Synthesis of serotonin from 5-hydroxytryptophan in the post-crush retina: inhibition of in vitro outgrowth by the intraocular administration of the precursor

Abstract

Serotonin is present in the retina of many species, in which plays roles as a neurotransmitter, as a modulator of regeneration, and as the precursor of melatonin. The turnover of serotonin in the goldfish retina is modified by the lesion of the optic nerve and, in postcrush goldfish retinal explants, serotonin inhibits the outgrowth. In the present study, the modification of the serotonergic system of the retina induced by the process of regeneration was explored. The addition of the precursor of serotonin, 5-hydroxytryptophan, to retinal explants, increased the levels of serotonin in a concentration-dependent manner. The concentration of serotonin differentially increased in control and postcrush explants cultured in the presence of 5-hydroxytryptophan for various periods of time, indicating a greater accumulation of the indoleamine at early periods of time in the control than in the postcrush tissue culture. This observation, together with the fact that serotonin concentration in postcrush retina cultured in the absence of 5-hydroxytryptophan and exposed to the precursor for 60 min increased less than in the control, indicates a saturation of the serotonergic system produced by the lesion. The addition of imipramine or citalopram, serotonin uptake blockers, did not significantly change the concentration of serotonin in the cultures, thus, the elevation of serotonin accumulation, especially in the post-crush tissue, might not be due to the transport from the medium. The intraocular injection of 5-hydroxytryptophan after the crush of the optic nerve resulted in a decrease in the outgrowth of retinal explants, supporting the in vivo role of serotonin during the regenerating process in situ. The lesion of the optic nerve did not affect the specific cells, since the number of serotonin-immunoreactive neurons in the retina were not modified by the crush. Taken together, retinal serotonin system is regulated after producing a lesion of the optic nerve, a modulation which has been demonstrated in vivo and in vitro. Thus, there is a reciprocal interaction, since serotonin influences outgrowth in the postcrush retina and the serotonergic system is modulated by the crush, indicating a mechanism of feed-back regulation.