Aberrant neuronal synaptic connectivity in CA1 area of the hippocampus from pilocarpine-induced epileptic rats observed by fluorogold.

In this study, we observed synaptic connectivity among neurons in CA1 region of pilocarpine-induced chronic seizures in rats. Twenty healthy male Sprague-Dawley rats were divided randomly into an epilepsy group (n = 10) and a control group (n = 10). Approximately 60 days after status epilepticus (SE) , Fluorogold (FG) was injected into the CA1 area of the hippocampus in vivo. Somatostatin (SS) expression was observed using immunofluorescence. The distribution of FG-positive and FG/SS double-labeled neurons was observed using a confocal microscope. FG-labeled pyramidal cells could be seen remotely from the FG-injected site in the CA1 area and in the subiculum in the experimental group. FG/SS double-labeled interneurons were distributed remotely from the FG-injected site in the CA1 area in the epileptic rats. These changes suggest aberrant neuronal connectivity in CA1 region, which may lead to the formation of aberrant excitatory and inhibitory circuitry, and may play an important role in the generation or compensation for temporal lobe epilepsy.

[Changes of expression of cation-chloride cotransporter genes in hippocampus of cortical dysplasia: experiment with rat].

OBJECTIVE: To investigate the roles of cation-chloride cotransporters-Na, K, 2Cl(-) cotransporter-1 (NKCC1) and K(+)-Cl(-) cotransporter-2 (KCC(2)) in the epileptogenesis of cortical dysplasia. METHODS: Six pregnant SD rats were given intraperitoneal injection of 1-3-bis-chloroethyl-nitrosourea (BCNU) on the embryonic day 17 (E17) and gave birth of 56 pups (experimental group) on the day P21. Five pregnant SD rats were given intraperitoneal injection of normal saline and gave birth of 48 pups (control group) on the day E21. Sixty days after birth the brains of 24 male pups in the experimental group and 22 male pups in the control group selected randomly were taken out to isolate the hippocampus. Cresyl-violet staining was applied to observe the histological alterations in the hippocampus. RT-PCR was used to detect the mRNA expression of NKCC1 and KCC2. RESULTS: Cresyl-violet staining revealed heterotopic cell clusters within the hippocampus. RT-PCR showed that the ratio of NKCC1 to beta-actin of the experimental group was 0.70 +/- 0.13, significantly higher than that of the control group (0.48 +/- 0.09, P < 0.01); while the ratio of KCC2 to beta-actin of the experimental group was 0.54 +/- 0.10, significantly lower than that of the control group (0.80 +/- 0.15, P < 0.01). CONCLUSION: The upregulation of NKCC1 mRNA and the concomitant downregulation of KCC2 mRNA may be deeply related to the mechanism of epileptogenicity in cortical dysplasias.

[Clinical and mutational analysis of KCNQ3 gene in a Chinese family with benign familial neonatal convulsions].

OBJECTIVE: To study the clinical and genetic characteristics of a Chinese family with benign familial convulsions (BFNC). METHODS: The clinical data of this family was analyzed. The blood samples were collected from 13 members of this family. By four microsatellite markers which are located in the gene loci of both K+ channel KCNQ2 and KCNQ3, the linkage analysis was performed in the family. With DNA direct sequencing and restriction endonuclease cutting analysis, the mutation analysis of KCNQ3 gene was made for the proband, other 12 family members and 76 unrelated normal individuals. RESULTS: There were 7 patients with BFNC observed in the three generation of family. The BFNC seizures of all patients disappeared during one month and no recurrence of seizures was found. The linkage analysis suggested the disease gene linked to KCNQ3 gene locus in the family. The mutation 988(C to T) of KCNQ3 gene was found in the proband by DNA-direct sequencing. Cosegregation of this mutation with BFNC was confirmed by restriction endonuclease cutting analysis. CONCLUSION: Chinese patients with BFNC can be caused by KCNQ3 gene mutation.