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Biomedical subjects

W J Thompson

Publications and source records attributed to W J Thompson.

At least 91 records · Page 5Linked to original sources
PubMed1988

Purification and partial characterization of membrane-associated type II (cGMP-activatable) cyclic nucleotide phosphodiesterase from rabbit brain.

Membrane-associated, Type II (cGMP-activatable) cyclic nucleotide phosphodiesterase (PDE) from rabbit brain, representing 75% of the total homogenate Type II PDE activity, was purified to apparent homogeneity. The enzyme was released from 13,000 x g particulate fractions by limited proteolysis with trypsin and fractionated using DE-52 anion-exchange, cGMP-Sepharose affinity and hydroxylapatite chromatographies. The enzyme showed 105 kDa subunits by SDS-PAGE and had a Stokes radius of 62.70 A as determined by gel filtration chromatography. Hydrolysis of cAMP or cGMP showed positive cooperativity, with cAMP kinetic behavior linearized in the presence of 2 microM cGMP. Substrate concentrations required for half maximum velocity were 28 microM for cAMP and 16 microM for cGMP. Maximum velocities were approx. 160 mumol/min per mg for both nucleotides. The apparent Kact for cGMP stimulation of cAMP hydrolysis at 5 microM substrate was 0.35 microM and maximal stimulation (3-5-fold) was achieved with 2 microM cGMP. Cyclic nucleotide hydrolysis was not enhanced by calcium/calmodulin. The purified enzyme can be labeled by cAMP-dependent protein kinase as demonstrated by the incorporation of 32P from [gamma-32P]ATP into the 105 kDa enzyme subunit. Initial experiments showed that phosphorylation of the enzyme did not significantly alter enzyme activity measured at 5 microM [3H]cAMP in the absence or presence of 2 microM cGMP or at 40 microM [3H]cGMP. Monoclonal antibodies produced against Type II PDE immunoprecipitate enzyme activity, 105 kDa protein and 32P-labeled enzyme. The 105 kDa protein was also photoaffinity labeled with [32P]cGMP. The purified Type II PDE described here is physicochemically very similar to the isozyme purified from the cytosolic fraction of several bovine tissues with the exception that it is predominantly a particulate enzyme. This difference may reflect an important regulatory mechanism governing the metabolism of cyclic nucleotides in the central nervous system.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1988

Relationships of secretagogue-induced cAMP accumulation and acid secretion in elutriated rat gastric parietal cells.

Centrifugal elutriation was adapted and analyzed as a method to separate rat gastric parietal cells from other fundic mucosal cells. Elutriated parietal cell fractions provided sufficient purity by morphological criteria, and fluorescence activated cell sorting analyses, yield, reproducibility and maintenance of functional responses. These characteristics allowed the study and comparison of the kinetics of histamine, isoproterenol, and forskolin-induced cyclic AMP and 14C-amino-pyrine accumulations in the presence of 1-methyl-3-isobutyl xanthine (IBMX) in parietal-cell-rich and parietal-cell-poor fractions. All three acid secretagogues studied produced the same maximal rate of acid secretion as judged by 14C-aminopyrine accumulation. Each secretagogue action peaked at different times and had different accumulation kinetics. For each agonist, cyclic AMP accumulation preceded secretion. However, the rate, extent, and temporal changes of cyclic AMP accumulation were independent of aminopyrine accumulation. HPLC separation and drug inhibition studies of cyclic nucleotide phosphodiesterase activities indicated the presence of multiple, high affinity (Type IV), but not lower affinity (Types I and II), enzyme forms in gastric mucosal cells. IBMX did not distinguish between the two forms, but SQ 65442 and RO 20-1724 were selective inhibitors. Inhibition constants of IBMX for phosphodiesterase hydrolysis agreed closely with its EC50 for histamine-stimulated acid secretion (2-8 microM). Elutriated parietal cells maintained their responses to selective receptor antagonists in the micromolar concentration range.

1-Methyl-3-isobutylxanthine↗
PubMed1988

Combination anchorage technique: an update of current mechanics.

