Copper modulates the large dense core vesicle secretory pathway in PC12 cells

Clare Duncan, Laura Bica, Peter J. Crouch, Aphrodite Caragounis, Grace E. Lidgerwood, Sarah J. Parker, Jodi Meyerowitz, Irene Volitakis, Jeffrey R. Liddell, Ravinarayan Raghupathi, Brett M. Paterson, Michael D. Duffield, Roberto Cappai, Paul S. Donnelly, Alexandra Grubman, James Camakaris, Damien J. Keating, Anthony R. White

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Copper (Cu) is an essential biometal involved in a number of cell functions. Abnormal Cu homeostasis has been identified as a major factor in a number of neurodegenerative disorders. However, little is known about how cells of brain origin maintain Cu homeostasis and in particular, how they respond to an elevated Cu environment. Understanding these processes is essential to obtaining a greater insight into the pathological changes in neurodegeneration and ageing. Although previous studies have shown that Cu in neurons can be associated with synaptic function, there is little understanding of how Cu modulates the regulated secretory vesicle pathways in these cells. In this study, we examined the effect of elevated intracellular Cu on proteins associated with the regulated secretory vesicle pathway in NGF-differentiated PC12 cells that exhibit neuronal-like properties. Increasing intracellular Cu with a cell-permeable Cu-complex (CuII(gtsm)) resulted in increased expression of synaptophysin and robust translocation of this and additional vesicular proteins from synaptic-like microvesicle (SLMV) fractions to chromogranin-containing putative large dense core vesicle (LDCV) fractions in density gradient preparations. The LDCV fractions also contained substantially elevated Cu levels upon treatment of cells with CuII(gtsm). Expression of the H+ pump, V-ATPase, which is essential for vesicle maturation, was increased in Cu-treated cells while inhibition of V-ATPase prevented translocation of synaptophysin to LDCV fractions. Cu treatment was found to inhibit release of LDCVs in chromaffin cells due to reduced Ca 2+-mediated vesicle exocytosis. Our findings demonstrate that elevated Cu can modulate LDCV metabolism potentially resulting in sequestration of Cu in this vesicle pool.
Original languageEnglish
Pages (from-to)700-714
Number of pages15
JournalMetallomics
Volume5
Issue number6
DOIs
Publication statusPublished - Jun 2013
Externally publishedYes

Cite this

Duncan, C., Bica, L., Crouch, P. J., Caragounis, A., Lidgerwood, G. E., Parker, S. J., ... White, A. R. (2013). Copper modulates the large dense core vesicle secretory pathway in PC12 cells. Metallomics, 5(6), 700-714. https://doi.org/10.1039/c3mt20231c
Duncan, Clare ; Bica, Laura ; Crouch, Peter J. ; Caragounis, Aphrodite ; Lidgerwood, Grace E. ; Parker, Sarah J. ; Meyerowitz, Jodi ; Volitakis, Irene ; Liddell, Jeffrey R. ; Raghupathi, Ravinarayan ; Paterson, Brett M. ; Duffield, Michael D. ; Cappai, Roberto ; Donnelly, Paul S. ; Grubman, Alexandra ; Camakaris, James ; Keating, Damien J. ; White, Anthony R. / Copper modulates the large dense core vesicle secretory pathway in PC12 cells. In: Metallomics. 2013 ; Vol. 5, No. 6. pp. 700-714.
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abstract = "Copper (Cu) is an essential biometal involved in a number of cell functions. Abnormal Cu homeostasis has been identified as a major factor in a number of neurodegenerative disorders. However, little is known about how cells of brain origin maintain Cu homeostasis and in particular, how they respond to an elevated Cu environment. Understanding these processes is essential to obtaining a greater insight into the pathological changes in neurodegeneration and ageing. Although previous studies have shown that Cu in neurons can be associated with synaptic function, there is little understanding of how Cu modulates the regulated secretory vesicle pathways in these cells. In this study, we examined the effect of elevated intracellular Cu on proteins associated with the regulated secretory vesicle pathway in NGF-differentiated PC12 cells that exhibit neuronal-like properties. Increasing intracellular Cu with a cell-permeable Cu-complex (CuII(gtsm)) resulted in increased expression of synaptophysin and robust translocation of this and additional vesicular proteins from synaptic-like microvesicle (SLMV) fractions to chromogranin-containing putative large dense core vesicle (LDCV) fractions in density gradient preparations. The LDCV fractions also contained substantially elevated Cu levels upon treatment of cells with CuII(gtsm). Expression of the H+ pump, V-ATPase, which is essential for vesicle maturation, was increased in Cu-treated cells while inhibition of V-ATPase prevented translocation of synaptophysin to LDCV fractions. Cu treatment was found to inhibit release of LDCVs in chromaffin cells due to reduced Ca 2+-mediated vesicle exocytosis. Our findings demonstrate that elevated Cu can modulate LDCV metabolism potentially resulting in sequestration of Cu in this vesicle pool.",
author = "Clare Duncan and Laura Bica and Crouch, {Peter J.} and Aphrodite Caragounis and Lidgerwood, {Grace E.} and Parker, {Sarah J.} and Jodi Meyerowitz and Irene Volitakis and Liddell, {Jeffrey R.} and Ravinarayan Raghupathi and Paterson, {Brett M.} and Duffield, {Michael D.} and Roberto Cappai and Donnelly, {Paul S.} and Alexandra Grubman and James Camakaris and Keating, {Damien J.} and White, {Anthony R.}",
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Duncan, C, Bica, L, Crouch, PJ, Caragounis, A, Lidgerwood, GE, Parker, SJ, Meyerowitz, J, Volitakis, I, Liddell, JR, Raghupathi, R, Paterson, BM, Duffield, MD, Cappai, R, Donnelly, PS, Grubman, A, Camakaris, J, Keating, DJ & White, AR 2013, 'Copper modulates the large dense core vesicle secretory pathway in PC12 cells' Metallomics, vol. 5, no. 6, pp. 700-714. https://doi.org/10.1039/c3mt20231c

