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Production of Haploid Zebrafish Embryos by In Vitro Fertilization
Published on: July 14, 2014
Description of the embryonic development in the convict cichlid (Amatitlania nigrofasciata)
Sayaka Matsuo1, Rie Kusakabe2, Shun Satoh3
1Graduate School of Marine Biosciences, Kitasato University, Sagamihara, Japan.
Abstract:
We provide a detailed description of embryonic development in the convict cichlid (Amatitlania nigrofasciata) from fertilization to hatching at 26°C, together with a practical staging table anchored to established teleost reference frameworks. Fertilized eggs were obtained by both natural spawning and artificial fertilization. Oocytes were ovoid and adhesive, surrounded by a chorion and a sticky mucous layer. Early development proceeded, in broad outline, through the teleost sequence of meroblastic discoidal cleavage, blastula, gastrula, segmentation and organogenesis. The first cleavage occurred at 1.75 h post fertilization (hpf), with subsequent cleavages at 30 min intervals, reaching the 64-cell stage at 4.25 hpf. Cleavage up to the 64-cell stage progressed on a timescale broadly comparable to that reported for other cichlids, whereas the interval from the 64-cell stage to early epiboly was relatively short in this species. The high, sphere and dome stages occurred at 8, 9 and 10 hpf, respectively, with epiboly initiating at the dome stage. At the dome stage, a marginal thickening interpreted as the presumptive embryonic shield became apparent. During early epiboly, the blastoderm showed pronounced spatial heterogeneity: it was consistently thicker and advanced more rapidly on the prospective embryonic axis side, yielding a readily detectable asymmetry. A morphologically distinct embryonic axis became visible at 40%-50% epiboly, and epiboly was completed at 28.5 hpf. Notably, somitogenesis began before epiboly completion (first somites at 85%-90% epiboly), indicating temporal overlap between late gastrulation and early segmentation. Major organ primordia became apparent during the overlapping segmentation/organogenesis interval, and hatching occurred around 70 hpf. Newly hatched larvae possessed three pairs of adhesive glands. This staging reference enables reproducible developmental sampling and should facilitate future comparative, mechanistic and experimental work using the convict cichlid.

