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NR4A1 Knockdown Suppresses Seizure Activity by Regulating Surface Expression of NR2B

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ABSTRACT

Nuclear receptor subfamily 4 group A member 1 (NR4A1), a downstream target of CREB that is a key regulator of epileptogenesis, has been implicated in a variety of biological processes and was previously identified as a seizure-associated molecule. However, the relationship between NR4A1 and epileptogenesis remains unclear. Here, we showed that NR4A1 protein was predominantly expressed in neurons and up-regulated in patients with epilepsy as well as pilocarpine-induced mouse epileptic models. NR4A1 knockdown by lentivirus transfection (lenti-shNR4A1) alleviated seizure severity and prolonged onset latency in mouse models. Moreover, reciprocal coimmunoprecipitation of NR4A1 and NR2B demonstrated their interaction. Furthermore, the expression of p-NR2B (Tyr1472) in epileptic mice and the expression of NR2B in the postsynaptic density (PSD) were significantly reduced in the lenti-shNR4A1 group, indicating that NR4A1 knockdown partly decreased surface NR2B by promoting NR2B internalization. These results are the first to indicate that the expression of NR4A1 in epileptic brain tissues may provide new insights into the molecular mechanisms underlying epilepsy.

No MeSH data available.


Related in: MedlinePlus

NR4A1 expression in a pilocarpine-induced epilepsy mouse models.The representative Western blot images show NR4A1 expression in the hippocampus (a) and the cortex (c) of mice with SRS (n = 12) or no SRS (n = 6). (b, d) Three such experiments were quantified from (a, c) by measuring the intensity of the NR4A1 proteins relative to the GAPDH control (*P < 0.05, compared to no SRS control group). The bars indicated the mean ± SD. (e) Representative hippocampal EEG recordings in the pilocarpine model during a spontaneous seizure in the chronic phase of epilepsy.
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f3: NR4A1 expression in a pilocarpine-induced epilepsy mouse models.The representative Western blot images show NR4A1 expression in the hippocampus (a) and the cortex (c) of mice with SRS (n = 12) or no SRS (n = 6). (b, d) Three such experiments were quantified from (a, c) by measuring the intensity of the NR4A1 proteins relative to the GAPDH control (*P < 0.05, compared to no SRS control group). The bars indicated the mean ± SD. (e) Representative hippocampal EEG recordings in the pilocarpine model during a spontaneous seizure in the chronic phase of epilepsy.

Mentions: To extend the results from the analysis of human tissues and exclude the possibility that altered NR4A1 expression may be caused by AEDs in patients with epilepsy, NR4A1 expression levels in the hippocampus and cortex of epileptic and control mice were detected by Western blotting. We observed significantly increased NR4A1 expression in both the hippocampus (3.72-fold, P < 0.05, Fig. 3a and 3b) and cortex (4.23-fold, P < 0.05, Fig. 3c and 3d) in epileptic mice compared with the controls (n = 6). Our results indicate that NR4A1, a downstream target of the CREB, may be involved in epileptic seizures.


NR4A1 Knockdown Suppresses Seizure Activity by Regulating Surface Expression of NR2B
NR4A1 expression in a pilocarpine-induced epilepsy mouse models.The representative Western blot images show NR4A1 expression in the hippocampus (a) and the cortex (c) of mice with SRS (n = 12) or no SRS (n = 6). (b, d) Three such experiments were quantified from (a, c) by measuring the intensity of the NR4A1 proteins relative to the GAPDH control (*P < 0.05, compared to no SRS control group). The bars indicated the mean ± SD. (e) Representative hippocampal EEG recordings in the pilocarpine model during a spontaneous seizure in the chronic phase of epilepsy.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5120300&req=5

f3: NR4A1 expression in a pilocarpine-induced epilepsy mouse models.The representative Western blot images show NR4A1 expression in the hippocampus (a) and the cortex (c) of mice with SRS (n = 12) or no SRS (n = 6). (b, d) Three such experiments were quantified from (a, c) by measuring the intensity of the NR4A1 proteins relative to the GAPDH control (*P < 0.05, compared to no SRS control group). The bars indicated the mean ± SD. (e) Representative hippocampal EEG recordings in the pilocarpine model during a spontaneous seizure in the chronic phase of epilepsy.
Mentions: To extend the results from the analysis of human tissues and exclude the possibility that altered NR4A1 expression may be caused by AEDs in patients with epilepsy, NR4A1 expression levels in the hippocampus and cortex of epileptic and control mice were detected by Western blotting. We observed significantly increased NR4A1 expression in both the hippocampus (3.72-fold, P < 0.05, Fig. 3a and 3b) and cortex (4.23-fold, P < 0.05, Fig. 3c and 3d) in epileptic mice compared with the controls (n = 6). Our results indicate that NR4A1, a downstream target of the CREB, may be involved in epileptic seizures.

View Article: PubMed Central - PubMed

ABSTRACT

Nuclear receptor subfamily 4 group A member 1 (NR4A1), a downstream target of CREB that is a key regulator of epileptogenesis, has been implicated in a variety of biological processes and was previously identified as a seizure-associated molecule. However, the relationship between NR4A1 and epileptogenesis remains unclear. Here, we showed that NR4A1 protein was predominantly expressed in neurons and up-regulated in patients with epilepsy as well as pilocarpine-induced mouse epileptic models. NR4A1 knockdown by lentivirus transfection (lenti-shNR4A1) alleviated seizure severity and prolonged onset latency in mouse models. Moreover, reciprocal coimmunoprecipitation of NR4A1 and NR2B demonstrated their interaction. Furthermore, the expression of p-NR2B (Tyr1472) in epileptic mice and the expression of NR2B in the postsynaptic density (PSD) were significantly reduced in the lenti-shNR4A1 group, indicating that NR4A1 knockdown partly decreased surface NR2B by promoting NR2B internalization. These results are the first to indicate that the expression of NR4A1 in epileptic brain tissues may provide new insights into the molecular mechanisms underlying epilepsy.

No MeSH data available.


Related in: MedlinePlus