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S C Guthrie

Publications and source records attributed to S C Guthrie.

11 recordsLinked to original sources

Gap junctional communication and development.

Embryonic development requires extensive interaction between cells; cell-to-cell communication through gap junctions may be one mechanism involved. Much correlative evidence suggests gap junctions are involved in cellular interactions during development. Recently, the biological role of junctions has been investigated using antibodies prepared against the major rat liver gap junction protein. Disrupting normal patterns of intercellular communication with such antibodies can drastically perturb development. Recent experiments emphasize, in particular, the importance of gap junctional communication for patterning processes.

Animals

Triphenylethylene antiestrogen binding sites (TABS) specificity.

The relative binding affinities (RBA) of various compounds for the triphenylethylene antiestrogen binding sites (TABS) were examined. The ability of tamoxifen to inhibit the binding of [3H]tamoxifen to salt extracted (0.4 M KCl) TABS from rat liver nuclei was used as a standard by which other compounds were compared (tamoxifen RBA, 100; Kd approximately 1 nM). Nafoxidine was the most effective triphenylethylene compound used (RBA 333; Kd approximately 0.3 nM) whereas the RBA of zuclomiphene and enclomiphene was not different from tamoxifen. MER-29 was the weakest inhibitor of the triphenylethylene derivatives (RBA 10; Kd approximately 10 nM). Trifluoperazine, chlorpromazine and the anti-calmodulin drugs W-13 and W-12 had RBA's of 25, 1, 1 and 0.1 respectively. The binding affinities of cholesterol and 7-ketocholesterol were significant (Kd approximately 22 nM) while the steroid hormones, estradiol, testosterone, progesterone and corticosterone displayed not observable affinity. Various compounds obtained from Merrill Dow Pharmaceuticals and the Eli Lilly Company which contained alklaminoethoxy side chains linked to aromatic ring structures had RBA's ranging from 1-0.3. We conclude, as other investigators have also concluded, that the similar binding affinities of various triphenylethylene antiestrogens for TABS and their divergent activities as antiestrogens makes it unlikely that TABS are directly involved in estrogen antagonism. The moderate but significant affinity of TABS for trifluoperazine and other drugs thought to be involved in calmodulin regulation indicates that TABS may be a linked in some way to calmodulin function. The binding of cholesterol and 7-ketocholesterol is also significant and may indicate that TABS are involved in cholesterol metabolism.

Animals

The estrogenic effects of clomiphene during the neonatal period in the rat.

The ability of clomiphene and its isomers to cause estrogenic responses during the neonatal period in the rat was examined. Rats were injected s.c. with clomiphene (CL), zuclomiphene (ZUC) or enclomiphene (ENC) on days 1,3, and 5 of life and the stimulation of the reproductive tract and estrogen receptor binding was observed. Uterine weight and DNA content were increased significantly by day 7 in animals treated with clomiphene or zuclomiphene. Uterine epithelial hypertrophy was present in all groups by day 10 and hyperplasia was present in the animals treated with ZUC and CL. The time of vaginal opening was greatly accelerated in all drug treated groups with the earliest day of opening occurring on day 7. Ovarian hemorrhage and blood in the periovarian sac occurred between days 12-14 and continued to be present through day 25. Drug treatment caused the estrogen receptor to accumulate in the nuclear fraction of the uterus and to be depleted from the cytosol fraction. We conclude that clomiphene administered to neonatal rats causes estrogenic stimulation of the reproductive tract in a fashion similar to other estrogens. This stimulation may account for the reproductive tract abnormalities which develop in rats treated with those drugs during the neonatal period.

Animals

Stimulation of vaginal growth and cornification in the absence of estrogen or nuclear binding of the estrogen receptor.

Mechanical stimulation of the vagina in ovariectomized rats results in vaginal epithelial proliferation and cornification which is identical to that seen with estradiol benzoate treatment. The estrogen receptor is localized primarily in the cytosol of mechanically stimulated vaginae, whereas, after estradiol benzoate treatment, the receptor is found in the nuclear compartment. Thus, it is possible to obtain a biological response ordinarily controlled by estrogen without the involvement of either the hormone or its receptor.

