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Role of Neurexin-1β 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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Effects of neurexin-1β and neuroligin-1 knockdown and overexpression on the interaction between them and cell death in cultured hippocampal neurons under oxyhemoglobin (OxyHb) treatment. Cultured primary hippocampal neurons were transfected with small interfering RNAs (siRNAs) or plasmids as indicated. A, Western blot (WB) analysis of neurexin-1β and neuroligin-1 knockdown and overexpression efficiency in cultured primary hippocampal neurons exposed to OxyHb. Quantification of relative protein levels of neurexin-1β and neuroligin-1 is shown below. Data are presented as mean±SEM. *P<0.05 compared with OxyHb+scramble siRNA group and #P<0.05, ##P<0.01 compared with OxyHb+empty vector group, &P<0.05, n=3. B, Immunoprecipitation analysis of the effect of neurexin-1β and neuroligin-1 knockdown and overexpression on the interaction between them in cultured primary hippocampal neurons exposed to OxyHb. Quantification of relative protein levels of neurexin-1β and neuroligin-1 is shown below. Data are presented as mean±SEM. *P<0.05 compared with OxyHb+scramble siRNA group and #P<0.05, ##P<0.01 compared with OxyHb+empty vector group, &P<0.05, n=3. C, TUNEL staining. Arrows point to TUNEL-positive cells. Scale bar=64 μm. Right, Percentage of TUNEL-positive cells are shown. Data are mean±SEM. **P<0.01, n=3.
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Figure 4: Effects of neurexin-1β and neuroligin-1 knockdown and overexpression on the interaction between them and cell death in cultured hippocampal neurons under oxyhemoglobin (OxyHb) treatment. Cultured primary hippocampal neurons were transfected with small interfering RNAs (siRNAs) or plasmids as indicated. A, Western blot (WB) analysis of neurexin-1β and neuroligin-1 knockdown and overexpression efficiency in cultured primary hippocampal neurons exposed to OxyHb. Quantification of relative protein levels of neurexin-1β and neuroligin-1 is shown below. Data are presented as mean±SEM. *P<0.05 compared with OxyHb+scramble siRNA group and #P<0.05, ##P<0.01 compared with OxyHb+empty vector group, &P<0.05, n=3. B, Immunoprecipitation analysis of the effect of neurexin-1β and neuroligin-1 knockdown and overexpression on the interaction between them in cultured primary hippocampal neurons exposed to OxyHb. Quantification of relative protein levels of neurexin-1β and neuroligin-1 is shown below. Data are presented as mean±SEM. *P<0.05 compared with OxyHb+scramble siRNA group and #P<0.05, ##P<0.01 compared with OxyHb+empty vector group, &P<0.05, n=3. C, TUNEL staining. Arrows point to TUNEL-positive cells. Scale bar=64 μm. Right, Percentage of TUNEL-positive cells are shown. Data are mean±SEM. **P<0.01, n=3.

Mentions: The efficiency of siRNA-mediated knockdown, as well as expression plasmid-mediated overexpression of neurexin-1β and neuroligin-1, in cultured hippocampal neurons was also verified by Western blot (Figure 4A). Consistently, the interaction between neurexin-1β and neuroligin-1 was also significantly decreased by the knockdown of neurexin-1β and neuroligin-1 and increased by the overexpression of neurexin-1β and neuroligin-1 (Figure 4B). In addition, TUNEL staining showed that neither the knockdown nor the overexpression of neurexin-1β and neuroligin-1 could affect oxyhemoglobin-induced neuron apoptosis. (Figure 4C; Figure V in the online-only Data Supplement).


Role of Neurexin-1β 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)

Effects of neurexin-1β and neuroligin-1 knockdown and overexpression on the interaction between them and cell death in cultured hippocampal neurons under oxyhemoglobin (OxyHb) treatment. Cultured primary hippocampal neurons were transfected with small interfering RNAs (siRNAs) or plasmids as indicated. A, Western blot (WB) analysis of neurexin-1β and neuroligin-1 knockdown and overexpression efficiency in cultured primary hippocampal neurons exposed to OxyHb. Quantification of relative protein levels of neurexin-1β and neuroligin-1 is shown below. Data are presented as mean±SEM. *P<0.05 compared with OxyHb+scramble siRNA group and #P<0.05, ##P<0.01 compared with OxyHb+empty vector group, &P<0.05, n=3. B, Immunoprecipitation analysis of the effect of neurexin-1β and neuroligin-1 knockdown and overexpression on the interaction between them in cultured primary hippocampal neurons exposed to OxyHb. Quantification of relative protein levels of neurexin-1β and neuroligin-1 is shown below. Data are presented as mean±SEM. *P<0.05 compared with OxyHb+scramble siRNA group and #P<0.05, ##P<0.01 compared with OxyHb+empty vector group, &P<0.05, n=3. C, TUNEL staining. Arrows point to TUNEL-positive cells. Scale bar=64 μm. Right, Percentage of TUNEL-positive cells are shown. Data are mean±SEM. **P<0.01, n=3.
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Figure 4: Effects of neurexin-1β and neuroligin-1 knockdown and overexpression on the interaction between them and cell death in cultured hippocampal neurons under oxyhemoglobin (OxyHb) treatment. Cultured primary hippocampal neurons were transfected with small interfering RNAs (siRNAs) or plasmids as indicated. A, Western blot (WB) analysis of neurexin-1β and neuroligin-1 knockdown and overexpression efficiency in cultured primary hippocampal neurons exposed to OxyHb. Quantification of relative protein levels of neurexin-1β and neuroligin-1 is shown below. Data are presented as mean±SEM. *P<0.05 compared with OxyHb+scramble siRNA group and #P<0.05, ##P<0.01 compared with OxyHb+empty vector group, &P<0.05, n=3. B, Immunoprecipitation analysis of the effect of neurexin-1β and neuroligin-1 knockdown and overexpression on the interaction between them in cultured primary hippocampal neurons exposed to OxyHb. Quantification of relative protein levels of neurexin-1β and neuroligin-1 is shown below. Data are presented as mean±SEM. *P<0.05 compared with OxyHb+scramble siRNA group and #P<0.05, ##P<0.01 compared with OxyHb+empty vector group, &P<0.05, n=3. C, TUNEL staining. Arrows point to TUNEL-positive cells. Scale bar=64 μm. Right, Percentage of TUNEL-positive cells are shown. Data are mean±SEM. **P<0.01, n=3.
Mentions: The efficiency of siRNA-mediated knockdown, as well as expression plasmid-mediated overexpression of neurexin-1β and neuroligin-1, in cultured hippocampal neurons was also verified by Western blot (Figure 4A). Consistently, the interaction between neurexin-1β and neuroligin-1 was also significantly decreased by the knockdown of neurexin-1β and neuroligin-1 and increased by the overexpression of neurexin-1β and neuroligin-1 (Figure 4B). In addition, TUNEL staining showed that neither the knockdown nor the overexpression of neurexin-1β and neuroligin-1 could affect oxyhemoglobin-induced neuron apoptosis. (Figure 4C; Figure V in the online-only Data Supplement).

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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Related in: MedlinePlus