Experience and advances in design and technique have greatly enhanced combination orthodontic techniques during the past 5 years. Improvements have made the delivery of treatment more effective and reduced the problems with appliance adjustment and esthetics. Some of these improvements are in the bracket itself, which has been modified for efficiency and esthetics. A modification of treatment technique by the addition of tandem wires, stabilizing arch wires, Dual Flex arch wires, and sectional wires have increased the movement capabilities and anchorage control. A technique of orthodontic mechanics has evolved that is called combination anchorage technique (CAT). The technique is designed to broaden the treatment effectiveness of the orthodontist by providing a combination of orthodontic technical capabilities. Use of the two different bracket slots provides a simple and efficient means to vary (1) anchorage (dynamic or static), (2) movement (tipping or bodily), (3) technique (light wire or straight wire), (4) resistance (one tooth or multiple teeth), and (5) treatment compensation (skeletal or dental).

Biomechanical Phenomena↗
PubMed1988

Synaptic rearrangements and alterations in motor unit properties in neonatal rat extensor digitorum longus muscle.

1. We have used in vitro intracellular recordings and measurements of the contractile properties of single motor units to examine the changes in muscle innervation occurring during the post-natal development of a fast-twitch muscle in the hindlimb of the rat, the extensor digitorum longus (EDL). 2. Intracellular recordings of end-plate potentials evoked in response to graded stimulation of the nerve supply to the muscle indicate that during the first day after birth, each muscle fibre receives synaptic input from at least two motoneurones and that some muscle fibres receive as many as six such inputs. With subsequent development, most of this polyneuronal innervation is eliminated: the first singly innervated fibres are encountered on day 3; by day 18 fewer than 5% of the fibres remain polyneuronally innervated. These results show that there are quantitative differences in post-natal synapse elimination in EDL compared to its well-studied counterpart, the soleus. Although the great majority of fibres in both muscles become singly innervated at about 18 days, the first singly innervated fibres appear at least a week earlier in the EDL. None the less, synapses are lost from EDL at about half the rate they are lost from soleus. 3. The number of motor units, determined by counting the number of twitch increments produced by graded stimulation of ventral root filaments teased to contain only a few EDL motor axons, remains unchanged from an average of forty-one from post-natal day 1 to day 17. In addition, the number of muscle fibres counted in muscle cross-sections stained with an anti-myosin antibody increases less than 10% from birth to adulthood. Therefore, synapse elimination in EDL occurs with a largely constant population of muscle fibres as well as motoneurones. 4. Measurements of tensions generated by single motor units indicate that the average size of a motor unit declines from 6.8% of the muscle fibres at day 1 to 2.3% at 17 days. This result indicates that each motoneurone, on average, comes to innervate threefold fewer muscle fibres. Motor units derived from each of the spinal segments innervating the muscle undergo equivalent reductions in motor unit size, indicating that there is no segmental disproportion to synapse elimination in this muscle. At all ages, there is a large diversity of motor unit sizes in the muscle. Synapse elimination therefore appears to maintain rather than decrease this diversity.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1988

The organization and development of compartmentalized innervation in rat extensor digitorum longus muscle.

1. We have examined the innervation of the rat extensor digitorum longus (EDL) muscle by the two extramuscular branches formed from the bifurcation of its muscle nerve. Observations of muscle contractions, recordings of end-plate potentials, and glycogen depletion of young adult muscles show that each branch innervates a separate region or 'compartment' in the muscle. The branch entering the muscle nearer the knee (the K branch) innervates fibres in the anteromedial half of the muscle whereas the branch entering closer to the foot (the F branch) innervates fibres located posterolaterally. Individual EDL motoneurones project either into the K or the F branch and therefore innervate fibres located in one compartment. The boundary between the compartments is usually sharply delineated. No obvious anatomical feature exists within the muscle which would explain the division of the muscle into two distinct regions. 2. The presence of a segmentotopic projection from the spinal cord to the muscle was investigated to evaluate its possible contribution to the compartmental pattern. The most posterior neurones of the EDL motor pool were found to project more frequently to the posterolateral F compartment; similarly, the most anterior neurones most frequently project to the anteromedial K compartment. However, each compartment is innervated by both anteriorly and posteriorly located motoneurones. The segmentotopic projection is too weak to explain the presence of neuromuscular compartments. 3. The post-natal period of synapse elimination appears to play at best a minor role in setting up the compartmentalized innervation. Glycogen depletion and intracellular recording in 1-2-day-old muscles show that each nerve branch innervates fibres in the same region of the muscle as in the adult. Most of the fibres in each compartment are polyneuronally innervated by axons in their own particular nerve branch, although fibres located near the boundary between the two compartments are innervated by axons from both nerve branches. This convergent innervation from the two branches disappears in concert with the elimination of polyneuronal innervation throughout the muscle. A random elimination of these convergent inputs appears adequate to explain the final compartmental pattern. 4. Our findings suggest that the compartmental pattern is primarily the consequence of te segregation of EDL motoneurones into two nerve branches which are directed into separate regions of the muscle.

