Studies on incorporation of 3H-thymidine in Arbacia eggs under hydrostatic pressure

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Abstract

Fertilized eggs of Arbacia punctulata were placed into 3H-thymidine-sea water solution (1–2 μC/ml) and subjected to pressures of 5000-15,000 lb/in2 at 20 °C for durations of 10–60 min. Immediately after the release of pressure, the eggs were fixed, embedded, sectioned and subjected to autoradiographic procedures. The uptake of 3H-thymidine was compared in the pressurized eggs and non-pressurized controls.

Eggs which were placed in 3H-thymidine but not pressurized showed incorporation of the isotope only in the interval from 15–30 min after insemination, corresponding to the time from syngamy to early streak. There was no evidence of 3H-thymidine incorporation into the pronuclei during the presyngamy period. However, eggs in which pronuclear fusion had been inhibited by a pressure of 5000 lb/in2 for 30–60 min did show isotope incorporation.

It was also found that whereas 5000 lb/in2 did not block DNA synthesis even in the face of cessation of morphological mitotic activity, pressures of 7500 lb/in2 and greater did inhibit DNA synthesis.

The results are discussed in terms of the effects of pressure on development and the independence of the DNA synthesis cycle and cell division.

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Work supported in part by grants from National Research Council of Canada and the National Cancer Institute of Canada.

2

Present address: Mary Fletcher Hospital, University of Vermont, Burlington, Vt, U.S.A.

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