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Role of Neurexin-1╬▓ and Neuroligin-1 in Cognitive Dysfunction After Subarachnoid Hemorrhage in Rats.

Shen H, Chen Z, Wang Y, Gao A, Li H, Cui Y, Zhang L, Xu X, Wang Z, Chen G - Stroke (2015)

Bottom Line: Both in vivo and in vitro experiments showed SAH-induced decrease in the expressions of neurexin-1β and neuroligin-1 and the interaction between neurexin-1β and neuroligin-1 in neurons.In addition, the interaction between neurexin-1β and neuroligin-1 was reduced by their knockdown and increased by their overexpression.The formation of excitatory synapses was inhibited by oxyhemoglobin treatment, which was significantly ameliorated by overexpression of neurexin-1β and neuroligin-1 and aggravated by the knockdown of neurexin-1β and neuroligin-1.

View Article: PubMed Central - PubMed

Affiliation: From the Department of Neurosurgery & Brain and Nerve Research Laboratory, The First Affiliated Hospital of Soochow University, Suzhou, China (H.S., Z.C., Y.W., A.G., H.L., Y.C., L.Z., X.X., Z.W., G.C.); and Department of Neurosurgery, Anhui Provincial Hospital Affiliated to Anhui Medical University, Anhui Province Key Laboratory of Brain Function and Brain Disease, Hefei, Anhui, China (Y.W.).

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Subarachnoid hemorrhage (SAH) downregulated neurexin-1╬▓ and neuroligin-1 expression and decreased the interaction between them. A, Time course of neurexin-1╬▓ and neuroligin-1 expression in the rat brain tissues after SAH. Upper, Representative Western blot (WB) bands of neurexin-1╬▓ and neuroligin-1 are shown. Bottom, Quantitative analysis of the relative protein level is shown, and the mean value of sham group was normalized to 1.0. Double-immunofluorescence analysis was performed with antibodies for neurexin-1╬▓ or neuroligin-1 (green) and neuron marker (NeuN, red), and nuclei were fluorescently labeled with DAPI (blue). Arrows point to neurexin-1╬▓ÔÇôpositive or neuroligin-1ÔÇôpositive neurons. Representative images of the expressions of neurexin-1╬▓ and neuroligin-1 in rat cortical neurons (B) and hippocampal neurons (C) at 72 hours after SAH. Scan bar was 64 ╬╝m in B and was 100 ╬╝m in C. Bottom, Quantification of the relative fluorescence intensity is shown. The mean fluorescence intensity of the sham group was normalized to 1.0. D, Immunoprecipitation (IP) analysis of the interaction between neurexin-1╬▓ and neuroligin-1 at 72 hours after SAH. WB analysis of cell lysates incubated with antibody of neurexin-1╬▓, which then underwent IP. All values are means┬▒SEM, *P<0.05 compared with sham group and &P<0.05 compared with SAH group evaluated at 72 hours. NS indicates no significant difference, n=6.
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Figure 1: Subarachnoid hemorrhage (SAH) downregulated neurexin-1╬▓ and neuroligin-1 expression and decreased the interaction between them. A, Time course of neurexin-1╬▓ and neuroligin-1 expression in the rat brain tissues after SAH. Upper, Representative Western blot (WB) bands of neurexin-1╬▓ and neuroligin-1 are shown. Bottom, Quantitative analysis of the relative protein level is shown, and the mean value of sham group was normalized to 1.0. Double-immunofluorescence analysis was performed with antibodies for neurexin-1╬▓ or neuroligin-1 (green) and neuron marker (NeuN, red), and nuclei were fluorescently labeled with DAPI (blue). Arrows point to neurexin-1╬▓ÔÇôpositive or neuroligin-1ÔÇôpositive neurons. Representative images of the expressions of neurexin-1╬▓ and neuroligin-1 in rat cortical neurons (B) and hippocampal neurons (C) at 72 hours after SAH. Scan bar was 64 ╬╝m in B and was 100 ╬╝m in C. Bottom, Quantification of the relative fluorescence intensity is shown. The mean fluorescence intensity of the sham group was normalized to 1.0. D, Immunoprecipitation (IP) analysis of the interaction between neurexin-1╬▓ and neuroligin-1 at 72 hours after SAH. WB analysis of cell lysates incubated with antibody of neurexin-1╬▓, which then underwent IP. All values are means┬▒SEM, *P<0.05 compared with sham group and &P<0.05 compared with SAH group evaluated at 72 hours. NS indicates no significant difference, n=6.

Mentions: The results of Western blot demonstrated that when compared with the sham group, the expressions of neurexin-1╬▓ and neuroligin-1 in the brain were reduced significantly from 3 hours after SAH, reached the lowest point at 72 hours, and then rebounded gradually, and the expressions were similar to that in the sham group at 1 and 2 weeks (Figure 1A). Immunofluorescence assay further verified the SAH-induced decrease in the expressions of neurexin-1╬▓ and neuroligin-1 in cortical and hippocampal neurons (Figure 1B and 1C), which is described in detail in Figures II and III in the online-only Data Supplement. In addition, SAH also decreased the interaction between neurexin-1╬▓ and neuroligin-1 at 72 hours after SAH (Figure 1D).


