Search PubMed⌕ Search

Biomedical subjects

R D Green

Publications and source records attributed to R D Green.

At least 37 records · Page 2Linked to original sources
PubMed1995

Determination and cellular localization of adenylyl cyclase isozymes expressed in embryonic chick heart.

Mammalian heart has been reported to express AC isozymes (types V and VI) that are inhibited by < microM [Ca2+]; avian heart has been reported to express adenylyl cyclase activity that is inhibited by < microM [Ca2]. We have used reverse transcription polymerase chain reaction (RT-PCR) to determine that type V and VI AC mRNAs are present in freshly isolated ventricular myocytes. Subsequent RNase protection assays revealed that that the type V signal is 4-5 times that for the type VI isozyme. In situ hybridization with high specific activity cRNA probes combined with immunocytochemistry with a chick anti-myosin antibody was used to probe the cellular origins of type V and type VI AC signals. These studies show that myocytes contain messages for both the type V and VI isozymes but that AC V is the major isoform. Interestingly, while the type V AC mRNA appears to be localized primarily, if not exclusively, in myocytes, the signal for type AC VI mRNA in non-myocytes is stronger than in myocytes.

Adenylyl Cyclases↗
PubMed1995

Isolation and characterization of an avian A1 adenosine receptor gene and a related cDNA clone.

We have isolated the gene for a chick A1 adenosine receptor along with a cDNA that codes for the same adenosine receptor. The cDNA clone was isolated from both adipose tissue and heart cDNA libraries and encodes a 324-amino acid protein with 80% identity with mammalian A1 adenosine receptors. Transient expression of the cDNA in human embryonic kidney (HEK) 293 cells shows that it encodes a protein that binds [3H]CCPA (2-chloro-N6-[cyclopentyl-2,3,4,5-3H]cyclopentyladenosine, a specific agonist radioligand, with a KD of 5.6 +/- 2.4 nM. Cyclic AMP measurements in HEK 293 cells co-transfected with the chick cDNA and a cDNA for a luteinizing hormone/choriogonadotropin receptor shows that A1 adenosine receptor agonists antagonize the cyclic AMP-elevating effect of bovine luteinizing hormone. Two partial genomic clones were isolated. The first contains 5'-untranslated sequence including a putative promoter region which does not contain a TATA box, an intron and the first third of the coding sequence of the A1 adenosine receptor cDNA. The coding sequence of this partial genomic clone terminates at a second intron. The second partial genomic clone contains the rest of the coding sequence and the 3'-untranslated elements in a single exon. Thus the chick A1 adenosine receptor gene contains one intron in the 5'-untranslated region and a minimum of one intron in the coding sequence.

Adenosine↗
PubMed1995

Real-time ultrasonic measurement of fat thickness and longissimus muscle area: I. Description of age and weight effects.

Serial ultrasonic measures of fat thickness (FTU) and longissimus muscle area (LMU) were taken on 180 feedlot steers representing 11 sire-breed groups at two 60-d intervals. Cattle then were slaughtered in four groups of 45 at 21-d intervals with ultrasonic measures taken at each interval. Analyses of variance indicated weight (WT) effects (P < .001) for all FTU measures and many of the LMU measures. Age was a source of variation (P < .05) for FTU and LMU over the first five measurement dates. Sire-breed (SBD) effects (P < .001) were prevalent for all FTU estimates and early LMU predictions. Some dam line and SBD x dam line interactions were detected (P < .1) for the FTU and LMU estimates. Age effects were described quadratically (P < .001) when FTU and LMU were regressed on age (R2 = 47 and 68%, respectively). When FTU and LMU were regressed on WT, effects were described quadratically (P < .001) with R2 values of 46 and 72%, respectively. Animals then were assigned to one of four biological types (BT) based on percentage of retail product (0-mm fat trim) using discriminant cluster analysis. Within BT, ultrasonic variables were regressed on age. Quadratic effects (P < .01) were detected for both FTU and LMU. Weight also contributed to variation observed in both ultrasonically measured variables for all BT (P < .001). These data indicate that changes in fat and muscle size as affected by age and weight are detectable using serially collected ultrasound measures. Furthermore, the age and weight relationships of predicted FTA and LMA varied across the four biological types. This suggests a need for derivation of biological type-specific age and(or) weight adjustment equations for cattle evaluation purposes.

