Protein synthesis during limb regeneration in the axolotl

Development ◽  
1982 ◽  
Vol 70 (1) ◽  
pp. 241-260
Author(s):  
J. M. W. Slack

A study has been made of limb regeneration in the axolotl using two-dimensional gel electrophoresis of proteins labelled with [35S]methionine. In the early stages of regeneration seven proteins are identified which are-specific to the mesenchyme and thirteen which are specific to the epidermis. There is very little change in the gel pattern until the onset of overt cytodifferentiation upon which the muscle and cartilage become substantially different both from each other and from the blastemal mesenchyme. The gel pattern of mesenchyme from the larval limb bud is almost identical to that of the blastemal mesenchyme, while the pattern of the limb-bud epidermis differs somewhat from the blastemal epidermis. A careful search has been made for differences which might be associated with ‘positional information’ by comparing forelimb with hindlimb, proximal with distal and anterior with posterior. The differences which have been found are all associated with differences in visible cellular composition or in rates of differentiation, rather than with position per se.It is concluded that positional codings cannot be detected by this technique. On the basis of biological experiments, six criteria are proposed by which to assess future searches for positional codings.

1984 ◽  
Vol 30 (3) ◽  
pp. 345-352 ◽  
Author(s):  
Robert L. Armstrong ◽  
Thomas P. West ◽  
Paul T. Magee

The uptake and incorporation of macromolecular precursors in germinating Saccharomyces cerevisiae ascospores were investigated. Addition of cycloheximide at various times during germination revealed that protein synthesis can occur within 20 min after the spores are shifted to glucose-containing media. The time of initiation of uptake and incorporation of several amino acids differed; this can be attributed to differing amino acid pool levels in the spores, as well as differing transport activities. Two-dimensional gel electrophoresis of proteins labeled with [35S]methionine for various 20-min periods after germination began showed at least one protein whose synthesis begins well after the bulk of the proteins.


1996 ◽  
pp. 203-219
Author(s):  
Chung Lee ◽  
Yi Qian ◽  
Julia A. Sensibar ◽  
Harold H. Harrison

2005 ◽  
Vol 15 (2) ◽  
pp. 111-121 ◽  
Author(s):  
Helena Lindmark-Månsson ◽  
Anna Timgren ◽  
Gun Aldén ◽  
Marie Paulsson

1981 ◽  
Vol 91 (2) ◽  
pp. 352-360 ◽  
Author(s):  
TW McKeithan ◽  
JL Rosenbaum

The alga polytomella contains several organelles composed of microtubules, including four flagella and hundreds of cytoskeletal microtubules. Brown and co-workers have shown (1976. J. Cell Biol. 69:6-125; 1978, Exp. Cell Res. 117: 313-324) that the flagella could be removed and the cytoskeletans dissociated, and that both structures could partially regenerate in the absence of protein synthesis. Because of this, and because both the flagella and the cytoskeletons can be isolated intact, this organism is particularly suitable for studying tubulin heterogeneity and the incorporation of specific tubulins into different microtubule-containing organelles in the same cell. In order to define the different species of tubulin in polytonella cytoplasm, a (35)S- labeled cytoplasmic fraction was subjected to two cycles of assembly and disassembly in the presence of unlabeled brain tubulin. Comparison of the labeled polytomella cytoplasmic tubulin obtained by this procedure with the tubulin of isolated polytomella flagella by two-dimensional gel electrophoresis showed that, whereas the β-tubulin from both cytoplasmic and flagellar tubulin samples comigrated, the two α-tubulins had distinctly different isoelectic points. As a second method of isolating tubulin from the cytoplasm, cells were gently lysed with detergent and intact cytoskeletons obtained. When these cytoskeletons were exposed to cold temperature, the proteins that were released were found to be highly enriched in tubulin; this tubulin, by itself, could be assembled into microtubules in vitro. The predominant α-tubulin of this in vitro- assembled cytoskeletal tubulin corresponded to the major cytoplasmic α-tubulin obtained by coassembly of labeled polytomella cytoplasmic extract with brain tubulin and was quite distinct from the α-tubulin of purified flagella. These results clearly show that two different microtubule-containing organelles from the same cell are composed of distinct tubulins.


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