The goldfish used in this study were obtained from a local pet shop.
To relate the retinal images to objects in the real world we measured the distance between the centre of the lens and the retina, for the goldfish used in this study (∼150 mm in length) this was ∼3.8 mm.
Furthermore, the size differences of the goldfish used as well as the range of frequencies tested might have been too small: the specimens were 45 to 48 mm SL and 110 to 120 mm SL, and the five test frequencies ranged from 100 to 2,000 Hz.
Total RNA was extracted from goldfish brain using the TRIzol® RNA isolation reagent (Invitrogen, Canada).
To determine whether extracellular carbonic anhydrase is present in the outer retina, we examined the distribution of the extracellular membrane-bound isoform of carbonic anhydrase (type XIV) in the goldfish retina using an antibody raised against the mouse carbonic anhydrase XIV [27], [28].
Furthermore, IHNV, VHSV, IPNV, KHV, NDV, AIV (H5, H7, H9), CO cell, EPC cell, FHM cell, eggs of sturgeon and salmon, tissues (muscle, pancreas, gill, spleen, fin, brain, kidney) of ornamental carps, common carps, goldfishes, and grass goldfish were used as control templates to verify the specificity of the method.
Fig. 2 Goldfish samples used for the visual sensory evaluation a Fish fed with A2, b Fish fed with A3, c Fish fed with C4.
Up to nine goldfish were used per episode, so they could be replaced when necessary.
For the goldfish sizes used in this study (∼150 mm in length), the distance between the lens centre and retina was ∼3.8 mm, this was estimated by freezing and bisecting eyes from 3 fish.
Adult male and female goldfish were used to optimize in vivo transfer methods and to determine if water-borne estrogenic chemicals activate ERE-TK-LUC injected into the adult brain.
