Role of nitric oxide in kainic acid-induced elevation of cochlear compound action potential thresholds.

Nitric oxide (NO) has specifically been found to mediate the effects of excitatory amino acids in the central nervous system (CNS). Excitatory amino acids are the primary neurotransmitters at the cochlear hair cell afferent nerve synapse. Recent studies in our laboratory demonstrate that NO synthase is an active enzyme in the spiral ganglion cells of the cochlea. Given our current understanding of neurotransmission in the cochlea, it is reasonable to postulate that the actions of NO in cochlear neuronal tissue are similar to the actions of NO in the CNS, and that NO acts as a neurotransmitter/neuromodulator in the cochlea. In addition, NO is implicated as a mediator of excitotoxicity in the CNS and may therefore play a similar role in excitotoxicity in the cochlea. To further elucidate the role of NO in cochlear excitotoxicity, this study investigated the effects of 7-nitroindazole (7-NI), a competitive inhibitor of neuronal nitric oxide synthase, with regard to kainic acid (KA)-induced elevation of compound action potential (CAP) thresholds. KA is a conformationally restricted analog of glutamate with well-known excitotoxic effects on SGC's and previously described inhibitory actions on cochlear CAP thresholds. In anesthetized gerbils, CAP thresholds were recorded before and after cochlear perfusions with control solutions of artificial perilymph solution and test solutions of KA. 7-NI was administered i.p. prior to KA perfusion in an effort to block its depolarizing and toxic effects. Results showed that cochlear perfusion with KA caused significant elevation (p < 0.05) of the mean CAP threshold. This threshold shift was significantly reduced (p < 0.05) in animals pretreated with 7-NI. These results indicate that NO is involved in the toxic effects on CAP thresholds elicited by KA in the cochlea.

Sodium nitroprusside/nitric oxide causes apoptosis in spiral ganglion cells.

OBJECTIVE: In the cochlea, excitatory amino acid receptor overstimulation induces toxicity in spiral ganglion neurons by an unknown mechanism. In the central nervous system, excitatory amino acid-induced toxicity is mediated by nitric oxide, which induces apoptosis in neurons. This study tested the hypothesis that cochlear nitric oxide-mediated toxicity is the result of induction of apoptosis in spiral ganglion neurons. METHODS: The cochleas of 15 gerbils randomly assigned to different groups were perfused for 30 minutes with a test solution of 1 mmol/L sodium nitroprusside, a nitric oxide donor, or a control solution of artificial perilymph. Animals were killed at varying times, including 2, 3, 4, 8, and 18 hours after perfusion. DNA fragmentation or in situ terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end-labeling analysis was done on cochleas for detection of apoptosis. RESULTS: Analysis by both techniques demonstrated marked apoptotic cell changes in spiral ganglion neurons of sodium nitroprusside-treated cochleas evident 4 to 8 hours after perfusion, as compared with minimal to no evidence of apoptosis in spiral ganglion neurons of control specimens. CONCLUSIONS: Exposure to high levels of nitric oxide induces apoptosis in spiral ganglion neurons. Because apoptosis is a delayed, potentially reversible cell death pathway, this may present an opportunity for intervention to prevent or attenuate hearing damage induced by excitotoxic stimuli.

Inhibition of nitric oxide synthase causes elevation of hearing thresholds.

OBJECTIVE: Nitric oxide mediates the effects of excitatory amino acids in the central nervous system. The excitatory amino acids are thought to be the neurotransmitters at the cochlear hair cell-afferent nerve synapse. Nitric oxide synthase is present in spiral ganglion cells. This study investigated the role of nitric oxide in cochlear neurotransmission. METHODS: In gerbils, cochlear compound action potential thresholds were recorded before and after cochlear perfusions with control solutions of artificial perilymph solution and test solutions of S-methyl-L-thiocitrulline (MTC), a competitive inhibitor of nitric oxide synthase. Cochleas were also preperfused with L-arginine before perfusion with a mixture of MTC/L-arginine (to overcome competitive inhibition by MTC with L-arginine, the natural substrate of nitric oxide synthase). RESULTS: Cochlear perfusion with MTC caused significant elevations of compound action potential threshold of 51 dB as opposed to insignificant elevations of only 10 dB in control animals. An insignificant threshold shift of 9 dB was observed when L-arginine was coperfused with MTC. CONCLUSIONS: Nitric oxide is involved in neurotransmission/neuromodulation in the cochlea. Because nitric oxide is both a mediator of neurotoxicity and an initiator of apoptosis in the central nervous system, nitric oxide may play a role in these processes in the cochlea.

Nitric oxide in the rat vestibular system.

Nitric oxide is known to function as a neurotransmitter in the central nervous system. It is also known to be involved in the central nervous system excitatory amino acid neurotransmission cascade. Activation of excitatory amino acid receptors causes an influx of calcium, which activates nitric oxide synthase. The resulting increase in intracellular nitric oxide activates soluble guanylate cyclase, leading to a rise in cyclic guanosine monophosphate. The excitatory amino acids glutamate and aspartate are found in the vestibular system and have been postulated to function as vestibular system neurotransmitters. Although nitric oxide has been investigated as a neurotransmitter in other tissues, no published studies have examined the role of nitric oxide in the vestibular system. Neuronal NADPH-diaphorase has been characterized as a nitric oxide synthase. This enzyme catalyzes the conversion of L-arginine to L-citrulline, producing nitric oxide during the reaction. We used a histochemical stain characterized by Hope et al. (Proc Natl Acad Sci 1991;88:2811) as specific for neuronal nitric oxide synthase to localize the enzyme in the rat vestibular system. An immunocytochemical stain was used to examine rat inner ear tissue for the presence of the enzyme's end product, L-citrulline, thereby demonstrating nitric oxide synthase activity. Staining of vestibular ganglion sections showed nitric oxide synthase presence and activity in ganglion cells and nerve fibers. These results indicate the presence of active nitric oxide synthase in these tissues and suggest modulation of vestibular neurotransmission by nitric oxide.

Nitric oxide synthase is an active enzyme in the spiral ganglion cells of the rat cochlea.

Nitric oxide (NO) mediates the effects of the excitatory amino acids in the central nervous system. Excitatory amino acids, in particular L-glutamate, are thought to be the neurotransmitter(s) present at the cochlear hair cell-afferent nerve synapse. To our knowledge, no studies to date have documented the presence of NO in the cochlea nor attempted to elucidate the role of NO in hearing. Rat cochlea frozen sections were examined for the presence of nitric oxide synthase (NOS) by NADPH diaphorase histochemistry. Vibratome sections of rat cochlea were examined by immunocytochemistry with an antibody to citrulline, an indication of NOS activity. Spiral ganglion cells in the rat cochlea were positive by NADPH diaphorase histochemistry and by anti-citrulline immunocytochemistry. These results indicate that NOS is present and that the enzyme actively produces nitric oxide in the spiral ganglion cells of the rat cochlea. Given our current understanding of neurotransmission in the cochlea, it is reasonable to postulate that the actions of NO in cochlear neuronal tissue are similar to the actions of NO in the CNS and that NO acts as a neurotransmitter/neuromodulator in the cochlea. In addition, because NO has been implicated as a mediator of excitotoxicity in the CNS, NO may play a role in neurotoxicity in the cochlea.