Role for a Novel Usher Protein Complex in Hair Cell Synaptic Maturation

The molecular mechanisms underlying hair cell synaptic maturation are not well understood. Cadherin-23 (CDH23), protocadherin-15 (PCDH15) and the very large G-protein coupled receptor 1 (VLGR1) have been implicated in the development of cochlear hair cell stereocilia, while clarin-1 has been suggested to also play a role in synaptogenesis. Mutations in CDH23, PCDH15, VLGR1 and clarin-1 cause Usher syndrome, characterized by congenital deafness, vestibular dysfunction and retinitis pigmentosa. Here we show developmental expression of these Usher proteins in afferent spiral ganglion neurons and hair cell synapses. We identify a novel synaptic Usher complex comprised of clarin-1 and specific isoforms of CDH23, PCDH15 and VLGR1. To establish the in vivo relevance of this complex, we performed morphological and quantitative analysis of the neuronal fibers and their synapses in the Clrn1−/− mouse, which was generated by incomplete deletion of the gene. These mice showed a delay in neuronal/synaptic maturation by both immunostaining and electron microscopy. Analysis of the ribbon synapses in Ames waltzerav3J mice also suggests a delay in hair cell synaptogenesis. Collectively, these results show that, in addition to the well documented role for Usher proteins in stereocilia development, Usher protein complexes comprised of specific protein isoforms likely function in synaptic maturation as well

Molecular Brain Research

2-Jan137-1498

Molecular Brain Research

2-JanThis is a erratum reprint - with original pagination of the original title - placed at the front of issue: Molecular Brain Research. 2000,.DEC 28. V.85:(1-2).137-1498

EIAV-Based Retinal Gene Therapy in the shaker1 Mouse Model for Usher Syndrome Type 1B: Development of UshStat

Usher syndrome type 1B is a combined deaf-blindness condition caused by mutations in the MYO7A gene. Loss of functional myosin VIIa in the retinal pigment epithelia (RPE) and/or photoreceptors leads to blindness. We evaluated the impact of subretinally delivered UshStat, a recombinant EIAV-based lentiviral vector expressing human MYO7A, on photoreceptor function in the shaker1 mouse model for Usher type 1B that lacks a functional Myo7A gene. Subretinal injections of EIAV-CMV-GFP, EIAV-RK-GFP (photoreceptor specific), EIAV-CMV-MYO7A (UshStat) or EIAV-CMV-Null (control) vectors were performed in shaker1 mice. GFP and myosin VIIa expression was evaluated histologically. Photoreceptor function in EIAV-CMV-MYO7A treated eyes was determined by evaluating α-transducin translocation in photoreceptors in response to low light intensity levels, and protection from light induced photoreceptor degeneration was measured. The safety and tolerability of subretinally delivered UshStat was evaluated in macaques. Expression of GFP and myosin VIIa was confirmed in the RPE and photoreceptors in shaker1 mice following subretinal delivery of the EIAV-CMV-GFP/MYO7A vectors. The EIAV-CMV-MYO7A vector protected the shaker1 mouse photoreceptors from acute and chronic intensity light damage, indicated by a significant reduction in photoreceptor cell loss, and restoration of the α-transducin translocation threshold in the photoreceptors. Safety studies in the macaques demonstrated that subretinal delivery of UshStat is safe and well-tolerated. Subretinal delivery of EIAV-CMV-MYO7A (UshStat) rescues photoreceptor phenotypes in the shaker1 mouse. In addition, subretinally delivered UshStat is safe and well-tolerated in macaque safety studies These data support the clinical development of UshStat to treat Usher type 1B syndrome

Chemical composition and antibacterial activity of red murta (Ugni molinae Turcz.) seeds: an undervalued Chilean resource

Murta (Ugni molinae Turcz) is an endemic Chilean specie mostly used in medical, cosmetic, and food industries. However, during the industrial processing of murta fruits, the biomass containing the seeds is discarded as an industrial byproduct that does not find significant uses yet. This work is a first approach to valorize murta biomass through the identification and quantification of principal chemical constituents and exploring their antibacterial properties. The proximal analysis revealed that murta seeds exhibited significant content of raw fiber (64%), crude fat (14%), crude protein (12%), and low levels of ashes (1.5%) and minerals (0.04-0.23%). Dietary fiber was mainly composed of lignin, cellulose, pectin, and hemicellulose. Polyunsaturated fatty acids (89.0%), monounsaturated fatty acids (7.7%), and saturated fatty acids (3.3%) were the main constituents of seed oils. The arginine, asparagine, glutamic acid, and glycine were the primary protein constituent amino acids. Tannin fractions, total polyphenolic content, and oxygen radical absorbance capacity as antioxidant activity were measured. The chromatographic and mass spectrometric analysis (HPLC-MS/MS) confirmed the presence of several phenolic compounds like phenolic acids, flavonols, flavones, proanthocyanidins, and high molecular weight polyphenols. The murta seed extract showed high antibacterial activity against both Gram positive (Staphylococcus aureus, Bacillus cereus, and Streptococcus pyogenes) and Gram negative (Escherichia coli, Salmonella typhi, and Pseudomonas aeruginosa) bacterial strains. Murta seeds could be considered as a new source of nutritional components and bioactive compounds for different nutraceutical and food applications.Grant CONICYT PIA/APOYO CCTE FB17000