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Changes in the sensitivity of chick fibroblasts to Ricinus lectin (RCA I) toxicity in relation to the stage of embryo development.

ABSTRACT: The toxic effect of Ricinus lectin RCA I, as estimated by the inhibition of [3H]leucine incorporation, was investigated on chick-embryo fibroblasts at different stages of development. There appeared to be a differential susceptibility of chick-embryo fibroblasts to lectin RCA I. Fibroblasts from 16-day embryos were the most sensitive to its toxic effect in terms of both concentration and time, and cells from 8-day embryos were the least sensitive. This differential sensitivity to the toxic effect of lectin RCA I was closely related to the binding of the lectin: fibroblasts from 16-day embryos had more binding sites (1.5 x 10(7)/cell) with a high affinity than did 12-day (0.45 x 10(7)/cell) or 8-day embryos (0.2 x 10(7)/cell). Studies on the specificity and the removal of bound lectin RCA I by D-galactose indicated that the lectin binding was necessary but not sufficient in itself to cause the toxic effect and that the lectin needed to enter the cells in order to be toxic. The amount of lectin RCA I needed to induce a 50-60% toxicity enters fibroblasts of 16-day embryos more rapidly than those of 12- and 8-day embryos.

SUBMITTER: Bernard B

PROVIDER: S-EPMC1161231 | biostudies-other | 1979 Jul

REPOSITORIES: biostudies-other

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Changes in lectin-induced deoxyribonucleic acid synthesis in cultures of chick-embryo fibroblasts at various stages of development.

Project description:Concanavalin A and Robinia pseudoacacia lectin decreased [(3)H]thymidine incorporation into acid-insoluble material of fibroblasts cultured from 6-10-day chick embryos. In contrast, these lectins stimulated [(3)H]thymidine incorporation in cells from 16-day embryos. These effects are due to neither [(3)H]thymidine permeability modification nor toxicity of the lectins. The specificity of lectin action was proved by blocking experiments with alpha-methyl mannopyranoside and with anti-(Robinia lectin) serum.

| S-EPMC1172487 | biostudies-other

NMR Based Cerebrum Metabonomic Analysis Reveals Simultaneous Interconnected Changes during Chick Embryo Incubation.

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| S-EPMC4618859 | biostudies-literature

Isolation and characterization of a vinculin cDNA from chick-embryo fibroblasts.

Project description:A chick-embryo fibroblast lambda gt11 cDNA library was screened with affinity-purified antibodies to chick gizzard vinculin. One recombinant was purified to homogeneity and the fusion protein expressed in Escherichia coli strain C600. The fusion protein was unstable, but polypeptides that reacted with vinculin antibodies, but not non-immune immunoglobulin, were detected by Western blotting. The recombinant contained a single EcoRI fragment of 2891 bp with a single open reading frame. The deduced protein sequence could be aligned with that of six CNBr-cleavage peptides and two tryptic peptides derived from chicken gizzard vinculin. AUG-247 has tentatively been identified as the initiation codon, as it is contained within the consensus sequence for initiation sites of higher eukaryotes. The cDNA lacks 3' sequence and encodes 74% of the vinculin sequence, presuming the molecular mass of vinculin to be 130,000 Da. Analysis of the deduced sequence showed no homologies with other protein sequences, but it does display a triple internal repeat of 112 amino acid residues covering residues 259-589. The sequences surrounding the seven tyrosine residues in the available sequence were aligned with the tyrosine autophosphorylation consensus sequence found in protein tyrosine kinases. Tyr-822 showed a good match to this consensus, and may represent one of the two major sites of tyrosine phosphorylation by pp60v-sre. Northern blots showed that the 2.89 kb vinculin cDNA hybridized to one size of mRNA (approx. 7 kb) in chick-embryo fibroblasts, chick smooth muscle and chick skeletal muscle. Southern blots revealed multiple hybridizing bands in genomic DNA.

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Dynamic Transcriptional Landscape of the Early Chick Embryo.

Project description:Defining the dynamic transcriptome of the early embryo at high resolution would assist greatly in understanding vertebrate development. Here, we describe the dynamic transcription landscape of early chick embryo development using advanced single-molecule long-read isoform sequencing (Iso-Seq) and RNA-Seq technology. Our transcriptomic profiling reflected the time course of chicken embryonic development from day 1 to day 8 of incubation, a period encompassing gastrulation, somitogenesis, and organogenesis. This analysis identified transcriptional isoforms, alternative splicing (AS) events, fusion transcripts, alternative polyadenylation (APA) sites, and novel genes. Our results showed that intron retention (IR) represented the most abundant AS type and displayed distinct features and dynamic modulation during development. Moreover, we constructed a high-resolution expression profile across embryonic development. Our combined expression dataset correlates distinct gene clusters with specific morphological changes, and provides the first framework for the molecular basis of early chicken embryogenesis. Analysis of gene expression in the developing chicken embryo highlighted the dynamic nature and complexity of the chicken transcriptome and demonstrated that dramatically increased IR events are associated with distinct gene sets.

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Context learning before birth: evidence from the chick embryo.

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Effects of chromic chloride on chick embryo fibroblast viability.

Project description:The objective of this study is to evaluate the effects of chromic chloride (CrCl3) on chick embryo fibroblast (CEF) viability. The cells were incubated with CrCl3 (0.02, 0.1, 0.5, 2.5, 12.5, and 62.5 ?M), and the viability was determined using MTT assay, morphological detection and flow cytometry. The results show that lower concentrations of CrCl3 (0.02, 0.1, and 0.5 ?M) did not damage CEF viability. At 0.1 ?M, CrCl3 can increase CEF viability (P < 0.05). However, at higher concentrations of CrCl3 (2.5, 12.5, and 62.5 ?M), the number of apoptotic and necrotic cells (P < 0.01) and intracellular reactive oxygen species (P < 0.01) increased. In addition, decreased mitochondrial membrane potential (P < 0.01) and enhanced intracellular calcium levels (P < 0.01) were observed after the exposure. Moreover, apoptotic morphological changes induced by these processes in CEF were confirmed using Hoechst 33258 staining. Cell death induced by higher concentrations of CrCl3 was caused by an apoptotic and a necrotic mechanism, whereas the main mechanism of oxidative stress and induced mitochondrial dysfunction was apoptotic death. The induced apoptotic death in CEF is concentration- and time-dependent.

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