Morpholino oligonucleotide (MO)-mediated knockdown of different GRKs in zebrafish larvae. a Nucleotide sequences of different GRK mRNAs in zebrafish. Blue and red underlining indicates GRK-MO1s- and GRK-MO2s-targeted sequences, respectively. The start codons are indicated by brackets. b Images showing whole bodies of 5 dpf larvae injected with control, GRK1b, GRK7a or GRK7b MO1s. Scale bar represents 1 mm. c Dorsal view of the pineal organs of 5 dpf larvae injected with control or one of the GRK MO1s. Landmarks indicate the pineal organs. Scale bar represents 100 μm. d Quantitative analyses of pineal size (c) in larvae injected with control MO or one of the GRK MO1s. The error bars indicate the standard error. Significance scores represent P values determined with the Wilcoxon rank sum test (NS, nonsignificant, P > 0.05)

The light responses of PP1 cells in zebrafish larvae were affected by GRK knockdown. a Quantification of GCaMP6s fluorescence intensity in larvae injected with control MO or one of the GRK MOs. The error bars indicate the standard error. b-g Calcium changes upon 405 nm laser flashes showing the effects of GRK knockdown on light responses in the PP1 cells of wild-type larvae (b-d) and SWS1-replacing larvae (e-g). b and e GRK1b MO-knockdown larvae. GRK1b-MO1, black open circle; GRK1b-MO2, black closed circle. c and f GRK7a MO-knockdown larvae. GRK7a-MO1, red open circle; GRK7a MO2-knockdown larvae, red closed circle. d and g GRK7b MO-knockdown larvae. GRK7b-MO1, blue open circle; GRK7b-MO2, blue closed circle. Gray open triangles in b-g indicate control MO-injected larvae. The number of animals tested (n) is indicated in parentheses after the MO names in each panel. The error bars indicate the standard error. The duration of each stimulus was ~ 450 ms

Immunoreactivity of the anti-GRK7 antibody in the PP1 cells of zebrafish larvae and adults. Double immunostaining for GRK7 (magenta) and PP1 (green) in the pineal organ and retina of larval and adult zebrafish using anti-PP1 and anti-GRK7 antibodies. a Zebrafish larva (5 dpf, whole-mount preparations); scale bars represent 20 μm. The rostral (R) and caudal (C) sides are shown. b Adult zebrafish (tissue sections); scale bars represent 50 μm. The merged image demonstrates that GRK7 is colocalized with PP1 (yellow arrowheads). The white dotted traces indicate the landmarks of the pineal organ. The rostral (R), caudal (C), dorsal (D), and ventral (V) sides are shown

Evaluation of the knockdown effect by immunohistochemistry using an anti-GRK7 antibody. Double immunostaining for GRK7 (magenta) and PP1 (green) in the knockdown zebrafish larvae. a Control MO knockdown, (b) GRK7a-MO1 knockdown, (c) GRK7a-MO2 knockdown, (d) GRK7b-MO1 knockdown and (e) GRK7b-MO2 knockdown. The white dotted traces indicate the landmarks of the pineal organ. The yellow arrowheads indicate PP1 cells in the pineal organ of larval zebrafish. The scale bar represents 25 μm. The rostral (R) and caudal (C) sides are shown. f Quantitative analyses of GRK7 immunoreactivity in different GRK MO- or control MO-injected larvae. The ratios of GRK7 immunoreactivity to PP1 immunoreactivity in PP1 cells are compared. The error bars indicate the standard error. Significance scores represent P values determined by Wilcoxon rank sum test (**, P < 0.01, ***, P < 0.001, NS, nonsignificant, P > 0.05)

ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.