Netrin-1 as a guidance molecule in the postnatal rat cochlea.

During synaptogenesis a number of growth factors and peptides control the guidance of auditory neuron (spiral ganglion neuron, SGN) axons to their target cells. Furthermore, evidence suggests that these factors exert their actions at discrete times and sites during development. This study demonstrates that the guidance molecule netrin-1 is expressed in the early postnatal rat cochlea, but shows decreasing expression with increasing age. These results suggest that netrin-1 may be involved in guiding axonal growth from SGNs for the onset of innervation, but is not required for maintenance of synaptic connections.

Delayed neurotrophin treatment supports auditory neuron survival in deaf guinea pigs.

As key factors in the development and maintenance of the auditory system, neurotrophins can prevent auditory neuron degeneration when applied within three to five days of deafening. We tested each of the neurotrophins BDNF, NT-3, NT-4/5 and NGF for their ability to support auditory neuron survival following a two-week period of deafness in guinea pigs, when approximately 15% auditory neuron degeneration has already occurred. Although delayed, the treatment with each neurotrophin prevented further degeneration with similar efficacy.

BDNF-induced survival of auditory neurons in vivo: Cessation of treatment leads to accelerated loss of survival effects.

Neurotrophic factors are important for the development and maintenance of the auditory system. They have also been shown to act as survival factors for auditory neurons in animal deafness models. Studies have demonstrated recently that these neurotrophic factors not only maintain survival of auditory neurons, but that these surviving neurons retain functionality. It remains to be determined, however, if a single administration of a neurotrophic factor is sufficient to maintain auditory neuron survival after loss of hair cells, or if sustained delivery is required. This study investigated the longevity of the survival effects of BDNF on auditory neurons in deafened guinea pigs. Briefly, the left cochleae of deafened guinea pigs were infused with BDNF for 28 days via a mini-osmotic pump, and neuronal survival was analyzed at various stages after the completion of treatment. BDNF treatment prevented the degeneration of auditory neurons that normally is seen after a loss of hair cells, supporting previous studies. Our results indicate, however, that cessation of BDNF treatment leads to an accelerated decline in auditory neuron survival as compared to that observed in deafened, untreated cochleae. These findings indicate that much work remains to be done to establish a technique for the long-term survival of auditory neurons in the deaf ear.

The transmembrane serine protease (TMPRSS3) mutated in deafness DFNB8/10 activates the epithelial sodium channel (ENaC) in vitro.

TMPRSS3 encodes a transmembrane serine protease that contains both LDLRA and SRCR domains and is mutated in non-syndromic autosomal recessive deafness (DFNB8/10). To study its function, we cloned the mouse ortholog which maps to Mmu17, which is structurally similar to the human gene and encodes a polypeptide with 88% identity to the human protein. RT-PCR and RNA in situ hybridization on rat and mouse cochlea revealed that Tmprss3 is expressed in the spiral ganglion, the cells supporting the organ of Corti and the stria vascularis. RT-PCR on mouse tissues showed expression in the thymus, stomach, testis and E19 embryos. Transient expression of wild-type or tagged TMPRSS3 protein showed a primary localization in the endoplasmic reticulum. The epithelial amiloride-sensitive sodium channel (ENaC), which is expressed in many sodium-reabsorbing tissues including the inner ear and is regulated by membrane-bound channel activating serine proteases (CAPs), is a potential substrate of TMPRSS3. In the Xenopus oocyte expression system, proteolytic processing of TMPRSS3 was associated with increased ENaC mediated currents. In contrast, 6 TMPRSS3 mutants (D103G, R109W, C194F, W251C, P404L, C407R) causing deafness and a mutant in the catalytic triad of TMPRSS3 (S401A), failed to undergo proteolytic cleavage and activate ENaC. These data indicate that important signaling pathways in the inner ear are controlled by proteolytic cleavage and suggest: (i) the existence of an auto-catalytic processing by which TMPRSS3 would become active, and (ii) that ENaC could be a substrate of TMPRSS3 in the inner ear.

Role of trophic factors in the development, survival and repair of primary auditory neurons.

1. Neurotrophic factors have been identified as crucial for the development of the auditory system and have also been proven to be important for continued survival and maintenance of auditory neural connections. 2. In addition, both in vitro and in vivo studies have demonstrated that these trophic molecules can prevent the secondary wave of auditory neuron degeneration normally seen following the loss of hair cells. 3. Furthermore, neurotrophic factors have been reported to enhance neuronal excitation and to improve the efficacy of synaptic transmission. 4. As such, these molecules are strong candidates to be used as therapeutic agents in conjunction with the cochlear implant, or even to repair and/or regenerate damaged or lost auditory nerve and sensory cells.