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NR4A1 Knockdown Suppresses Seizure Activity by Regulating Surface Expression of NR2B

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.


Immunofluorescence labeling of NR4A1 in the mouse models.NR4A1 (green) and MAP2 (blue) (not GFAP, red) are co-expressed in the hippocampus (a) and the cortex (b) of mice. The scale bar = 100 μm. (original magnification ×200).
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f2: Immunofluorescence labeling of NR4A1 in the mouse models.NR4A1 (green) and MAP2 (blue) (not GFAP, red) are co-expressed in the hippocampus (a) and the cortex (b) of mice. The scale bar = 100 μm. (original magnification ×200).

Mentions: NR4A1 immunofluorescence staining was principally observed in the cytomembrane and cytoplasm in the mouse hippocampus (Fig. 2a) and cortex (Fig. 2b). The neuron marker MAP2 (blue) and NR4A1 (green) were co-expressed in neurons, but not with glial fibrillary acidic protein (GFAP, a marker of astrocytes, red). Furthermore, NR4A1 (green) were not co-expressed with Aldehyde Dehydrogenase 1 Family Member L1 (Aldh1L1, a marker of pan-astrocyte, red) (Supplementary Figure 2). It showed that NR4A1 can not distribute in astrocytes, which is consistent with the results from Figs 1a and 2.


NR4A1 Knockdown Suppresses Seizure Activity by Regulating Surface Expression of NR2B
Immunofluorescence labeling of NR4A1 in the mouse models.NR4A1 (green) and MAP2 (blue) (not GFAP, red) are co-expressed in the hippocampus (a) and the cortex (b) of mice. The scale bar = 100 μm. (original magnification ×200).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f2: Immunofluorescence labeling of NR4A1 in the mouse models.NR4A1 (green) and MAP2 (blue) (not GFAP, red) are co-expressed in the hippocampus (a) and the cortex (b) of mice. The scale bar = 100 μm. (original magnification ×200).
Mentions: NR4A1 immunofluorescence staining was principally observed in the cytomembrane and cytoplasm in the mouse hippocampus (Fig. 2a) and cortex (Fig. 2b). The neuron marker MAP2 (blue) and NR4A1 (green) were co-expressed in neurons, but not with glial fibrillary acidic protein (GFAP, a marker of astrocytes, red). Furthermore, NR4A1 (green) were not co-expressed with Aldehyde Dehydrogenase 1 Family Member L1 (Aldh1L1, a marker of pan-astrocyte, red) (Supplementary Figure 2). It showed that NR4A1 can not distribute in astrocytes, which is consistent with the results from Figs 1a and 2.

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.