scholarly journals Analysis of chick somite myogenesis by in situ confocal microscopy of desmin expression.

1994 ◽  
Vol 42 (2) ◽  
pp. 265-272 ◽  
Author(s):  
W H Borman ◽  
D E Yorde
Keyword(s):  
Confocal Microscopy ◽  
Intermediate Filament ◽  
Intermediate Filament Protein ◽  
Chick Embryos ◽  
Whole Mount ◽  
Somite Formation ◽  
Filament Protein ◽  
The Relationship

We explored the relationship in chick embryos between somitogenesis and the onset of somite myogenesis by immunodetection of the muscle-specific intermediate filament protein desmin. Early somite desmin expression was detected by whole-mount in situ confocal microscopy. No detectable somite desmin was observed in embryos of 15 somites (Stage 12) or younger. In embryos having between 16 and 26 somites (Stages 12-15), desmin could be detected in somites positioned increasingly more caudal in the embryo. Finally, in embryos of 27 somites (Stage 16) and older, somite desmin expression was consistently present in all but the caudal-most six somites. Although the rate of somite formation is fairly constant, the rate of observed somite desmin expression progressing caudally in the embryo is greater initially than the rate of segmentation. After an embryo has formed about 27 somites, the rate of desmin appearance parallels the rate of segmentation at a distance of about six somites. This result suggests that very early somite myogenesis is not linked to somitogenesis.

A whole-mount immunocytochemical analysis of the expression of the intermediate filament protein vimentin in Xenopus

Development ◽  
1989 ◽  
Vol 105 (1) ◽  
pp. 61-74 ◽  
Author(s):  
J.A. Dent ◽  
A.G. Polson ◽  
M.W. Klymkowsky
Keyword(s):  
Neural Tube ◽  
Intermediate Filament ◽  
Mesenchymal Cells ◽  
Neural Tube Closure ◽  
Otic Vesicle ◽  
Intermediate Filament Protein ◽  
Western Blots ◽  
A6 Cells ◽  
Whole Mount ◽  
Filament Protein

We have developed a whole-mount immunocytochemical method for Xenopus and used it to map the expression of the intermediate filament protein vimentin during early embryogenesis. We used two monoclonal antibodies, 14h7 and RV202. Both label vimentin filaments in Xenopus A6 cells, RV202 reacts specifically with vimentin (Mr, 55 × 10(3] on Western blots of A6 cells and embryos. 14h7 reacts with vimentin and a second, insoluble polypeptide of 57 × 10(3) Mr found in A6 cells. The 57 × 10(3) Mr polypeptide appears to be an intermediate filament protein immunochemically related to vimentin. In the whole-mount embryo, we first found vimentin at the time of neural tube closure (stage 19) in cells located at the lateral margins of the neural tube. By stage 26, these cells, which are presumably radial glia, are present along the entire length of the neural tube and in the tail bud. Cells in the optic vesicles express vimentin by stage 24. Vimentin-expressing mesenchymal cells appear on the surface of the somites at stage 22/23; these cells appear first on anterior somites and on progressively more posterior somites as development continues. Beginning at stage 24, vimentin appears in mesenchymal cells located ventral to the somites and associated with the pronephric ducts; these ventral cells first appear below the anterior somites and later appear below more posterior somites. The dorsal fin mesenchyme expresses vimentin at stage 26. In the head, both mesodermally-derived and neural-crest-derived mesenchymal tissues express vimentin by stage 26. These include the mesenchyme of the branchial arches, the mandibular arch, the corneal epithelium, the eye, the meninges and mesenchyme surrounding the otic vesicle. By stage 33, vimentin-expressing mesenchymal cells are present in the pericardial cavity and line the vitelline veins. Vimentin expression appears to be a marker for the differentiation of a subset of central nervous system cells and of head and body mesenchyme in the early Xenopus embryo.

