Effects of Aldosterone on RNA and Protein Synthesis in the Toad Bladder

Rousseau, Guy; Crabbé, J.

doi:10.1111/j.1432-1033.1972.tb01727.x

Cited by 23 publications

(7 citation statements)

References 47 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…(5) Well-defined labeling of all of the classes of cytoplasmic RNA was achieved with a 30-min [3H]uridine pulse and a chase of 90-150 min. Some of the earlier studies (13,14,23) used continuous labeling which may have obscured the increment in labeling of the cytoplasmic 9S-12S class imparted by the steroid. We recently found that pulse labeling 150 mim after administration of the hormone resulted in lesser incorporation of [3H]uridine into 9S-12S RNA than a similar pulse during the first 30 mim of exposure to the steroid (unpublished observations).…”

Section: Discussionmentioning

confidence: 99%

“…Precise definition of regulation of transcription by aldosterone, however, has not yet been achieved. Recent attempts to demonstrate direct effects on the synthesis of discrete classes of RNA were unsuccessful (13,14). …”

mentioning

confidence: 99%

“…A variety of findings support this hypothesis including: the presence of nuclear receptors in target tissues (2), effects of inhibitors of RNA and protein synthesis (3)(4)(5)(6), and effects on specific mitochondrial enzymes (7), RNA polymerase activity (5,8), chromatin template activity (9), and incorporation of precursors into total or nuclear RNA (10)(11)(12)(13). Precise definition of regulation of transcription by aldosterone, however, has not yet been achieved.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Kinetics of RNA Labeling in Toad Bladder Epithelium: Effects of Aldosterone and Related Steroids

Rossier

Wilce

Edelman

1974

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

View full text Add to dashboard Cite

Sucrose gradient analysis revealed an aldosterone-dependent increase in the incorporation of [3H]uridine into a nonmethylated, rapidly labeled RNA that sedimented at 9-12 S. This effect was antagonized by an anti-mineralocorticoid, spironolactone (SC 9420), and was not elicited either by a glucocorticoid, cortisol, or by the inactive isomer, 17-a-isoaldosterone. These findings are in accord with the inference that mRNA is induced by aldosterone during the latent period and that the induced mRNA mediates the action on sodium transport.The action of aldosterone appears to be mediated by the following sequence of events: (1) binding of aldosterone to a specific cytoplasmic receptor and attachment of this complex to gene sites, (2) enhanced synthesis of RNA and of specific protein(s), (3) augmentation of sodium transport by the induced protein(s) (1, 2). A variety of findings support this hypothesis including: the presence of nuclear receptors in target tissues (2), effects of inhibitors of RNA and protein synthesis (3-6), and effects on specific mitochondrial enzymes (7), RNA polymerase activity (5, 8), chromatin template activity (9), and incorporation of precursors into total or nuclear RNA (10-13). Precise definition of regulation of transcription by aldosterone, however, has not yet been achieved. Recent attempts to demonstrate direct effects on the synthesis of discrete classes of RNA were unsuccessful (13,14). MATERIALS AND METHODSColombian, female toads (Bufo marinus) (from Tarpon Zoo, Florida) were partially immersed in saline (0.075 M), at room temperature, for 48 hr before use. After double pithing and perfusion of the circulation with about 200 ml of oxygenated incubation medium through a heart puncture, everted hemibladders (mucosal side outside) were mounted as sacs on plastic canulas (15). The sacs were filled with 5 ml and immersed in 90 ml of the incubation medium. Both sides were oxygenated with 97% 02-3% CO2 and the temperature of the bath was maintained at 250 (4±0.2). Potential difference (P.D.) and short-circuit current (SCC) were measured at 30-min intervals as described previously (15). Four hours after mounting, aldosterone (final concentration = 70 nM) was added to the mucosal and serosal solutions of the test hemibladders and diluent to the control. The instituted by addition of 1000-fold excess of unlabeled precursors. The tissue was washed twice with ice-cold incubation medium and all subsequent steps carried out at 0°-4°. Epithelial cells were removed by scraping with a glass slide and scrapings from 5 to 10 paired hemibladders were pooled into experimental and control groups.Isolation of Nuclear and Cytoplasmic RNA. Epithelial nuclei were isolated and extracted by the procedure of Penman (16): The pooled cells were homogenized in a Teflon-glass PotterElvehjem homogenizer at top speed with 15 strokes in 2 ml of a hypotonic medium (1 mM NaCl, 1 mM Tris HCl, p11 7.4, 1.5 mM MgCl2). Cytoplasmic RNA was extracted twice from the supernatant fractions with phenol containing 0.5% sod...

show abstract

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Kinetics of RNA Labeling in Toad Bladder Epithelium: Effects of Aldosterone and Related Steroids

Rossier

Wilce

Edelman

1974

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

View full text Add to dashboard Cite

show abstract

“…In concurrence with our results. Rousseau and Crabbe [20] found an increased intracellular radioactive uridine pool already 30 min after hormone applica tion. Rossier et al [19] ascertained a slight in crease after first 90 min.…”

Section: Intracellular Uridine Pool and Rna Synthesismentioning

confidence: 99%

“…After binding to a cytosolic receptor protein, the activated aldosterone/receptor complex is transferred to the nucleus [7,9]. Between the introduction of aldosterone and the first ap pearance of a physiological effect, namely the increase of Na+ transport [25], a latent period exists, ranging from 45 to 90 min [7,15,18], For this period, few biochemical data on the am phibian urinary bladder [8,20,26] and mam malian kidney [2,5,6,22] are available. In contrast, for the early and late physiological response (2-24 h), more information exists in dicating the translation of aldosterone-dependent mRNA into specific proteins, the al dosterone-induced proteins [7, 15.…”