Adipose Tissue↗
PubMed1995

Real-time ultrasonic measurement of fat thickness and longissimus muscle area: II. Relationship between real-time ultrasound measures and carcass retail yield.

Feedlot steers (n = 180) representing 11 sire-breed groups were ultrasonically measured for fat thickness (FTU) and longissimus muscle area (LMU) at two 60-d intervals during the feeding period and four 21-d intervals corresponding to serial slaughter dates to predict carcass retail yield parameters. Two fat trim levels, 8 and 0 mm, were used to calculate percentage of trimmable fat (FAT8P and FAT0P) and retail product percentage (RPD8P and RPD0P) for each carcass. Regression techniques were used to evaluate best-fit equations that explained variation in retail product components. When FAT8P, FAT0P, RPD8P, and RPD0P were regressed on USDA yield grade (YG), R2 values ranged from 75 to 76% (P < .001). Comparatively, when live animal predictors of YG components (FTU, LMU, and final live weight) were used as the independent variables, equations predicting retail yield had R2 values of 61 to 65% (P < .01). Equations using final FTU as the independent variable explained 58 to 64% (P < .001) of the variation in FAT8P, FAT0P, RPD8P, and RPD0P. Equations with FTU, LMU, and either WT, AGE, marbling, or quality grade resulted in R2 values similar to those with only FTU, indicating the strong influence of fat on retail yields. These results indicate that ultrasonic predictors explained about 10% less variation in retail product percentage than did carcass measures.

Adipose Tissue↗
PubMed1994

Chromosomal localization of the callipyge gene in sheep (Ovis aries) using bovine DNA markers.

A mutation causing muscular hypertrophy, with associated leanness and improved feed efficiency, has been recently identified in domestic sheep (Ovis aries). Preliminary results indicate that an autosomal dominant gene may be responsible for this economically advantageous trait. We have exploited the conservation in sequence and chromosomal location of DNA markers across Bovidae to map the corresponding callipyge locus to ovine chromosome 18 using a battery of bovine chromosome 21 markers. Chromosomal localization of the ovine callipyge locus is the first step toward positional cloning of the corresponding gene.

Animals↗
PubMed1994

Adenosine receptor-mediated inhibition of cardiac adenylyl cyclase activity may involve multiple receptor subtypes.

We previously reported that the adenosine receptor agonist N6-phenylisopropyladenosine (R-PIA) inhibits adenylyl cyclase in detergent-permeabilized embryonic chick ventricular myocytes in the presence of the adrenergic receptor agonist, isoproterenol (Ma and Green 1992). The slope of the dose response curve of this inhibition is very shallow (nH 0.3-0.4). The present studies on detergent-permeabilized chick myocytes evaluate the mechanisms underlying this shallow inhibition curve. We find that in contrast to R-PIA, two additional adenosine receptor agonists, N6-cyclopentyladenosine (CPA) and 2-chloro-N6-cyclopentyladenosine (CCPA), inhibit cardiac adenylyl cyclase activity in a monophasic, dose-dependent manner (nH approximately 1). Two A1 adenosine receptor antagonists, 8-cyclopentyl-1,3,-dipropylxanthine (CPX) and 3-(4-amino)phenethyl-1-propyl-8-cyclopentylxanthine (BW-A884U) affect the R-PIA responses differently. BW-A884U shifts the R-PIA dose response curve to the right in a parallel fashion while CPX both shifts the R-PIA response curve and increases its steepness. Cardiac A1 adenosine receptors were further characterized using one antagonist ([3H]CPX) and two agonist ([3H]R-PIA and [3H]CCPA) radioligands. [3H]CPX binds to the adenosine receptors in detergent-permeabilized ventricular myocytes with a Kd value of 3.3 +/- 0.2 nM and a BMAX value of 30.1 +/- 2.4 fmol/mg protein (means +/- SEM; N = 4). [3H]R-PIA detects more sites than [3H]CCPA (22.8 +/- 4.0 and 8.3 +/- 1.3 fmol/mg protein, respectively; GTP-free conditions). CPA and CCPA inhibit [3H]R-PIA binding in a shallow, dose-dependent manner (nH approximately 0.4), while R-PIA and CPA inhibit [3H]CCPA binding with a nH approximately 1.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenylyl Cyclase Inhibitors↗
PubMed1994

