Figure 5.

Supply of purified GS enzyme in vitro or in ovo reduces the extent of LDH release. (A) E17 retina was excised, cut into pieces, and organ-cultured in the absence (−) or presence of purified GS enzyme in the indicated unit amounts. The level of LDH in the culture medium was measured after 4 h. Each bar represents the mean ± SD of two separate experiments, each performed in triplicate (n = 6). (B) E17 eggs were injected with carrier (−), with purified GS enzyme in the indicated unit amounts (1 to 8 units per egg), with ovalbumin (20 μg per egg; OVA), or with heat-inactivated GS enzyme (2 units per egg; IGS). The injected eggs were incubated for 2 h before the retina was excised. Retinal explants were cut into pieces and organ-cultured for 4 h, and the level of LDH in the culture medium was then measured. Each bar represents the mean ± SD of two separate experiments, each performed in quadruplicate (n = 8). (C) E15, E16, and E17 eggs were injected with purified GS enzyme (2 units per egg). Retinas were excised from the injected E17 eggs immediately (−) or after 2 or 4 h of incubation. Injected E16 and E15 eggs were incubated for an additional 24 or 48 h, respectively, before the retinal tissues were excised. Tissue explants were cut into pieces and organ-cultured for 4 h, and the level of LDH in the culture medium was then measured. Each bar represents the mean ± SD of two separate experiments, each performed in quadruplicate (n = 8).