Experimental yolk sac dysfunction as a model for studying nutritional disturbances in the embryo during early organogenesis

Teratology ◽  
1990 ◽  
Vol 41 (4) ◽  
pp. 405-413 ◽  
Author(s):  
R. L. Brent ◽  
D. A. Beckman ◽  
M. Jensen ◽  
T. R. Koszalka
Development ◽  
1992 ◽  
Vol 115 (1) ◽  
pp. 289-303 ◽  
Author(s):  
A. Orr-Urtreger ◽  
M.T. Bedford ◽  
M.S. Do ◽  
L. Eisenbach ◽  
P. Lonai

The alpha receptor of PDGF (Pdgfra) is expressed in primitive endoderm and mesoderm derivatives throughout embryogenesis. In the early primitive streak stage the gene is transcribed in the visceral and parietal endoderm. Later it is expressed in the presomitic mesoderm, yolk sac and amnion. During somitogenesis its transcription localizes to the heart and the somites. Subsequently, it is transcribed in the dermatome, the sclerotome, the developing limb and in various mesenchymal tissues of visceral organs. Its wild-type expression pattern correlates well with the phenotype of homozygous mutant Patch (Ph) embryos, where the Pdgfra gene is deleted. The Ph phenotype is first detectable at the primitive streak stage with convoluted and hypertrophic visceral yolk sac, deformed neural plate and disorganized or missing mesoderm. Most Ph/Ph embryos die before the 11th day of gestation. Those that survive till early organogenesis are very small, have hypertrophic yolk sacs, small and undifferentiated somites, convoluted neural tubes, large heart and pericardium, rudimentary limb buds and branchial arches. Our observations together suggest that the alpha PDGF receptor may be required for the normal development of visceral endoderm and mesoderm derivatives.


Development ◽  
1985 ◽  
Vol 86 (1) ◽  
pp. 271-281
Author(s):  
Stuart J. Freeman ◽  
Nigel A. Brown

The effects of two proteinase inhibitors, leupeptin and pepstatin on the development of 9·5-day rat conceptuses in vitro has been studied. All cultures were of 48 h duration and the inhibitors were present throughout the entire period. When pepstatin was added to the culture medium (5–25 μg/ml) conceptuses developed and grew to an extent that did not differ from untreated controls. However, leupeptin (l–4 μg/ml) caused severe growth retardation and abnormal development of conceptuses. The effects of the two inhibitors on the hydrolysis of 125I-labelled BSA and haemoglobin by homogenates of 10·5-day yolk sac indicated the biochemical basis for the differential toxic effects of the two inhibitors on development. Leupeptin was highly inhibitory of the degration of both substrates whereas pepstatin caused no inhibition of 125I-labelled BSA hydrolysis, and only a slight inhibition of haemoglobin hydrolysis. These observations demonstrate that cathepsin D, a lysosomal aspartic proteinase that is specifically inhibited by pepstatin is not involved in yolk-sac-mediated protein utilization by early organogenesis-phase conceptuses and that lysosomal cysteine proteinases, specifically inhibited by leupeptin, are of paramount importance in this yolk sac function.


Author(s):  
D.A. Beckman ◽  
YP. Wang ◽  
G. Yang ◽  
M. Zeng ◽  
YP. Wang ◽  
...  

The human secondary yolk sac persists for a short but critical period of development during early organogenesis. Although it is established that the yolk sac is the site of origin of several cell lines, a locus of hematopoiesis and secretes several proteins, the yolk sac may have other as yet less well defined functions. Here we focus on ultrastructural evidence of the role of the human yolk sac as an organ of exchange with the extraembryonic coelom and the yolk-sac cavity. Yolk sacs of 6 to 10 weeks developmental age were fixed and processed for light microscopy, TEM and SEM by standard procedures. Numerous microvilli, coated and uncoated vesicles, and lysosomes in the mesothelial layer together suggest a high capacity for pinocytosis of coelomic fluid and lysosomal digestion of internalized substrate. It is not known whether this process is a significant nutritional source for the yolk sac and embryo as it is in the rodent. Both SEM and TEM observations provide evidence of cilia on dispersed cells in the endodermal layer lining the yolk sac cavity. To our knowledge, there is only one other brief report of cilia on the human yolk sac.


Placenta ◽  
1991 ◽  
Vol 12 (1) ◽  
pp. 37-46 ◽  
Author(s):  
D.A. Beckman ◽  
J.E. Pugarelli ◽  
T.R. Koszalka ◽  
R.L. Brent ◽  
J.B. Lloyd

Author(s):  
William P. Jollie

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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