Health needs of homeless clients accessing nursing care at a free clinic.

Homeless people need emotional support, caring professionals, and resources that work within their frame of reality. Each homeless community will have different demographics and health needs. The common denominators are the obstacles encountered when trying to access health care: bureaucracy, transportation, accessibility, present-time orientation, limited resources, frustration, and anxiety. Nurse-managed free clinics are an effective means of serving the needs of homeless and indigent populations. Through these clinics, clients can receive continuity of care, proper follow-up, referrals, and teaching to aid in the prevention of illness. A free clinic was established in a northwestern Georgia community to meet the health needs of homeless and indigent persons. This article describes the demographics, attitudes, values, perceived support systems, and health needs of clients accessing care at this nursing clinic.

Adult↗
PubMed1993

Calcium entry via L-type calcium channels acts as a negative regulator of adenylyl cyclase activity and cyclic AMP levels in cardiac myocytes.

It is well established that the inotropic effect of beta-adrenergic agonists is mediated by the stimulation of adenylyl cyclase activity and the subsequent phosphorylation of specific proteins by cAMP-dependent protein kinase. The L-type calcium channel is believed to be one of the proteins phosphorylated; the phosphorylation of calcium channels is believed to increase calcium entry into myocytes, which is, at least in part, responsible for the positive inotropic effect. The present studies show that the cAMP-elevating effect of isoproterenol is increased as extracellular calcium is lowered and that calcium channel blockers potentiate the cAMP-elevating effect of isoproterenol in the presence in extracellular calcium. This effect is not dependent on effects on cAMP catabolism and is not specific for beta-adrenergic receptors, because the cAMP-elevating effect of forskolin is similarly affected. Measurements of adenylyl cyclase activity in cardiac membranes show that submicromolar Ca2+ concentrations directly inhibit adenylyl cyclase activity. These results demonstrate that increased entry of Ca2+ via L-type calcium channels in response to beta-adrenergic receptor stimulation acts as a negative regulator of the effect of beta receptor stimulation on adenylyl cyclase activity.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗
PubMed1992

Evaluation of ultrasonic estimates of carcass fat thickness and longissimus muscle area in beef cattle.

Yearling crossbred feedlot steers (n = 495) and heifers (n = 151) were ultrasonically measured at the 12-13th rib interface 24 h before slaughter to evaluate the accuracy of ultrasonic measurements of fat thickness (BFU) and longissimus muscle area (LMAU) for prediction of actual carcass measures. Isonification was with an Aloka 210DX ultrasound unit equipped with a 12.5-cm, 3.0-MHz, linear array transducer by two technicians. Carcass fat thickness (BFC) and longissimus muscle area (LMAC) were measured 48 h postmortem. Differences between ultrasonic and actual carcass measures were expressed in actual (BFDIFF and LMADIFF) and in absolute (magnitude of BFDIFF and magnitude of LMADIFF) terms for backfat and longissimus muscle area, respectively. When expressed as percentages of the actual carcass measures, the average absolute differences indicated error rates of 20.6% for backfat and 9.4% for longissimus muscle area. Average actual differences (BFDIFF and LMADIFF) indicated that underprediction occurred more often than overprediction for both measures. The BFU was within .25 cm of BFC 70% of the time, and LMAU was within 6.5 cm2 of LMAC 53% of the time. Ultrasound measurements BFU and LMAU more accurately predicted BFC and LMAC in thinner and more lightly muscled cattle, respectively. Simple correlation coefficients between ultrasonic and carcass measures were .75 (P less than .01) for BF and .60 (P less than .01) for LMA. Analyses of variance of absolute differences between ultrasonic and carcass measures indicated no significant differences to exist between technicians. Predictive accuracy of ultrasonic measures did not change as the level of experience of technicians increased during the study.(ABSTRACT TRUNCATED AT 250 WORDS)

