Differential regulation of Ca2+ homeostasis in ovine large and small luteal cells.
This study was undertaken to characterize differences in Ca2+ homeostasis between small and large ovine luteal cells. Although increasing extracellular pH (pHex) resulted in increases in intracellular calcium ([Ca2+]in) in both cell types, the large cells exhibited a greater sensitivity, suggesting that distinct [Ca2+]in regulatory mechanisms with distinct pH optima are operating in the two cell types. The sarcoplasmic/endoplasmic reticulum Ca(2+)-ATPase inhibitors thapsigargin (TG) and cyclopiazonic acid (CPA) increased [Ca2+]in in both cell types. Treatment of small cells with CPA resulted in transient increases in [Ca2+]in, whereas CPA produced sustained increases in [Ca2+]in in large cells. In small cells, pretreatment with CPA prevented further increases in [Ca2+]in in response to TG and vice versa. In large cells, TG pretreatment precluded further increases in [Ca2+]in with either prostaglandin F2 alpha (PGF2 alpha) or CPA. In contrast, after CPA pretreatment, PGF2 alpha or TG induced further increases in [Ca2+]in in large cells. This suggests that a TG-sensitive, CPA-insensitive Ca2+ pool is present in large cells but not in small cells. Neither Na+ removal nor KCl addition affected [Ca2+]in in either cell type, indicating that neither the Na+/Ca2+ exchanger nor voltage-dependent Ca2+ channels are involved in Ca2+ homeostasis in these cells. Addition of the calcium antagonist, LaCl3 (La3+), produced a gradual increase in [Ca2+]in in large cells but no changes in [Ca2+]in in small cells. Additionally, treatment with increasing concentrations of 4-bromo-A23187 resulted in titratable increases in [Ca2+]in that are greater in large than small cells, suggesting that small cells possess a higher Ca(2+)-buffering capacity than large cells. Progesterone secretion by large cells was significantly inhibited at alkaline pHex. In the presence of PGF2 alpha, progesterone secretion exhibited a distinct pH optimum of 7.0. In contrast, pHex did not affect secretion of progesterone in small cells under any of the conditions tested. TG, CPA, and La3+ all reduced secretion of progesterone in large, but not small, cells. These results demonstrate that ovine large and small luteal cells differ in regulation of [Ca2+]in homeostasis, and that treatments that increase [Ca2+]in decrease progesterone secretion in large cells but have no effect in small cells.(ABSTRACT TRUNCATED AT 400 WORDS)
Department of Biochemistry, University of Arizona, Tucson 85724.
R
R
Martínez-Zaguilán
J A
JA
Wegner
R J
RJ
Gillies
P B
PB
Hoyer
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000032:analysis / Q000502:physiology; D002352:Carrier Proteins / Q000502:physiology; D003338:Corpus Luteum / Q000166:cytology* / Q000378:metabolism* / Q000502:physiology; D015237:Dinoprost / Q000494:pharmacology; D005260:Female; D006706:Homeostasis / Q000187:drug effects / Q000502:physiology*; D006863:Hydrogen-Ion Concentration; D007211:Indoles / Q000494:pharmacology; D011374:Progesterone / Q000378:metabolism; D012756:Sheep / Q000502:physiology*; D019831:Sodium-Calcium Exchanger; D013729:Terpenes / Q000494:pharmacology; D019284:Thapsigargin
D015220:Calcium Channels; D002352:Carrier Proteins; D007211:Indoles; D019831:Sodium-Calcium Exchanger; D013729:Terpenes; D011374:Progesterone; D019284:Thapsigargin; D015237:Dinoprost; D002118:Calcium; C000543:cyclopiazonic acid