PURPOSE: The regulation of Ca(2+) entry and removal is a fine-tuned process which remains not well understood in mouse retinal ganglion cells (RGCs). The latter are known to be sensitive to dysfunctions of mitochondria, organelles playing a pivotal role in Ca(2+) reuptake. METHODS: We first described the Ca(2+) signals of RGCs in response to varied drugs with Fura-2 imaging, and secondly tested the role of optic atrophy 1 or OPA1, the gene responsible for Autosomal Dominant Optic Atrophy, on mitochondrial ability to capture intracellular Ca(2+) in cells transfected with the OPA1 small interfering ribonucleic acids (siRNAs). RESULTS: In control RGCs, K(+)-evoked [Ca(2+)](i) increase was blocked by the Ca(2+) channel antagonists (Ni(2+)+ Cd(2+)) and GABA(A) receptor agonist muscimol-induced [Ca(2+)](i) responses were attenuated by the GABA(A) receptor antagonists, picrotoxin and gabazine. We also prove the presence of NMDA and AMPA/Kainate (glutamate receptor agonists) responsive receptors in this model. Application of cyclopiazonic acid, an inhibitor of Ca(2+)-ATPase pumps of the intracellular Ca(2+) stores, induced an increase in [Ca(2+)](i) while ryanodine or caffeine had no effect on resting [Ca(2+)](i). Spontaneous Ca(2+) oscillations in contacting neurons highlighted the importance of cross-talks between RGCs during maturation. The mitochondrial respiration uncoupler, carbonyl cyanide 3-chlorophenylhydrazone (CCCP), induced robust raises of intracellular Ca(2+) after K(+) application, with a more pronounced effect in cells silenced for OPA1, which could lead to cell death. CONCLUSIONS: Our results indicate an important role of OPA1 in mitochondrial dependent Ca(2+) homeostasis and cell survival in RGCs, suggesting a possible patho-physiological mechanism involved in inherited optic neuropathies.
PURPOSE: The regulation of Ca(2+) entry and removal is a fine-tuned process which remains not well understood in mouse retinal ganglion cells (RGCs). The latter are known to be sensitive to dysfunctions of mitochondria, organelles playing a pivotal role in Ca(2+) reuptake. METHODS: We first described the Ca(2+) signals of RGCs in response to varied drugs with Fura-2 imaging, and secondly tested the role of optic atrophy 1 or OPA1, the gene responsible for Autosomal Dominant Optic Atrophy, on mitochondrial ability to capture intracellular Ca(2+) in cells transfected with the OPA1 small interfering ribonucleic acids (siRNAs). RESULTS: In control RGCs, K(+)-evoked [Ca(2+)](i) increase was blocked by the Ca(2+) channel antagonists (Ni(2+)+ Cd(2+)) and GABA(A) receptor agonist muscimol-induced [Ca(2+)](i) responses were attenuated by the GABA(A) receptor antagonists, picrotoxin and gabazine. We also prove the presence of NMDA and AMPA/Kainate (glutamate receptor agonists) responsive receptors in this model. Application of cyclopiazonic acid, an inhibitor of Ca(2+)-ATPase pumps of the intracellular Ca(2+) stores, induced an increase in [Ca(2+)](i) while ryanodine or caffeine had no effect on resting [Ca(2+)](i). Spontaneous Ca(2+) oscillations in contacting neurons highlighted the importance of cross-talks between RGCs during maturation. The mitochondrial respiration uncoupler, carbonyl cyanide 3-chlorophenylhydrazone (CCCP), induced robust raises of intracellular Ca(2+) after K(+) application, with a more pronounced effect in cells silenced for OPA1, which could lead to cell death. CONCLUSIONS: Our results indicate an important role of OPA1 in mitochondrial dependent Ca(2+) homeostasis and cell survival in RGCs, suggesting a possible patho-physiological mechanism involved in inherited optic neuropathies.
Authors: Allison R O'Rourke; Angus Lindsay; Michael D Tarpey; Samantha Yuen; Preston McCourt; D'anna M Nelson; Benjamin J Perrin; David D Thomas; Espen E Spangenburg; Dawn A Lowe; James M Ervasti Journal: FEBS J Date: 2018-01-08 Impact factor: 5.542
Authors: Y E Kushnareva; A A Gerencser; B Bossy; W-K Ju; A D White; J Waggoner; M H Ellisman; G Perkins; E Bossy-Wetzel Journal: Cell Death Differ Date: 2012-11-09 Impact factor: 15.828
Authors: D Nguyen; M V Alavi; K-Y Kim; T Kang; R T Scott; Y H Noh; J D Lindsey; B Wissinger; M H Ellisman; R N Weinreb; G A Perkins; W-K Ju Journal: Cell Death Dis Date: 2011-12-08 Impact factor: 8.469