Section: Introductionmentioning

confidence: 99%

Action of Aldosterone on Cultured Renal Collecting Duct Cells during the Latent Period

Zwanzig¹,

Minuth

Groß³

1990

Kidney Blood Press Res

View full text Add to dashboard Cite

The effects of aldosterone on protein synthesis in the latent period were investigated on cultured renal collecting duct cells from neonatal rabbit kidneys. Tissue was incubated with radioactively labelled uridine and amino acids and then precipitated with trichloroacetic acid in order to determine the intracellular precursor pool and identify new synthesis of RNA and protein. During the latent period, aldosterone increased the intracellular radioactive uridine pool and total radioactive RNA content already 20 and 60 min after its application; conversely 40 min after aldosterone introduction, no stimulation was found. Further experiments revealed that the intracellular radioactive amino acid pool was generally increased by aldosterone after 20, 40 and 60 min, while a distinct increased radioactive protein content was found to be induced by aldosterone only after 40 min. This indicates that aldosterone increases the uptake of RNA and protein precursors and the new synthesis of RNA and proteins. These events seem to to be regulated not continously but intermittently. The induced proteins possibly take part in the mediation of the early hormone response. Experiments with the aldosterone antagonist, spironolactone, provide evidence for the specificity of the described hormone effects. The results after application of the Na⁺ channel blocker, amiloride, and the Na⁺/K⁺-ATPase inhibitor, G-strophanthin, indicate that the aldosterone effects are controlled by Na⁺ channels and Na⁺ pumps and therefore by the intracellular Na⁺ content. The inhibitory effect of cycloheximide on the aldosterone-induced protein synthesis indicates the role of these proteins on the hormone-stimulated Na⁺ transport.

show abstract

Lymphoid cells in the turtle bladder

et al. 1973

View full text Add to dashboard Cite

Lymphoid nodules and diffuse lymphocytic infiltration are common in turtle urinary bladders. Nodules, usually located immediately beneath the epithelial layer, were found in 15 of 29 different hemibladders surveyed. A whole cross-section of each hemibladder was examined in the survey. The nodules consisted of aggregates of dense lymphocytes without germinal centers. Erosion of the epithelial layer over the nodule was found in approximately onethird of the cases. Scattered lymphocytes were abundant within the epithelial layer and were found in smears from the mucosal surface. Pyroninophilic cells resembling plasma cells were found in all regions of the bladder. Electron micrographs showed lymphocytes and plasma cells in the epithelial layer above the basement membrane.In the course of a study of the morphology of the urinary bladder of the water turtle, we became impressed with the abundance of lymphoid cells in this tissue. Both lymphoid nodules and diffuse lymphocytic infiltration were common. Previous descriptions of the turtle bladder have not mentioned this finding (LeFevre et al., '71; Rosen, '70a,b). Since knowledge of the nature and extent of lymphoid activity in the turtle bladder is necessary for a full understanding of the functions of this tissue, we investigated the lymphoid cells of the bladder in some detail. The findings and their implications are the topics of the present paper.A preliminary report of this work has appeared elsewhere (LeFevre et al., '72).Turtles (Pseudemys scripta) were obtained from Mogul-Ed Co. (Oshkosh, Wisc.) and were kept in tanks with access to fresh running water until used. The urinary bladder is bilobed and after removal from the turtle was divided into hemibladders. Parasitized and abnormal bladders were discarded. The hemibladders were rinsed, fixed in 10% formalin containing 0.24 M sodium acetate and embedded in paraffin. Sections of whole hemi- MATERIALS AND METHODSANAT. REC., 176: 111-120.bladders were stained with hematoxylin and eosin. Tissue to be examined for pyroninophilic cells was fixed in fresh Carnoy's solution and stained with methyl green-pyronin (Elias, '69 ) .Smears were made by touching the unbroken mucosal surface of a fresh hemibladder to a glass slide and staining with Wright-Giemsa.Small pieces of tissue were fixed in 3% glutaraldehyde in 0.1 M phosphate buffer (pH 7.2) and embedded in Epon. Sections were cut at 0.5 with a glass knife and stained with 1% methylene blue. Sections for electron microscopy were fixed in 3% glutaraldehyde and postfixed in 1% OsO+ both in 0.1 M phosphate buffer (pH 7.2). The sections were embedded in Epon and stained with uranyl acetate. Electron micrographs were taken with a Philips EM 300. OBSERVATIONS Lymphoid nodulesA survey of three groups of turtles obtained in different months was made to determine the frequency of lymphoid nodules in turtle bladders. Random sections of 29 different hemibladders were prepared and the whole circumference of each was ex-

show abstract

Effects of Aldosterone on RNA and Protein Synthesis in the Toad Bladder

Cited by 23 publications

References 47 publications

Kinetics of RNA Labeling in Toad Bladder Epithelium: Effects of Aldosterone and Related Steroids

Kinetics of RNA Labeling in Toad Bladder Epithelium: Effects of Aldosterone and Related Steroids

Action of Aldosterone on Cultured Renal Collecting Duct Cells during the Latent Period

Lymphoid cells in the turtle bladder

Contact Info

Product

Resources

About