolic activity associated with a porcine genetic obesity.

To determine the age at which phenotypic expression of adipose tissue metabolic activities or adipocyte hypertrophy were detectable in a porcine obesity model, adipose tissue samples were obtained from genetically obese and lean pigs at 0, 7, 14, 21, 28, 42 and 56-d postpartum (weaning at 28 d). Adipocyte size was determined on osmium-fixed tissue slices using particle counter methods. Rates of glucose metabolism to CO2, total lipids and glyceride-fatty acids, as well as the basal and epinephrine plus theophylline-stimulated lipolytic rates were measured in tissue slices in vitro. Larger adipocytes were discernible in obese than lean pigs at birth and all subsequent postpartal ages. Expressed per cell, adipose tissue from obese pigs had greater glucose metabolism rates in vitro than that from lean pigs at 42 and 56 d; the rates were the same in obese and lean pigs at 28 d. Adipose tissue lipolytic rates in obese compared with lean pigs tended to be greater, were greater and were similar at 28, 42 and 56 d, respectively. These metabolic rates in vitro suggest that the early manifestation of the obesity evident as cell hypertrophy at birth and during early postnatal development is not the result of increased rates of adipose tissue glucose metabolism or decreased rates of adipose tissue lipid degradation.

Overview of the effects of beta-adrenergic receptor agonists on animal growth including mechanisms of action.

The beta-adrenergic receptors (beta-AR) are present on the surface of almost every type of mammalian cell. These receptors are stimulated physiologically by the neurotransmitter, norepinephrine and the adrenal medullary hormone, epinephrine. There are three subtypes of beta-AR, namely, beta1-AR, beta2-AR, and beta3-AR; the pharmacological and physiological responses of an individual cell result from the particular mixture of the three beta-AR subtypes present on that cell. Species-specific structure (amino acid sequence) also causes modification of the function of a given beta-AR subtype. Knowledge of the beta-AR subtypes present in various cell types, coupled with knowledge of receptor structure (sequence), will allow an understanding of the complexity of physiological function regulated by beta-AR. Oral administration of some beta-AR agonists increases muscle and decreases fat accretion in cattle, pigs, poultry, and sheep. The large number of physiological functions controlled by beta-AR suggests that the mechanism(s) for the observed changes in carcass composition may be extremely complex. Any proposed mechanism must begin with the possibility of direct effects of the agonist on skeletal muscle and adipocyte beta-AR. However, many other mechanisms, such as modification of blood flow, release of hormones, or central nervous system control of feed intake may contribute to the overall effects observed with a given beta-AR agonist in a given species. Furthermore, the pharmacodynamic properties of a particular agonist are complex and expected to vary among species as well as within the same species at different ages or when fed different diets.

Evidence of classic beta 3-adrenergic receptors in porcine adipocytes.

Adipocytes from several mammalian species have predominant beta 3-adrenergic receptors (beta 3-AR). Attempts to classify porcine adipocyte beta-adrenergic receptors (beta-AR) into subtypes have not been successful. Selectivity of agonists and antagonists for stimulation of lipolysis and for ligand binding is considerably more restrictive than for the classic rat and guinea pig beta-AR subtypes. The unique pattern for activity of agonists and antagonists in porcine beta-AR precludes analogy to classic receptors and consequently there is no conclusive evidence regarding porcine beta-AR subtypes. Porcine adipocyte membranes were used in ligand binding experiments designed specifically to demonstrate beta 3-AR. Equilibrium saturation curves with dihydroalprenolol, CGP 12,177, or iodocyanopindolol indicated saturation at low concentrations with a single binding site. Equilibrium competitive ligand binding with iodocyanopindolol as radioligand and isoproterenol or propranolol as competitive ligands indicated both ligands totally inhibited radioligand binding; propranolol was more potent than isoproterenol. Nonradioactive CGP 12,177 also competed with iodocyanopindolol. Ligand binding experiments provided no evidence of a low-affinity beta-AR (binding at high concentrations of ligand), the beta 3-AR. Positive evidences of a beta 3-AR were that CGP 12,177, a beta 1- and beta 2-adrenergic receptor antagonist but a beta 3-AR agonist, partially stimulated porcine adipocyte lipolysis. Furthermore, transcripts for a beta 3-AR, as well as a beta 1- and beta 2-AR, have been demonstrated in RNA from porcine adipocytes in other studies. The beta-AR subtypes expressed and functional in porcine adipocytes remain unknown. The multiple ligand binding sites cannot be attributed to classic beta-AR subtypes. The porcine beta-AR may be a single unique receptor to impart atypical binding properties, or more likely, multiple subtypes, each different enough from classic subtypes to impart the unique properties observed.

