Download Molecular Aspects of Early Development by William R. Jeffery, Craig R. Tomlinson (auth.), George M. PDF

By William R. Jeffery, Craig R. Tomlinson (auth.), George M. Malacinski, William H. Klein (eds.)

The early embryo has emerged because the point of interest for research of the rules of gene expression for a number of purposes. First, the truth that embryogenesis is below genetic regulate has been preferred from the earliest days of classical embryology. while experimental thoughts turned to be had it was once as a result logical that they need to be utilized to the embryo. With each one new enhance in technique, curiosity in embryonic gene expression reports has elevated. moment, many embryos supply designated possibilities for the research of particular elements of the rules of gene expression. a number of phenomena--eg. , regulate of translation--can be very with ease studied in a number of marine invertebrate embryos. these embryos comprise huge shops of maternally inherited mRNA that are translated in a hugely ordered model in the course of particular levels of submit fertilization improvement. Marine invertebrate eggs might be comfortably artifically inseminated and classified with radioactive precursors. Their research is resulting in vital insights into the mechanisms which keep watch over gene expression at post-transcriptional degrees. 3rd, fresh advances in either transmission and recombinant DNA genetics, specifically in organisms equivalent to Drosophila, are delivering targeted possibilities for the research of regulatory mechanisms which function on the point of the genome. particular genes were pointed out, remoted, and--in a few instances--sequenced. the chance is now on hand to check the rules of the expression of unmarried genes in a vertical fashion--from the first series of the gene to the tissues and organs that are the goods of morphogenesis.

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The proteins are stained with silver. The association of mRNA with the egg cytoskeleton has recently been tested by Jeffery (1983a). He reasoned that i f mRNA is attached to cytoskeletal elements, it should be resistant to extraction by non-ionic detergents such as Triton X-100. Because these detergents disrupt membranous systems, and would also be expected to destroy any local ionic microenvironments, the retention of mRNA in detergent extracted eggs should exclude these possibilities. When Styela eggs are extracted with Triton X-100, about 60% of the egg poly(A) is left in the detergent-insoluble fraction although only 20% of the total proteins, 30% of the total RNA and less than 8% of the lipid (most of this is due to the retention of lipid pigment granules bound to the filamentous lattice in the residue) is present (Jeffery and Meier, 1983).

Most of the translation products of RNA from the various fractions were qualitatively similar to those of W. R. JEFFERY ET AL. 24 A I ~ IEF ~ 8 sos T • 16: Two-dimensional polyacrylamide gel electrophoresis of SS-methionine labeled translation products synthesized in a wheat germ lysate directed by RNA from the isolated yellow crescent (A) or the supernatant (B) egg (ectoplasm and endoplasm) fraction of Styela plicata eggs. A major egg polypeptide undetected in the translation products of yellow crescent RNA is shown by the arrow.

These proteins were labeled in intact embryos before separation of the tissues. The separation techniques employed can induce a stress response which leads to alterations in protein synthesis; some of the induced polypeptides are of the same sizes as those induced in heat shocked embryos (our unpublished observations). The relative rates of synthesis of all of these tissue-enriched polypeptides increases substantially during development, though some are detectibly synthesized in eggs. Some of these tissue-specific proteins are weIl represented by mass in the egg; it is not yet known whether they are nonrandomly distributed in the egg cytoplasm or segregated during early cleavage.

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