Biochemistry and physiology of zebrafish photoreceptors

All vertebrates share a canonical retina with light-sensitive photoreceptors in the outer retina. These photoreceptors are of two kinds: rods and cones, adapted to low and bright light conditions, respectively. They both show a peculiar morphology, with long outer segments, comprised of ordered stacks of disc-shaped membranes. These discs host numerous proteins, many of which contribute to the visual transduction cascade. This pathway converts the light stimulus into a biological signal, ultimately modulating synaptic transmission. Recently, the zebrafish (Danio rerio) has gained popularity for studying the function of vertebrate photoreceptors. In this review, we introduce this model system and its contribution to our understanding of photoreception with a focus on the cone visual transduction cascade

Detection of Zebrafish Retinal Proteins by Infrared Western Blotting

The zebrafish retina is a canonical vertebrate retina. Since the past few years, with the continually growing genetic toolbox and imaging techniques, zebrafish plays a crucial role in retinal research. This protocol describes a method to quantitatively evaluate the expression of Arrestin3a (Arr3a) and G-protein receptor kinase7a (Grk7a) in the adult zebrafish retina at protein levels by infrared fluorescence western blot. Our protocol can be easily adapted to measure protein levels in additional zebrafish tissues

Comparison of nitrogen fertigation management strategies for center-pivot irrigated maize in the sub-humid area of China

In the sub-humid region of Northeast China, increasing use of center-pivot irrigation systems has caused increased interest in sprinkler fertigation technology in maize production to improve nitrogen (N) use efficiency and protect the environment. However, the lack of fertigation strategies for maize cultivation restrains the adoption of sprinkler fertigation technology. A field experiment was carried out in a sub-humid region of Northeast China on maize to determine the effect of different fertigation management strategies on plant growth, grain yield and nitrate content in the soil during the maize growing season. Three N rates (200, 160, and 120 kg N ha-1) and three fertigation schedules were tested. After a uniform nitrogen fertilizer application at an early stage, the N treatments applied 100% of the remaining amount of fertilizer at the stage of vegetative (V) 14 (T1); applied 66.7% and 33.3% of the remaining amount of fertilizer at the stage of V14 and reproductive 2 (R2), respectively (T2); and applied 75% and 25% of the remaining amount of fertilizer at the stage of V14 and R2, respectively (T3). The N rates and fertigation schedules were combined to make nine treatments: T1N200, T1N160, T1N120, T2N200, T2N160, T2N120, T3N200, T3N160, and T3N120, each having three replications. Full irrigation was applied in order to minimize water stress. All treatments received the same irrigation depth in each fertigation event. Results showed that maize grain yield and above-ground biomass production increased with the increasing of N rates; N200T1 produced a higher yield (12,710 kg ha-1) than the other fertigation treatments. However, there was no significant difference in yield between the N rates of 160 and 200 kg ha-1(P＜0.05), while partial factor productivity decreased with increased N application. Furthermore, the amount of the mineral nitrogen (NO3–N) accumulated in the 0- to 100-cm layer after harvest increased as the N rates increased. At the high N level, the residual NO3–N in the soil in T1 was 65% and 51% less than that in T2 and T3, which decreased the risk of NO3–N leaching out of the 0- to 100-cm soil layer. Based on this research, the recommended management practice of fertigation via center-pivot irrigation systems is to apply 160 kg ha-1 of nitrogen (N160) to maize through two in-season fertigation events (T1), which can obtain relatively high production, meanwhile reducing the risk of nitrogen leaching in the sub-humid region of China

The Binding Properties and Physiological Functions of Recoverin

Recoverin (Rcv) is a low molecular-weight, neuronal calcium sensor (NCS) primarily located in photoreceptor outer segments of the vertebrate retina. Calcium ions (Ca2+)-bound Rcv has been proposed to inhibit G-protein-coupled receptor kinase (GRKs) in darkness. During the light response, the Ca2+-free Rcv releases GRK, which in turn phosphorylates visual pigment, ultimately leading to the cessation of the visual transduction cascade. Technological advances over the last decade have contributed significantly to a deeper understanding of Rcv function. These include both biophysical and biochemical approaches that will be discussed in this review article. Furthermore, electrophysiological experiments uncovered additional functions of Rcv, such as regulation of the lifetime of Phosphodiesterase-Transducin complex. Recently, attention has been drawn to different roles in rod and cone photoreceptors.This review article focuses on Rcv binding properties to Ca2+, disc membrane and GRK, and its physiological functions in phototransduction and signal transmission

