FIGURE SUMMARY
Title

Genetically Blocking the Zebrafish Pineal Clock Affects Circadian Behavior

Authors
Ben-Moshe Livne, Z., Alon, S., Vallone, D., Bayleyen, Y., Tovin, A., Shainer, I., Nisembaum, L.G., Aviram, I., Smadja-Storz, S., Fuentes, M., Falcón, J., Eisenberg, E., Klein, D.C., Burgess, H.A., Foulkes, N.S., Gothilf, Y.
Source
Full text @ PLoS Genet.
Fig. 1

Generation of Tg(aanat2:EGFP-ΔCLK) fish.

(A) Schematic representation of the transgenic construct used for the generation of Tg(aanat2:EGFP-ΔCLK) fish. The pT2-aanat2:EGFP-2A-Δclocka-5×MYC plasmid consists of two arms from the Tol2 transposable element (black), aanat2 regulatory regions (gray), EGFP coding sequence (CDS; green), 2A peptide sequence (orange), ΔCLK CDS (blue), 5×Myc tags (purple) and SV40 poly(A) signal (yellow). (B and B') EGFP expression is restricted to the pineal gland of Tg(aanat2:EGFP-ΔCLK) larvae. Dorsal views of the head region of 7-day post-fertilization (dpf) larvae, anterior to the top; confocal z-stack projection (B) and a single confocal plane (B'). (C) Immunostaining with anti-Myc antibody confirms that ΔCLK is specifically expressed in the pineal gland of Tg(aanat2:EGFP-ΔCLK) larvae. Dorsal view of the head region of a 5-dpf larva, anterior to the top; confocal z-stack projection.
Fig. 2

Clock-controlled aanat2 mRNA rhythm is abolished in the pineal gland of Tg(aanat2:EGFP-ΔCLK) larvae.

(A) Tg(aanat2:EGFP-ΔCLK) larvae (ΔCLK) exhibit arrhythmic pineal aanat2 mRNA levels under DD compared with a robust rhythm in their WT siblings (p<0.0001, two-way ANOVA), indicating that the pineal gland molecular clock is disrupted. Each value represents the mean optical density ± SE of the pineal aanat2 whole-mount ISH signal (n = 10−15 larvae). Different letters represent statistically different values (p<0.05, Tukey's test). (B) Representative whole-mount ISH aanat2 signals (8 dpf, dorsal view) in the pineal glands of WT sibling larvae (upper panel) and Tg(aanat2:EGFP-ΔCLK) larvae (ΔCLK; bottom panel). Black and gray bars represent subjective night and day, respectively. CT, circadian time.
Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ PLoS Genet.
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