Adrenalectomy

Characterization of a triphenylethylene-antiestrogen-binding site on rat serum low density lipoprotein.

Several laboratories have reported the presence of an intracellular, high affinity [dissociation constant (Kd), 1-2 nM] antiestrogen-specific binding site in estrogen target tissues. In this report, we describe the binding properties and characteristics of an additional triphenylethylene antiestrogen-binding site in the serum of rats. By using saturation and competitive binding analyses with [3H]tamoxifen as the radio-labeled ligand, we have determined that rat serum contains a relatively high affinity (Kd, 28 nM), protease-sensitive binding site that is specific for antiestrogens of the triphenylethylene type (clomiphene and nafoxidine). Since this serum site has little affinity for the benzythiophene antiestrogens (LY 117018 and 156758), we have chosen the name of triphenylethylene-antiestrogen binding site (TABS). The concentration of serum TABS (picomoles of [3H]tamoxifen bound per ml serum) is roughly the same in males and females but is significantly greater in both sexes at days 5, 10, and 15 of age (400-500 pmol/ml) than in newborn or adult animals (100-200 pmol/ml). Preliminary characterization studies indicated that the serum TABS is probably a serum lipoprotein. Separation of rat serum lipoproteins on potassium bromide density gradients revealed that the serum TABS migrated with rat low density lipoprotein (LDL). Furthermore, rat LDL purified by density gradient centrifugation has an affinity and specificity for [3H]tamoxifen that is similar to the TABS found in whole serum. In contrast, the rat liver intracellular TABS does not have these density gradient characteristics. These data suggest that the rat serum TABS is LDL; however, the role of this serum lipoprotein in the mechanism of action of antiestrogens remains to be determined.

Animals

An endogenous ligand for the triphenylethylene antiestrogen binding site.

We have isolated an endogenous ligand which acts as a competitive inhibitor of the binding of 3H-tamoxifen to triphenylethylene antiestrogen-binding sites (TABS) prepared from liver and from serum low density lipoproteins (LDL). This ligand is present in boiled ethanol extracts of rat liver and may represent an "endogenous antiestrogen". "Endogenous antiestrogen" is used here as an operational term, since it has not been shown that TABS are involved in the mechanism of action of the triphenylethylene antiestrogens.

Animals

Antibodies to gap-junctional protein selectively disrupt junctional communication in the early amphibian embryo.

Antibodies to the major protein of rat liver gap junctions, molecular weight 27,000 (27K), have been microinjected into one identified cell of 8-cell stage Xenopus embryos. This treatment selectively disrupts both dye transfer and electrical coupling between the progeny cells. These results provide evidence that the 27K protein is an integral component of the cell-to-cell junctional channel. The disruption of junctional communication at early stages results in specific developmental defects, suggesting that blocking intercellular communication can have a pronounced influence on embryonic development.

Animals

Patterns of junctional communication in the early amphibian embryo.

It has long been recognized that cells in early embryos can communicate with each other via a direct cell-to-cell pathway, probably mediated by gap junctions. Low electrical resistance pathways, detected electrophysiologically, have been identified in all species examined so far. However, studies in various embryos on the transfer of molecules larger than small ions (for example, fluorescent dyes in the molecular weight range 350-500) have given conflicting results. In all these studies the ability to transfer dyes from cell to cell was determined without reference to the position of the injected cell in the embryo. In the experiments reported here, cell-cell transfer of the fluorescent dye, Lucifer yellow (molecular weight (Mr) 450) was re-examined in the early Xenopus laevis embryo by injecting the dye into identified cells, as the position of the injected cell within the embryo may be important. At the 32-cell stage, we found that dye transfer often occurred between animal pole blastomeres which were not sisters, as well as between sister cells, and also that Lucifer yellow was indeed transferred via gap junctions. The cell-cell transfer was not uniform within the animal pole; transfer was maximal near the dorsal side and minimal at the ventral side. This pattern may reflect differences in permeability or numbers of gap junctions across the embryo, and could be related to early events in development.

Animals