Publisher Correction: The ciliopathy protein TALPID3/KIAA0586 acts upstream of Rab8 activation in zebrafish photoreceptor outer segment formation and maintenance

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper

Charge Measurement of Cosmic Ray Nuclei with the Plastic Scintillator Detector of DAMPE

One of the main purposes of the DArk Matter Particle Explorer (DAMPE) is to measure the cosmic ray nuclei up to several tens of TeV or beyond, whose origin and propagation remains a hot topic in astrophysics. The Plastic Scintillator Detector (PSD) on top of DAMPE is designed to measure the charges of cosmic ray nuclei from H to Fe and serves as a veto detector for discriminating gamma-rays from charged particles. We propose in this paper a charge reconstruction procedure to optimize the PSD performance in charge measurement. Essentials of our approach, including track finding, alignment of PSD, light attenuation correction, quenching and equalization correction are described detailedly in this paper after a brief description of the structure and operational principle of the PSD. Our results show that the PSD works very well and almost all the elements in cosmic rays from H to Fe are clearly identified in the charge spectrum.Comment: 20 pages, 4 figure

Mutation of vsx genes in zebrafish highlights the robustness of the retinal specification network

Genetic studies in human and mice have established a dual role for Vsx genes in retina development: an early function in progenitors' specification, and a later requirement for bipolar-cells fate determination. Despite their conserved expression patterns, it is currently unclear to which extent Vsx functions are also conserved across vertebrates, as mutant models are available only in mammals. To gain insight into vsx function in teleosts, we have generated vsx1 and vsx2 CRISPR/Cas9 double knockouts (vsxKO) in zebrafish. Our electrophysiological and histological analyses indicate severe visual impairment and bipolar cells depletion in vsxKO larvae, with retinal precursors being rerouted toward photoreceptor or Müller glia fates. Surprisingly, neural retina is properly specified and maintained in mutant embryos, which do not display microphthalmia. We show that although important cis-regulatory remodelling occurs in vsxKO retinas during early specification, this has little impact at a transcriptomic level. Our observations point to genetic redundancy as an important mechanism sustaining the integrity of the retinal specification network, and to Vsx genes regulatory weight varying substantially among vertebrate species

Megalencephalic leukoencephalopathy with subcortical cysts protein 1 regulates glial surface localization of GLIALCAM from fish to humans

Megalencephalic leukoencephalopathy with subcortical cysts (MLC) is a leukodystrophy characterized by myelin vacuolization and caused by mutations in MLC1 or GLIALCAM. Patients with recessive mutations in either MLC1 or GLIALCAM show the same clinical phenotype. It has been shown that GLIALCAM is necessary for the correct targeting of MLC1 to the membrane at cell junctions, but its own localization was independent of MLC1 in vitro. However, recent studies in Mlc1−/− mice have shown that GlialCAM is mislocalized in glial cells. In order to investigate whether the relationship between Mlc1 and GlialCAM is species-specific, we first identified MLC-related genes in zebrafish and generated an mlc1−/− zebrafish. We have characterized mlc1−/− zebrafish both functionally and histologically and compared the phenotype with that of the Mlc1−/− mice. In mlc1−/− zebrafish, as in Mlc1−/− mice, Glialcam is mislocalized. Re-examination of a brain biopsy from an MLC patient indicates that GLIALCAM is also mislocalized in Bergmann glia in the cerebellum. In vitro, impaired localization of GlialCAM was observed in astrocyte cultures from Mlc1−/− mouse only in the presence of elevated potassium levels, which mimics neuronal activity. In summary, here we demonstrate an evolutionary conserved role for MLC1 in regulating glial surface levels of GLIALCAM, and this interrelationship explains why patients with mutations in either gene (MLC1 or GLIALCAM) share the same clinical phenotyp