Adipose tissue lipolytic rate in genetically obese and lean swine.

In vitro lipolytic rate was determined in adipose tissue from genetically obese and lean pigs. There were about 20 pigs/genetic strain at 25 and 80 kg and 10 pigs/strain at 50 kg body weight. When expressed on a cellular basis, the in vitro adipose tissue basal (no exogenous hormone) lipolytic rate was similar in obese and lean pigs at 25 and 50 kg body weight. At 80 kg body weight the basal rate was greater in obese than in lean pigs. The in vitro adipose tissue epinephrine-stimulated lipolytic rate expressed on a cell basis was greater at 25 kg, was similar at 50 kg body weight and tended (P less than or equal to .1) to be greater at 80 kg in obese compared with lean pigs. The in vitro sensitivity of lipolysis to epinephrine was slightly greater in lean compared with obese pigs. The data obtained in vitro indicate that obese pigs do not have low adipose tissue lipolytic rates compared with lean pigs. Consequently, adipose tissue lipolysis does not appear to be a major metabolic factor leading to the excessive fat accretion in these obese pigs.

Acute changes in blood flow in pigs infused with beta-adrenergic agonists.

Previous results indicate that clenbuterol decreases carcass adipose tissue accretion when administered to pigs but does not appear to stimulate the adipose tissue beta-adrenergic receptor. Clenbuterol increases plasma free fatty acid concentration when acutely infused in vivo, suggesting an indirect affect. One possible indirect effect is that clenbuterol could change blood flow to adipose tissue. Blood flow was measured with radiolabeled microspheres in tissues from pigs before and after infusion of a beta-adrenergic agonist for 30 min. High probability levels (up to P less than .2) were used to indicate trends due to extreme variability. Infusion of isoproterenol increased heart rate, plasma free fatty acid concentration and blood flow at many adipose tissue sites and at a few skeletal muscle sites. Infusion of isoproterenol decreased blood pressure. Infusion of clenbuterol increased heart rate and tended to increase blood flow to several skin and adipose tissue sites slightly. The results suggest that increased adipose tissue blood flow may contribute to the accelerated release of free fatty acids when clenbuterol is infused acutely in vivo.

Influence of infused beta-adrenergic agonists on porcine blood metabolites and catecholamines.

Anesthetized pigs were infused sequentially with increased concentrations of beta-adrenergic agonists. At selected times during infusion, blood pressure, heart rate and plasma concentrations of free fatty acids (FFA), glycerol, glucose, lactate, norepinephrine, epinephrine and dopamine were measured. Azaperone, a drug used to calm the pigs before anesthesia, caused hypotension and bradycardia but did not affect plasma metabolites. Infusion of norepinephrine, epinephrine, isoproterenol or clenbuterol produced changes in plasma metabolites and plasma catecholamines. These changes during norepinephrine infusion were attributed to the infused agonist, whereas those during epinephrine infusion might have resulted to some extent from release of norepinephrine. Plasma isoproterenol was not quantified because it interfered with the assay of epinephrine and dopamine so that it was not possible to distinguish between infused isoproterenol and release of endogenous epinephrine and dopamine. Infusion of clenbuterol caused a small increase in plasma norepinephrine so that some of the increase in plasma FFA, glycerol and lactate during clenbuterol infusion may result from release of endogenous norepinephrine.

Lipoprotein and hormone-sensitive lipases in porcine adipose tissue.

