Beneficial and Detrimental Pressure-Related Effects on Inner Neurons in the Adult Porcine In Vitro Retina.

Abstract

To explore pressure-related effects in the adult porcine retina in vitro. Retinal explants were subjected to 0, 10, 30, or 60 mmHg of pressure for 24 or 48 hours in culture. Overall tissue damage in sections was assessed by lactate dehydrogenase media levels, hematoxylin and eosin staining, and TUNEL staining. Inner retinal neurons were evaluated by protein kinase C alpha (rod bipolar cells), CHX10 (overall bipolar cell population), parvalbumin (amacrine cells), and RBPMS (ganglion cells) immunohistochemistry. All retinas kept in culture displayed increased pyknosis and apoptosis compared with directly fixed controls. The 10-mmHg explants displayed attenuation of overall tissue damage compared with the 0-, 30-, and 60-mmHg counterparts. No difference in the number of rod bipolar cells was seen in the 10-mmHg explants compared with directly fixed controls, whereas significantly fewer cells were detected in the remaining pressure groups. No difference in the number of ganglion cells in the 0-, 10-, and 60-mmHg groups was seen compared with directly fixed controls after 24 hours, whereas a lower number was found in the 30-mmHg counterpart. A decline of ganglion cells was found in the 0-, 10-, and 60-mmHg group after 48 hours, but no further decrease was seen in the 30-mmHg group. No differences were detected in overall bipolar and amacrine cells in the pressure groups after 24 hours compared with directly fixed controls. A moderate amount of pressure attenuates culture-related retinal neurodegeneration. Rod bipolar cells are specifically vulnerable to excessive pressure. These findings are relevant for glaucoma-related research.