Electrophysiological and electrochemical responses of NMDA in the cerebellum: interactions with nonadrenergic pathway.

In the present experiments, we measured N-methyl-D-aspartate (NMDA)-induced norepinephrine (NE) release and extracellular action potentials in the cerebellar cortex of urethane-anesthetized rats. The overflow of NE was measured using a Nafion coated-carbon fiber electrode and in vivo chronoamperometry. We found that both NMDA and quisqualate evoked NE release. Our previous work demonstrated that the electrophysiological activity of cerebellar Purkinje neurons could be either excited or inhibited by local NMDA application. It was reported that bicuculline antagonized NMDA-induced inhibition in Purkinje neurons, suggesting that a GABAergic mechanism was activated during NMDA application. We and others previously found that GABA-mediated electrophysiological depressions were enhanced by NE acting via beta-adrenergic receptors while these responses were decreased by activation of alpha-adrenergic receptors. Since NMDA evokes overflow of endogenous NE, the electrophysiological depression induced by NMDA may contain an NE-mediated modulatory component. In this study, we first examined the interaction of NMDA with beta-adrenergic agonist. We found that local application of isoproterenol facilitated NMDA- or GABA-mediated electrophysiological depressions of the Purkinje neurons. Applications of phenoxybenzamine, which antagonized the alpha-adrenergic response of synaptically released NE, also facilitated NMDA-elicited depression. In contrast, the depression induced by GABA, which did not induce NE overflow, was not potentiated by phenoxybenzamine. The facilitation of NMDA-induced depression by phenoxybenzamine was antagonized by the beta-adrenergic antagonist timolol. Taken together, these data suggested that the nonadrenergic pathway is involved in NMDA-induced electrophysiological responses in the cerebellum. NMDA may induce neuronal depression through modulation of GABAergic inhibition via NMDA-evoked NE release onto cerebellar Purkinje neurons.

Phagocyte-derived free radicals stimulated by ingestion of iron-rich Staphylococcus aureus: a spin-trapping study.

Phagocytic cells generate superoxide (O2-) and hydrogen peroxide (H2O2), creating the substrates for hydroxyl radical (HO.) in the presence of redox active metals. Previously it was shown that HO. is not a physiologic product of human neutrophils or monocytes but can be generated in the presence of high concentrations of iron. This study was undertaken to determine whether bacterial iron could be used for the generation of HO. The growth of Staphylococcus aureus under iron-rich conditions increased bacterial iron concentration and phagocytosis of iron-rich bacteria allowed neutrophils to accumulate threefold more iron than ingestion of iron-starved organisms. Neither neutrophils nor monocytes ingesting iron-rich S. aureus generated iron-catalyzed HO. at levels detectable by spin-trapping techniques. No differences in the killing of iron-rich organisms by neutrophils was noted. The results suggest that HO. does not play a role in the killing of S. aureus by human neutrophils, regardless of their ability to deliver iron to the cell.