title: ATP-induced apoptosis of human granulosa luteal cells cultured in vitro.
abstract: OBJECTIVE
  To investigate the role of extracellular adenosine triphosphatase (ATP) as an inducer of apoptotic cell death in human granulosa cells and to elucidate its underlying mechanism.
  
  
  METHODS
  Prospective study.
  
  
  METHODS
  Gynecologic clinic and human reproduction research laboratory.
  
  
  METHODS
  Twenty-five patients undergoing IVF or IVF-ET.
  
  
  METHODS
  ATP treatment of granulosa luteal cells subjected to primary culture.
  
  
  METHODS
  Apoptosis was assessed by the annexin V binding assay and the terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick-end labeling assay. The receptor type that binds ATP, thus mediating ATP-induced apoptosis, was determined by using the calcium imaging and patch-clamp techniques. Mitochondrial depolarization was assessed by staining with the membrane potential-sensitive dye 5,5',6,6'-tetrachloro-1,1',3,3'- tetraethylbenzimidazolyl carbocyanide iodide (JC-1).
  
  
  RESULTS
  ATP elevated [Ca(2+)](i) by mobilizing intracellularly stored Ca(2+). An ionic membrane current evoked by ATP was mediated by the Ca(2+)-activated K(+) channel. ATP induced a mitochondrial depolarization with a concomitant increase in cellular apoptosis. Treatment with hCG eliminated both ATP-induced mitochondrial depolarization and apoptosis.
  
  
  CONCLUSIONS
  Extracellular ATP recognized by P(2Y) type purinoceptor triggers apoptosis in human granulosa luteal cells, and the downstream apoptotic cascade may act at least in part through mitochondria. The antiapoptotic effect of hCG has possible clinical implications in the treatment of disorders involving granulosa cell degeneration (such as follicular atresia).
authors.affiliation: Department of Molecular Science and Technology, Ajou University, Suwon, South Korea.
authors.forename: Dong-Wook
authors.initials: DW
authors.lastname: Park
authors.forename: Taesup
authors.initials: T
authors.lastname: Cho
authors.forename: Mi Ran
authors.initials: MR
authors.lastname: Kim
authors.forename: Young Ah
authors.initials: YA
authors.lastname: Kim
authors.forename: Churl K
authors.initials: CK
authors.lastname: Min
authors.forename: Kyung Joo
authors.initials: KJ
authors.lastname: Hwang
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D006063:Chorionic Gonadotropin / Q000494:pharmacology; D005260:Female; D006801:Humans; D007425:Intracellular Membranes / Q000378:metabolism; D008184:Luteal Cells / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology*; D008564:Membrane Potentials / Q000187:drug effects; D008928:Mitochondria / Q000502:physiology; D009994:Osmolar Concentration; D018408:Patch-Clamp Techniques; D011446:Prospective Studies
chemical_list: D006063:Chorionic Gonadotropin; D000255:Adenosine Triphosphate; D002118:Calcium