Adipose Tissue↗
PubMed1992

Ultrasonic prediction of carcass merit in beef cattle: evaluation of technician effects on ultrasonic estimates of carcass fat thickness and longissimus muscle area.

The objective of this study was to determine technician effects of live animal ultrasonic estimates of fat thickness (FTU) and longissimus muscle area (LMAU). Steers (n = 36) representing four breed-types (Brown Swiss, Average Zebu-cross Mexican, Corriente Mexican, and typical British crossbred) of commercial slaughter cattle were isonified to estimate accuracy and repeatability of fat thickness (FT) and longissimus muscle area (LMA) measurements by two experienced technicians. Repeated measures of FTU and LMAU were taken by technicians on two consecutive days with an Aloka 500V ultrasound unit equipped with a 3.5-MHz, 172-mm scanning width, linear-array transducer. Ultrasonic estimates of fat thickness and LMAU were taken at the 12th and 13th rib interface 48 h before slaughter; carcass fat thickness (FTC) and longissimus muscle area (LMAC) were measured 48 h postmortem. Means for FTU, FTC, LMAU, and LMAC were .91 +/- .36 cm, .82 +/- .40 cm, 70.7 +/- 9.43 cm2, and 72.4 +/- 8.9 cm2, respectively. Ultrasound and carcass measures of FT and LMA were different (P less than .01) among breed-types but were not different (P greater than .10) between technicians or for technician x breed-type interactions. Pooled simple correlation coefficients (P less than .01) were .87 and .86 between FTU and FTC and .76 and .82 between LMAU and LMAC for Technicians 1 and 2, respectively. Repeatabilities estimated by intraclass correlation methods were .91 +/- .03 and .81 +/- .06 for images repeated over 2 d and .95 +/- .02 and .83 +/- .05 for images repeated by two technicians for FT and LMA, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Adipose Tissue↗
PubMed1992

Modulation of cardiac cyclic AMP metabolism by adenosine receptor agonists and antagonists.

The mechanism(s) underlying adenosine receptor-mediated modulation of cardiac cAMP levels has been investigated using detergent-permeabilized embryonic chick ventricular myocytes. The beta-adrenergic receptor agonist isoproterenol (ISO) stimulated adenylyl cyclase activity in detergent-permeabilized cells by 5-10-fold, with an EC50 value of 0.3 microM. Three adenosine receptor agonists, (R)-N6-phenylisopropyladenosine, N6-(3-iodo-4-aminobenzyl)adenosine, and 5'-N-ethylcarboxamidoadenosine, inhibited ISO (10 microM)-stimulated adenylyl cyclase activity in a concentration-dependent manner. The maximum inhibition of the ISO-stimulated adenylyl cyclase activity by (R)-N6-phenylisopropyladenosine (10 microM) was 30-40%. This inhibition was antagonized by the adenosine receptor antagonists xanthine amine congener and 8-cyclopentyl-1,3-dipropylxanthine and was abolished by pertussis toxin treatment, suggesting that the inhibition of adenylyl cyclase activity is mediated by A1 adenosine receptors acting via a pertussis toxin-sensitive guanine nucleotide-binding protein (G protein). Because the adenosine receptor agonists had no detectable effect on phosphodiesterase activity, the adenosine receptor-mediated inhibition of adenylyl cyclase activity appears to account for the cAMP-lowering effect of adenosine receptor agonists seen in intact cardiac myocytes. Moreover, two A1 adenosine receptor antagonists, 8-cyclopentyl-1,3-dipropylxanthine and 3-(4-amino)phenethyl-1-propyl-8-cyclopentylxanthine, stimulated basal adenylyl cyclase activity in the absence of an adenosine receptor agonist; this stimulation was abolished by pretreatment of the cells with pertussis toxin. We postulate that "precoupled" A1 adenosine receptor-G protein complexes, present in the cardiac myocytes, exert a tonic inhibitory influence on adenylyl cyclase activity and that some adenosine receptor antagonists remove this tonic inhibition by destabilizing these precoupled receptor-G protein complexes.

