Glia re‐sealed particles freshly prepared from adult rat brain are competent for exocytotic release of glutamate

Glial subcellular re‐sealed particles (referred to as gliosomes here) were purified from rat cerebral cortex and investigated for their ability to release glutamate. Confocal microscopy showed that the glia‐specific proteins glial fibrillary acidic protein (GFAP) and S‐100, but not the neuronal proteins 95‐kDa postsynaptic density protein (PSD‐95), microtubule‐associated protein 2 (MAP‐2) and β‐tubulin III, were enriched in purified gliosomes. Furthermore, gliosomes exhibited labelling neither for integrin‐αM nor for myelin basic protein, which are specific for microglia and oligodendrocytes respectively. The Ca2+ ionophore ionomycin (0.1–5 µm) efficiently stimulated the release of tritium from gliosomes pre‐labelled with [3H]d‐aspartate and of endogenous glutamate in a Ca2+‐dependent and bafilomycin A1‐sensitive manner, suggesting the involvement of an exocytotic process. Accordingly, ionomycin was found to induce a Ca2+‐dependent increase in the vesicular fusion rate, when exocytosis was monitored with acridine orange. ATP stimulated [3H]d‐aspartate release in a concentration‐ (0.1–3 mm) and Ca2+‐dependent manner. The gliosomal fraction contained proteins of the exocytotic machinery [syntaxin‐1, vesicular‐associated membrane protein type 2 (VAMP‐2), 23‐kDa synaptosome‐associated protein (SNAP‐23) and 25‐kDa synaptosome‐associated protein (SNAP‐25)] co‐existing with GFAP immunoreactivity. Moreover, GFAP or VAMP‐2 co‐expressed with the vesicular glutamate transporter type 1. Consistent with ultrastructural analysis, several ∼30‐nm non‐clustered vesicles were present in the gliosome cytoplasm. It is concluded that gliosomes purified from adult brain contain glutamate‐accumulating vesicles and can release the amino acid by a process resembling neuronal exocytosis.

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