OBJECTIVE@#To investigate functions of
proteins and signaling pathways involved in epileptogenesis during the chronic stage of
temporal lobe epilepsy in
mouse models.@*
METHODS@#
Kainic acid-induced
temporal lobe epilepsy models were conducted, when reaching stage 4 using racine scale, the
mice of experimental group were supposed to be successfully established.
Pentobarbital sodium was injected to stop epileptic seizure in case of
death. Twenty-eight days after the
kainic acid injection, when the experimental group generally turned into chronic spontaneous seizures,
mice hippocampal
tissues were extracted from the control and the experimental groups respectively for phosphoproteomic. Enriched phosphorylated
proteins were detected using
mass spectrometry, only the
proteins whose density was greater than 106 were analyzed by matching the
Gene Ontology (GO) database, Kyoto
Encyclopedia of
Genes and
Genomes (KEGG) database and STRING database to detect
proteins involved in epileptogenesis in
protein functions, signaling pathways and
protein-
protein interaction respectively. After that,
literatures were reviewed about the key
proteins.@*RESULTS@#(1) Total of 12 697
phosphorylation sites of enriched
proteins were detected by
mass spectrometry, and there were 159 sites whose
phosphorylation levels were significantly different from the control (P<0.001). (2) GO database showed that 35.7% of the 159 sites were about "catalytic activity", 39.5% were about "binding" and 20.8% were about "
cell communication", and the 159
proteins also participated in many
biological processes, such as "primary
metabolic process" "response to stimulus" "developmental process" "
localization" and "
phosphate-containing compound
metabolic process". (3) KEGG database showed that the 159
protein sites mainly involved in 10 signaling pathways glutamatergic
synapse,
Ras signaling pathway,
African trypanosomiasis,
Cocaine addiction, Circadian entrainment,
Amyotrophic lateral sclerosis (ALS),
Long-term potentiation,
Endocytosis,
Gap junction,
Nicotine addiction. (4) STRING database showed that the
protein-protein interaction network formed by the 159
proteins was focused on Grin1/Dlg3, Arhgef 2/Arhgap33/Tiam1 and Sptnb1/3/4/Add3/Ank2
protein group respectively. (5)
Phosphorylation levels of Grin1, Arhgef 2, Arhgap33, Tiam1, Sptbn1/2/4 and Ank2 in experimental group were significantly higher than in the control (P<0.001).@*CONCLUSION@#Phosphoproteomic illustrated integral distribution of phosphorylated
proteins at the chronic stage of
temporal lobe epilepsy in the
mouse model.
Literatures showed that most key
proteins were closely related to epileptogenesis, suggesting that some
proteins or signaling pathways may
play a
role in epileptogenesis, such as
dopamine and Kir3.1.