XIF3, a Xenopus peripherin gene, requires an inductive signal for enhanced expression in anterior neural tissue

Development ◽  
1989 ◽  
Vol 107 (4) ◽  
pp. 701-714 ◽  
Author(s):  
C.R. Sharpe ◽  
A. Pluck ◽  
J.B. Gurdon
Keyword(s):  
Cdna Clone ◽  
Intermediate Filament ◽  
Neural Induction ◽  
Neural Tissue ◽  
Intermediate Filament Protein ◽  
Enhanced Expression ◽  
Tailbud Stage ◽  
Motor Neurones ◽  
Filament Protein

A full-length cDNA clone for the Xenopus intermediate filament gene XIF3 has been isolated. It is very similar in sequence to the rat intermediate filament cDNA clone 73 that is thought to encode the neuronal intermediate filament protein ‘peripherin’. By analysing dissected embryos, we show that XIF3 is expressed predominantly in anterior and dorsal structures and most strongly in the brain of the tailbud (stage 26) embryo. In situ hybridization shows XIF3 transcripts to be localized in neural tissue and especially in regions that most probably correspond to the motor neurones of the neural tube and to some cranial nerve ganglia. New XIF3 transcripts are first found at the start of gastrulation at a low level throughout the ectoderm and are not localized to the presumptive neurectoderm. Expression subsequently increases by about 10-fold in neural tissue, and requires an interaction of the mesoderm with overlying ectoderm. Because new transcripts are found predominantly in neural tissue of the head, this response can be used as a marker of anterior neural induction.

scholarly journals Chromosomal localisation of the mouse and human peripherin genes

1992 ◽  
Vol 59 (2) ◽  
pp. 125-129 ◽  
Author(s):  
Anne Moncla ◽  
Françoise Landon ◽  
Marie-Genevieve Mattei ◽  
Marie-Madeleine Portier
Keyword(s):  
Intermediate Filament ◽  
Major Part ◽  
Cdna Probe ◽  
Chromosome 15 ◽  
Intermediate Filament Protein ◽  
Mouse Cdna ◽  
F Region ◽  
Filament Protein ◽  
Chromosomal Localisation

SummaryUsing a mouse cDNA probe encoding for the major part of peripherin, a type III intermediate filament protein, we have assigned, by in situ hybridization, the mouse and human peripherin genes, Prph, to the E–F region of chromosome 15 and to the q12–q13 region of chromosome 12, respectively. These regions are known as homologous chromosomal segments containing other intermediate filament genes (keratins) and also other genes which could be co-ordinately regulated.

Immunocytochemical localization of desmin and vimentin in chick embryonic myocardial cells

1990 ◽  
Vol 48 (3) ◽  
pp. 448-449
Author(s):  
Yukiko Sugi
Keyword(s):  
Sodium Methoxide ◽  
Intermediate Filament Protein ◽  
Chick Embryos ◽  
Immunocytochemical Localization ◽  
Myocardial Cells ◽  
Immunofluorescence Study ◽  
Immunofluorescent Localization ◽  
Rabbit Igg ◽  
Semithin Sections ◽  
Filament Protein

In cultured skeletal muscle cells of chick, one intermediate filament protein, vimentin, is primarily formed and then synthesis of desmin follows. Coexistence of vimentin and desmin has been immunocytochemically confirmed in chick embryonic skeletal musclecells. Immunofluorescent localization of vimentin and desmin has been described in developing myocardial cells of hamster. However, initial localization of desmin and vimentin in early embryonic heart has not been reported in detail. By quick-freeze deep-etch method a loose network of intermediate filaments was revealed to exist surrounding myofibrils. In this report, immunocytochemical localization of desmin and vimentin is visualized in early stages of chick embryonic my ocardium.Chick embryos, Hamburger-Hamilton (H-H) stage 8 to hatch, and 1 day old postnatal chicks were used in this study. For immunofluorescence study, each embryo was fixed with 4% paraformaldehyde and embedded in Epon 812. De-epoxinized with sodium methoxide, semithin sections were stained with primary antibodies (rabbit anti-desmin antibody and anti-vimentin antibody)and secondary antibody (RITC conjugated goat-anti rabbit IgG).

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