Clostridium difficile toxin B,an inhibitor of the small GTPases Rho,Rac and Cdc42, influences spiral ganglion neurite outgrowth

OBJECTIVE: Neurotrophins and extracellular matrix (ECM) molecules are involved in neurite guidance during the development of spiral ganglion (SG) neurons. Several intracellular signaling molecules can be activated by ECMs and neurotrophins via their cognate receptors. In other systems these include the Rho small GTPases, which influence reorganization of the actin cytoskeleton that is required for axon growth. The aim of this study was to determine whether neurotrophin-3 (NT-3)-mediated SG neurite outgrowth on laminin-1 (LN) is dependent on the activation of the small GTPases Rho/Rac/Cdc42. MATERIAL AND METHODS: SG explants from postnatal day 4 rats were cultured on LN with and without NT-3 and increasing concentrations of Clostridium difficile Toxin B, an inhibitor of Rho GTPases. After fixation and immunocytochemical labeling, neurite growth was evaluated. RESULTS: Treatment with C. difficile Toxin B without NT-3 led to a dose-dependent decrease in the length and number of processes on LN. In contrast, C. difficile Toxin B had no significant effect on NT-3-mediated stimulation of neurite growth on LN, in terms of either number or length. CONCLUSION: The results suggest that the Rho GTPases (Rho, Rac and Cdc42) are not involved in the pathways linking NT-3 signals to neurite outgrowth, but appear to be involved in LN signaling in these neurons. However, NT-3 can override or bypass LN signaling to promote neurite extension.     

Rescue of auditory hair cells from ototoxicity by CEP-11 004, an inhibitor of the JNK signaling pathway

The hair cells (HCs) are the most vulnerable elements in the cochlea and damage to them is the most common cause of sensorineural hearing loss (SNHL). Understanding the intracellular events that lead to the death of HCs is a key to developing protective strategies. Recently, it has been shown that the c-Jun-N-terminal kinase (JNK) pathway is activated in HCs in response to aminoglycosides and CEP-1347, an inhibitor of the JNK signaling pathway protected HCs from ototoxicity. We have studied another inhibitor (CEP-11 004) of this signaling pathway in its ability to protect HCs from aminoglycoside ototoxicity in vitro. Organ of Corti explants from p5 rat basal turns were maintained in tissue culture and treated with CEP-11 004 for 12 hours. They were then treated with CEP-11 004 plus gentamicin for 72 hours. Significantly less HC death was observed compared to gentamicin alone. CEP-11 004 alone had no effect on HCs. We conclude that the JNK signaling pathway plays a role in aminoglycoside ototoxicity signaling.     

Blockade of synaptic transmission from hair cells to auditory afferent nerves by 6-Cyano-2,3-dihydroxy-7-nitroquinoxaline,a selective non-NMDA receptor antagonist

Summary 6-Cyano-2,3-dihydroxy-7-nitroquinoxaline (CNQX) is a new, potent and selective competitive antagonist for the non-N-methyl-d-aspartate (non-NMDA) type of excitatory amino acids receptors. We studied the effect of CNQX on the compound auditory nerve action potential (CAP), cochlear microphonics (CM), summating potential (SP) and endocochlear potential (EP). CNQX in doses of 10–20 M reduced the CAP magnitude and increased the N₁ latency without affecting the CM, SP, or EP. Parallel shifts of CAP amplitude- and latency-intensity functions were observed. The CAP suppressed by 10 M CNQX was completely reversed by a 10-min washout with artificial perilymph. As 10 M and 20 M CNQX seem to exert a selective antagonism for non-NMDA receptors, results indicate that non-NMDA receptors play a major role in synaptic transmission from hair cells to auditory afferent nerves.