Lipoprotein lipase (LPL) is an adipocyte enzyme that cleaves fatty acids from circulating lipoproteins. Fatty acids enter the cell to be oxidized or esterified. Hormone-sensitive lipase (HSL) is an adipocyte enzyme that cleaves fatty acids from intracellular triacylglycerol. The HSL is activated by phosphorylation. Assays for the two lipases are complex because the hydrophobic substrate, triacylglycerol, must be presented as a gum-based suspension or as a detergent-based emulsion to a relatively hydrophilic enzyme. A convenient, stable glycerol/phospholipid suspension of the substrate was used for measurement of porcine adipose tissue LPL and HSL in vitro. This substrate was excellent for LPL. It produced rates five times those using a more complex and less convenient gum-based substrate suspension. The LPL activity was released by heparin, had a pH optimum of approximately 8.5, was activated by serum, and was inhibited by NaCl and protamine. This LPL assay measures enzyme capacity. The same substrate was used to measure an adipose tissue lipase activity that had a pH optimum below 7, was not activated by serum, and was not inhibited by NaCl or protamine. These are all characteristics of HSL. Despite the convenience, this substrate was not appropriate for porcine adipose tissue HSL because the rates were only 30 to 50% of those produced with a more complex, less convenient gum-based substrate suspension. Furthermore, incubation of enzyme or tissue slices with insulin, or agents that elevate cAMP concentration, did not modulate this lipase activity, as expected. These incubations poorly modulated LPL activity.

Effect of sex on lipogenic activity in swine adipose tissue.

Adipose tissue was obtained from male, female and male pigs castrated either at birth, 2 or 4 mo of age. Pigs were biopsied at 11 and 20 wk of age to obtain adipose tissue samples. Lipogenic capacity was assessed by measurement of in vitro glucose incorporation into lipids of adipose tissue slices. At 20 wk of age, males were less fat than females or males castrated either at birth or 20 mo of age. Adipocyte volume at 20 wk of age was smaller in males, females and recently castrated males (4 mo) than males castrated at birth or 2 mo of age. Lipogenic rate at 20 wk of age was lower in males than in castrated pigs; females had intermediate lipogenic rates. The results provide a partial metabolic explanation for the difference in subcutaneous fat deposition in the sexually diverse groups. There was no effect of estradiol-17 beta or testosterone in vitro on adipose tissue lipogenesis, suggesting that sex hormone effects in vivo may not be involved in short-term regulation of lipogenesis.

Effect of limited feed intake on growth of subcutaneous adipose tissue layers and on carcass composition in swine.

Little is known about the function of individual backfat layers in swine. Our objective in this study was to evaluate differential growth of each backfat layer in pigs fed different amounts of the same diet. Relationships of growth of backfat layers to deposition of carcass protein and ether extract were also studied. Feed intake levels were ad libitum, 92.5% of ad libitum, and 85% of ad libitum for 36 barrows averaging 95 d of age and 37.8 kg of weight at the start of the study. Chemical composition of the carcass soft tissue was determined at the end of the 84-d study. Daily gain tended to decrease (P less than .10) and conversion of feed to gain improved (P less than .15) as feed intake decreased. The daily depositions (g/d) of protein and ether extract were 62.9 and 205 for the ad libitum group, 61.1 and 176 for the 92.5% intake group, and 59.3 and 153 for the 85% intake group, respectively. The compositional effect of limiting feed intake for 84 d was to produce less carcass soft tissue of greater lean content. Overall backfat depths were 31.5, 29.6, and 27.8 mm for the ad libitum, 92.5, and 85% intake groups, respectively. The outer backfat layer differed among groups; the 92.5 and 85% groups were thinner than the ad libitum group. Relative to the ad libitum group, the change in depth of the middle backfat layer was 86 and 75% for the 92.5 and 85% intake groups (P less than .01). The inner backfat layer was not affected significantly by level of feed intake.(ABSTRACT TRUNCATED AT 250 WORDS)

Beta-adrenergic receptor binding in crude porcine adipose tissue plasma membranes.

Methods have been detailed to prepare a crude membrane fraction from isolated porcine adipose tissue cells. Adipocytes were obtained after incubation of 5 g of adipose tissue slices with 4,500 units of a selected lot of collagenase in a total volume of 15 mL at 37 degrees C for 90 min. There was no bovine serum albumin present during cell isolation because albumin did not enhance cell yield or yield of lipolytic activity. Isolated cells were lysed by exposure to hypotonic conditions in the presence of 7.5 mM ethylene glycol tetraacetic acid (EGTA) and .8 mM phenylmethylsulfonyl fluoride (PMSF). A 30,000 x g centrifugal pellet was used as the crude membrane preparation. Binding of tritiated dihydroalprenolol (DHA), a beta-adrenergic antagonist, was measured in the presence of 7.5 mM EGTA and .2 mM PMSF, because these protease inhibitors improved specific binding by approximately 50% to greater than 150 fmol/mg of protein and decreased non-specific binding to less than 10% at 2.5 nM DHA.

