Increased deafferentation-induced cell death in chick brainstem auditory neurons following blockade of mitochondrial protein synthesis with chloramphenicol

Abstract

Second-order auditory neurons in nucleus magnocellularis (NM) of the chick brainstem undergo a series of rapid metabolic changes following unilateral cochlea removal, culminating in the death of 25% of NM neurons. Within hours of cochlea removal, ipsilateral NM neurons show marked increases in histochemical staining for the mitochondrial enzymes succinate dehydrogenase and cytochrome oxidase (CO). We have shown previously in an ultrastructural study that these increases in oxidative capacity are mediated in part by a rapid increase in mitochondrial volume within deafferented neurons. In neurons that are destined to die as a result of deafferentation, mitochondria are smaller, stain poorly for CO, and often contain vacuoles associated with oxidative dysfunction. Our present set of experiments is designed to test the hypothesis that increases in oxidative metabolism are necessary for NM neuronal survival following removal of afferent input. We used chloramphenicol (CAP), a mitochondrial protein synthesis inhibitor, to block the characteristic increase in CO activity and mitochondrial proliferation following cochlea removal. We then studied the effects of CAP on NM neuronal survival following deafferentation. When CAP was administered continuously at 600 mg/kg/d for 5 d following cochlea removal, deafferentation-induced neuronal death in NM was significantly increased from 22% to 36%. Higher-dose (1200 mg/kg/d) pulses of CAP were administered for the first 6, 12, or 24 hr following cochlea removal. After 5 d survival, greater increases in neuronal cell death were found in animals treated for the first 12 or 24 hr (65% neuronal death). CAP administration for the first 6 hr had no significant effect on neuronal survival. The effects of CAP on neuronal cell death in NM are not likely to be due to systemic effects of the drug, but instead to a specific change in mitochondrial function. Our results suggest that enhanced oxidative enzyme function plays an important role in the survival of NM neurons during the first 24 hr after deafferentation. The nature of the signal(s) eliciting mitochondrial enhancement and the means by which it influences NM survival are not known.