Cooperation between glutathione depletion and protein synthesis inhibition against naturally occurring neuronal death
Section snippets
Animals and products
White Leghorn chick embryos were purchased from Pulfer Geflügefarm (Thörishaus, Switzerland), BSO and cycloheximide from Sigma, l-[4,5-3H]leucine from Amersham Life Science, and Reagents for biochemical assays from Sigma or Fluka (Buchs, Switzerland).
The embryos received an injection into the right eye of BSO, cycloheximide, or vehicle (0.9% NaCl), the volume injected was adjusted to 5 μl, as described previously.[8]Injections were performed after 12 embryonic days of incubation (E12, E0 being
Effects of intraocular injections on glutathione retinal concentrations
Intraocular injection of BSO at E12, E12+8h, or E12+16h did not modify the survival of the embryos as compared to NaCl-treated embryos (i.e. 80% of the embryos survived until E13). The concentration of GSH in NaCl-treated embryos ranged between 15.77 and 23.65 nmol/mg of protein; comparable concentrations have been reported in bovine,[54]guinea-pig,[32]and human retinas.[47]BSO induced a dose-dependent reduction of GSH retinal concentrations (Table 1). When injected at E12, the lowest dose of
The nature of the cells counted as pyknotic
The present report describes the effects on naturally-occurring neuronal death of BSO, a GSH depleting agent, and of cycloheximide, a protein synthesis inhibitor. The data confirm and extend our previous report of the synergist effects of the two drugs on axotomized ganglion cells.[9]
The ganglion cell layer of the chick embryo contains the ganglion cells, and the displaced amacrine cells. Displaced amacrine cells represent 22 to 24% of the neurons in E10–E15 chick embryos[34]and 30–35% after
Conclusions
We have previously shown that axotomy-induced ganglion cell death can be reduced by antioxidants[8]and cycloheximide,8, 9suggesting that ganglion cells die through mechanisms involving oxidative stress and protein synthesis.6, 31Moreover, axotomy-induced ganglion cell death could be nearly totally inhibited by combinations of cycloheximide and BSO,[9]suggesting cell death-regulating interactions between oxidative processes and the protein synthesis machinery.[58]Our present data show that, as
Acknowledgements
This study was supported by the Swiss National Foundation for Scientific Research, grants 31-40709.94 and 31-50598.97 and by the Swiss Federal Office for Education and Science (EU Bio4 CT 96-0649). We thank L. Grollimund and N. Turrian for their skilful technical support.
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