title: Granulosa cells regulate intracellular pH of the murine growing oocyte via gap junctions: development of independent homeostasis during oocyte growth.
abstract: Oocytes grow within ovarian follicles in which the oocyte is coupled to the surrounding granulosa cells by gap junctions. It was previously found that small growing oocytes isolated from juvenile mice and freed of their surrounding granulosa cells (denuded) lacked the ability to regulate their intracellular pH (pH(i)), did not exhibit the pH(i)-regulatory HCO(3)(-)/Cl(-) and Na(+)/H(+) exchange activities found in fully-grown oocytes, and had low pH(i). However, both exchangers became active as oocytes grew near to full size, and, simultaneously, oocyte pH(i) increased by approximately 0.25 pH units. Here, we show that, in the more physiological setting of the intact follicle, oocyte pH(i) is instead maintained at approximately 7.2 throughout oocyte development, and the growing oocyte exhibits HCO(3)(-)/Cl(-) exchange, which it lacks when denuded. This activity in the oocyte requires functional gap junctions, as gap junction inhibitors eliminated HCO(3)(-)/Cl(-) exchange activity from follicle-enclosed growing oocytes and substantially impeded the recovery of the oocyte from an induced alkalosis, implying that oocyte pH(i) may be regulated by pH-regulatory exchangers in granulosa cells via gap junctions. This would require robust HCO(3)(-)/Cl(-) exchange activity in the granulosa cells, which was confirmed using oocytectomized (OOX) cumulus-oocyte complexes. Moreover, in cumulus-oocyte complexes with granulosa cells coupled to fully-grown oocytes, HCO(3)(-)/Cl(-) exchange activity was identical in both compartments and faster than in denuded oocytes. Taken together, these results indicate that growing oocyte pH(i) is controlled by pH-regulatory mechanisms residing in the granulosa cells until the oocyte reaches a developmental stage where it becomes capable of carrying out its own homeostasis.
authors.affiliation: Hormones, Growth and Development Program, Ottawa Health Research Institute, Ottawa, ON K1Y 4E9, Canada.
authors.forename: Greg
authors.initials: G
authors.lastname: Fitzharris
authors.forename: Jay M
authors.initials: JM
authors.lastname: Baltz
mesh_terms: D000818:Animals; D001693:Biological Transport, Active; D027963:Chloride-Bicarbonate Antiporters / Q000378:metabolism; D005260:Female; D017629:Gap Junctions / Q000502:physiology*; D006107:Granulosa Cells / Q000502:physiology*; D006706:Homeostasis; D006863:Hydrogen-Ion Concentration; D051379:Mice; D009865:Oocytes / Q000254:growth & development / Q000502:physiology*
chemical_list: D027963:Chloride-Bicarbonate Antiporters