Institute of Neuroscience, Lobachevsky State University of Nizhny Novgorod, 23 Gagarin Ave, 603022 Nizhny Novgorod, Russia.
Institute of Cell Biophysics of the Russian Academy of Sciences, Federal Research Center "Pushchino Scientific Center for Biological Research of the Russian Academy of Sciences", Institutskaya st. building 3, 142290 Pushchino, Russia.
Int J Mol Sci. 2024 Sep 30;25(19):10537. doi: 10.3390/ijms251910537.
Since genetic mutations during brain development play a significant role in the genesis of epilepsy, and such genetically determined epilepsies are the most difficult to treat, there is a need to study the mechanisms of epilepsy development with deletions of various transcription factors. We utilized heterozygous mice (Sip1) with a neuronal deletion of the transcription factor Sip1 (Smad interacting protein 1) in the cerebral cortex. These mice are characterized by cognitive impairment and are prone to epilepsy. It is known that the brain-derived neurotrophic factor (BDNF) has a neuroprotective effect in various neurodegenerative diseases. Therefore, we created and applied an adeno-associated construct carrying the BDNF sequence selectively in neurons. Using in vitro and in vivo research models, we were able to identify a key gen, the disruption of whose expression accompanies the deletion of Sip1 and contributes to hyperexcitation of neurons in the cerebral cortex. Overexpression of BDNF in cortical neurons eliminated epileptiform activity in neurons obtained from heterozygous Sip1 mice in a magnesium-free model of epileptiform activity (in vitro). Using PCR analysis, it was possible to identify correlations in the expression profile of genes encoding key proteins responsible for neurotransmission and neuronal survival. The effects of BDNF overexpression on the expression profiles of these genes were also revealed. Using BDNF overexpression in cortical neurons of heterozygous Sip1 mice, it was possible to achieve 100% survival in the pilocarpine model of epilepsy. At the level of gene expression in the cerebral cortex, patterns were established that may be involved in the protection of brain cells from epileptic seizures and the restoration of cognitive functions in mice with Sip1 deletion.
由于基因突变在大脑发育过程中起着重要作用，导致了癫痫的发生，而这种由遗传决定的癫痫是最难治疗的，因此需要研究各种转录因子缺失引起的癫痫发展机制。我们利用大脑皮层神经元缺失转录因子 Sip1（Smad 相互作用蛋白 1）的杂合子小鼠（Sip1）进行研究。这些小鼠表现出认知障碍，易患癫痫。已知脑源性神经营养因子（BDNF）在各种神经退行性疾病中具有神经保护作用。因此，我们创建并应用了一种携带 BDNF 序列的腺相关载体，选择性地在神经元中表达。利用体外和体内研究模型，我们能够确定一个关键基因，其表达的破坏伴随着 Sip1 的缺失，并导致大脑皮层神经元的过度兴奋。在无镁癫痫样活动模型（体外）中，BDNF 在皮质神经元中的过表达消除了来自杂合 Sip1 小鼠的神经元中的癫痫样活动。通过 PCR 分析，可以确定编码负责神经递质传递和神经元存活的关键蛋白的基因表达谱之间的相关性。还揭示了 BDNF 过表达对这些基因表达谱的影响。通过在杂合 Sip1 小鼠的皮质神经元中过表达 BDNF，可以在匹鲁卡品癫痫模型中实现 100%的神经元存活。在大脑皮层的基因表达水平上，建立了可能参与保护脑细胞免受癫痫发作和恢复 Sip1 缺失小鼠认知功能的模式。