Animals↗
PubMed1988

Subcellular distribution of high-affinity type IV cyclic AMP phosphodiesterase activity in rabbit ventricular myocardium: relations to the effects of cardiotonic drugs.

Rabbit ventricular myocardium contains distinct cytosolic and particulate cyclic AMP (cAMP) phosphodiesterase activities that exhibit characteristics ascribed to a high-affinity type IV cAMP phosphodiesterase activity found in several tissues. The particulate activity associated with sarcoplasmic reticulum vesicles has an apparent Km for cAMP of about 0.3 microM and a Vmax of 2.45 +/- 0.55 nmol/min/mg. Cyclic GMP (cGMP) inhibits hydrolysis measured at 0.25 microM cAMP with an IC50 value of 0.28 microM. In comparison, a ventricular cytosolic high-affinity cAMP phosphodiesterase activity obtained by anion exchange chromatography (Peak III) has an apparent Km of 0.93 microM and a Vmax of 17 +/- 1 nmol/min/mg. Hydrolysis of 0.25 microM cAMP by this cytosolic activity is weakly inhibited by cGMP with an IC50 value of 142 microM. Particulate enzyme activity is 60-fold more sensitive to inhibition by milrinone than is the cytosolic form (Ki = 0.18 versus 11 microM, respectively); the pyridazinone imazodan is a 12-fold more potent inhibitor of the particulate activity than of the cytosolic form (Ki = 1.5 versus 18 microM, respectively). Inhibition of both cytosolic and particulate enzyme activities appears competitive in nature. Solubilization of particulate activity did not significantly alter its affinity for substrate or sensitivity to inhibition by cGMP. In the presence of a submaximally activating concentration of forskolin (0.4 microM), selective phosphodiesterase inhibitors potentiated the activation of protein kinase in isolated ventricular septal slices. Under these conditions, changes in cAMP-dependent protein kinase activity ratios correlated more closely with contractile responses than did changes in intracellular content of cAMP.(ABSTRACT TRUNCATED AT 250 WORDS)

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1988

Effects of a potent carbonic anhydrase inhibitor on bone resorption in organ culture.

We have examined the effects of a potent inhibitor of carbonic anhydrase (CA), 5-(3-hydroxybenzoyl)2-thiophenesulfonamide (HTS), and compared them with the effects of acetazolamide (AZ) and ethoxzolamide (EX) on bone resorption in organ cultures in fetal rat long bones under control unstimulated conditions and in response to PTH, prostaglandin E2 (PGE2), 1,25-dihydroxyvitamin D [1,25-(OH)2D3], and interleukin-1 (IL-1). The relative potencies of HTS, EX, and AZ for inhibition of CA and bone resorption were similar. HTS inhibited control resorption at 10(-5) to 3 X 10(-5) M, while EX at 10(-4) M inhibited and AZ was ineffective. In the presence of PTH, the inhibitory effect of HTS was seen at concentrations as low as 3 X 10(-6) M and was maximal at 3 X 10(-5) M. EX was inhibitory at 10(-5) M and maximal at 10(-4) M, while AZ at 10(-4) M only partially inhibited PTH-stimulated bone resorption. The responses to PGE2 (10(-7) M), 1,25-(OH)2D3 (10(-9) M), and IL-1 (50 U/ml) were inhibited by HTS at 10(-5) M, while AZ at 10(-4) M was ineffective against PGE2 and 1,25-(OH)2D3. The effect of HTS did not appear to be due to nonspecific toxicity, since after 2 days of treatment at 3 X 10(-5) M and 3 days of recovery, the bone resorptive response to PTH was completely restored. Moreover, HTS at 3 X 10(-5) M did not inhibit the incorporation of labeled proline or thymidine into fetal rat long bones. HTS was more potent as an inhibitor when the CO2 concentration in the gas phase was reduced from 5% to 2% and the pH was increased from 7.2 to 7.5, consistent with an effect on CA-mediated hydrogen ion generation.

Acetazolamide↗
PubMed1988

Effects of intracisternal bead devices on lacteal secretion components, plaque formation, and bacterial infection during the nonlactating period.

