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Analysis of zebrafish cryptochrome2 and 4 expression in UV cone photoreceptors.

Gene Expr Patterns. 2020 Jan;35:119100. Epub 2020 Feb 21
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摘要


Cryptochromes (Cry) are ancient flavoproteins known to regulate circadian rhythms. In plants and some animals, Cry is sensitive to blue light due to its ability to bind the chromophore flavin adenine dinucleotide (FAD). Cry is also suggested to function in magnetoreception, since it can create light-dependent radical pairs with FAD that are sensitive to magnetic fields (Ritz2000; Liedvogel et al., 2007; Solov'yov et al., 2014). Cry is expressed in the visual system of various animals and specifically co-localizes with both short- and long-wavelength cone photoreceptors in birds (Bischof et al., 2011; Günther et al., 2018). However, magnetoreception is not limited to birds and the expression of cry genes in the photoreceptors of other vertebrates is unknown. Here, we use zebrafish to examine the retinal expression pattern of cry family genes. Zebrafish have seven cry genes and while most are known regulators of the circadian clock, relatively little is known about cry2 and cry4 (Haug et al., 2015; Liu et al., 2015). Therefore, we explored cry2 and cry4 expression in the larval and adult zebrafish retina. We demonstrate that cry4 is predominantly expressed in the short-wavelength ultraviolet (UV)-sensitive cone photoreceptors, while cry2 is expressed in UV cones and additional retinal photoreceptors during the day. Using Nitroreductase (NTZ)-mediated cell ablation and qRT-PCR, we find that cry4 expression significantly decreases when UV cones are ablated, but not when the neighboring short-wavelength sensitive blue cones are ablated. cry2 expression decreases after UV cone ablation but is still significantly detectable, while blue cone ablation does not alter cry2 expression. This study provides a more detailed annotation of cry2 and cry4 expression in the zebrafish retina and highlights the feasibility of a well-established ablation paradigm to test if photoreceptors are required for magnetoreception in fish. Although evidence of magnetoreception in adult zebrafish has gained considerable evidence over the last decade (Shcherbakov et al., 2005; Takebe et al., 2012; Krylov et al., 2016; Myklatun et al., 2018) the mediating mechanism(s) remain unknown. Additionally, despite limited evidence that larval zebrafish are magnetoreceptive, many other larval fish have a characterized magnetic sense; sockeye salmon fry, larval coral reef fish, larval medaka and larval Atlantic haddock have been shown to be responsive to magnetic fields (Quinn; 1980; Bottesch et al., 2016; O'Connor and Muheim. 2017; Myklatun et al., 2018; Cresci, et al. 2019). If cry-cone interactions are conserved within fish, our findings may suggest one potential mechanism, such that UV cones appear poised for light-dependent magnetoreception via photoreceptor subtype-specific expression of cry.

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