Adenosine receptors and the regulation of adenylate cyclase.
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Biomedical subjects
Publications and source records attributed to D M Cooper.
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A broad array of immunological tests was performed in children with serologically confirmed Mycoplasma pneumoniae infection. In a group of 21 children studied 0 to 6 wk after onset of of illness, neutrophil chemotaxis was 1.46 +/- 0.43 mm/3 hr (mean +/- S.D.) in patients compared to 1.79 +/- 0.28 mm/3 hr in controls (P less than 0.05). In a subgroup of seven children studied 0 to 2 wk after onset of 2,635 cpm in patients compared to 26,454 +/- 3,345 in controls for phytohaemagglutinin (P less than 0.02) and 5,321 +/- 535 in patients less than 0.02). There was, however, no impairment in response to soluble concanavalin A: 18,715 +/- 1,446 in patients compared to 25,193 +/- 2,564 in controls (P greater than 0.1). Follow-up studies on five of these seven children showed that lymphocyte proliferative responses to phytohaemagglutinin and pokeweed mitogen of all five children returned to normal values some weeks later. In another group of 28 children studied 13 to 18 wk after onset of illness, four had subnormal IgG values. Of the three children available for followup studies, all had normal values for serum IgG 2 to 5 months later.
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Treatment of membranes from HeLa cells, rat adipocytes, and rat liver with organic mercurials results in complex effects on adenylate cyclase activity that are not mimicked by the reversible sulfhydryl reagent, tetrathionate. At low concentrations (0.1 mM or less 1 mercurials inactivate the enzyme; inactivation is reversed by the thiol-reducing agent, dithiothreitol. Treatment with higher concentrations of organic mercurials (1 mM and above) results in a time-dependent, irreversible change in the ability of guanine nucleotides and fluoride ion to stimulate adenylate cyclase activity. The irreversible changes are blocked by treatment of membranes with cholera toxin and NAD, suggesting that the GTP-regulatory component is the site of mercurial action. This is further suggested by the lack of irreversible effects of mercurials on adenylate cyclase activity in membranes from mouse lymphoma cells that lack this component. Irreversible effects of mercurials on the adipocyte cyclase system also include enhancement of basal activity and potentiation of the inhibitory effects of GTP on cyclase activity; the latter effects of GTP are mediated through a process independent from that mediating stimulation of activity by GTP. It is concluded that the GTP-regulatory proteins responsible for the modulation of adenylate cyclase activity by hormones and neurotransmitters contain the sites of action of organic mercurials. Their possible mode of action is discussed.
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Cell surface adenosine receptors mediate either stimulation or inhibition of adenylate cyclase activity [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1], and the receptors that mediate these different responses can be discriminated with selected adenosine analogs. 5'-N-Ethylcarboxamide-adenosine is a more potent agonist at stimulatory receptors (Ra) than is N6-phenylisopropyladenosine, whereas the reverse potency order is seen with inhibitory receptors (Ri). The potency of adenosine is intermediate between the potencies of these two analogs. The relative potencies of adenosine receptor agonists are maintained in physiological responses in intact cells, such as steroidogenesis and inhibition of lipolysis. As with adrenergic receptors, subclasses of adenosine receptors differ functionally and pharmacologically.
The ceca of newly hatched chicks obtained either from commercial hatcheries or from a carefully controlled experimental hatchery have been shown to contain high numbers of a variety of microorganisms. Fecal streptococci, clostridia, enterobacteria, pediococci, and occasionally Pseudomonas aeruginosa have all been isolated, but never lactobacilli. After the chick has been on feed for 1 day, the numbers of lactobacilli in the crops and ceca are quite variable; by the 3rd day, however, large numbers are present throughout the alimentary tract. Lactobacilli could be established after 1 day by incorporating them in the feed or by treating the chicks with 0.5 ml of a broth culture. Spraying the eggs before hatching did not lead to their early establishment. Lactobacilli given alone to the newly hatched chick failed to prevent the establishment of Salmonella. However, when the chicks were given a complex mixture of facultative anaerobes and anaerobes which had been isolated from an adult bird, cecal colonization with Salmonella was prevented. The possible importance of the facultative anaerobes, particularly Streptococcus faecalis, in lowering the redox potential to encourage the growth of the anaerobes is discussed.
We evaluated lung function in 17 children with thalassemia major in stable condition receiving blood transfusions at regular intervals and subcutaneous desferoxamine daily. Total lung capacity (TLC) was below 2 SD of normal values for height in 7 of the 17 children and arterialized capillary PO2 was below the normal range in 15. We studied lung mechanics in 4 children with reduced TLC and found static and dynamic compliance below 2 SD of normal values for height in 3, and lung recoil at TLC above normal values and specific upstream conductance (Gus/TLC) above 2 SD of normal values in all 4. Although these alterations in lung function have been described in patients with pulmonary fibrosis, we found no fibrosis in autopsy specimens of lung from 8 other patients with thalassemia. The rate constant of carbon monoxide diffusion (kCO) was above the predicted mean in 14 of 15 children. These findings can be explained by a decrease in the growth of airspace relative to the vascular bed and major airways during childhood.
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GTP evoked both an activatory and an inhibitory response from adipocyte adenylate cyclase. This paper describes the persistence of the bimodal response under a variety of assay conditions. Additionally, manipulations are described which eliminate one or other of these actions. Treatment of adipocyte plasma membranes with cholera toxin A1 peptide and NAD+ abolishes the inhibitory phase of GTP action while preserving the activating phase. Treatment of the membranes with p-hydroxymercuriphenylsulfonic acid eliminates the activatory phase while maintaining the inhibitory processes mediated by GTP in adipocytes normally coexist and operate through different pathways since either phase can be abolished leaving the other intact. Adenosine and its purine-modified analogs inhibit fat cell adenylate cyclase in the GTP inhibitory phase (Londos, C., Cooper, D. M. F., Schlegel, W., and Rodbell, M. (1978) Proc. Natl. Acad. Sci. U.S.A. 75, 5362-5366). When this effect of GTP is abolished by either cholera toxin or Gpp(NH)p pretreatment, the inhibitory action of adenosine analogs is also lost. These data suggest a central role for GTP in mediating both activation and inhibition of adenylate cyclase by agents which act through cell surface receptors.
This short review summarizes some of the data concerning the regulation of adrenocortical adenylate cyclase by ACTH and other putative effectors, such as guanosine and nucleotides, divalent cations and adenosine. The available information on ACTH-sensitive adenylate cyclase of the adrenal cortex is discussed in comparison to other cyclase systems and the possible biochemical mechanisms of action of ACTH on the adrenal cortex.
An isolator is described, fabricated in glass-reinforced plastic, suitable for housing 25-30 chicks to 6 weeks of age, or for fewer older birds, for use in experiments involving infectious agents.
Direct effects of adenosine on adipocyte and hepatic adenylate cyclase have been demonstrated in an assay system where adenosine is not generated. The substrate used, 2'-deoxy ATP may, on metabolism, only give rise to 2'-deoxyadenosine, which does not act at adenosine receptors. With a slight modification of existing assay techniques this assay system has been used to detect a hitherto undiscovered adenosine receptor on liver plasma membranes, which is antagonised by methylxanthines and which stimulates adenylate cyclase activity in a GTP-dependent manner. The potent inhibitory effects of purine-modified adenosine analogs on fat cell adenylate cyclase are reproduced by adenosine in this assay system. An application of this approach to the study of adenylate cyclase not only simplifies detection of the role of adenosine, but also yields insights into the interaction between guanine nucleotides and hormones.
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