Effects of placing intracisternal bead devices (ICB) into teat cisterns of 6 dairy cows, from the end of lactation through parturition, were studied. Lacteal secretion samples were collected weekly from each mammary quarter during the nonlactating period to monitor composition changes in ICB-fitted and nonfitted quarters. In quarters remaining uninfected (n = 15), there were significantly higher mean somatic cell counts (P less than 0.05), percentage of neutrophils (P less than 0.019), and cell viability (P less than 0.038), but significantly lower percentage of macrophages (P less than 0.013) in ICB-fitted quarters compared with those in nonfitted quarters. The ICB had no significant effect on mean weekly values for percentage of lymphocytes, pH, lactoferrin, citrate, citrate/lactoferrin molar ratio, serum albumin, alpha-lactalbumin, and N-acetyl-beta-D-glucosaminidase. In infected quarters (n = 9), pH of mammary secretions was significantly (P less than 0.004) higher in ICB-fitted quarters, but concentrations of lactoferrin (P less than 0.004), alpha-lactalbumin (P less than 0.013), and N-acetyl-beta-D-glucosaminidase (P less than 0.028) were significantly lower, compared with those in nonfitted quarters. Coagulase-negative staphylococci comprised approximately 90% of all infections. Over the nonlactating period, 16.4 and 41.5% of samples from nonfitted and ICB-fitted quarters, respectively, contained coagulase-negative staphylococci. Microscopic examination of ICB from uninfected quarters revealed a thin coating of plaque with adhering neutrophils, macrophages, and multinucleated giant cells. Microscopic examination of plaque on devices from ICB-fitted quarters harboring coagulase-negative staphylococci revealed numerous adherent cocci and neutrophils.

Animals↗
PubMed1987

Increase in liver protein phosphatase-1 in spontaneously diabetic Chinese hamsters.

Two broad-specifically protein phosphatases, termed protein phosphatase-1 (PrP-1) and protein phosphatase-2A (PrP-2A), accounting for all the hepatic activity regulating glycogen phosphorylase, were measured in spontaneously diabetic Chinese hamsters exhibiting persistent glycosuria. When compared with genetically related inbred sublines free of glycosuria, diabetic animals demonstrated approximately 25% increase in PrP-1 activity measured either in crude tissue extracts or in cytosols fractionated by ion-exchange chromatography. No significant alteration in total PrP-2A activity was observed in the diabetic animals. These findings indicate that a specific change in hepatic PrP-1 is associated with genetically acquired diabetes in Chinese hamsters. In contrast to reported data using animals with experimentally induced diabetes mellitus, hepatic PrP-1 was increased in the spontaneously diabetic Chinese hamsters. The data suggests that distinct alterations in PrP-1 and associated metabolic consequences are exhibited by different types of diabetes.

Animals↗
PubMed1987

Selective innervation of types of fibres in developing rat muscle.

The technique of glycogen depletion has been used to identify the types of muscle fibres innervated by individual motor neurones in the neonatal rat. This analysis shows that neonatal motor units are highly biased in their fibre type composition, even at times when the fibres receive extensive polyneuronal innervation. This finding suggests that the innervation of muscle fibres is somehow sorted according to type during early development. This sorting does not appear to occur during the removal of the polyneuronal innervation because little, if any, increase in the bias of unit compositions occurs as the number of synapses present in the muscle is reduced 2- to 3-fold. To determine whether the sorted innervation might be explained by a selective synaptogenesis, a study was made of the type compositions of units formed by reinnervation of neonatal soleus muscle. Glycogen depletion of single units 2 weeks following crush of the soleus nerve at postnatal day 2 showed that most of them (10/12) had biased type compositions which could not be explained by a random reinnervation. The location of fibres in the reinnervated motor units suggests that the regenerating axons innervated a novel set of fibres. The differentiation of fibres into types was apparently not changed during their reinnervation. These results imply that regenerating motor neurones in the neonatal rat selectively reinnervate muscle fibre types. These and other studies further imply that the organization of fibres into motor units during normal development does not occur, as is widely believed, by a random innervation of naive fibres and their subsequent differentiation under the influence of innervation.

Animals↗
PubMed1987

Histological response of bovine mammary tissue to an intracisternal bead device.