Role of Neurexin-1╬▓ and Neuroligin-1 in Cognitive Dysfunction After Subarachnoid Hemorrhage in Rats.

Shen H, Chen Z, Wang Y, Gao A, Li H, Cui Y, Zhang L, Xu X, Wang Z, Chen G - Stroke (2015)

Subarachnoid hemorrhage (SAH) downregulated neurexin-1╬▓ and neuroligin-1 expression and decreased the interaction between them. A, Time course of neurexin-1╬▓ and neuroligin-1 expression in the rat brain tissues after SAH. Upper, Representative Western blot (WB) bands of neurexin-1╬▓ and neuroligin-1 are shown. Bottom, Quantitative analysis of the relative protein level is shown, and the mean value of sham group was normalized to 1.0. Double-immunofluorescence analysis was performed with antibodies for neurexin-1╬▓ or neuroligin-1 (green) and neuron marker (NeuN, red), and nuclei were fluorescently labeled with DAPI (blue). Arrows point to neurexin-1╬▓ÔÇôpositive or neuroligin-1ÔÇôpositive neurons. Representative images of the expressions of neurexin-1╬▓ and neuroligin-1 in rat cortical neurons (B) and hippocampal neurons (C) at 72 hours after SAH. Scan bar was 64 ╬╝m in B and was 100 ╬╝m in C. Bottom, Quantification of the relative fluorescence intensity is shown. The mean fluorescence intensity of the sham group was normalized to 1.0. D, Immunoprecipitation (IP) analysis of the interaction between neurexin-1╬▓ and neuroligin-1 at 72 hours after SAH. WB analysis of cell lysates incubated with antibody of neurexin-1╬▓, which then underwent IP. All values are means┬▒SEM, *P<0.05 compared with sham group and &P<0.05 compared with SAH group evaluated at 72 hours. NS indicates no significant difference, n=6.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4542569&req=5

Figure 1: Subarachnoid hemorrhage (SAH) downregulated neurexin-1╬▓ and neuroligin-1 expression and decreased the interaction between them. A, Time course of neurexin-1╬▓ and neuroligin-1 expression in the rat brain tissues after SAH. Upper, Representative Western blot (WB) bands of neurexin-1╬▓ and neuroligin-1 are shown. Bottom, Quantitative analysis of the relative protein level is shown, and the mean value of sham group was normalized to 1.0. Double-immunofluorescence analysis was performed with antibodies for neurexin-1╬▓ or neuroligin-1 (green) and neuron marker (NeuN, red), and nuclei were fluorescently labeled with DAPI (blue). Arrows point to neurexin-1╬▓ÔÇôpositive or neuroligin-1ÔÇôpositive neurons. Representative images of the expressions of neurexin-1╬▓ and neuroligin-1 in rat cortical neurons (B) and hippocampal neurons (C) at 72 hours after SAH. Scan bar was 64 ╬╝m in B and was 100 ╬╝m in C. Bottom, Quantification of the relative fluorescence intensity is shown. The mean fluorescence intensity of the sham group was normalized to 1.0. D, Immunoprecipitation (IP) analysis of the interaction between neurexin-1╬▓ and neuroligin-1 at 72 hours after SAH. WB analysis of cell lysates incubated with antibody of neurexin-1╬▓, which then underwent IP. All values are means┬▒SEM, *P<0.05 compared with sham group and &P<0.05 compared with SAH group evaluated at 72 hours. NS indicates no significant difference, n=6.
Mentions: The results of Western blot demonstrated that when compared with the sham group, the expressions of neurexin-1╬▓ and neuroligin-1 in the brain were reduced significantly from 3 hours after SAH, reached the lowest point at 72 hours, and then rebounded gradually, and the expressions were similar to that in the sham group at 1 and 2 weeks (Figure 1A). Immunofluorescence assay further verified the SAH-induced decrease in the expressions of neurexin-1╬▓ and neuroligin-1 in cortical and hippocampal neurons (Figure 1B and 1C), which is described in detail in Figures II and III in the online-only Data Supplement. In addition, SAH also decreased the interaction between neurexin-1╬▓ and neuroligin-1 at 72 hours after SAH (Figure 1D).

Bottom Line: Both in vivo and in vitro experiments showed SAH-induced decrease in the expressions of neurexin-1β and neuroligin-1 and the interaction between neurexin-1β and neuroligin-1 in neurons.In addition, the interaction between neurexin-1β and neuroligin-1 was reduced by their knockdown and increased by their overexpression.The formation of excitatory synapses was inhibited by oxyhemoglobin treatment, which was significantly ameliorated by overexpression of neurexin-1β and neuroligin-1 and aggravated by the knockdown of neurexin-1β and neuroligin-1.

View Article: PubMed Central - PubMed

Affiliation: From the Department of Neurosurgery & Brain and Nerve Research Laboratory, The First Affiliated Hospital of Soochow University, Suzhou, China (H.S., Z.C., Y.W., A.G., H.L., Y.C., L.Z., X.X., Z.W., G.C.); and Department of Neurosurgery, Anhui Provincial Hospital Affiliated to Anhui Medical University, Anhui Province Key Laboratory of Brain Function and Brain Disease, Hefei, Anhui, China (Y.W.).

Show MeSH
Related in: MedlinePlus