Post-embryonic development of sheepshead minnow Cyprinodon variegatus: a staging tool based on externally visible anatomical traits

The sheepshead minnow Cyprinodon variegatus has become a favoured model for laboratory studies because of their small size, rapid development, and tolerance of laboratory conditions. Here, we analyse sheepshead minnow post-embryonic development with the goal of providing a generally useful method for staging fish after embryogenesis. Groups of three females and two males were placed in breeding chambers designed for this experiment. More than 100 eggs were collected and maintained in seawater. Embryos were selected under a dissection microscope and placed in incubation dishes (50 per dish) at 26 °C. On day six, embryos hatched and larvae were transferred to 1 L beakers. To define a simplified normalization table for sheepshead minnow development, we measured each fish for its standard length and examined the fish for four externally evident traits: pigmentation pattern, caudal fin morphology, anal fin morphology, and dorsal fin morphology. The four traits were chosen, because they are easily visualized with standard laboratory equipment such as the stereomicroscope and camera. We have provided criteria for staging sheepshead minnows in studies of post-embryonic development. Our data suggest that dorsal and anal fin morphology may serve as a useful phenotype for defining metamorphic climax stages throughout post-embryonic development in C. variegatus. The staging systems we propose should facilitate detailed anatomical and developmental analyses in relation to ecotoxicological studies on potential disruption of the thyroid axis by xenobiotics and endocrine-disrupting compounds.