Effect of TrkA and p75NTR on the Survival of Neurons in the Cultured Organotypic Hippocampal Slice

ABSTRACT

BACKGROUND: Nerve growth factor (NGF) promotes the survival and differentiation of vertebrate neurons, and their actions are mediated by two classes of cell surface receptors tyrosine kinase A receptor (TrkA) and p75 neurotrophic receptor (p75NTR). We evaluated the role of NGF receptors in neuronal survival and the physical interactions between them. METHODS: Organotypic hippocampal slices were obtained from 5 to 7-day-old rat pups and were grown for 14 days in vitro. The expression of the TrkA and p75NTR was evaluated by the western blot and immunohistochemical methods. The neuroprotective effect of NGF on the blocking of antibody-induced neuronal cell death was tested by the application of NGF (0, 50 and 150 ng/ml) to the culture media in the presence of 200 ng/ml of blocking antibodies against TrkA and p75NTR. Functional interactions between the two receptors were examined using the immunoprecipitation method. RESULTS: TrkA and p75NTR were co-expressed in the principal neurons of the hippocampal slice culture, and the expression level was increased time dependently until 14 days of culture. The blocking antibody against each receptor induced neuronal damage in time and dose-dependent manners. NFG delayed or prevented the blocking antibody from inducing neuronal damage. Results from the immunoprecipitation experiment showed physical interactions between the two NGF receptors. CONCLUSIONS: Our results indicate that the co-expressed NGF receptors, TrkA and p75NTR, might have protective roles in the survival of neuronal cells through the cooperative interactions between them.