Figure - PMC

Skip to main content

View full-text article in PMC

. Author manuscript; available in PMC: 2016 Nov 1.

Published in final edited form as: Anesthesiology. 2015 Nov;123(5):1067–1083. doi: 10.1097/ALN.0000000000000857

Fig. 6 — The expression of cyclin dependent kinase 1 (CDK1) was significantly increased in human embryonic stem cell (hESC)-derived neurons following exposure to propofol and inhibition of CDK1 attenuated the propofol-induced cell death, dynamin-related protein 1 (Drp1) activation and mitochondrial fission. (A) CDK1 is responsible for phosphorylating Drp1 at the serine 616 position which, in turn, activates Drp1 and induces mitochondrial fission. The expression of CDK1 was significantly increased following exposure to 6 hours of 20 μg/mL propofol when compared to control treated cells as assessed by Western blot. (B) Blockade of CDK1 by pretreatment of the cells with Roscovitine attenuated the propofol-induced cell death as assessed by TUNEL staining, indicating that CDK1 is involved in propofol-induced neurotoxicity. (C) Pretreatment of the cells with Roscovitine also attenuated the propofol-induced activation of Drp1 (pDrp1 Serine 616) as assessed by Western blot, further confirming the role of CDK1 and Drp1 in propofol-induced neuronal cell death. This data is presented as a percent of the control group following normalization to COX IV. (D) Finally, pretreatment of the cells with Roscovitine attenuated the propofol-induced mitochondrial fission as assessed by immunostaining for the mitochondrial-specific marker, TOM20 (translocase of outer mitochondrial membranes, 20 kDa). This finding confirms the role of CDK1 in propofol-induced mitochondrial fission. (*P < 0.05 vs. control, **P < 0.01 vs. all other groups, #P < 0.05 vs. all other groups, n = 5/group). TUNEL = terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate in situ nick end labeling. DMSO = dimethyl sulfoxide (the vehicle control).

Fig. 6 — The expression of cyclin dependent kinase 1 (CDK1) was significantly increased in human embryonic stem cell (hESC)-derived neurons following exposure to propofol and inhibition of CDK1 attenuated the propofol-induced cell death, dynamin-related protein 1 (Drp1) activation and mitochondrial fission. (A) CDK1 is responsible for phosphorylating Drp1 at the serine 616 position which, in turn, activates Drp1 and induces mitochondrial fission. The expression of CDK1 was significantly increased following exposure to 6 hours of 20 μg/mL propofol when compared to control treated cells as assessed by Western blot. (B) Blockade of CDK1 by pretreatment of the cells with Roscovitine attenuated the propofol-induced cell death as assessed by TUNEL staining, indicating that CDK1 is involved in propofol-induced neurotoxicity. (C) Pretreatment of the cells with Roscovitine also attenuated the propofol-induced activation of Drp1 (pDrp1 Serine 616) as assessed by Western blot, further confirming the role of CDK1 and Drp1 in propofol-induced neuronal cell death. This data is presented as a percent of the control group following normalization to COX IV. (D) Finally, pretreatment of the cells with Roscovitine attenuated the propofol-induced mitochondrial fission as assessed by immunostaining for the mitochondrial-specific marker, TOM20 (translocase of outer mitochondrial membranes, 20 kDa). This finding confirms the role of CDK1 in propofol-induced mitochondrial fission. (*P < 0.05 vs. control, **P < 0.01 vs. all other groups, #P < 0.05 vs. all other groups, n = 5/group). TUNEL = terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate in situ nick end labeling. DMSO = dimethyl sulfoxide (the vehicle control).