Intrinsic Optical Signaling Denoting Neuronal Damage in Response to Acute Excitotoxic Insult by Domoic Acid in the Hippocampal Slice

Abstract

Using the seafood contaminant domoic acid (an AMPA/kainate receptor agonist), we demonstrate a distinct excitotoxic sequence of events leading to acute neuronal damage in the hippocampal slice as measured by (1) loss of the evoked CA1 field potential, (2) irreversible changes in light transmittance, (3) histopathology, and (4) lucifer yellow injection of single CA1 pyramidal neurons. Change in light transmittance (LT) through the submerged slice indirectly measures altered cell volume, both neuronal and glial. At 37°C, a 1-min superfusion of 10 μM domoate induced a prolonged reversible increase in LT, primarily in the dendritic regions of CA1 and dentate granule cells (GC), but not in the CA3 region. Spectral analysis (400–800 nm) revealed a wide-band transmittance increase, indicating cell swelling as a major source of the intrinsic signal. The evoked field potential recorded in the CA1 cell body region (PYR) was lost as LT peaked, but completely recovered upon return to the baseline LT level. Increasing domoate exposure to 10 min elicited a different and distinct LT sequence in CA1 and dentate regions. An initial LT increase in dendritic regions evolved into an irreversibledecreasein LT. At the same time, LT irreversiblyincreasedin cell body regions (CA1 PYR and GC) and the evoked field potential was irretrievably lost. Also, there was histological damage to cell body and dendritic regions of CA1 and granule cells. Injection of lucifer yellow into single CA1 neurons in slices displaying the irreversible LT sequence revealed extensive dendritic beading, whereas CA1 cells in control slices displayed a smoothly contoured arbor. Consistent with acute neuronal damage, the optical changes generated by domoate did not require extracellular Ca2+, and lowering the temperature protected the slice from irreversible damage to CA1 and GC regions. Although glial changes may also occur, we conclude that imaging light transmittance reveals dynamic and compartmentalized excitotoxic changes in neuronal volume. Beading of the dendritic arbor increases light scatter, thereby decreasing LT and highlighting damaged dendritic regions.