Methods: Retinas were obtained from Sprague-Dawley (SD) rats at the 14th postnatal day. The cells were maintained in B27-supplemented medium
supplemented with 2% fetal bovine serum (FBS),1 mMglutamine and antibiotics. The culture media were with or
200μM AIP were added to the medium after2mMglutamate treatment for 24 hours. Expression
of RGCs markers thy-1 and axon marker neuronal class III β-tubulin and
neurofilament light chain (NFL) were studied by immunohistochemistry, and numbers of both surviving RGCs and
their axons were quantified using a microscope.
Results: We successfully
established an ex vivo model of rats’ retinal explants of RGCs. The RGCs shrank
the third day of culture, and were even much smaller and fewer the fifth day (P<0.05).
The axons of RGCs around the papilla were greatest and in good order toward
papilla the first day of culture. Their number reduced and they twisted the
third day, and the fifth day, their number was significantly reduced and
twisted (P<0.05). The expression of both neuronal class
III β-tubulin and neurofilament light chain (NFL) were reduced after treatment with glutamate while the
AIP relieved the damage caused by glutamate.
Conclusion: Our study demonstrated that both the AIP has the
neuroprotective effect of preventing RGCs from glutamate-caused injury in a retinal
explant model. This study also showed a significant change in RGCs death and
axonal loss with time in ex vivo model of retinal explants, which is an
essential characteristic for cell death and survival study. To our knowledge, the rat
retinal explants of RGCs model we discuss is a novel one for the study of RGCs
Key words：RGCs, glutamate, retinal