Protein Kinase C Regulates [3H]d-Aspartate Release in Auditory Brain Stem Nuclei

Abstract

We previously found that unilateral cochlear ablation altered transmitter release from glutamatergic synaptic endings in several brain stem auditory nuclei. To determine if this release activity could be regulated by protein kinase C (PKC), which has been associated with regulation of transmitter release, the electrically evoked release of [3H]d-aspartate ([3H]d-Asp) was quantified in vitro as an index of exocytosis from glutamatergic presynaptic endings in the major subdivisions of the cochlear nucleus (CN) and in the main nuclei of the superior olivary complex (SOC). Treating dissected tissues with a PKC activator, such as phorbol 12,13-diacetate (PDA) or phorbol 12,13-dibutyrate (PDBu) (3 μM), elevated the evoked release of [3H]d-Asp by 1.5- to 3.3-fold. The PKC inhibitor Ro31-8220 (50 nM) did not alter the evoked release but blocked the stimulatory effects of PDA and PDBu. These findings suggested that PKC could positively regulate transmitter release from glutamatergic presynaptic endings in brain stem auditory pathways. Seven days after unilateral cochlear ablation, when cochlear nerve endings had degenerated in the ipsilateral CN, PDBu elevated the evoked release bilaterally in each CN subdivision and SOC nucleus, implying that PKC could regulate glutamatergic release in the noncochlear pathways remaining in the ipsilateral CN and in the other pathways after unilateral hearing loss. After 145 postlesion days, Ro31-8220 blocked endogenous elevations in the evoked release in the ipsilateral SOC but did not alter the elevated or upregulated release in the other tissues. This suggested that the elevations of glutamatergic release activity in the ipsilateral SOC that appeared after unilateral cochlear ablation depended on endogenous activation of PKC.