Adenylyl Cyclases↗
PubMed1991

Apparent heterogeneity of cardiac A1 adenosine receptors as revealed by radioligand binding experiments on N-ethylmaleimide-treated membranes.

While G protein-coupled receptors are often studied by analyzing antagonist radioligand: "cold" agonist inhibition curves using an independent site model, it is now clear that KL and KH values determined in these analyses are not reliable estimates of the affinities of the agonists for "free" and G protein-coupled forms of the receptor. Thus, such experiments cannot be used to contrast the characteristics of a given type of receptor in different tissues, i.e., to probe for the existence of receptor subtypes. Since treatment with N-ethylmaleimide treatment blocks receptor: Gi/Go protein interactions, such analyses on N-ethylmaleimide-pretreated membranes should allow direct assessment of the affinities of competing ligands for the free receptor or for multiple receptor subtypes. As A1 adenosine receptors couple to Gi, and perhaps to Go, we have performed A1 adenosine receptor radioligand "competition" studies first on control, then on N-ethylmaleimide-pretreated bovine cardiac and cerebral cortical membranes. Results of experiments with the antagonist radioligand [3H]xanthine amine congener appeared to be confounded by ligand binding to A2 adenosine receptors present in the cardiac membrane preparations. Further experiments utilized the A1-specific radioligand [3H]1,3-dipropyl-8-cyclopentylxanthine. These experiments confirmed once more that the KL values determined by computer analysis of "competition" curves performed on control membranes are not reliable estimates of the affinities of the competing ligand for free receptors.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine↗
PubMed1991

Output/input differences among nonpregnant, lactating Bos indicus-Bos taurus and Bos taurus-Bos taurus F1 cross cows.

Nonpregnant F1 crossbred cows, progeny of either Hereford (H) or Angus (A) dams and sired by Brahman (Bm), Sahiwal (Sw), Pinzgauer (Pz), H, or A sires, were fed to maintain initial weight while rearing Charolais (C)-sired progeny for a period of 126 d in drylot commencing at about 48 d postpartum. Cow-calf pairs were assigned to equalize cow age, calf sex, and breed of cow's dam among three replicate pens of approximately 12 pairs each. Cows and calves were weighed every 2 wk and feed intake was adjusted to minimize change in cow weight. Metabolizable energy (ME) consumption for zero cow weight change was estimated by regression. Milk production was estimated by weight-suckle-weigh at 58, 85, 125, and 170 d of lactation. Calf gain (GAIN, kg) relative to cow weight (CWT1, kg) was higher (P less than .01) for calves from Bm-X (139.5/585) and Sw-X (132.2/534) than for calves from Pz-X (127.2/552) and HA-X (116.9/547) cows. Estimated mean daily production of milk was 7.40, 7.15, 7.28, and 6.37 kg for the Bm-X, Sw-X, Pz-X, and HA-X, respectively. Total cow ME intake (TMEcow) for breed groups ranked (P less than .05) with cow size and milk production, and calf creep-feed intake (FMEcalf) was inversely related to estimated milk intake. Proportion of total feed ME (TMEcow+calf) consumed by calves was higher (P less than .05) for HA-X cows (18%) than for the others (14%). Total efficiency of calf gain in weight (GAIN/TMEcow+calf) was 11% greater (P less than .05) for crossbred cows of Bos indicus X Bos taurus (Bm-X, Sw-X) than for Bos taurus X Bos taurus (Pz-X, HA-X) cows (35 vs 32 g/Mcal) in the 126-d lactation period.

