Lampbrush Chromosomes by Professor Dr. Harold Garnet Callan (auth.)

By Professor Dr. Harold Garnet Callan (auth.)

In 1965 i used to be requested via Dr. Konrad Springer even if i'd examine writing a monograph on "Lampbrush chromosomes and their physiological meaning", and even supposing I permitted in precept I refused to jot down there after which, or to satisfy a closing date. i wished to proceed with my very own examine, and that i had different duties that left me with little loose time, yet a way more vital attention used to be that during the Nineteen Sixties the topic used to be beset by means of a number of unresolved questions. I felt that to write down a overview then will be untimely, packed with speculations lots of which might be refuted, and certainly have been refuted, in the subsequent decade or . Had I written at the moment the only virtue over the current could were that few biologists have been learning lampbrush chromo­ somes, and the printed literature was once accordingly scanty. i'm pleased that I insisted on hold up, and am thankful for Springer Verlag's sufferer reputation of my choice. the 1st bankruptcy of this monograph describes the historical past of study on lampbrush chromosomes from their discovery in the direction of the top of the nineteenth century till the early Nineteen Sixties. through then a number of evidence touching on their constitution and chemistry have been firmly validated, together with the facts lampbrush chromatid is unineme; it features a unmarried uninterrupted DNA duplex. This uncovered a big challenge, the C-value paradox; grades of complexity of association in eukaryotes are unrelated to their genome sizes.

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The simpler saline is likely to be that used for nuclear isolation, and the dispersal saline a derivative thereof, so the same syringe and filter can often be used for both. Filter the isolation saline first, then remove the Swinnex holder, remove the plunger from the syringe, replace the Swinnex holder, pour the dispersal saline into the syringe and re-insert the plunger. Now proceed as before, discard the first few drops of dispersal saline, dispense the rest into an Erlenmeyer flask, and cap it.

In the oocytes of a great many amphibian species, for example Xenopus, Necturus, Cynops sinensis, Ambystoma mexicallum, A. macrodactylum and all plethodontid salamanders, the rigid gel phase within the germinal vesicle remains compact even after isolation in 100 mM KCI plus NaCl, so also with these species the nuclear membrane can be removed at leisure prior to the transfer of its contents to a dispersing saline. The problem with these species is not how to avoid gel dispersal, but how to achieve it.

In my limited experience the method is not successful with T. cristatus oocytes whose nuclear gels are intrinsically fluid, but succeeds if they are rigid, in other words the "5:1 plus P0 4" maintains rather than confers rigidity. In the oocytes of a great many amphibian species, for example Xenopus, Necturus, Cynops sinensis, Ambystoma mexicallum, A. macrodactylum and all plethodontid salamanders, the rigid gel phase within the germinal vesicle remains compact even after isolation in 100 mM KCI plus NaCl, so also with these species the nuclear membrane can be removed at leisure prior to the transfer of its contents to a dispersing saline.

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