Effects of Ethanol on Development of Locus Coeruleus Brain Stem Transplantsin oculo

Abstract

In this investigation, we studied the effects of ethanol (EtOH) on the development of noradrenergic (NE) neurons of the locus coeruleus. Fetal brainstem tissue from embryonic days 15-17 was grafted into the anterior chamber of the eye of adult rats. Two different experimental groups were exposed to 16% EtOH in the drinking water during different developmental windows. The first group received EtOH 24 h after transplantation and during the whole experimental period of 7 weeks (continuous EtOH), and the second group only during the last 5 weeks of the experimental period (delayed EtOH). The control group received water ad libitum. After 7 weeks, all the animals were sacrificed and morphological evaluations were performed. Immunohistochemical analysis showed that axon bundle formation and NE fiber outgrowth into the host iris was significantly reduced in the continuous EtOH-treated group compared to controls. We also studied the morphology of TH-positive neurons and processes in the intraocular transplants. A significant decrease in TH-positive staining intensity was observed in the continuous EtOH-treated group compared to controls. Moreover, we found a significant decrease in cell size and neuronal survival in both EtOH-treated groups compared to controls. The present results suggest that chronic EtOH exposure during development leads to an altered axonal outgrowth and decreased cell sizes and number of NE neurons in intraocular brain stem grafts. Furthermore, we found that NE neurons are more sensitive to EtOH exposure during the last prenatal days and the first postnatal week of development, compared to a later developmental period.