INCORPORATION OF FLUORESCENT LIPIDS INTO LIVING RABBIT HIPPOCAMPAL AND CEREBRAL SLICES

Incorporation of exogenously applied fluorescent lipids into living ce lls was exploited to probe cellular structure and function in living h ippocampal and cerebellar slices as assessed by fluorescent imaging te chniques and intracellular recording. Nitrobenzoxadiole-phosphatidylch oline (NBD-PC) and BODIPY phorbol ester, in vitro substrates of phosph olipase activity and protein kinase C, respectively, were incorporated and distributed into specific cell populations. In the hippocampal sl ice, both probes labeled the somata and proximal dendrites of pyramida l and granule cells but were hetrogeneously distributed across the dif ferent hippocampal fields. Changes in fluorescent properties of NBD-PC in individual pyramidal cell and granule cell somata were quantified upon challenge with a muscarinic agonist known to modulate phospholipa se A2 activity. In the cerebellar slice, both probes labeled Purkinje cell bodies and dendrites but only NBD-PC labeled stellate and granule cells. The cellular and functional specificity of these fluorescent l ipid probes shows great promise for monitoring biochemical events in c omplex neuronal systems with significant spatial and temporal resoluti on. (C) 1994 Academic Press, Inc.