In vitro development of rat embryos obtained from diabetic mothers

Rat embryos of 9.5 or 10 days of gestation were removed from control or streptozotocin-diabetic mothers and cultured in normal rat serum (180 mg% glucose) or in diabetic serum (600 mg% glucose). The development of control embryos in normal serum was adequate. Embryos from normal mothers cultured in diabetic serum showed signs of developmental retardation. The development of embryos obtained from diabetic mothers was severely impaired, regardless of the gestational age or the culture medium. These results suggest that a diabetic maternal milieu produces irreversible effects in the embryo very early in gestation.     

Effects of puromycin on rat embryos in vitro

Somite-staged rat embryos were exposed to varying concentrations of puromycin for 48 h in vitro. Medium concentrations below 0.92 microM had no significant effects, while concentrations above 1.84 microM were lethal. Between these extremes, there were concentration dependent increases in the incidence of malformations in a close relationship to growth retardation.     

Steroid metabolism by mouse preimplantation embryos in vitro

Summary Mouse preimplantation embryos were incubated with radioactive pregnenolone, progesterone or dehydroepiandrosterone for various periods of time. These substrates were not converted to metabolites even after incubation of 120 h. We suggest that preimplantation mouse embryo does not possess enzyme activities for steroid metabolism.Acknowledgments. The authors would like to thank Prof. L. Saxén for giving the animals and the facilities of his tissue culture laboratory.     

Effects of puromycin on rat embryos in vitro

Summary Somite-staged rat embryos were exposed to varying concentrations of puromycin for 48 h in vitro. Medium concentrations below 0.92 M had no significant effects, while concentrations above 1.84 M were lethal. Between these extremes, there were concentration dependent increases in the incidence of malformations in a close relationship to growth retardation.     

Steroid metabolism by mouse preimplantation embryos in vitro.

Mouse preimplantation embryos were incubated with radioactive pregnenolone, progesterone or dehydroepiandrosterone for various periods of time. These substrates were not converted to metabolites even after incubation of 120 h. We suggest that preimplantation mouse embryo does not possess enzyme activities for steroid metabolism.     

A method for karyotyping mouse blastocyst embryos developing from in vivo and in vitro fertilized eggs

A method for karyotyping blastocyst-staged mouse embryos is described. The use of this protocol results in the recovery of a high percentage (greater than 70%) of readable karyotypes and can be completed rapidly.     

A method for karyotyping mouse blastocyst embryos developing from in vivo and in vitro fertilized eggs

Summary A method for karyotyping blastocyst-staged mouse embryos is described. The use of this protocol results in the recovery of a high percentage (>70%) of readable karyotypes and can be completed rapidly.This research was supported by NIH grants HD-00827 and HD-09753.     

Scanning electron microscopy study of spinal cord nerve cells from chick embryos cultured in vitro with poly-L-lysine]

Chick embryo nerve cells from the lumbo-sacral spinal cord have been isolated by trypsinisation and cultivated in Rose chambers on polylysine-L as a substrate. The cells are analysed by scanning electron microscopy. The mode of adherence of these cells to the substrate and the modifications of the surface of the neuroblasts during their transformation into neurocytes are studied.     

The effect of 5-bromodeoxyuridine on mouse embryos during neurulation in vitro

Summary Mouse embryos explanted at various stages during neurulation were cultured for 20–28 h in the presence of 25–900 g/ml of 5-bromodeoxyuridine (BUdR). BUdR strongly inhibited closure of the cranial neural tube, which was found to be stage-dependent. When mouse embryos were exposed to BUdR after development of the concave curvature in the neuroepithelium of the midbrain to the upper hindbrain regions, they became insensitive to the drug-induced open cranial neural tube. Histological observations showed that BUdR interfered with interkinetic migration and cytokinesis of the neuroepithelial cells. These cellular abnormalities were not dependent on the morphological development of the cranial neural folds. The³H-BUdR experiment confirmed that the label was mostly incorporated into the DNA fraction.Acknowledgment. This work was supported by Grant-in-Aids for scientific research No. 557469 and 58480391 from the Ministry of Education, Japan.     

The effect of 5-bromodeoxyuridine on mouse embryos during neurulation in vitro

Mouse embryos explanted at various stages during neurulation were cultured for 20-28 h in the presence of 25-900 micrograms/ml of 5-bromodeoxyuridine (BUdR). BUdR strongly inhibited closure of the cranial neural tube, which was found to be stage-dependent. When mouse embryos were exposed to BUdR after development of the concave curvature in the neuroepithelium of the midbrain to the upper hindbrain regions, they became insensitive to the drug-induced open cranial neural tube. Histological observations showed that BUdR interfered with interkinetic migration and cytokinesis of the neuroepithelial cells. These cellular abnormalities were not dependent on the morphological development of the cranial neural folds. The 3H-BUdR experiment confirmed that the label was mostly incorporated into the DNA fraction.     

Allophenic mice produced from embryos aggregated with antibody

One difficulty in the production of allophenic mice by aggregation of preimplantation embryos is that they frequently roll apart before the bonds between the blastomeres have had time to form. One solution to the problem, described here, is to pretreat one of the embryos with rabbit anti-mouse serum just prior to pushing them together. Blastocyst formation is unhampered by antibody treatment, and numerous allophenic mice have already been produced with this new procedure.     

Spontaneous peristaltic activity of veins of chick embryos and newly hatched chickens explantedin vitro

Résumé Les veines du système porte et la veine ombilicale gauche d'embryons de poulet dans la dernière semaine d'incubation et de poussins à l'éclosion ont été cultivéesin vitro. Les auteurs y ont démontré la présence de contractions péristaltiques spontanées. Ils pensent que cette activité contractile rythmique favorisein vivo la circulation dans ces territories veineux.     

DNA in wheat seeds from European archaeological sites

We have used hybridization analysis to detect ancient DNA in wheat seeds collected from three archaeological sites in Europe and the Middle East. One of these samples, carbonizedT. spelta dated to the first millennium BC, has yielded PCR products after amplification with primers directed at the leader regions of the HMW (high molecular weight) glutenin alleles. Sequences obtained from these products suggest that the DNA present in the Danebury seeds is chemically damaged, as expected for ancient DNA, and also indicate that it should be possible to study the genetic variability of archaeological wheat by ancient DNA analysis. Finally, we describe a PCR-based system that enables tetraploid and hexaploid wheats to be distinguished.     

Effect of the calmodulin inhibitor R24571 (calmidazolium) on rat embryos cultured in vitro

Summary The possible effects of inhibition of the calcium-binding protein, calmodulin, on mammalian morphogenesis have been investigated by culturing rat embryos in vitro from 9 1/2 to 11 1/2 days of development in the presence of R24571 (calmidazolium), a specific inhibitor of calmoldulin. Embryos cultured in 10^–2 mM R24571 for 48 h show inhibited development and exhibit a range of morphogenetic abnormalities including assymetry and neural tube defects. Embryos exposed to R24571 for the first 24 h of a 48 h culture are more severely affected than embryos exposed to R24571 for the last 24 h.Acknowledgments. We wish to thank Mr C. Veltkamp and Mr B. lewis for their expert help with the scanning electron microscopy and photography.