A modified fluorometric method for the determination of microgram quantities of DNA from cell or tissue cultures

Setaro, Frank; Morley, Colin G.D.

doi:10.1016/0003-2697(76)90043-9

Cited by 305 publications

(53 citation statements)

References 7 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To estimate dry lung weight, previously weighed fetal lungs were dried to constant weight in a vacuum oven at 86°C. To quantify DNA, frozen lungs were homogenized by ultrasonication, and DNA content was measured using the fluorometric method of Setaro and Morley (43). Protein in lung tissue was measured with a commercially available kit (Sigma, St. Louis, MO) using the bicinchoninic acid (BCA) modification of the Lowry method (44).…”

Section: Methodsmentioning

confidence: 99%

Effects of oligohydramnios on lung growth and maturation in the fetal rat

Kitterman

Chapin

Vanderbilt

et al. 2002

American Journal of Physiology-Lung Cellular and Molecular Phys

View full text Add to dashboard Cite

Oligohydramnios (OH) retards fetal lung growth by producing less lung distension than normal. To examine effects of decreased distension on fetal lung development, we produced OH in rats by puncture of uterus and fetal membranes at 16 days of gestation; fetuses were delivered at 21 or 22 days of gestation. Controls were position-matched littermates in the opposite uterine horn. OH lungs had lower weights and less DNA, protein, and water, but no differences in saturated phosphatidylcholine, surfactant proteins (SP)-A and -B, and mRNA for SP-A, -B, -C, and -D. To evaluate effects on epithelial differentiation, we used RTI 40 and RTII70, proteins specific in lung to luminal surfaces of alveolar type I and II cells, respectively. At 22 days of gestation, OH lungs had less RTI40 mRNA (P Ͻ 0.05) and protein (P Ͻ 0.001), but RTII70 did not differ from controls. With OH, type I cells (in proportion to type II cells) covered less distal air space perimeter (P Ͻ 0.01). We conclude that OH, which retards lung growth, has little effect on surfactant and impedes formation of type I cells relative to type II cells. fetal lung development; lung distension; pulmonary epithelial differentiation; pulmonary hypoplasia; pulmonary surfactant IN LATE GESTATION, the fetal lung undergoes marked changes in preparation for the transition to extrauterine life. These changes include growth, enlargement of distal potential air spaces, thinning of the septa, maturation of the surfactant system, and differentiation of distal pulmonary epithelium into mature alveolar type I and type II cells. Several studies have shown that fetal lung growth is controlled primarily by mechanical factors, especially distension of the lung (for reviews, see Refs. 26 and 29) and that maturation of the surfactant system is controlled primarily by endocrine factors (for reviews, see Refs. 3 and 48). In contrast, relatively little is known about factors that regulate differentiation of the alveolar epithelium.In 1977, Alcorn and associates (2) reported that tracheal ligation in fetal sheep, which increased lung distension, caused accelerated lung growth and a qualitative reduction in the number of alveolar type II cells; conversely, chronic drainage of tracheal fluid, which inhibited lung distension, retarded lung growth and increased the number of type II cells. However, they reported neither quantitative data for cell counts nor measurements of indicators of surfactant, which is produced by type II cells. Subsequently, other investigators have studied tracheal ligation in fetal sheep and have confirmed that an increase in lung distension results in a lower number of type II cells (6,11,37) and a higher percentage of type I cells (18). Tracheal ligation also results in lower concentrations in the lung of surfactant protein (SP)-A and saturated phosphatidylcholine (SatPC), the major surface-active lipid in pulmonary surfactant (28), as well as mRNA for SP-A, -B, and -C (32). Conversely, transection of the cervical spinal cord (which abolishes fetal breathing movemen...

show abstract

Section: Methodsmentioning

confidence: 99%

Effects of oligohydramnios on lung growth and maturation in the fetal rat

Kitterman

Chapin

Vanderbilt

et al. 2002

American Journal of Physiology-Lung Cellular and Molecular Phys

View full text Add to dashboard Cite

show abstract

“…When the cells were harvested, DNA content was determined by use of the microftuorometric method of Setaro and Morley (19), and DNA synthesis was measured as we have previously reported (16).…”

Section: Animalsmentioning

confidence: 99%

Hepatocyte proliferation and gene expression induced by triiodothyronine in vivo and in vitro

Francavilla¹

1994

Hepatology

View full text Add to dashboard Cite

Subcutaneous injections of hormone triiodothyronine in rats resulted in peak blood levels at 24 hr with return to baseline by 96 hr. The injections stimulated a liver regeneration response that resembled in timing and in magnitude of DNA synthesis (peak, 24 hr) that induced by 40% hepatic resection. The principal proliferation was of hepatocytes. Although there were some temporal differences from the gene expression of transforming growth factor-a, transforming growth factor-~, and c-Ha-ras that are known to follow partial hepatectomy, the overall profile of these changes was similar to those after partial resection. The effect was liver specific and could be reproduced three times with no diminution in response in the same animal with injections at 10-day intervals. No response was detected in kidney or intestine. This effect in intact animals contrasted with the minimal ability of triiodothyronine to stimulate hepatocytes in culture. However, when the culture medium was enriched with epidermal growth factor, there was a dose-related response to triiodothyronine. The totality of these experiments provides a preliminary basis for the creation with pharmacological techniques of an in vivo hyperplastic hepatic condition permissive of transfection of new genes, as an alternative to partial hepatectomy. Although triiodothyronine was the test agent used, other hepatic growth factors singly or in combination could be candidates for this purpose.

show abstract

“…DNA was prepared from 0.1-0.2 ml of the lung homogenate by precipitation with 10% trichloracetic acid and extraction with methanol and absolute alcohol. DNA was then assayed by the fluorometric method of Setaro and Morley (20).…”

mentioning

confidence: 99%

Ontogeny of Surfactant Apoproteins in the Rat

et al. 1989

View full text Add to dashboard Cite

ABSTRACT. Content of the 26-38-kD surfactant apoprotein (SP-A) was determined in lung homogenates from fetal (17-21 d gestation), postnatal (1-28 d of life), and adult male and female rats by a double sandwich ELISA. Expression of mRNA for SP-A as well as the hydrophobic apoproteins, SP-B and SP-C, were also determined in lung homogenates from fetal and adult rats of both sexes by Northern blot analysis. SP-A was undetectable in fetal

show abstract

A modified fluorometric method for the determination of microgram quantities of DNA from cell or tissue cultures

Cited by 305 publications

References 7 publications

Effects of oligohydramnios on lung growth and maturation in the fetal rat

Effects of oligohydramnios on lung growth and maturation in the fetal rat

Hepatocyte proliferation and gene expression induced by triiodothyronine in vivo and in vitro

Ontogeny of Surfactant Apoproteins in the Rat

Contact Info

Product

Resources

About