Adipose Tissue↗
PubMed1991

Life-cycle biological efficiency of Bos indicus x Bos taurus and Bos taurus crossbred cow-calf production to weaning.

A deterministic model was developed that accounted for all biological inputs and outputs for a theoretical herd of F1 females at age equilibrium mated to produce three-way terminal-cross calves and for the required proportion of straightbred cows needed to produce replacements. Two Bos indicus x Bos taurus vs two Bos taurus x Bos taurus types of crossbred cows were compared in the production environment of south-central Nebraska. The four types of F1 females were from Hereford (H) or Angus (A) dams and by H or A (HA), Pinzgauer (Pz), Brahman (Bm), or Sahiwal (Sw) sires. The crossbred females were assumed mated to Red Poll (R) sires for their first calving and to Simmental (S) sires thereafter. Two evaluations of efficiency for each of the four breeding systems were total cow and calf feed energy input 1) per unit of only weaned calf weight output (CALFEFF, Mcal/kg) and 2) per unit of weaned calf plus .55 x cull cow weight output (TVALEFF, Mcal/kg). Results for a terminal age of 7 yr in systems using HA, Pz, Bm, and Sw crossbred cows, respectively, were 64.9, 64.5, 60.9, and 59.3 Mcal/kg for CALFEFF and 45.7, 46.4, 44.1, and 43.7 Mcal/kg for TVALEFF. Changing terminal age to 11 yr reduced CALFEFF about 6% but increased TVALEFF about 7%, because total inputs increased more than output value (10 vs 3%) from 7 to 11 yr terminal ages. These results suggest differences in efficiency among these breed crosses favoring the Bos indicus crossbred cows by over 4% in this particular environment.

Animal Feed↗
PubMed1990

Embryo-transfer twinning and performance efficiency in beef production.

Effects of twinning on efficiency of beef production were estimated from results of bilateral transfer of two Angus x Hereford (AxH) embryos into each of 241 heifers and 84 cows (H, A, HxA or Holstein x H) over 4 yr. Calves were weaned at 180 d and fed either 220 d in a feedlot (1977) or 170 d on forage and 140 d in a feedlot (1978 to 1980). Effects of parity, twinning and sex of calf were estimated as covariates within year-breed of dam. Pregnancy at 45 to 60 d of gestation was 68% in heifers (H) and 74% in cows (C), with 40% single (S) and 60% twin (T) births. Dystocia was 28% in H vs 10% in C (P less than .05), and tended to be less (P greater than .05) for T than S in H. More placentas were retained (P less than .05) for T than for S in both H (35 vs 12%) and C (24 vs 4%). Twin gestations averaged 3 d shorter and subsequent calving intervals 13 d longer (P less than .05), but total calf mortality was slightly higher (P greater than .05). Abortions were 4% in H only. Twinning females lost maternal weight during late gestation (P less than .05) when crowding limited voluntary feed intake, while fetal requirements were 60% higher (P less than .01). Twins increased milk output 25% (P less than .05), but 11% higher feed intake maintained cow weight during lactation. Twinning reduced birth weight 13% and weaning weight 17% (P less than .05), but 400-d feedlot weight only 9% because of compensating feedlot gain. Twins gained 18% faster than S during postweaning 170-d forage feeding, but 5% slower in feedlot to 8% lighter 490-d weight (P less than .05). Assuming 40% higher veterinary and labor costs for twins, estimated integrated herd costs per unit of age-constant output value would be lower for T than for S production by about 24% for marketing either at weaning or at 400 d.

