Changes in calcium currents and GABAergic spontaneous activity in cultured rat hippocampal neurons after a neurotropic influenza A virus infection.
In order to study mechanisms by which a neurotropic strain of influenza A virus (A/WSN/33) may affect neuronal function or cause nerve cell death, hippocampal cultures from embryonic rats were infected with this virus. Approximately 70% of the neurons in the infected cultures became immunopositive for viral antigens and showed reduced voltage-dependent Ca(2+) currents in whole-cell patch clamp recordings, but no changes in other membrane properties or in cytosolic Ca(2+) concentration were seen. These immunopositive neurons underwent apoptosis 3-4 days after infection. Ca(2+) channel inhibitors had no significant effect on neuronal survival. The immunonegative population of neurons survived, but displayed increased frequency of miniature inhibitory postsynaptic currents of gamma-amino-butyric acid origin compared with controls. The frequency of alpha-amino-hydroxy-5-methylisoxazole-4-propionic acid hydrobromide (AMPA) receptor-mediated miniature excitatory postsynaptic currents was not altered. Viral nucleoproteins, overexpressed using the Semliki Forest virus system, were localized to the dendritic spines as shown by double immunolabeling with actinin, but did not by themselves cause neuronal death or changes in synaptic transmission as measured by AMPA-mediated excitatory postsynaptic currents. Our results show that an influenza A virus infection can cause selective neurophysiological changes in hippocampal neurons and that these can persist even after the viral antigens have been cleared.
Brask
J
J
Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden.
Owe-Larsson
B
B
Hill
R H
RH
Kristensson
K
K
D000818:Animals; D002118:Calcium / Q000502:physiology*; D002470:Cell Survival; D002478:Cells, Cultured; D004553:Electric Conductivity; D019706:Excitatory Postsynaptic Potentials / Q000502:physiology; D006624:Hippocampus / Q000166:cytology / Q000503:physiopathology*; D009980:Influenza A virus / Q000378:metabolism; D007425:Intracellular Membranes / Q000378:metabolism; D009474:Neurons / Q000502:physiology*; D009698:Nucleoproteins / Q000378:metabolism; D009976:Orthomyxoviridae Infections / Q000473:pathology / Q000503:physiopathology*; D009994:Osmolar Concentration; D051381:Rats; D017207:Rats, Sprague-Dawley; D014764:Viral Proteins / Q000378:metabolism; D005680:gamma-Aminobutyric Acid / Q000502:physiology*
D009698:Nucleoproteins; D014764:Viral Proteins; D005680:gamma-Aminobutyric Acid; D002118:Calcium