Localization of the action of cholera toxin on adenyl cyclase in mucosal epithelial cells of rabbit intestine

Parkinson, David K.; Ebel, H.; DiBona, Donald R.; Sharp, Geoffrey W. G.

doi:10.1172/jci107039

Cited by 101 publications

(33 citation statements)

References 28 publications

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“…Minimal staining is found in the brush border area. This localization of cyclic AMP is consistent with the distribution of adenylate cyclase from mucosal epithelial cells of rabbit intestine (11). Cyclic AMP is also found in the laminal propria (Fig.…”

Section: Resultssupporting

confidence: 87%

“…Further studies exploring the regulation of guanylate cyclase and cyclic GMP action in small intestinal transport mechanisms are indicated by the present immunocytochemical studies. The predominant localization of cyclic AMP in the basal and lateral sides of the cells of the villus and not at the brush border is consistent with the finding of adenylate cyclase at these sites in rabbit intestine (11).…”

Section: Discussionsupporting

confidence: 88%

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Immunohistochemical localization of 3': 5'-cyclic AMP and 3': 5'-cyclic GMP in rat liver, intestine, and testis.

Ong¹,

Whitley²,

Stowe³

et al. 1975

Proc. Natl. Acad. Sci. U.S.A.

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Cyclic GMP and cyclic AMP have been localized in rat liver, small intestine, and testis by a fluorescent immunocytochemical procedure. In liver, cyclic AMP is distributed along sinusoids predominantly, and increased fluorescence is seen in sinusoidal areas after glucagon administration. Cyclic GMP is located in nuclear elements and on the plasma membranes of hepatocytes. In jejunum, cyclic AMP is found predominantly at the basal and lateral sides of brush border cells and in the lamina propria, while cyclic GMP is located to the brush border membrane, smooth muscle, and nuclear elements. In testis, cyclic AMP is found in cytoplasm of cells at the perimeter of the seminiferrous tubules and in interstitial cells, while cyclic GMP is visualized on the plasma membrane of the cells lining the tubules. Cyclic GMP is also seen on chromosomes of premeiotic spermatocytes and in sperm. These data provide histological evidence implicating diverse roles for the nucleotides in these tissues. The nuclear localization of cyclic GMP in all of these tissues suggests a role for the nucleotide in nucleus-directed events.Accumulating evidence suggests that cyclic GMP is involved in a number of cellular events ranging from the action of acetylcholine at muscarinic receptors (1-3) to growth regulation in lymphocytes (4) and fibroblasts (5). In these studies increased tissue concentrations of cyclic GMP have been demonstrated after hormonal stimulation. Despite this evidence, the role of cyclic GMP in cell function has been difficult to determine, partly because exogenously administered cyclic GMP in most tissues acts as a weaker cyclic AMP (6).To gain insight into the relative roles of cyclic AMP and cyclic GMP in cell function, we have recently applied the technique of cyclic nucleotide immunocytochemistry to studies in canine thyroid tissue, and contrasted the localization of cyclic GMP with that of cyclic AMP. We utilized canine thyroid tissue for these studies because thyrotrophin stimulating hormone increases cyclic AMP, without affecting cyclic GMP levels, and cholinergic compounds increase cyclic GMP while cyclic AMP is unchanged. We found cyclic GMP to be located in the area of the follicular plasma membrane bordering the colloid, and showed an increase in cytoplasmic fluorescence after acetylcholine treatment. In contrast, cyclic AMP was found throughout the cytoplasm of the follicular cells, and increased cytoplasmic fluorescence was seen after administration of thyrotrophin stimulating hormone. The distinct differences in localization of cyclic AMP and cyclic GMP suggested that the specific roles and the intracellular localization of these two nucleotides may be related (7).We have continued to apply this immunocytochemical procedure for the localization of cyclic AMP and cyclic GMP in a variety of rat tissues. In general, the two nucleotides have distinctly different staining patterns in individual cells. Cyclic AMP is found mostly in cytoplasm and sometimes on plasma membranes, but cyclic GMP is usually distinctl...

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Section: Resultssupporting

confidence: 87%

Section: Discussionsupporting

confidence: 88%

Immunohistochemical localization of 3': 5'-cyclic AMP and 3': 5'-cyclic GMP in rat liver, intestine, and testis.

Ong¹,

Whitley²,

Stowe³

et al. 1975

Proc. Natl. Acad. Sci. U.S.A.

