No description available.

Human Usher syndrome is a severe and congenital form of syndromic deafness that affects 1 person in 25,000 people in the world population. Normally the stereocilia, microvillar protrusions of the apical membrane of inner ear hair cells, are organized into coherent bundles. This precise organization is critical for mechanosensing, i.e. for hearing. Mutation in any of the five known Usher syndrome genes is sufficient to alter the precise organization of stereocilia, a condition that results in deafness. To date, however, the molecular mechanisms responsible for the splaying of stereocilia and genesis of the disease are not well understood. Here, I identified Drosophila melanogaster genes related to human Usher syndrome and characterized some of them (Cad99C, DSANS and crinkled) during Drosophila development, in the processes of microvilli morphogenesis in the follicular and wing imaginal disc epithelia. Cadherin Cad99C is a transmembrane protein with putative cell adhesion properties. Similar to its human ortholog Protocadherin 15, Drosophila Cad99C localizes to microvillar protrusions in the follicular epithelium. In this epithelium, Cad99C is required for the proper morphogenesis and organization of microvilli into bundles, similar to human Protocadherin 15. Further, overexpression of the full-length Cad99C or of a deleted version, devoid of the cytoplasmic region, promotes microvilli bundling. This finding suggests that Cad99C establishes adhesive interactions between microvilli via its extracellular region. Interestingly, morphological alteration of follicle cell microvilli associates with defective deposition of the vitelline membrane, an extracellular matrix that protects the embryo from osmotic stresses. These findings suggest that microvilli are normally required for the even deposition of the extracellular matrix. In order to test whether Cad99C is involved in microvilli morphogenesis and bundling in other tissues, I analyzed the function of Cad99C in a larval tissue, the wing imaginal disc. Cad99C overexpression, but not Cad99C removal, is sufficient to alter microvilli morphology and organization in the columnar epithelium of the wing imaginal disc. Likely, other molecules can compensate for Cad99C loss of function in this tissue. To possibly get some insights on the molecular function of other Usher syndrome proteins, I analyzed the function of Drosophila SANS and crinkled in the follicular epithelium, where both these genes are expressed. crinkled is the ortholog of myosinVIIa, that encodes a motor protein of the actin cytoskeleton. DSANS is related to human SANS and encodes a cytoplasmic protein of unknown function. It has been puzzling how removal of SANS, a cytoplasmic protein, could impair adhesion and bundling of stereocilia. To study the function of DSANS, I generated null mutant flies and observed that, in the absence of DSANS, delivery of Cad99C to microvilli is impaired. Cad99C localization is however unperturbed in crinkled mutant follicle cells. By immunostaining, DSANS immunoreactivity was detected diffusively in the cytoplasm and in dot-like structures, possibly corresponding to vesicles. In conclusion, DSANS is a cytoplasmic protein that is required for the efficient delivery of Cad99C to microvilli protrusions. Taken together, the analysis that I here performed of Drosophila Usher syndrome related genes indicates two novel molecular mechanisms of function for the corresponding human Usher syndrome proteins. First, human Protocadherin 15, like Drosophila Cad99C, could be involved in establishing adhesive interactions between microvilli protrusions of the inner ear (stereocilia). Removal of Protocadherin 15 would then cause splaying of stereocilia due to lack of inter-stereocilia adhesive links. Second, the analysis here performed suggests that SANS is involved in the efficient delivery of Protocadherin 15 to stereocilia. Mutations in SANS would then lead to splaying of stereocilia and deafness due to poor localization of Protocadherin 15 to stereocilia.

