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.

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.

scholarly journals Histochemical differences in expression of X-linked glucose-6-phosphate dehydrogenase between ectoderm- and endoderm-derived embryonic and extra-embryonic tissues.

1991 ◽  
Vol 39 (5) ◽  
pp. 569-574 ◽  
Author(s):  
H Sobis ◽  
A Verstuyf ◽  
M Vandeputte
Keyword(s):  
X Chromosome ◽  
Yolk Sac ◽  
Wild Type ◽  
X Chromosomes ◽  
Tissue Sections ◽  
C3h Mice ◽  
Visceral Yolk Sac ◽  
Enzyme Deficient ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Derivatives Of

We examined the activity of X-linked glucose-6-phosphate dehydrogenase (G6PD) in concepti of the enzyme-deficient mutant and wild-type C3H mice. By using different crosses between the G6PD-deficient homozygous, heterozygous, or wild-type females and hemizygous or wild-type males, we confirmed the inactivation of one of the two X chromosomes in female concepti by a histochemical method. With this technique, a dual (G6PD + or -) cell population could be observed in the tissue sections. We demonstrate that the paternal X chromosome is inactivated in the endoderm of parietal and visceral yolk sac and in the trophoblast, whereas in the embryo and in the yolk sac mesoderm this inactivation is random. Our results confirm biochemical observations showing that only the maternal X chromosome is expressed in all derivatives of trophectoderm and primitive endoderm, whereas derivatives of the primitive ectoderm show random X chromosome expression.

The role of the visceral yolk sac in mediating protein utilization by rat embryos cultured in vitro

Development ◽  
1981 ◽  
Vol 66 (1) ◽  
pp. 223-234
Author(s):  
Stuart J. Freeman ◽  
Felix Beck ◽  
John B. Lloyd
Keyword(s):  
Serum Proteins ◽  
Yolk Sac ◽  
Rat Serum ◽  
Pregnant Rats ◽  
Digestion Product ◽  
Background Levels ◽  
Visceral Yolk Sac ◽  
Chorioallantoic Placenta

Conceptuses from 9·5-day pregnant rats have been cultured for 48 h in heat-inactivated homologous serum. Embryonic development was normal. The protein contents of embryos and visceral yolk sacs after different periods of culture were recorded. When 125-labelled polyvinylpyrrolidone or [3H]dextran were added to the culture serum, radioactivity was accumulated by the yolk sac, but only background levels were detected in the embryo itself. The amount of radioactivity found in the yolk sac varied with the length of the interval before harvesting during which 125 I-labelled PVP or [3H]dextran was present. When formaldehyde-denatured 125 I-labelled bovine serum albumin was added to the culture serum, little radioactivity accumulated in the yolk sac and only background levels were found in the embryo. Trichloroacetic acid-soluble radioactivity steadily appeared in the culture serum, however. When conceptuses were cultured in glucose- and vitamin-supplemented dialysed serum from rats injected 2 h previously with [3H]leucine, radioactivity was found in both embryos and yolk sacs. The amount of radioactivity in these tissues increased with duration of exposure to 3H-labelled serum proteins. After short exposures little of the yolk sac and embryonic radioactivity was acid-insoluble, but this proportion increased with duration of exposure. These results are interpreted as follows. Intact macromolecules cannot enter the cells of the embryo itself, but are captured by pinocytosis into the cells of the visceral yolk-sac endoderm. Indigestible macromolecules such as 125 I-labelled polyvinylpyrrolidone and [3H]- dextran accumulate within the yolk-sac lysosomes, but proteins are digested there by the lysosomal enzymes. The radiolabelled digestion product of 125 I-labelled bovine albumin is [125 I]iodotyrosine, which cells cannot utilize and so is excreted into the culture serum. The labelled digestion product of the 3H-labelled rat serum proteins is [3H]leucine, which is used for protein synthesis in both embryo and yolk sac. The experiments provide direct evidence for the long-suspected role of the yolk sac in mediating embryonic nutrition in the period of development prior to the establishment of a functional chorioallantoic placenta.

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.

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.

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