Homeostatic plasticity and burst activity are mediated by hyperpolarization-activated cation currents and T-type calcium channels in neuronal cultures.

Homeostatic plasticity stabilizes neuronal networks by adjusting the responsiveness of neurons according to their global activity and the intensity of the synaptic inputs. We investigated the homeostatic regulation of hyperpolarization-activated cyclic nucleotide-gated (HCN) and T-type calcium (CaV3) channels in dissociated and organotypic slice cultures. After 48 h blocking of neuronal activity by tetrodotoxin (TTX), our patch-clamp experiments revealed an increase in the depolarizing voltage sag and post-inhibitory rebound mediated by HCN and CaV3 channels, respectively. All HCN subunits (HCN1 to 4) and T-type Ca-channel subunits (CaV3.1, 3.2 and 3.3) were expressed in both control and activity-deprived hippocampal cultures. Elevated expression levels of CaV3.1 mRNA and a selective increase in the expression of TRIP8b exon 4 isoforms, known to regulate HCN channel localization, were also detected in TTX-treated cultured hippocampal neurons. Immunohistochemical staining in TTX-treated organotypic slices verified a more proximal translocation of HCN1 channels in CA1 pyramidal neurons. Computational modeling also implied that HCN and T-type calcium channels have important role in the regulation of synchronized bursting evoked by previous activity-deprivation. Thus, our findings indicate that HCN and T-type Ca-channels contribute to the homeostatic regulation of excitability and integrative properties of hippocampal neurons

Organotipikus agyszelet tenyészetek optimalizálása és mikroszkópos vizsgálata

INST: L_200Az eredeti 3D szöveti szerkezetet megőrző organotipikus szelettenyészetek alkalmazásával áttekintőbb képet kaphatunk a CNS-ben zajló sejtélettani folyamatokról. Az optimális tenyésztési beállításokkal a szeleteket in vitro hetekig tartottuk fenn. IHC jelöléssel igazoltam, hogy megőrződik a szövetet jellemző citoarchitektúra, patch clamp méréssel, hogy a sejtek komplex hálózati aktivitással rendelkeznek. A szeletekbe részecske-mediált génbeviteli módszerekkel sikeresen juttattunk be DNS-t