Placement of an intracisternal bead device into teat cisterns of dairy cows at drying off was studied. Histological analysis of teat cisternal tissues from quarters fitted with devices demonstrated significant elevation in leukocyte infiltration and hypertrophy of the epithelial lining. Tissue changes ranged from degeneration and sloughing of surface cells of double-layered epithelia to marked thickening and lysis of cisternal epithelium, leaving only the basement membrane intact. Quantification of mammary parenchymal components from quarters fitted with devices demonstrated no differences in percentage area occupied by alveolar epithelium, lumen, and intraalveolar stroma from unfitted control quarters. However, quantification of leukocyte numbers demonstrated slightly greater leukocytosis into tissues of quarters fitted with devices in comparison with controls. Devices were successful in stimulating a localized leukocytosis into teat cisternal tissues, which may serve to amplify the leukocyte response to mastitis-causing organisms once they penetrate the teat duct.

Animals↗
PubMed1987

Reinnervation of muscle fiber types in the newborn rat soleus.

We have examined the selectivity of reinnervation of fiber types in rat soleus muscle denervated by crush of the soleus nerve 2 d after birth. The fibers innervated by single, regenerated motor axons were identified by use of glycogen depletion approximately 2 weeks following denervation. The types of fibers were determined by immunohistochemistry employing anti-myosin antibodies and, in some cases, by myofibrillar ATPase staining. Two distinct types of fibers are present in soleus at 2 d and through the next 16 d of normal postnatal development. These fiber types are retained in a denervated muscle for the period of time required for reinnervation. Although 40% of the fibers are lost from the muscle during reinnervation, we find no evidence for interconversion of muscle fiber types. Nonetheless, 10 of the 12 single motor units examined had fiber type compositions that were markedly biased toward one or the other of these 2 types; the bias in these units could not be explained by chance reinnervation. On the basis of the topographical distribution of the muscle fibers in each of these units, the motor axons reinnervated a novel set of fibers. We interpret these findings to mean that neonatal soleus motor neurons reinnervate fiber types in a selective manner. This selective innervation may explain the bias in the fiber type composition of normal motor units during early postnatal development.

Animals↗
PubMed1986

Bronchodilator activity of a nonxanthine phosphodiesterase inhibitor; 2,4-diamino-5-cyano-6-bromopyridine (compound I).

The ability of 2,4-diamino-5-cyano-6-bromopyridine (compound 1) to inhibit bronchiolar smooth muscle constriction was examined in isolated rings of rabbit primary bronchi and intrapulmonary bronchioles. After carbachol-induced constriction these tissue were significantly relaxed by either compound I or 1-methyl-3-isobutylxanthine (MIX) in a similar dose-dependent manner with 50 to 80% relaxation occurring at 100 microM of either compound. Compound I also attenuated the constrictor response of bronchial rings to histamine and significantly reduced the tension generated by horseradish peroxidase in sensitized tissues responding to this antigen. In addition, both compound I and MIX were found to inhibit the soluble cyclic AMP phosphodiesterase activity of rabbit bronchioles. Finally, both compound I and MIX caused a nearly 2-fold, time-dependent increase in cyclic AMP levels in isolated rabbit intrapulmonary bronchioles. The similarities of both the in vitro tissue responses to these compounds and the phosphodiesterase inhibitory properties suggest that the ability of compound I to reduce constrictor-induced tension generation in bronchial smooth muscle is related to the inhibition of cyclic nucleotide phosphodiesterases and the consequent elevation of cyclic AMP.

1-Methyl-3-isobutylxanthine↗
PubMed1985

Characterization of a Ca2+-calmodulin-stimulated cyclic GMP phosphodiesterase from bovine brain.

