Effect of Gametes Aging on Their Activation and Fertilizability in Zebrafish (Danio rerio)

Abstract

The zebrafish represents an important model organism for biological research. In this context, in vitro collection and fertilization of zebrafish gametes are basic and widely used techniques for many topical research works. In this work, the fertilization ability and normal embryo development of gold-type zebrafish sperm and eggs were re-evaluated after being stored for different times at 8 degrees C in a modified medium (Hanks' saline supplemented with 1.5 g BSA and 0.1 g ClNa; 320 mOsm, pH 7.4). Results obtained indicated that the temporal limits usually recommended for zebrafish sperm to fertilize fresh eggs (2 h) could be extended for up to 24 h without significant differences compared with fresh sperm. In contrast, the rapid egg aging observed (even less than 1 h) recommends minimizing as far as possible the egg storage time before fertilization. These results suggest a possible strain difference in the fertilization response.

Electroactivation of zebrafish (Danio rerio) eggs

“…The procedure carried out was described in detail by Cardona-Costa et al (2009). Briefly, eggs were in vivo extracted at the artificial «dawn» time from those aquariums where fish displayed reproductive behaviour, from females previously anaesthetized with clove oil solution of 100 µL L -1 (Grush et al, 2004;Mylonas et al, 2005).…”

Epigenetic reprogramming of somatic cells by nuclear transplant in zebrafish

“…Unfortunatelly, the egg culture medium used in the present work is only able to preserve the non activated eggs in good conditions for 1 hour (Cardona-Costa et al, 2009a). A recent study has detected that the Chinook salmon ovarian fluid (CSOF) can preserve non activated eggs for longer periods of time (Siripattarapravat et al, 2009a), but they commercial availability is, at this time, practically null.…”

“…Nevertheless, they observed that MPF levels The third experiment evaluated the activation ability of eggs stored (aged) for up to 2 h. Under our storage conditions (Cardona-Costa et al, 2009a), this was the maximum time in which eggs were still able to be fertilized.…”

“…using rainbow trout or coho salmon ovarian fluid (Allende and Weinberg, 1994) or Hank's saline supplemented with 0.5% BSA (Hatta et al, 1994). However, Cardona-Costa et al (2009a), in a modified egg medium advise to not extend this time above 60 min. Recently, it has shown that zebrafish eggs keep their fertilizability cultured in Chinook salmon ovarian fluid (CSOF) for 5h (Siripattarapravat et al, 2009a).…”

“…The procedure carried out was described in detail by Cardona-Costa et al (2009a). Briefly, gametes were extracted at the same artificial "dawn" time from those aquariums where fish displayed reproductive behaviour.…”