Separation of germ layers in presomite rat embryos

Levak-Švajger, B.; Švajger, Anton; Škreb, N.

doi:10.1007/bf01897519

Cited by 82 publications

(12 citation statements)

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“…After separating the embryonic region from all extraembryonic tissues, the node and the primitive streak were removed to prevent contamination of cells from one germ layer with those from another. The embryos were incubated in 2.5% pancreatin and 0.5% trypsin in PBS for 8 min on ice (Levak-Svajger et al, 1969;Hogan and Tilly, 1981), and then placed in DME with 10% FCS again. Embryonic ectoderm, mesoderm, and endoderm (most likely a mixture of cells of primitive endoderm and epiblast origin; Lawson et al, 1986) were separated from each other with tungsten needles and processed separately.…”

Section: A6ab1 and A6bp1 In Mouse Development Experimental Proceduresmentioning

confidence: 99%

Expression patterns of laminin receptor splice variants α6Aβ1 and α6Bβ1 suggest different roles in mouse development

Þorsteinsdóttir

Roelen

Freund

et al. 1995

Developmental Dynamics

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The a6pl integrin is a receptor for laminins and is present from early stages of mouse embryogenesis. In the present study we determined the temporal and spatial expression of the two cytoplasmic splice variants of the a6 integrin subunit, a6A and a6B, in the early-and midgestation mouse postimplantation embryo using RT-PCR, in situ hybridization, and immunofluorescence. Our results show that a6B is present in the embryo at all stages studied and is expressed before a6A. a6A expression begins in 8.5 day p.c. embryos and is initially exclusively localized to the developing heart. In 8.5 (and 9.5) day p.c. embryos a6A mRNA and protein are present in a gradient in the myocardium of the heart tube from strongest expression in the sinus venosus and in the common atrial chamber to a weakening expression along the ventricle and bulbus cordis. In 10.5 day p.c. embryos this gradient is less evident and in 12.5 day p.c. embryos a6A mRNA and protein are present in comparable amounts between atria and ventricles. Neither a6A nor a6B is present in endocardial cushion tissue. By day 12.5 p.c. (w6A expression is also present in the developing epidermis, dental primordia, lens, gonads, and in a few epithelia such as those of the digestive tract. a6B expression is always much more widespread than a6A expression. For example, only a6B is present in the myotome of the somites of 9.5 day p.c, embryos, in the developing central and peripheral nervous systems, and in the nephrogenic system at all stages studied, except after the differentiation of the gonads when a6A is also present. Furthermore, a6B is the only splice variant present on endothelial cells. We also examined the distribution of the f34 integrin subunit to determine whether the a6P4 integrin was present during these stages of development. p4 protein was absent in early postimplantation stages but was present in the epidermis and digestive tract of 12.5 day p.c. embryos. These results show a differential distribution of a6A and a6B during mouse development and thus strongly suggest a different function of these splice variants during embryogenesis. Our results point to a possible role for the 0 1995 WILEY-LISS, INC. a6Apl integrin in the development of the myocardium of the developing heart, but not in the migration of endocardial cushion cells, while a6Bp1 could be important in the developing nephrogenic and nervous systems. o l W 5 Wiley-Liss, Inc.

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Section: A6ab1 and A6bp1 In Mouse Development Experimental Proceduresmentioning

confidence: 99%

Expression patterns of laminin receptor splice variants α6Aβ1 and α6Bβ1 suggest different roles in mouse development

Þorsteinsdóttir

Roelen

Freund

et al. 1995

Developmental Dynamics

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“…Embryos were dissected from their deciduainto PB1 (Whittingham & Wales, 1969) containing04% polyvinylpyrollidone (PVP) instead of albumin. The extra-embryonic endoderm layer (which has formed yolk-sac endoderm by 8 days) was loosened from the embryonic and extra-embryonic ectoderm regions by digestion in 0 -5 % trypsin (Sigma) and 2-5 % pancreatin (BDH) in calcium-and magnesium-free phosphate-buffered saline (PBS) for 10 min at 4 °C (Levak-Svajger, Svajger & Skreb, 1969). Further enzyme digestion was prevented by washing embryos in PB1-PVP containing 10% foetal calf serum (FCS) at 4 °C.…”

Section: (Ii) Dissections and Sample Collectionsmentioning

confidence: 99%

X-chromosome activity in female mouse embryos heterozygous forPgk-1and Searle's translocation, T(X; 16) 16H

McMahon

Monk

1983

Genet. Res.

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To investigate whether the preferential expression of genes on the translocated X chromosome in female mice carrying the X-autosome translocation T(X; 16)16H (Searle's translocation) is due to non-random inactivation or to cell selection, we examined tissues of mouse embryos heterozygous for the X-linked gene coding for phosphoglycerate kinase (Pgk-1). From the cross Tl6RPgk-l b /+ Pgk-1» 9 x + Pgk-l a /Y

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“…1). The egg cylinders were incubated in a mixture of 0.5% trypsin and 2.5% pancreatin in calcium-and magnesium-free Tyrode's solution for 10-20 minutes at 4°C [21]. The layers were then separated by a combination of mouth pipetting with a finely drawn pipette and cutting with tungsten needles.…”

Section: Dissectionsmentioning

confidence: 99%

Non‐random X‐chromosome expression early in mouse development

et al. 1981

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We have used a sensitive electrophoretic technique for estimating the activity, or ratio, of two allozymes of the X-chromosome-linked enzyme phosphoglycerate kinase (PGK-I), in order to investigate the randomness of X-chromosome expression in the derivatives of the three primary cell lineages of the early mouse conceptus. The maternally derived Pgk-f allele is preferentially expressed in the derivatives of the primitive endoderm and trophectoderm lineages at 6% days post coitum in Pgk-l"/Pgk-l* heterozygous conceptuses, and in the one informative S%-day heterozygous conceptus analysed. This evidence for preferential expression of the maternally derived X chromosome (Xm), so soon after the time of X-chromosome inactivation, favors the possibility that the preferential expression of Xm is a consequence of primary nonrandom X-chromosome inactivation, rather than a secondary selection phenomenon. The majority of embryos analysed at 4% and 5% days pc produced only a single PGK-I band, corresponding to the allozyme produced by the Pgk-l allele on X", although 50% of these embryos should have been heterozygous females. Possible explanations are discussed.

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Separation of germ layers in presomite rat embryos

Cited by 82 publications

References 0 publications

Expression patterns of laminin receptor splice variants α6Aβ1 and α6Bβ1 suggest different roles in mouse development

Expression patterns of laminin receptor splice variants α6Aβ1 and α6Bβ1 suggest different roles in mouse development

X-chromosome activity in female mouse embryos heterozygous forPgk-1and Searle's translocation, T(X; 16) 16H

Non‐random X‐chromosome expression early in mouse development

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