. The Biological bulletin. Biology; Zoology; Marine biology. 358 S. B. ZIMMERMAN, T. H. MURAKAMI, AND A. M. ZIMMERMAN Feulgen staining. The eggs were fixed in acetic acid-alcohol fixative (ll3) for 24 hours. The eggs were washed three times with distilled water, then placed on subbed slides and air-dried. The cells were hydrolyzed in 1 N HCl at 60° C. for 10 minutes. The staining period in Schiff's reagent was 20 minutes. Three washings, 10 minutes each, in SOg water followed by 3 washings in distilled water removed excess stains. In one experimental series, the eggs were counter- stained with
. The Biological bulletin. Biology; Zoology; Marine biology. 358 S. B. ZIMMERMAN, T. H. MURAKAMI, AND A. M. ZIMMERMAN Feulgen staining. The eggs were fixed in acetic acid-alcohol fixative (ll3) for 24 hours. The eggs were washed three times with distilled water, then placed on subbed slides and air-dried. The cells were hydrolyzed in 1 N HCl at 60° C. for 10 minutes. The staining period in Schiff's reagent was 20 minutes. Three washings, 10 minutes each, in SOg water followed by 3 washings in distilled water removed excess stains. In one experimental series, the eggs were counter- stained with 1% light green alcoholic solution prior to dehydration. The eggs were rapidly dehydrated in 80, 95 and 100 per cent alcohol, cleared in xylol, and mounted in Permount. Results As was previously established (Marsland et al, 1960; Zimmerman and Mars- land, 1960) cleavage furrows can be observed in fertilized eggs of Arbacia within a few minutes after the release of pressure, following pressure-centrifugation treat-. A B Figure 2. (A) Puromycin-trcated Arbacia egg stained with Schiff's reagent. Cells were incubated in 5 X 10"* M puromycin, IS minutes before insemination and fixed 2i hours after insemination. The Feulgen-positive material is accumulated over the nuclear material. (B) Furrow induction in puromycin-treated Arbacia egg. The cells were exposed to 10-* M puromycin, 15 minutes after insemination and pressure-centrifugation treatment was initiated 30 minutes after insemination. ment. The induced furrows are always at right angles to the centripetal axis at the equator or in varying positions from the equator approaching the centripetal pole. Only those eggs that showed a furrow induction frequency of 50% or greater, in which the furrows were maintained for at least 10 minutes after the release of pressure, were used in the experiments (Fig, 1), In an attempt to elucidate the biochemical activities involved in furrow forma- tion, the cells were treated with a variet
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