Abstract
Certain viruses can infect neurons and cause persistent infections with restricted expression of viral proteins. To study the consequences of such viral proteins on synaptic functions, the effects of two influenza A virus proteins, the nonstructural protein 1 (NS1) and the nucleoprotein (NP), were analyzed in cultures of rat hippocampal neurons. Transduction of the NS1 and NP proteins into the neurons was performed by applying the 11–amino acid peptide transduction domain (PTD) of human immunodeficiency virus (HIV) TAT coupled to the viral proteins. Neurons exposed to the NS1 and NP fusion proteins (NS1-PTD and NP-PTD, respectively) for 4 h were immunopositive for these proteins as diffuse cytoplasmic and nuclear distribution. After exposure for 48 h to NP-PTD, a punctate pattern of the immunolabel appeared in dendritic spinelike processes. Electrophysiologically, a reduction in both the frequency of spontaneous excitatory synaptic activity and in the amplitude of the miniature excitatory postsynaptic currents were recorded after exposing the hippocampal neurons to NP-PTD between 17 and 22 days in culture. These changes may reflect disturbances in postsynaptic functions. No such alterations in synaptic activities were recorded after exposure to NS1-PTD or to green fluorescent protein–PTD, which was used as a control. Based on these findings the authors hypothesize that the viral NP, by its localization to dendritic spinelike structures, interferes with the expression or anchoring of postsynaptic glutamate receptors and thereby disturbs synaptic functions. Thus a persistent viral infection in the brain may be associated with functional disturbances at the synaptic level.
Johan Brask and Ashok Chauhan contributed equally to this work.
The present address of Ashok Chauhan is Department of Neurology, Johns Hopkins Hospital, Baltimore, Maryland, USA.
This study was supported by grants from The Stanley Medical Research Institute and Swedish Research Council (04480). Received; revised; accepted