Beta-adrenergic receptor subtype transcripts in porcine adipose tissue.

Expression of transcripts for beta 1-, beta 2-, and beta 3-adrenergic receptors (beta 1-AR, beta 2-AR, and beta 3-AR, respectively) was investigated in porcine tissues. Northern blot analysis of total RNA indicated hybridization of the rat beta 1-AR probe to transcripts of approximately 3.0-kb in heart and lung and 2.9-kb in perirenal, omental, and subcutaneous adipose tissue. Hybridization of the hamster beta 2-AR probe with total RNA revealed transcripts of approximately 1.8-kb in the heart, 2.0-kb in the lung, and 2.0-kb in perirenal, omental, and subcutaneous adipose tissue; a minor transcript of 1.8-kb was also observed in the perirenal tissue. Northern blot analysis of poly (A+) RNA from porcine adipose tissue with the mouse beta 3-AR probe revealed no detectable signals in heart and lung but detected a major 2.2-kb transcript and a minor 1.9-kb transcript in subcutaneous adipose tissue. The expression of porcine beta 1-AR and beta 2-AR transcripts was compared in perirenal, omental, and subcutaneous adipose depots. The expression of beta 1-AR transcripts was four to five times greater in the perirenal depot than in the omental depot and two to three times greater in the subcutaneous depot than in the omental depot. The expression of beta 2-AR mRNA seemed to be the same in all three depots.

Variables in estimation of adipocyte size and number with a particle counter.

Porcine adipose tissue slices were fixed with osmium tetroxide and cells released by treatment with urea. Cell size and number were determined by an instrumental method using a particle counter. Storage of adipose tissue samples as frozen slices or in osmium, in post-osmium saline, in urea or in post-urea Triton all tended to produce less acceptable results than obtained with fresh tissue slices. Various storage conditions either tended to diminish cell size, to produce small particles or to cause aggregation. Repeatabilities of cell number, cell diameter and cell volume from multiple samples obtained from one anatomical location within an animal (perfect repeatability = 1) were .39, .52 and .65, respectively. Repeatabilities of instrument determinations were greater than .98 for cell number, cell diameter and cell volume. Cell number may be estimated from particle counts or indirectly from average size. Particle count number and that calculated from mean cell diameter were similar, whereas cell numbers estimated from mean cell volumes were smaller. Different adipose tissue depots and backfat layers had divergent cell size, making extrapolation to whole-animal cell number complex.

Distribution and quantification of beta1-, beta2-, and beta3-adrenergic receptor subtype transcripts in porcine tissues.

Distribution and concentration of beta-adrenergic receptor (betaAR) subtype transcripts (beta1AR, beta2AR, and beta3AR) were investigated in porcine and rat tissues. Reverse transcription (RT) coupled with PCR indicated the presence of beta1AR and beta2AR transcripts in porcine left ventricle, lung, longissimus muscle, and subcutaneous adipose tissue. The RT-PCR indicated that beta3AR transcripts were present in porcine subcutaneous adipose tissue, and transcripts were suggested in left ventricle and lung, but they were not detected in longissimus muscle. Quantitative ribonuclease protection assays indicated that the porcine beta1AR, beta2AR, and beta3AR transcript concentrations (amol/microg of total RNA) were, respectively as follows: heart left ventricle = 6.1, 2.4, and .02; lung = 3.8, 1.9, and .01; liver = 1.7, 2.1, and undetectable; skeletal muscle = 1.7, 1.1, .02; subcutaneous adipose tissue = 1.3, .4, .14. The proportions of porcine betaAR subtypes (beta1AR:beta2AR:beta3AR) were as follows: heart left ventricle = 72:28:.25; lung = 67:33:.2; liver = 45:55:0; skeletal muscle = 60:39:.7; and subcutaneous adipose = 73:20:7. Normalization of data to beta-actin transcript concentration did not change these relationships. Rat perigonadal adipose tissue beta1AR and beta3AR transcript concentrations were, respectively, .59 and 1.84 amol/microg of total RNA. The rat perigonadal adipose tissue concentration of beta3AR transcript was tenfold that of porcine subcutaneous adipose tissue. The predominant betaAR transcript in rat adipose tissue was beta3AR, and the predominant betaAR transcript in pig adipose tissue was beta1AR.