Parkinson’s disease cybrids exhibit mitochondrial respiration deficiency & altered expression of mitochondrial integrity proteins

Nilufar Ali; Ambili Appukuttan; Debashis Dutta; Raghavendra Singh; Joy Chakraborty; Emily Banerjee; P. Ganguly; Kochupurackal Mohanakumar

Parkinson’s disease cybrids exhibit mitochondrial respiration deficiency & altered expression of mitochondrial integrity proteins

Nilufar Ali
, Ambili Appukuttan
, Debashis Dutta
, Raghavendra Singh
, Joy Chakraborty
, Emily Banerjee
, P. Ganguly
, Kochupurackal Mohanakumar

Biological Sciences

Research output: Contribution to conference › Poster › peer-review

Abstract

Mitochondrial transgenic cells or cybrids prepared from Indian population could express mitochondrial defects from Parkinson’s disease (PD) patients. The present study examined respiratory potential of control and PD cybrids, and compared with ρ0 and the parent cell line, SH-SY5Y. We also evaluated altered protein expression in these cell lines. The cybrids were created by fusing platelets from PD patients or controls with SH-SY5Y derived ρ0 cells. Mitochondrial proteome was studied using 2D gel electrophoresis followed by MALDI-TOF-TOF analysis. Status of certain proteins was confirmed using PCR, immunoblot and immunocytochemistry. Respiration was monitored using an oxygraph. MTT assay and trypan blue staining provided cell viability status. State 3 respiration was found to be significantly reduced in PD cybrids in comparison to both SH-SY5Y and control cybrids. Analysis of mitochondrial proteome revealed a significant alteration in prohibitin in PD cybrids, which was confirmed by immunoblot assay. Detailed immunocytochemical analysis evolved differential localization of this protein in these cell lines. Dynamin related protein 1 (DRP1) was found to be over-expressed in PD cybrids as compared to other cell lines. Significantly reduced survivability was observed for PD cybrids, as compared to control cybrids and SH-SY5Y cells. PD cybrids established from Indian population clearly revealed mitochondrial dysfunction, and changes in its protein levels. It has become evident from the present study that prohibitin is localized predominantly in the nucleus of these cells and it may be unrelated to mitochondrial respiration. These results warrant investigation of prohibitin in relation to cell death and survivability, following differentiation into neurons, since this protein is known to be associated with cell proliferation.

Original language	American English
State	Published - Jun 2013
Event	24th Biennial Meeting of the International Society for Neurochemistry and the American Society for Neurochemistry Cancun, Mexico, 20–24th, April 2013 - Cancun, Mexico Duration: 20 Apr 2013 → 24 Apr 2013

Conference

Conference	24th Biennial Meeting of the International Society for Neurochemistry and the American Society for Neurochemistry Cancun, Mexico, 20–24th, April 2013
Abbreviated title	ISN-ASN
Country/Territory	Mexico
City	Cancun
Period	20/04/13 → 24/04/13

Cite this

Ali, N., Appukuttan, A., Dutta, D., Singh, R., Chakraborty, J., Banerjee, E., Ganguly, P., & Mohanakumar, K. (2013). Parkinson’s disease cybrids exhibit mitochondrial respiration deficiency & altered expression of mitochondrial integrity proteins. Poster session presented at 24th Biennial Meeting of the International Society for Neurochemistry and the American Society for Neurochemistry Cancun, Mexico, 20–24th, April 2013, Cancun, Mexico.

Ali, Nilufar ; Appukuttan, Ambili ; Dutta, Debashis et al. / Parkinson’s disease cybrids exhibit mitochondrial respiration deficiency & altered expression of mitochondrial integrity proteins. Poster session presented at 24th Biennial Meeting of the International Society for Neurochemistry and the American Society for Neurochemistry Cancun, Mexico, 20–24th, April 2013, Cancun, Mexico.

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title = "Parkinson{\textquoteright}s disease cybrids exhibit mitochondrial respiration deficiency \& altered expression of mitochondrial integrity proteins",

abstract = "Mitochondrial transgenic cells or cybrids prepared from Indian population could express mitochondrial defects from Parkinson{\textquoteright}s disease (PD) patients. The present study examined respiratory potential of control and PD cybrids, and compared with ρ0 and the parent cell line, SH-SY5Y. We also evaluated altered protein expression in these cell lines. The cybrids were created by fusing platelets from PD patients or controls with SH-SY5Y derived ρ0 cells. Mitochondrial proteome was studied using 2D gel electrophoresis followed by MALDI-TOF-TOF analysis. Status of certain proteins was confirmed using PCR, immunoblot and immunocytochemistry. Respiration was monitored using an oxygraph. MTT assay and trypan blue staining provided cell viability status. State 3 respiration was found to be significantly reduced in PD cybrids in comparison to both SH-SY5Y and control cybrids. Analysis of mitochondrial proteome revealed a significant alteration in prohibitin in PD cybrids, which was confirmed by immunoblot assay. Detailed immunocytochemical analysis evolved differential localization of this protein in these cell lines. Dynamin related protein 1 (DRP1) was found to be over-expressed in PD cybrids as compared to other cell lines. Significantly reduced survivability was observed for PD cybrids, as compared to control cybrids and SH-SY5Y cells. PD cybrids established from Indian population clearly revealed mitochondrial dysfunction, and changes in its protein levels. It has become evident from the present study that prohibitin is localized predominantly in the nucleus of these cells and it may be unrelated to mitochondrial respiration. These results warrant investigation of prohibitin in relation to cell death and survivability, following differentiation into neurons, since this protein is known to be associated with cell proliferation.",

author = "Nilufar Ali and Ambili Appukuttan and Debashis Dutta and Raghavendra Singh and Joy Chakraborty and Emily Banerjee and P. Ganguly and Kochupurackal Mohanakumar",

year = "2013",

month = jun,

language = "American English",

note = "24th Biennial Meeting of the International Society for Neurochemistry and the American Society for Neurochemistry Cancun, Mexico, 20–24th, April 2013, ISN-ASN ; Conference date: 20-04-2013 Through 24-04-2013",

}

Ali, N, Appukuttan, A, Dutta, D, Singh, R, Chakraborty, J, Banerjee, E, Ganguly, P & Mohanakumar, K 2013, 'Parkinson’s disease cybrids exhibit mitochondrial respiration deficiency & altered expression of mitochondrial integrity proteins', 24th Biennial Meeting of the International Society for Neurochemistry and the American Society for Neurochemistry Cancun, Mexico, 20–24th, April 2013, Cancun, Mexico, 20/04/13 - 24/04/13.

