MicroRNAs (miRNAs) inhibit the translation of target mRNAs and affect, directly or indirectly, the expression of a large portion of the protein-coding genes. This study focuses on miRNAs that are expressed in the mouse cochlea and vestibule, the 2 inner ear compartments. A conditional knock-out mouse for Dicer1 demonstrated that miRNAs are crucial for postnatal survival of functional hair cells of the inner ear. We identified miRNAs that have a role in the vertebrate developing inner ear by combining miRNA transcriptome analysis, spatial and temporal expression patterns, and bioinformatics. Microarrays revealed similar miRNA profiles in newborn-mouse whole cochleae and vestibules, but different temporal and spatial expression patterns of six miRNAs (miR-15a, miR-18a, miR-30b, miR-99a, miR-182, and miR-199a) may reflect their roles. Two of these miRNAs, miR-15a-1 and miR-18a, were also shown to be crucial for zebrafish inner ear development and morphogenesis. To suggest putative target mRNAs whose translation may be inhibited by selected miRNAs, we combined bioinformatics-based predictions and mRNA expression data. Finally, we present indirect evidence that Slc12a2, Cldn12, and Bdnf mRNAs may be targets for miR-15a. Our data support the hypothesis that inner ear tissue differentiation and maintenance are regulated and controlled by conserved sets of cell-specific miRNAs in both mouse and zebrafish.cochlea ͉ deafness ͉ Dicer ͉ mouse ͉ zebrafish
In eukaryotes, most integral membrane proteins are synthesized, integrated into the membrane, and folded properly in the endoplasmic reticulum (ER). We screened the mutants affecting rhabdomeric expression of rhodopsin 1 (Rh1) in the Drosophila photoreceptors and found that dPob/EMC3, EMC1, and EMC8/9, Drosophila homologs of subunits of ER membrane protein complex (EMC), are essential for stabilization of immature Rh1 in an earlier step than that at which another Rh1-specific chaperone (NinaA) acts. dPob/EMC3 localizes to the ER and associates with EMC1 and calnexin. Moreover, EMC is required for the stable expression of other multi-pass transmembrane proteins such as minor rhodopsins Rh3 and Rh4, transient receptor potential, and Na+K+-ATPase, but not for a secreted protein or type I single-pass transmembrane proteins. Furthermore, we found that dPob/EMC3 deficiency induces rhabdomere degeneration in a light-independent manner. These results collectively indicate that EMC is a key factor in the biogenesis of multi-pass transmembrane proteins, including Rh1, and its loss causes retinal degeneration.DOI:
http://dx.doi.org/10.7554/eLife.06306.001
Two kinds of retinal cDNA fragments (OlGRK-R and -C) encoding the putative G-protein-coupled receptor kinases (GRKs) were isolated from medaka, Oryzias latipes. OlGRK-R appears to be closely related to the rhodopsin kinase (RK) found in the outer segments of mammalian photoreceptors, but the deduced amino acid sequence of OlGRK-C shows less than 50% identity to those of GRKs known to date, suggesting that OlGRK-C is a novel GRK subtype (GRK7). The mRNA of OlGRK-R is detectable in rods, and that of OlGRK-C is found in all four types of cone photoreceptor. The C-terminal of OlGRK-R has a consensus sequence for farnesylation, whereas, surprisingly, OlGRK-C has a consensus sequence for geranylgeranylation. Our result are consistent with the concept that lower vertebrates have rod-and cone-specific opsin kinases.z 1998 Federation of European Biochemical Societies.
Five cDNA fragments (KFH-R, -V, -G, -B and -Rh) encoding the putative visual pigments of killifish were isolated and sequenced. Judging from the deduced amino acid sequences, each cDNA falls into a different group of the five major families of vertebrate visual pigment genes. In situ hybridization localized the mRNA of KFH-R and -G to the principle and accessory members, respectively, of double cones. Visual pigment genes KFH-Rh, -B and -V were expressed in the rods, and the long and short single cones, respectively. It is suggested that the relationships between the cell types and their respective visual pigment gene groups may be a common pattern among teleost fishes.
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