title: Calretinin and calbindin D-28k delay the onset of cell death after excitotoxic stimulation in transfected P19 cells.
abstract: In some neurological diseases, injury to neurones reflects an over-stimulation of their receptors for excitatory amino acids. This response may disturb the Ca(2+)-homeostasis and lead to a pronounced and sustained increase in the intracellular concentration of this ion. On the basis of data derived from correlative studies, calcium-binding proteins have been postulated to play a protective role in these pathologies. We tested, directly, the capacity of the three calcium-binding proteins calretinin (CR), calbindin D-28k (CB) and parvalbumin (PV) to buffer [Ca(2+)], and to protect cells against excitotoxic death. We used P19 murine embryonic carcinoma cells, which can be specifically induced (by retinoic acid) to transform into nerve-like ones. The differentiated cells express functional glutamate-receptors and are susceptible to excitotoxic shock. Undifferentiated P19-cells were stably transfected with the cDNA for CR, CB or PV, induced to differentiate, and then exposed to NMDA, a glutamate-receptor agonist. The survival rates of clones expressing CR, CB or PV were compared with those of untransfected P19-cells using the lactate-dehydrogenase assay. CR- and CB-expressing cells were protected from death during the first 2 h of exposure to NMDA. This protection was, however, transient, and did not suffice to rescue P19-cells after prolonged stimulation. Two of the three PV-transfected clones raised were vulnerable to NMDA-induced excitotoxicity; the third, which expressed the lowest level of PV, was protected to a similar degree as that found for the CR- and CB-transfected clones. Our results indicate that in the P19-cell model, CR and CB can help to delay the onset of cell death after excitotoxic stimulation.
authors.affiliation: Institute of Histology and General Embryology, University of Fribourg, CH-1705, Fribourg, Switzerland.
authors.forename: C
authors.initials: C
authors.lastname: D'Orlando
authors.forename: B
authors.initials: B
authors.lastname: Fellay
authors.forename: B
authors.initials: B
authors.lastname: Schwaller
authors.forename: V
authors.initials: V
authors.lastname: Salicio
authors.forename: A
authors.initials: A
authors.lastname: Bloc
authors.forename: V
authors.initials: V
authors.lastname: Gotzos
authors.forename: M R
authors.initials: MR
authors.lastname: Celio
mesh_terms: D000818:Animals; D064032:Calbindin 2; D064026:Calbindins; D002118:Calcium / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects / Q000502:physiology*; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D048429:Cell Size / Q000187:drug effects / Q000502:physiology; D002490:Central Nervous System / Q000187:drug effects / Q000378:metabolism* / Q000503:physiopathology; D002493:Central Nervous System Diseases / Q000188:drug therapy* / Q000378:metabolism / Q000503:physiopathology; D018690:Excitatory Amino Acid Agonists / Q000494:pharmacology; D018846:Excitatory Amino Acids / Q000378:metabolism; D005998:Glycine / Q000494:pharmacology; D007150:Immunohistochemistry; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D051379:Mice; D016202:N-Methylaspartate / Q000494:pharmacology; D014411:Neoplastic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D009474:Neurons / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D018696:Neuroprotective Agents / Q000378:metabolism*; D009498:Neurotoxins / Q000378:metabolism*; D010320:Parvalbumins / Q000235:genetics / Q000378:metabolism; D016194:Receptors, N-Methyl-D-Aspartate / Q000187:drug effects / Q000378:metabolism; D064030:S100 Calcium Binding Protein G / Q000235:genetics / Q000378:metabolism; D014212:Tretinoin / Q000494:pharmacology
chemical_list: C576511:Calb2 protein, mouse; D064032:Calbindin 2; D064026:Calbindins; D002135:Calcium-Binding Proteins; D018690:Excitatory Amino Acid Agonists; D018846:Excitatory Amino Acids; D018696:Neuroprotective Agents; D009498:Neurotoxins; D010320:Parvalbumins; D016194:Receptors, N-Methyl-D-Aspartate; D064030:S100 Calcium Binding Protein G; D014212:Tretinoin; D016202:N-Methylaspartate; D007770:L-Lactate Dehydrogenase; D002118:Calcium; D005998:Glycine