Studies on the teratogenic properties of trypan blue and its components in mice

Development ◽  
1964 ◽  
Vol 12 (1) ◽  
pp. 1-14
Author(s):  
Aeleta N. Barber ◽  
Jack C. Geer
Keyword(s):  
Convenient Method ◽  
Trypan Blue ◽  
Yolk Sac ◽  
Teratogenic Effect ◽  
Time Limit ◽  
Direct Access ◽  
Maximal Effect ◽  
Visceral Yolk Sac ◽  
The Stability

Since the discovery of the teratogenicity of trypan blue by Gillman, Gilbert, Gilhnan and Spence (1948) the dye has offered a very concise and convenient method for studying teratogenesis in mammals. Dijkstra & Gillman (1960) fractionated the dye and found that the purple component stimulated the endothelial system in rats, thereby raising the speculation that the varied effects of the dye were caused by contamination. According to our previous experiments (1957, 1963) and those of Wilson, Beaudoin & Free (1959) one aspect of the problem has remained fairly constant, namely, the narrow time limit of the maximal effect of the dye and the stability of the pattern of malformations. Wilson and his associates suggest that the teratogenic effect is due to direct access of the dye to the embryo before the visceral yolk-sac is completely formed.

Effects of trypan blue and Niagara blue 2 B on the in vitro absorption of ions by the rat visceral yolk sac

Development ◽  
1969 ◽  
Vol 22 (1) ◽  
pp. 115-125
Author(s):  
Marten M. Kernis ◽  
E. Marshall Johnson
Keyword(s):  
Trypan Blue ◽  
Vascular System ◽  
Yolk Sac ◽  
Visceral Yolk Sac

Prior to 1927, when Brunschwig proposed that the rat visceral yolk sac is functionally a placenta, it was customary to consider the chorio-allantoic placenta as the primary organ for bringing nourishment to and taking waste from the developing embryo and growing fetus. Eight years later, Everett (1935) predicted that the visceral yolk sac is functionally at least as important as the chorio-allantoic placenta, while Noer & Mossman (1947) suggested that the yolk sac functions differently from the placenta and is therefore complementary in its role. More recently, Halliday (1955) indicated that the proximal yolk sac of the rat is capable of absorbing antibodies by day 17 of gestation. This was confirmed by Brambell & Halliday (1956), who were able to demonstrate that the vitelline epithelium and its underlying vascular system are partly responsible for antibodies penetrating into the embryo.

Differentiation of alkaline phosphatase and glucose-6-phosphate dehydrogenase in rat yolk-sac

Development ◽  
1968 ◽  
Vol 19 (2) ◽  
pp. 137-143
Author(s):  
E. Marshall Johnson ◽  
Ralph Spinuzzi
Keyword(s):  
Vitamin B12 ◽  
Trypan Blue ◽  
Intrinsic Factor ◽  
Yolk Sac ◽  
Early Gestation ◽  
Apical Cytoplasm ◽  
Visceral Yolk Sac ◽  
Chorioallantoic Placenta ◽  
Pass Through ◽  
Glucose 6 Phosphate Dehydrogenase

During early gestation in the rat, at the time of rapid embryonic differentiation and prior to the full formation of the chorioallantoic placenta, the function of ‘placental’ nutrition was attributed to the yolk-sac by Brunschwig (1927). Such a paraplacental function of the yolk-sac would assume that nutrients pass through the parietal wall into the yolk-sac cavity and thence into the embryo via the visceral yolk-sac epithelium and its underlying vitelline vessels. Supporting this concept were the findings of Everett (1935) who demonstrated in 13-day embryos that toluidine blue was able to pass into the omphalomesenteric vessels more rapidly than it could reach the umbilical veins via the chorioallantoic placenta. Furthermore, the visceral entodermal cells appeared to exert some selectivity in that trypan blue did not pass into the embryo but was localized in the apical cytoplasm. More recently, Padykula, Deren ' Wilson (1966) demonstrated that the rat yolk-sac concentrated both vitamin B12 and vitamin B12 plus intrinsic factor throughout most of gestation.

Hepoxilin A, hydroxyepoxide metabolite of arachidonic acid, stimulates transport of 45Ca across the guinea pig visceral yolk sac

1984 ◽  
Vol 62 (12) ◽  
pp. 1466-1469 ◽  
Author(s):  
L. O. Derewlany ◽  
C. R. Pace-Asciak ◽  
I. C. Radde
Keyword(s):  
Arachidonic Acid ◽  
Guinea Pig ◽  
Calcium Transport ◽  
Yolk Sac ◽  
Maximal Effect ◽  
Isolated Pancreatic Islets ◽  
Ussing Chambers ◽  
Visceral Yolk Sac

