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PDT‐induced death of sensory neurons and glial cells in the isolated crayfish stretch receptor after proteolytic treatment

To study the involvement of neuroglial interactions in photodynamic damage of crayfish stretch receptor, which consists of only two neurons surrounded by satellite glial cells (SGCs), we attempted to proteolytically uncouple neurons and glia and then compare the responses of these cells to photosensitization when intercellular communications were intact or impaired. After incubation of isolated stretch receptors with pronase or collagenase they were photosensitized with Photosens, a mixture of sulfonated alumophthalocyanines AlPcSn (n = 2, 3, and 4; mean n = 3.1). In the next 6 hr the preparations were double fluorochromed with propidium iodide and Hoechst‐33342 to visualize necrotic and apoptotic cells. Proteolytic treatment shortened bioelectric neuron response and precipitated its functional inactivation; however, it did not significantly impair neuron morphology and did not induce its necrosis either in the darkness or under photosensitization. Photodynamic treatment induced necrosis of neurons and SGC and apoptosis of glial cells. Pronase but not collagenase increased percent of necrotic and apoptotic SGCs in the darkness and thus reduced the number of glial cells around the neuron; however, both pronase and collagenase prevented photodynamically induced apoptosis of glial cells. The involvement of neuron‐to‐glia signaling interactions in this phenomenon is suggested. © 2005 Wiley‐Liss, Inc.