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“…Radioactive materials were from the Radiochemical Centre, Amersham. Arginine-vasopressin (synthetic, grade IV) was from Sigma, PTH ( [20] ) and thus contain a mixture of guanylate cyclase activities localized in basal-lateral membranes and partiall); developed microviUi.…”

Section: Methodsmentioning

confidence: 99%

Properties of guanylate cyclase and levels of cyclic GMP in rat small intestinal villous and crypt cells

Jonge

1975

FEBS Letters

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“…This secretory mechanism is probably mediated by an activation of adenylate cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1.] activity and a concomitant elevation in cyclic AMP (cAMP) levels within the intestinal mucosa (4)(5)(6). It is assumed that the toxin exerts its effect on the intestinal microvillous membrane, since experimental studies indicate that the Vibrio cholera organism does not invade the intestinal mucosal barrier (7) and that the enterotoxin released by the organism does not penetrate the luminal epithelial surface in detectable quantities (8).…”

mentioning

confidence: 99%

Specific Binding of Cholera Toxin to Isolated Intestinal Microvillous Membranes

Walker

Field²,

1974

Proc. Natl. Acad. Sci. U.S.A.

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A sucrose density gradient assay was used to demonstrate the specificity and saturation of the binding of [l25Icholera toxin to isolated intestinal microvillous membranes from rat small intestine. When the toxin is first complexed to antitoxin and then exposed to intestinal membranes, the binding of cholera toxin is inhibited. To emphasize the physiologic importance of these observations, similar concentrations of [l2bl]cholera toxin were shown to stimulate the accumulation of cyclic AMP in mucosal homogenates and to increase the secretion of fluid into intestinal loops, whereas the same concentrations of toxin mixed with antitoxin had no effect on cyclic AMP accumulation. These studies suggest that cholera toxin attaches to brush border binding sites before exerting its biologic effect and that local intestinal antibody protection against cholera toxin may be due to inhibition of toxin attachment to these binding sites.Cholera toxin (CT) affects the small intestine by stimulating the active secretion of chloride ion into the intestinal lumen (1-3). This secretory mechanism is probably mediated by an activation of adenylate cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1.] activity and a concomitant elevation in cyclic AMP (cAMP) levels within the intestinal mucosa (4-6). It is assumed that the toxin exerts its effect on the intestinal microvillous membrane, since experimental studies indicate that the Vibrio cholera organism does not invade the intestinal mucosal barrier (7) and that the enterotoxin released by the organism does not penetrate the luminal epithelial surface in detectable quantities (8). Adenylate cyclase is a membrane-associated enzyme (9) and the toxin binds with specific affinity to cell membranes with adenylate cyclase activity (10, 11). Because of the polar nature of the intestinal epithelial cell, it is not yet understood whether the toxin attaches to a specific receptor site on intestinal microvillous membranes and affects adenylate cyclase activity at that location or whether it modifies adenylate cyclase activity elsewhere on the cell surface. Indirect evidence obtained with animal studies supports the association between intestinal binding of CT and its biologic action (2, 6, 12). Using secretion as an indicator for intestinal CT binding, the toxin has been noted to attach rapidly and efficiently to intestinal epithelial cells. Once exposure has occurred, the biologic effect cannot be reversed with specific antibodies (13) or by using chloreagenoid (natural toxoid) to displace the membrane attachment of the toxin (12).Specific immunization by either oral exposure or parenteral exposure to a toxoid or attentuated toxin represents a potential means of controlling the effects of CT on the small intestine. In experimental studies, both means of immunization have proven effective in producing a significant and prolonged protection from the biologic effect of CT on small intestinal secretion (14, 15). However, the mechanism whereby antibodies interfere with toxin activity has...

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Localization of the action of cholera toxin on adenyl cyclase in mucosal epithelial cells of rabbit intestine

Cited by 101 publications

References 28 publications

Immunohistochemical localization of 3': 5'-cyclic AMP and 3': 5'-cyclic GMP in rat liver, intestine, and testis.

Immunohistochemical localization of 3': 5'-cyclic AMP and 3': 5'-cyclic GMP in rat liver, intestine, and testis.

Properties of guanylate cyclase and levels of cyclic GMP in rat small intestinal villous and crypt cells

Specific Binding of Cholera Toxin to Isolated Intestinal Microvillous Membranes

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