Abortion, Veterinary↗
PubMed1990

Analysis of agonist-antagonist interactions at A1 adenosine receptors.

Previous work from our laboratory using sucrose gradient centrifugation and the antagonist radioligand [3H]xanthine amine congener led us to propose that A1 adenosine receptors are coupled to a GTP-binding protein (G protein) in the absence of an agonist and that adenosine receptor antagonists bind to free uncoupled receptors with high affinity and coupled receptors with low affinity and cause a destabilization of receptor-G protein complexes [Mol. Pharmacol. 36:412-419 (1989)]. Because agonists form high affinity ternary complexes composed of the agonist, receptor, and G protein, this hypothesis would imply that interactions between adenosine receptor agonists and antagonists, while competitive, would appear to be "noncompetitive" in nature. Interactions between unlabeled and radiolabeled A1 receptor agonist and antagonist ligands have been investigated using bovine cerebral cortical membranes to further probe this point. The availability of both 3H- and 125I-radioligands allowed us to use both single- and dual-isotope experimental designs. Radioligand antagonist-agonist competition curves along with saturation analyses using filtration and centrifugation to isolate bound radioligand suggested that agonists bind to two sites or receptor states with high affinity and to one site with low affinity. Agonist radioligand saturation curves with or without unlabeled antagonist suggested that antagonists do not bind to all states of the receptor with equal affinity. The computer program EQUIL was used to define models capable of simultaneously fitting all parts of complex experiments in which 125I-N6-aminobenzyladenosine saturation isotherms with or without 8-cyclopentyl-1,3-dipropylxanthine ([3H]CPX) and a saturation isotherm of [3H]CPX were performed. The data were not compatible with two-independent site models or with ternary complex models involving one receptor and one G protein. The data were fit by a model involving one receptor and two G proteins and by a model involving two receptors and one G protein. Both models suggest that 1) a high percentage of the receptor(s) is coupled to a G protein in the absence of an agonist and 2) agonists stabilize whereas antagonists destabilize precoupled receptor-G protein complexes. Because of this, competitive interactions between A1 agonists and antagonists appear noncompetitive in nature.

Animals↗
PubMed1989

Density gradient profiles of A1 adenosine receptors labeled by agonist and antagonist radioligands before and after detergent solubilization.

A1 adenosine receptors in bovine cerebral cortex have been solubilized and subjected to sedimentation analysis using sucrose density gradient centrifugation. Because the receptors bound both agonists and antagonists with high affinity after solubilization, receptors labeled with an agonist or an antagonist radioligand could be studied before solubilization, after solubilization but before sucrose gradient centrifugation, or after sucrose gradient centrifugation. In each instance the agonist radioligand 125I-N6-p-aminobenzyladenosine (125I-ABA)-labeled receptor migrated as a single symmetrical peak that was located in the same area of the gradient. In contrast, the location of the receptor labeled with the antagonist [3H]xanthine amine congener [( 3H]XAC) varied in the different types of samples. When membranes were incubated with radioligands before solubilization, the peak of antagonist-labeled receptor was symmetrical and was located at a lower density than the peak of agonist-labeled receptor. In addition, receptors incubated with antagonist before solubilization migrated with an apparent lower density than receptors labeled with antagonist either after solubilization or after density gradient centrifugation. Treatments with agents that alter receptor/G protein interactions also resulted in a shift of antagonist-labeled receptors to lower density. These results suggest that the receptors that migrate to the lower density fractions of the gradients are free receptors, whereas those that migrate to the higher density fractions are coupled to a G protein. It is hypothesized that a large proportion of A1 receptors exist in the membrane coupled to a G protein and that this is the species labeled by the agonist radioligand 125I-ABA. It is, furthermore, hypothesized that the antagonist radioligand [3H]XAC preferentially binds to the free uncoupled A1 receptors.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine↗