Table 1 A. limnaeus stages selected for qPCR analysis
Stage | Age | WS a | Abbreviation | Description | Pooled individuals | N |
|---|---|---|---|---|---|---|
Early cleavage | 3 hpf | 3-5 | EC | 1-4 blastomeres | 100 | 3 |
Early hollow blastula | 1 dpf | 12 | EHB | Presence of a segmentation cavity containing blastomeres and covered by enveloping layer cells | 100 | 4 |
50% epiboly | 2 dpf | 17 | 50% EP | Half of the yolk surface covered by periblast and enveloping layer. In between the two layers are embryonic blastomeres that have become ameboid and migrated away from the central blastula | 100 | 4 |
Dispersed blastomere phases | 4 dpf | 20-21 | DBP | Yolk surface completely covered by periblast, enveloping layer, and randomly distributed embryonic blastomeres | 100 | 4 |
Reaggregation phases | 8 dpf | 22-26 | RP | Embryonic blastomeres remain distributed across yolk, but a subpopulation are beginning to migrate towards a small area of the yolk where the future embryo will form. No discernable embryonic axis present | 100 | 4 |
Solid neural keel | 10 dpf | 28 | SNK | Presence of a solid neural keel, head fold, and Kupffer’s vesicle. No somites present | 50 | 4 |
Diapause II | 32 dpf | 33 | DII | Presence of optic cups and associated lenses, otic vesicles, functional heart, and 38 to 40 pairs of somites. Heart rate of 0 to 10Â bpm | 100 | 3 |
Three-quarter overgrowth | 9 dpd | 39 | 3/4 OG | Embryo occupies about three-fourths of the perimeter of the yolk. Eyes are heavily pigmented with gold colored material. Presence of incompletely developed gut, liver, and swimbladder | 40 | 4 |
Diapause III | 24 dpd | 43 | DIII | Fully formed larva that has completed embryonic development, but has not yet hatched | 40 | 4 |
Adult liver | Adult | N/A | Liver | Whole liver from an adult female | 1 | 3 |