Regulation of the hyperpolarization-activated cationic current Ih in mouse hippocampal pyramidal neurones by vitronectin, a component of extracellular matrix.
Because the hyperpolarization-activated cation-selective current I(h) makes important contributions to neural excitability, we examined its long-term regulation by vitronectin, an extracellular matrix component commonly elevated at injury sites and detected immunochemically in activated microglia. Focusing on mouse hippocampal pyramidal neurones in organotypic slice cultures established at postnatal day 0 or 1 and examined after 3-4 days in vitro, we observed differences in the amplitude and activation rate of I(h) between neurones in naive and vitronectin-exposed slices (10 microg ml(-1) added to serum-free medium), and between neurones in slices derived from wild-type and vitronectin-deficient mice. The potassium inward rectifier I(K(ir)), activated at similar voltages to I(h), was not affected by vitronectin. In CA1, differences in I(h) amplitude primarily reflected changes in maximum conductance (G(max)): a 23.3% increase to 3.18 +/- 0.64 nS from 2.58 +/- 0.96 nS (P < 0.05) in vitronectin-exposed neurones, and a 17.9% decrease to 2.24 +/- 0.26 nS from 2.73 +/- 0.64 nS (P < 0.05) in neurones from vitronectin-deficient slices. The voltage of one-half maximum activation (V(1/2)) was not significantly affected by vitronectin exposure (-78.1 +/- 2.3 mV versus -80.0 +/- 4.9 mV in naive neurones; P > 0.05) or vitronectin deficiency (-83.8 +/- 3.1 mV versus -82.0 +/- 2.9 mV in wild-type neurones; P > 0.05). In CA3 neurones, changes in I(h) reflected differences in both G(max) and V(1/2): in vitronectin-exposed neurones there was a 35.4% increase in G(max) to 1.30 +/- 0.49 nS from 0.96 +/- 0.26 nS (P < 0.01), and a +3.0 mV shift in V(1/2) to -89.8 mV from -92.8 mV (P < 0.05). The time course of I(h) activation could be fitted by the sum of two exponential functions, fast and slow. In both CA1 and CA3 neurones the fast component amplitude was preferentially sensitive to vitronectin, with its relatively larger contribution to total current in vitronectin-exposed cells contributing to the acceleration of I(h) activation. Further, HCN1 immunoreactivity appeared elevated in vitronectin-exposed slices, while HCN2 levels appeared unaltered. We suggest that vitronectin-stimulated increases in I(h) may potentially affect excitability under pathological conditions.
Division of Neurosciences, Beckman Research Institute of the City of Hope, 1450 East Duarte Road, Duarte, CA 91010, USA.
Dmitry V
DV
Vasilyev
Michael E
ME
Barish
D000818:Animals; D054815:Cyclic Nucleotide-Gated Cation Channels; D005109:Extracellular Matrix / Q000502:physiology*; D006624:Hippocampus / Q000187:drug effects / Q000502:physiology; D064428:Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels; D007473:Ion Channels / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D009419:Nerve Tissue Proteins / Q000502:physiology*; D009924:Organ Culture Techniques; D015221:Potassium Channels; D017966:Pyramidal Cells / Q000187:drug effects* / Q000502:physiology; D019096:Vitronectin / Q000172:deficiency / Q000235:genetics / Q000502:physiology*
D054815:Cyclic Nucleotide-Gated Cation Channels; C578666:Hcn1 protein, mouse; D064428:Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels; D007473:Ion Channels; D009419:Nerve Tissue Proteins; D015221:Potassium Channels; D019096:Vitronectin