Copper modulates the large dense core vesicle secretory pathway in PC12 cells. / Duncan, Clare; Bica, Laura; Crouch, Peter J.; Caragounis, Aphrodite; Lidgerwood, Grace E.; Parker, Sarah J.; Meyerowitz, Jodi; Volitakis, Irene; Liddell, Jeffrey R.; Raghupathi, Ravinarayan; Paterson, Brett M.; Duffield, Michael D.; Cappai, Roberto; Donnelly, Paul S.; Grubman, Alexandra; Camakaris, James; Keating, Damien J.; White, Anthony R.

In: Metallomics, Vol. 5, No. 6, 06.2013, p. 700-714.

Research output: Contribution to journalArticleResearchpeer-review

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T1 - Copper modulates the large dense core vesicle secretory pathway in PC12 cells

AU - Duncan, Clare

AU - Bica, Laura

AU - Crouch, Peter J.

AU - Caragounis, Aphrodite

AU - Lidgerwood, Grace E.

AU - Parker, Sarah J.

AU - Meyerowitz, Jodi

AU - Volitakis, Irene

AU - Liddell, Jeffrey R.

AU - Raghupathi, Ravinarayan

AU - Paterson, Brett M.

AU - Duffield, Michael D.

AU - Cappai, Roberto

AU - Donnelly, Paul S.

AU - Grubman, Alexandra

AU - Camakaris, James

AU - Keating, Damien J.

AU - White, Anthony R.

PY - 2013/6

Y1 - 2013/6

N2 - Copper (Cu) is an essential biometal involved in a number of cell functions. Abnormal Cu homeostasis has been identified as a major factor in a number of neurodegenerative disorders. However, little is known about how cells of brain origin maintain Cu homeostasis and in particular, how they respond to an elevated Cu environment. Understanding these processes is essential to obtaining a greater insight into the pathological changes in neurodegeneration and ageing. Although previous studies have shown that Cu in neurons can be associated with synaptic function, there is little understanding of how Cu modulates the regulated secretory vesicle pathways in these cells. In this study, we examined the effect of elevated intracellular Cu on proteins associated with the regulated secretory vesicle pathway in NGF-differentiated PC12 cells that exhibit neuronal-like properties. Increasing intracellular Cu with a cell-permeable Cu-complex (CuII(gtsm)) resulted in increased expression of synaptophysin and robust translocation of this and additional vesicular proteins from synaptic-like microvesicle (SLMV) fractions to chromogranin-containing putative large dense core vesicle (LDCV) fractions in density gradient preparations. The LDCV fractions also contained substantially elevated Cu levels upon treatment of cells with CuII(gtsm). Expression of the H+ pump, V-ATPase, which is essential for vesicle maturation, was increased in Cu-treated cells while inhibition of V-ATPase prevented translocation of synaptophysin to LDCV fractions. Cu treatment was found to inhibit release of LDCVs in chromaffin cells due to reduced Ca 2+-mediated vesicle exocytosis. Our findings demonstrate that elevated Cu can modulate LDCV metabolism potentially resulting in sequestration of Cu in this vesicle pool.

AB - Copper (Cu) is an essential biometal involved in a number of cell functions. Abnormal Cu homeostasis has been identified as a major factor in a number of neurodegenerative disorders. However, little is known about how cells of brain origin maintain Cu homeostasis and in particular, how they respond to an elevated Cu environment. Understanding these processes is essential to obtaining a greater insight into the pathological changes in neurodegeneration and ageing. Although previous studies have shown that Cu in neurons can be associated with synaptic function, there is little understanding of how Cu modulates the regulated secretory vesicle pathways in these cells. In this study, we examined the effect of elevated intracellular Cu on proteins associated with the regulated secretory vesicle pathway in NGF-differentiated PC12 cells that exhibit neuronal-like properties. Increasing intracellular Cu with a cell-permeable Cu-complex (CuII(gtsm)) resulted in increased expression of synaptophysin and robust translocation of this and additional vesicular proteins from synaptic-like microvesicle (SLMV) fractions to chromogranin-containing putative large dense core vesicle (LDCV) fractions in density gradient preparations. The LDCV fractions also contained substantially elevated Cu levels upon treatment of cells with CuII(gtsm). Expression of the H+ pump, V-ATPase, which is essential for vesicle maturation, was increased in Cu-treated cells while inhibition of V-ATPase prevented translocation of synaptophysin to LDCV fractions. Cu treatment was found to inhibit release of LDCVs in chromaffin cells due to reduced Ca 2+-mediated vesicle exocytosis. Our findings demonstrate that elevated Cu can modulate LDCV metabolism potentially resulting in sequestration of Cu in this vesicle pool.

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DO - 10.1039/c3mt20231c

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Duncan C, Bica L, Crouch PJ, Caragounis A, Lidgerwood GE, Parker SJ et al. Copper modulates the large dense core vesicle secretory pathway in PC12 cells. Metallomics. 2013 Jun;5(6):700-714. https://doi.org/10.1039/c3mt20231c