The effect of hepoxilin A, a newly isolated hydroxyepoxide metabolite of arachidonic acid, on calcium transport across the visceral yolk sac membrane of the guinea pig was investigated in vitro in Ussing chambers. While 1-14C-labelled hepoxilin A itself was not transported across the membrane, it increased the rate of transport of calcium toward the side to which hepoxilin A was added. The degree of increase in calcium transport was similar whether hepoxilin A was added to the maternal side or to the fetal side of the membrane. The observed effect was dependent on the concentration of hepoxilin A over a narrow range (0.5–1.0 × 10−6 M). It was also dependent on the time of incubation reaching maximal effect by 25 min. We have recently observed that hepoxilin A is formed from platelet-derived 12-hydroperoxyeicosatetraenoic acid (12-HPETE) through hernin and hemoglobin catalysis as well as during perifusion of 12-HPETE through isolated pancreatic islets. The present study suggests that hepoxilin A, if formed in vivo, could play a role in the mobilization of calcium.

Visualization of Antibody Transport in Rat Visceral Yolk-Sac Placenta

1982 ◽  
Vol 40 ◽  
pp. 246-247
Author(s):  
William P. Jollie
Keyword(s):  
Phosphate Buffer ◽  
Yolk Sac ◽  
Female Rats ◽  
Thin Sections ◽  
Visceral Yolk Sac ◽  
Antibody Complex ◽  
Cytochemical Reaction ◽  
Hind Foot ◽  
Antigen Antibody ◽  
Yolk Sac Placenta

A technique has been developed for visualizing antibody against horseradish peroxidase (HRP) in rat visceral yolk sac, the placental membrane across which passive immunity previously has been shown to be transferred from mother to young just prior to birth. Female rats were immunized by injecting both hind foot pads with 1 mg HRP emulsified in complete Freund's adjuvant. They were given a booster of 0.5mg HRP in 0.1 ml normal saline i.v. after one week, then bred and autopsied at selected stages of pregnancy, viz., 12, 1 7 and 22 days post coitum, receiving a second booster, injected as above, five days before autopsy. Yolk sacs were removed surgically and fixed immediately in 2% paraformaldehye, 1% glutaraldehye in 0.1 M phosphate buffer with 0.01% CaCl2 at pH 7.4, room temperature, for 3 hr, rinsed 3X in 0.1 M phosphate buffer plus 5% sucrose, then exposed to 1 mg HRP in 1 ml 0.1 M phosphate buffer at pH 7.4 for 1 hr. They were refixed in aldehydes, as above, for 1 5 min (to assure binding of antigen-antibody complex). Following buffer washes, the tissues were incubated in 3 mg diaminobenzidine tetrahydrochloride and 0.01% H2O2 in 0.05 M Tris-HCl buffer for 30 min. After brief buffer washes, they were postfixed in 2% OsO4. in phosphate buffer at pH 7.4, 4°C for 2 hr, dehydrated through a graded series of ethanols, and embedded in Durcupan. Thin sections were observed and photographed without contrast-enhancement with heavy metals. Cytochemical reaction product marked the site of HRP (i.e., antigen) which, in turn, was present only where it was bound with anti-HRP antibody.

The activities of thiol proteases in the rat visceral yolk sac increase during late gestation

Placenta ◽  
1991 ◽  
Vol 12 (2) ◽  
pp. 143-151 ◽  
Author(s):  
John D. Grubb ◽  
Thomas R. Koszalk ◽  
Joseph J. Drabick ◽  
Robert M. Metrione
Keyword(s):  
Yolk Sac ◽  
Late Gestation ◽  
Visceral Yolk Sac ◽  
Thiol Proteases

Effect of yolk-sac antibody on rat embryos grown in culture

Development ◽  
1972 ◽  
Vol 27 (3) ◽  
pp. 543-553
Author(s):  
D. A. T. New ◽  
R. L. Brent
Keyword(s):  
Direct Contact ◽  
Culture Medium ◽  
Gamma Globulin ◽  
Yolk Sac ◽  
Amniotic Cavity ◽  
Pregnant Rats ◽  
Rat Embryos ◽  
Visceral Yolk Sac ◽  
Primary Action

Rat embryos, explanted with their embryonic membranes during the early stages of organogenesis ( days gestation), were grown in culture in roller tubes. Yolk-sac antibody (sheep anti rat yolk-sac gamma globulin), known to be teratogenic when injected into pregnant rats, was added to the culture medium. At concentrations of 0·1 mg/ml or more the antibody caused gross retardation of growth and differentiation. Injection of antibody into the amniotic cavity so that it had direct contact with the embryo, or between the amnion and yolk sac so that it was in contact with the mesodermal surface of the yolk sac, had little or no effect on development of the embryo or its membranes. These in vitro experiments indicate that yolk-sac antibody has an effect on development independent of any immunological reaction of the mother, and the primary action is probably on the visceral yolk-sac endoderm.

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