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Biomedical subjects

J R Sheppard

Publications and source records attributed to J R Sheppard.

At least 55 records · Page 3Linked to original sources
PubMed1977

Catecholamine hormone receptor differences identified on 3T3 and simian virus-transformed 3T3 cells.

Identification and characterization of hormone receptors on the cell surface is an effective tool for studying the plasma membrane. Using the direct binding of a radiolabeled antagonist, (-)[3H]alprenolol, to crude membrane preparations, and a physiological response (cellular cyclic AMP levels), I demonstrated a catecholamine (beta-adrenergic) hormone receptor site coupled to a catecholamine responsive adenylate cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1] on 3T3 and simian virus 40 (SV40)-transformed 3T3 cells. At a concentration of 1 muM, epinephrine and isoproterenol elevate cellular cyclic AMP levels 8- and 12-fold, respectively, in both cell lines. Norepinephrine was also a potent agonist on 3T3 cells (8-fold stimulation), but SV3T3 cells showed a lesser (2-fold) response to this hormone. The specificity of the physiological response (as well as the direct binding studies using the alprenolol radiolabel) is indicated by the increased effectiveness of (-) compared to (+) stereoisomers, rapid and reversible kinetics (steady state within 2 min), high affinity (Kd approximately 30 nM) and saturability (indicating a finite number of hormone receptors). These hormone receptor studies indicate the 3T3 cells have a beta1 adrenergic receptor while the SV3T3 cells have a receptor with beta2 qualities. In addition, the number of beta-adrenergic hormone receptors appear to be increased in the normal 3T3 cells by approximately 2-fold over the SV3T3 cells (300 versus versus 120 femtomol/mg of protein).

Adenylyl Cyclases↗
PubMed1975

Adenosine 3',5'-monophosphate analogs promote a circular morphology of cultured Schwannoma cells.

Dibutyryl adenosine 3',5'-monophosphate (dibutyrl cycle AMP) and 8-bromo cyclic AMP stimulated cells cultured from a rat Schwannoma to change their morphology from irregularly shaped to flattened circular and hollow circular forms within 30 minutes. The change in shape was specifically effected by analogs of cyclic AMP and cyclic AMP elevating agents, was reversible after removal of these additives, and was prevented by vinblastine and cytochalasin b, but was not affected by actinomycin D or cycloheximide.

Bromine↗
PubMed1975

Cyclic AMP, membrane transport and cell division. I. Effects of various chemicals on cyclic AMP levels and rate of transport of neucleosides, hypoxanthine and deoxyglucose in several lines of cultured cells.

Nutrient transport rates and cyclic AMP levels have been implicated in the regulation of cell proliferation. In the present study, however, changes in intracellular cyclic AMP level in several lines of cultured cells (normal 3T3 and SV40 and polyomavirus-transformed 3T3 cells; 3T6, C6 GLIOMA, MOUSE L, and Novikoff rat hepatoma cells) by treatment with papaverine, prostaglandine E1 or isoproterenol did not correlate with the inhibition of the uridine, hypoxanthine or deoxyglucose transport rates by these chemicals. Transport inhibitions by above chemicals or Persantin or Cytochalasin B occurred in most cell lines in the absence of any measurable change in intracellular cyclic AMP concentration. Furthermore, treatment of several cell lines with 1 mM dibutyryl cyclic AMP had no immediate effect on the transport of uridine, thymidine or deoxyglucose, although the transport capacity of the cells for uridine and thymidine, but not that for deoxyglucose, decreased progressively with time of treatment. We also observed that the uridine transport system of all cell lines derived from 3T3 cells and the hypoxanthine transport system of L cells exhibited high degrees of resistance to inhibition by the various chemicals. On the other hand, deoxyglucose transport was inhibited to about the same extent by these chemicals in all the cell lines investigated.

Animals↗
PubMed1971

Cyclic membrane changes in animal cells: transformed cells permanently display a surface architecture detected in normal cells only during mitosis.

A membrane site exposed only during mitosis in normal 3T3 cells, but exposed in all polyomavirus transformed 3T3 (Py3T3) cells, has been detected The site was identified by the specific binding of a wheat-germ agglutinin conjugated with fluorescein isothiocyanate. The receptor site has been studied previously. It binds wheat-germ agglutinin, causing the selective agglutination of Py3T3 cells, but not normal 3T3 cells. A short treatment with low concentrations of trypsin exposes cryptic sites in 3T3 cell membranes; this permits agglutination of these cells. The experiments reported here indicate that these sites normally are exposed during a brief period of the cell cycle for 3T3 cells. Specific hapten inhibitors of agglutination also inhibit the binding of the conjugated agglutinin and prevent fluorescence.

Animals↗
PubMed1971

Restoration of contact-inhibited growth to transformed cells by dibutyryl adenosine 3':5'-cyclic monophosphate.

Spontaneously transformed and virally-transformed cells are restored to contact-inhibited growth by the addition of dibutyryl cyclic AMP to the nutrient medium. Theophylline, an inhibitor of the phosphodiesterase that degrades cyclic nucleotides, must also be present for maximal effect. Once the transformed cells reach a saturation density in the presence of the additives, release from the contact-inhibited state occurs upon removal of the dibutyryl cyclic AMP and theophylline from the medium. Agglutinability of the transformed cells by wheat-germ agglutinin (a monitor of architectural changes in the plasma membrane) is decreased by dibutyryl cyclic AMP-theophylline treatment, but increases again upon removal of the additives.

Agglutination↗