Parkinson’s disease cybrids exhibit mitochondrial respiration deficiency & altered expression of mitochondrial integrity proteins. / Ali, Nilufar; Appukuttan, Ambili; Dutta, Debashis et al.
2013. Poster session presented at 24th Biennial Meeting of the International Society for Neurochemistry and the American Society for Neurochemistry Cancun, Mexico, 20–24th, April 2013, Cancun, Mexico.

Research output: Contribution to conference › Poster › peer-review

TY - CONF

T1 - Parkinson’s disease cybrids exhibit mitochondrial respiration deficiency & altered expression of mitochondrial integrity proteins

AU - Ali, Nilufar

AU - Appukuttan, Ambili

AU - Dutta, Debashis

AU - Singh, Raghavendra

AU - Chakraborty, Joy

AU - Banerjee, Emily

AU - Ganguly, P.

AU - Mohanakumar, Kochupurackal

PY - 2013/6

Y1 - 2013/6

N2 - Mitochondrial transgenic cells or cybrids prepared from Indian population could express mitochondrial defects from Parkinson’s disease (PD) patients. The present study examined respiratory potential of control and PD cybrids, and compared with ρ0 and the parent cell line, SH-SY5Y. We also evaluated altered protein expression in these cell lines. The cybrids were created by fusing platelets from PD patients or controls with SH-SY5Y derived ρ0 cells. Mitochondrial proteome was studied using 2D gel electrophoresis followed by MALDI-TOF-TOF analysis. Status of certain proteins was confirmed using PCR, immunoblot and immunocytochemistry. Respiration was monitored using an oxygraph. MTT assay and trypan blue staining provided cell viability status. State 3 respiration was found to be significantly reduced in PD cybrids in comparison to both SH-SY5Y and control cybrids. Analysis of mitochondrial proteome revealed a significant alteration in prohibitin in PD cybrids, which was confirmed by immunoblot assay. Detailed immunocytochemical analysis evolved differential localization of this protein in these cell lines. Dynamin related protein 1 (DRP1) was found to be over-expressed in PD cybrids as compared to other cell lines. Significantly reduced survivability was observed for PD cybrids, as compared to control cybrids and SH-SY5Y cells. PD cybrids established from Indian population clearly revealed mitochondrial dysfunction, and changes in its protein levels. It has become evident from the present study that prohibitin is localized predominantly in the nucleus of these cells and it may be unrelated to mitochondrial respiration. These results warrant investigation of prohibitin in relation to cell death and survivability, following differentiation into neurons, since this protein is known to be associated with cell proliferation.

AB - Mitochondrial transgenic cells or cybrids prepared from Indian population could express mitochondrial defects from Parkinson’s disease (PD) patients. The present study examined respiratory potential of control and PD cybrids, and compared with ρ0 and the parent cell line, SH-SY5Y. We also evaluated altered protein expression in these cell lines. The cybrids were created by fusing platelets from PD patients or controls with SH-SY5Y derived ρ0 cells. Mitochondrial proteome was studied using 2D gel electrophoresis followed by MALDI-TOF-TOF analysis. Status of certain proteins was confirmed using PCR, immunoblot and immunocytochemistry. Respiration was monitored using an oxygraph. MTT assay and trypan blue staining provided cell viability status. State 3 respiration was found to be significantly reduced in PD cybrids in comparison to both SH-SY5Y and control cybrids. Analysis of mitochondrial proteome revealed a significant alteration in prohibitin in PD cybrids, which was confirmed by immunoblot assay. Detailed immunocytochemical analysis evolved differential localization of this protein in these cell lines. Dynamin related protein 1 (DRP1) was found to be over-expressed in PD cybrids as compared to other cell lines. Significantly reduced survivability was observed for PD cybrids, as compared to control cybrids and SH-SY5Y cells. PD cybrids established from Indian population clearly revealed mitochondrial dysfunction, and changes in its protein levels. It has become evident from the present study that prohibitin is localized predominantly in the nucleus of these cells and it may be unrelated to mitochondrial respiration. These results warrant investigation of prohibitin in relation to cell death and survivability, following differentiation into neurons, since this protein is known to be associated with cell proliferation.

M3 - Poster

T2 - 24th Biennial Meeting of the International Society for Neurochemistry and the American Society for Neurochemistry Cancun, Mexico, 20–24th, April 2013

Y2 - 20 April 2013 through 24 April 2013

ER -

Ali N, Appukuttan A, Dutta D, Singh R, Chakraborty J, Banerjee E et al.. Parkinson’s disease cybrids exhibit mitochondrial respiration deficiency & altered expression of mitochondrial integrity proteins. 2013. Poster session presented at 24th Biennial Meeting of the International Society for Neurochemistry and the American Society for Neurochemistry Cancun, Mexico, 20–24th, April 2013, Cancun, Mexico.

Parkinson’s disease cybrids exhibit mitochondrial respiration deficiency & altered expression of mitochondrial integrity proteins

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