Endocytosis of tropomyosin-related kinase (Trk) receptors is critical for neurotrophin signal transduction and biological functions. However, the mechanism governing endocytosis of tropomyosin-related kinase B (TrkB) and the specific contributions of TrkB endocytosis to downstream signaling are unknown. In this study, we report that blocking clathrin, dynamin or AP2 in cultured neurons of the central nervous system inhibited brain-derived neurotrophic factor (BDNF)-induced activation of Akt but not ERK. Treating neurons with the clathrin inhibitor MDC or a peptide that blocks dynamin function specifically abrogated Akt pathway activation in response to BDNF but did not affect the response of other downstream effectors, or the upregulation of immediate early genes NPY and ARC. Similar effects were found in neurons expressing AP2 RNAi or a dominant negative form of dynamin that inhibits clathrin-mediated endocytosis. In PC12 cells, ERK but not Akt activation required TrkA endocytosis following stimulation with nerve growth factor (NGF) whereas the opposite was true when TrkA expressing neurons were stimulated with NGF in central nervous system. Thus, the specific effects of internalized Trk receptors likely depend on the presence of cell-type specific modulators of neurotrophin signaling and not on differences inherent to Trk receptors themselves. Endocytosis-dependent activation of Akt in neurons was found to be critical for BDNF-supported survival and dendrite outgrowth. Together these results demonstrate the functional requirement of clathrin- and dynamin-dependent endocytosis in generating the full intracellular response of neurons to BDNF in central nervous system.