title: High level over-expression of different NCX isoforms in HEK293 cell lines and primary neuronal cultures is protective following oxygen glucose deprivation.
abstract: In this study we have assessed sodium-calcium exchanger (NCX) protein over-expression on cell viability in primary rat cortical neuronal and HEK293 cell cultures when subjected to oxygen-glucose deprivation (OGD). In cortical neuronal cultures, NCX2 and NCX3 over-expression was achieved using adenoviral vectors, and following OGD increased neuronal survival from ≈20% for control vector treated cultures to ≈80% for both NCX isoforms. In addition, we demonstrated that NCX2 and NCX3 over-expression in cortical neuronal cultures enables neurons to maintain intracellular calcium at significantly lower levels than control vector treated cultures when exposed to high (9mM) extracellular calcium challenge. Further assessment of NCX activity during OGD was performed using HEK293 cell lines generated to over-express NCX1, NCX2 or NCX3 isoforms. While it was shown that NCX isoform expression differed considerably in the different HEK293 cell lines, high levels of NCX over-expression was associated with increased resistance to OGD. Taken together, our findings show that high levels of NCX over-expression increases neuronal and HEK293 cell survival following OGD, improves calcium management in neuronal cultures and provides additional support for NCX as a therapeutic target to reduce ischemic brain injury.
authors.affiliation: Centre for Neuromuscular and Neurological Disorders, University of Western Australia and Australian Neuromuscular Research Institute, Western Australia, Australia. jane.cross@anri.uwa.edu.au
authors.forename: Jane L
authors.initials: JL
authors.lastname: Cross
authors.forename: Sherif
authors.initials: S
authors.lastname: Boulos
authors.forename: Kate L
authors.initials: KL
authors.lastname: Shepherd
authors.forename: Amanda J
authors.initials: AJ
authors.lastname: Craig
authors.forename: Sharon
authors.initials: S
authors.lastname: Lee
authors.forename: Anthony J
authors.initials: AJ
authors.lastname: Bakker
authors.forename: Neville W
authors.initials: NW
authors.lastname: Knuckey
authors.forename: Bruno P
authors.initials: BP
authors.lastname: Meloni
mesh_terms: D000818:Animals; D020520:Brain Infarction / Q000235:genetics / Q000378:metabolism* / Q000503:physiopathology; D020013:Calcium Signaling / Q000235:genetics; D005260:Female; D005947:Glucose / Q000172:deficiency*; D057809:HEK293 Cells; D006801:Humans; D020925:Hypoxia-Ischemia, Brain / Q000235:genetics / Q000378:metabolism* / Q000503:physiopathology; D009410:Nerve Degeneration / Q000235:genetics / Q000378:metabolism* / Q000503:physiopathology; D010100:Oxygen / Q000378:metabolism*; D061251:Primary Cell Culture; D020033:Protein Isoforms / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D019831:Sodium-Calcium Exchanger / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology
chemical_list: D020033:Protein Isoforms; C110927:Slc8a2 protein, rat; C491102:Slc8a3 protein, rat; D019831:Sodium-Calcium Exchanger; C110374:sodium-calcium exchanger 1; D005947:Glucose; D010100:Oxygen