A calmodulin-stimulated form of cyclic nucleotide phosphodiesterase from bovine brain has been extensively purified (1000-fold). Its specific activity is approximately 4 mumol min-1 (mg of protein)-1 when 1 microM cGMP is used as the substrate. This form of calmodulin-sensitive phosphodiesterase activity differs from those purified previously by showing a very low maximum hydrolytic rate for cAMP vs. cGMP. The purification procedure utilizing ammonium sulfate precipitation, ion-exchange chromatography on DEAE-cellulose, gel filtration on Sephacryl S-300, isoelectric focusing, and affinity chromatography on calmodulin-Sepharose and Cibacron blue-agarose results in a protein with greater than 80% purity with 1% yield. Kinetics of cGMP and cAMP hydrolysis are linear with Km values of 5 and 15 microM, respectively. Addition of calcium and calmodulin reduces the apparent Km for cGMP to 2-3 microM and increases the Vmax by 10-fold. cAMP hydrolysis shows a similar increase in Vmax with an apparent doubling of Km. Both substrates show competitive inhibition with Ki's close to their relative Km values. Highly purified preparations of the enzyme contain a major protein band of Mr 74 000 that best correlates with enzyme activity. Proteins of Mr 59 000 and Mr 46 000 contaminate some preparations to varying degrees. An apparent molecular weight of 150 000 by gel filtration suggests that the enzyme exists as a dimer of Mr 74 000 subunits. Phosphorylation of the enzyme preparation by cAMP-dependent protein kinase did not alter the kinetic or calmodulin binding properties of the enzyme. Western immunoblot analysis indicated no cross-reactivity between the bovine brain calmodulin-stimulated gGMP phosphodiesterase and the Mr 60 000 high-affinity cAMP phosphodiesterase present in most mammalian tissues.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1985

Activity and synapse elimination at the neuromuscular junction.

The neuromuscular junction undergoes a loss of synaptic connections during early development. This loss converts the innervation of each muscle fiber from polyneuronal to single. During this change the number of motor neurons remains constant but the number of muscle fibers innervated by each motor neuron is reduced. Evidence indicates that a local competition among the inputs on each muscle fiber determines which inputs are eliminated. The role of synapse elimination in the development of neuromuscular circuits, other than ensuring a single innervation of each fiber, is unclear. Most evidence suggests that the elimination plays little or no role in correcting for errant connections. Rather, it seems that connections are initially highly specific, in terms of both which motor neurons connect to which muscles and which neurons connect to which particular fibers within these muscles. A number of attempts have been made to determine the importance of neuromuscular activity during early development for this rearrangement of synaptic connections. Experiments reducing neuromuscular activity by muscle tenotomy, deafferentation and spinal cord section, block of nerve impulse conduction with tetrodotoxin, and the use of postsynaptic and presynaptic blocking agents have all shown that normal activity is required for normal synapse elimination. Most experiments in which complete muscle paralysis has been achieved show that activity may be essential for the occurrence of synapse elimination. Furthermore, experiments in which neuromuscular activity has been augmented by external stimulation show that synapse elimination is accelerated. A plausible hypothesis to explain the activity dependence of neuromuscular synapse elimination is that a neuromuscular trophic agent is produced by the muscle fibers and that this production is controlled by muscle-fiber activity. The terminals on each fiber compete for the substance produced by that fiber. Inactive fibers produce large quantities of this substance; on the other hand, muscle activity suppresses the level of synthesis of this agent to the point where only a single synaptic terminal can be maintained. Inactive muscle fibers would be expected to be able to maintain more nerve terminals. The attractiveness of this scheme is that it provides a simple feedback mechanism to ensure that each fiber retains a single effective input.

Animals↗
PubMed1985

Elevated leukocyte phosphodiesterase as a basis for depressed cyclic adenosine monophosphate responses in the Basenji greyhound dog model of asthma.

The BG dog manifests various characteristics of human asthma, including airway hyperreactivity to low concentrations of methacholine. Studies have suggested that airway hyperreactivity in asthma is related to inadequate intracellular cAMP responses. We studied cAMP characteristics in MNL from 19 BG and 14 mongrel dogs. beta-Adrenergic receptors were assessed by 125I CYP in the presence and absence of propranolol. The responses of cAMP to ISO were measured by radioimmunoassay. Adenylate cyclase activity was determined in homogenized MNL preparations by cAMP generation. PDE activity was quantitated by radioenzyme assay. Mongrel dog leukocyte ISO-stimulated cAMP levels doubled, whereas there were negligible increases in MNL from BG dogs. Basal PDE levels were higher in BG dogs than in mongrel dogs. The PDE inhibitor Ro 20-1724 restored ISO-stimulated cAMP responses in MNL of BG dogs. Adenylate cyclase activity was not lower in MNL homogenates from BG dogs than in mongrel dogs. Cells from both BG and mongrel dogs demonstrated similar receptor numbers and affinities of saturable, specific beta-adrenergic binding over a 10 pM to 400 pM range. Our results suggest that depressed cAMP responses in BG dogs are due to high PDE activity rather than to a defect in the beta-adrenergic receptor adenylate cyclase system.

Adenylyl Cyclases↗