Zinc modulates synaptic transmission by differentially regulating synaptic glutamate homeostasis in hippocampus

A subset of presynaptic glutamatergic vesicles in the brain co-releases zinc (Zn²⁺ ) with glutamate into the synapse. However, the role of synaptically released Zn²⁺ is still under investigation. Here, we studied the effect of Zn²⁺ on glutamate homeostasis by measuring the evoked extracellular glutamate level (EGL) and the probability of evoked action potential (P_EAP ) at the Zn²⁺ -containing or zincergic mossy fiber-CA3 synapses of the rat hippocampus. We found that the application of Zn²⁺ (ZnCl₂ ) exerted bidirectional effects on both EGL and P_EAP : facilitatory at low concentration (~1 µM) while repressive at high concentration (~50 µM). To determine the action of endogenous Zn²⁺ , we also used extracellular Zn²⁺ chelator to remove the synaptically released Zn²⁺ . Zn²⁺ chelation reduced both EGL and P_EAP , suggesting that endogenous Zn²⁺ has mainly a facilitative role in glutamate secretion on physiological condition. We revealed that calcium/calmodulin-dependent protein kinase II was integral to the mechanism by which Zn²⁺ facilitated the release of glutamate. Moreover, a glutamate transporter was the molecular entity for the action of Zn²⁺ on glutamate uptake by which Zn²⁺ decreases glutamate availability. Taken together, we show a novel action of Zn²⁺ , which is to biphasically regulate glutamate homeostasis via Zn²⁺ concentration-dependent synaptic facilitation and depression. Thus, co-released Zn²⁺ is physiologically important for enhancing weak stimulation, but potentially mitigates excessive stimulation to keep synaptic transmission within optimal physiological range.