Neurotrophic factors and depolarization do not enhance viability and process regrowth on a purified set of chick embryo retinal ganglion cells.

As retinal ganglion cells (RGCs) develop, the amount of membrane glycoprotein Thy-1 increases, consequently, the RGC binding to Thy-1 antibody enlarges. This phenomenon sustained Thy-1 panning purification of two sets: loose bound cells (LBCs); and tight bound cells (TBCs). Thy-1, neurofilament and growth associated protein-43 characterized both sets as RGCs, but the expression of RA4 antigen and gangliotetraosylgangliosides distinguished LBCs as immature and TBCs as mature. The cell composition in 4-day TBC cultures remained unchanged as immunocharacterization indicated. These cells survived and regrew their processes in plain medium, however, trophic factors (TFs) and depolarization did not enhance their development. This peculiar behavior might be due to their neural maturity, whereas in LBC cultures, TFs produced positive effects.

Retinal ganglion cells are autonomous circadian oscillators synthesizing N-acetylserotonin during the day.

Retinal ganglion cells send visual and circadian information to the brain regarding the environmental light-dark cycles. We investigated the capability of retinal ganglion cells of synthesizing melatonin, a highly reliable circadian marker that regulates retinal physiology, as well as the capacity of these cells to function as autonomous circadian oscillators. Chick retinal ganglion cells presented higher levels of melatonin assessed by radioimmunoassay during both the subjective day in constant darkness and the light phase of a light-dark cycle. Similar changes were observed in mRNA levels and activity of arylalkylamine N-acetyltransferase, a key enzyme in melatonin biosynthesis, with the highest levels of both parameters during the subjective day. These daily variations were preceded by the elevation of cyclic-AMP content, the second messenger involved in the regulation of melatonin biosynthesis. Moreover, cultures of immunopurified retinal ganglion cells at embryonic day 8 synchronized by medium exchange synthesized a [3H]melatonin-like indole from [3H]tryptophan. This [3H]indole was rapidly released to the culture medium and exhibited a daily variation, with levels peaking 8 h after synchronization, which declined a few hours later. Cultures of embryonic retinal ganglion cells also showed self-sustained daily rhythms in arylalkylamine N-acetyltransferase mRNA expression during at least three cycles with a period near 24 h. These rhythms were also observed after the application of glutamate. The results demonstrate that chick retinal ganglion cells may function as autonomous circadian oscillators synthesizing a melatonin-like indole during the day.