Human Usher syndrome is a severe and congenital form of syndromic deafness that affects 1 person in 25,000 people in the world population. Normally the stereocilia, microvillar protrusions of the apical membrane of inner ear hair cells, are organized into coherent bundles. This precise organization is critical for mechanosensing, i.e. for hearing. Mutation in any of the five known Usher syndrome genes is sufficient to alter the precise organization of stereocilia, a condition that results in deafness. To date, however, the molecular mechanisms responsible for the splaying of stereocilia and genesis of the disease are not well understood. Here, I identified Drosophila melanogaster genes related to human Usher syndrome and characterized some of them (Cad99C, DSANS and crinkled) during Drosophila development, in the processes of microvilli morphogenesis in the follicular and wing imaginal disc epithelia. Cadherin Cad99C is a transmembrane protein with putative cell adhesion properties. Similar to its human ortholog Protocadherin 15, Drosophila Cad99C localizes to microvillar protrusions in the follicular epithelium. In this epithelium, Cad99C is required for the proper morphogenesis and organization of microvilli into bundles, similar to human Protocadherin 15. Further, overexpression of the full-length Cad99C or of a deleted version, devoid of the cytoplasmic region, promotes microvilli bundling. This finding suggests that Cad99C establishes adhesive interactions between microvilli via its extracellular region. Interestingly, morphological alteration of follicle cell microvilli associates with defective deposition of the vitelline membrane, an extracellular matrix that protects the embryo from osmotic stresses. These findings suggest that microvilli are normally required for the even deposition of the extracellular matrix. In order to test whether Cad99C is involved in microvilli morphogenesis and bundling in other tissues, I analyzed the function of Cad99C in a larval tissue, the wing imaginal disc. Cad99C overexpression, but not Cad99C removal, is sufficient to alter microvilli morphology and organization in the columnar epithelium of the wing imaginal disc. Likely, other molecules can compensate for Cad99C loss of function in this tissue. To possibly get some insights on the molecular function of other Usher syndrome proteins, I analyzed the function of Drosophila SANS and crinkled in the follicular epithelium, where both these genes are expressed. crinkled is the ortholog of myosinVIIa, that encodes a motor protein of the actin cytoskeleton. DSANS is related to human SANS and encodes a cytoplasmic protein of unknown function. It has been puzzling how removal of SANS, a cytoplasmic protein, could impair adhesion and bundling of stereocilia. To study the function of DSANS, I generated null mutant flies and observed that, in the absence of DSANS, delivery of Cad99C to microvilli is impaired. Cad99C localization is however unperturbed in crinkled mutant follicle cells. By immunostaining, DSANS immunoreactivity was detected diffusively in the cytoplasm and in dot-like structures, possibly corresponding to vesicles. In conclusion, DSANS is a cytoplasmic protein that is required for the efficient delivery of Cad99C to microvilli protrusions. Taken together, the analysis that I here performed of Drosophila Usher syndrome related genes indicates two novel molecular mechanisms of function for the corresponding human Usher syndrome proteins. First, human Protocadherin 15, like Drosophila Cad99C, could be involved in establishing adhesive interactions between microvilli protrusions of the inner ear (stereocilia). Removal of Protocadherin 15 would then cause splaying of stereocilia due to lack of inter-stereocilia adhesive links. Second, the analysis here performed suggests that SANS is involved in the efficient delivery of Protocadherin 15 to stereocilia. Mutations in SANS would then lead to splaying of stereocilia and deafness due to poor localization of Protocadherin 15 to stereocilia.

Children with sensorineural hearing loss may (re)gain hearing with a cochlear implant—a device that transforms sounds into electric pulses and bypasses the dysfunctioning inner ear by stimulating the auditory nerve directly with an electrode array. Many implanted children master the acquisition of spoken language successfully, yet we still have little knowledge of the actual input they receive with the implant and specifically which language sensitive cues they hear. This would be important however, both for understanding the flexibility of the auditory system when presented with stimuli after a (life-) long phase of deprivation and for planning therapeutic intervention. In rhythmic languages the general stress pattern conveys important information about word boundaries. Infant language acquisition relies on such cues and can be severely hampered when this information is missing, as seen for dyslexic children and children with specific language impairment. Here we ask whether children with a cochlear implant perceive differences in stress patterns during their language acquisition phase and if they do, whether it is present directly following implant stimulation or if and how much time is needed for the auditory system to adapt to the new sensory modality. We performed a longitudinal ERP study, testing in bimonthly intervals the stress pattern perception of 17 young hearing impaired children (age range: 9–50 months; mean: 22 months) during their first 6 months of implant use. An additional session before the implantation served as control baseline. During a session they passively listened to an oddball paradigm featuring the disyllable “baba,” which was stressed either on the first or second syllable (trochaic vs. iambic stress pattern). A group of age-matched normal hearing children participated as controls. Our results show, that within the first 6 months of implant use the implanted children develop a negative mismatch response for iambic but not for trochaic deviants, thus showing the same result as the normal hearing controls. Even congenitally deaf children show the same developing pattern. We therefore conclude (a) that young implanted children have early access to stress pattern information and (b) that they develop ERP responses similar to those of normal hearing children.

Children with sensorineural hearing loss may (re)gain hearing with a cochlear implant—a device that transforms sounds into electric pulses and bypasses the dysfunctioning inner ear by stimulating the auditory nerve directly with an electrode array. Many implanted children master the acquisition of spoken language successfully, yet we still have little knowledge of the actual input they receive with the implant and specifically which language sensitive cues they hear. This would be important however, both for understanding the flexibility of the auditory system when presented with stimuli after a (life-) long phase of deprivation and for planning therapeutic intervention. In rhythmic languages the general stress pattern conveys important information about word boundaries. Infant language acquisition relies on such cues and can be severely hampered when this information is missing, as seen for dyslexic children and children with specific language impairment. Here we ask whether children with a cochlear implant perceive differences in stress patterns during their language acquisition phase and if they do, whether it is present directly following implant stimulation or if and how much time is needed for the auditory system to adapt to the new sensory modality. We performed a longitudinal ERP study, testing in bimonthly intervals the stress pattern perception of 17 young hearing impaired children (age range: 9–50 months; mean: 22 months) during their first 6 months of implant use. An additional session before the implantation served as control baseline. During a session they passively listened to an oddball paradigm featuring the disyllable “baba,” which was stressed either on the first or second syllable (trochaic vs. iambic stress pattern). A group of age-matched normal hearing children participated as controls. Our results show, that within the first 6 months of implant use the implanted children develop a negative mismatch response for iambic but not for trochaic deviants, thus showing the same result as the normal hearing controls. Even congenitally deaf children show the same developing pattern. We therefore conclude (a) that young implanted children have early access to stress pattern information and (b) that they develop ERP responses similar to those of normal hearing children.

No description available.