help button home button Endocrine Society Endocrinology
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Purchase Article
Right arrow View Shopping Cart
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow Request Copyright Permission
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Flores, J. A.
Right arrow Articles by Veldhuis, J. D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Flores, J. A.
Right arrow Articles by Veldhuis, J. D.
Right arrowPubmed/NCBI databases
*Compound via MeSH
*Substance via MeSH
Hazardous Substances DB
*CALCIUM COMPOUNDS
*CALCIUM, ELEMENTAL
*MANGANESE COMPOUNDS
*MANGANESE, ELEMENTAL
Endocrinology Vol. 139, No. 8 3606-3612
Copyright © 1998 by The Endocrine Society

ARTICLES

Luteinizing Hormone (LH) Stimulates Both Intracellular Calcium Ion ([Ca2+]i) Mobilization and Transmembrane Cation Influx in Single Ovarian (Granulosa) Cells: Recruitment as a Cellular Mechanism of LH-[Ca2+]i Dose Response1

Jorge A. Flores, Claudia Aguirre, Om P. Sharma2 and Johannes D. Veldhuis

Division of Endocrinology and Metabolism, Department of Internal Medicine, University of Virginia Health Sciences Center, and National Science Foundation Center for Biology Timing, Charlottesville, Virginia 22908

Address all correspondence and requests for reprints to: Dr. Jorge A. Flores, Biology Department, West Virginia University, Brooks Hall, P.O. Box 6057, Morgantown, West Virginia 26506-6057.

The gonadotropic hormones, LH and FSH, activate adenylyl cyclase in their respective target cells and thereby initiate many biochemical responses. In addition to stimulating cAMP production, both LH and FSH promote agonist-specific increases in the cytoplasmic concentration of free calcium ions ([Ca2+]i) in gonadal cells. Here, we have applied single cell fluorescence video microscopy with the Ca2+-sensitive dye fura-2 to investigate the mechanism(s) by which LH induces a rise in the [Ca2+]i in individual (swine) granulosa cells collected from single Graafian follicles. Stimulation with LH induced a rapid onset, biphasic, spike- and plateau-like [Ca2+]i signal in responsive granulosa cells. The cellular mechanisms mediating this biphasic LH-stimulated increase in [Ca2+]i were examined by external Ca2+ removal and via the manganese (Mn2+) quench technique, which showed that LH triggers initial intracellular Ca2+ mobilization followed by delayed transmembrane Ca2+ influx. Single cell Ca2+ assessment of the LH dose-response mechanism(s) revealed that higher concentrations of LH progressively recruit a larger number of responding individual granulosa cells. Further analyses disclosed a marked [Ca2+]i response heterogeneity among individual granulosa cells harvested from the same Graafian follicle. In addition, the percentage of cells responding to LH [but not to an alternative putative agonist of the phospholipase C (PLC) pathway, viz. endothelin-1] with a biphasic [Ca2+]i rise increased with maturational development of the follicle. Pretreatment of granulosa cells with a specific PLC inhibitor, U-73122 (but not with its inactive congener U-73343), significantly reduced the percentage of cells responding to a LH challenge from 78% to 25% (P < 0.0001) and prolonged the time required to achieve a half-maximal value of the [Ca2+]i transient, viz. from 22 ± 1.5 sec (n = 27 cells) to 39 ± 7.2 sec (n = 12 cells; P = 0.002). In cell population studies, LH stimulated in a concentration- and time-dependent manner the accumulation of inositol phosphate in porcine granulosa cells.

In summary, the present single cell investigations in mature granulosa cells demonstrate that LH drives initial intracellular Ca2+ mobilization followed by transmembrane divalent cation influx. The PLC inhibitor U-73122 antagonizes this action of LH. By analyzing [Ca2]i responses in individual living granulosa cells, we further show that, despite within-follicle diversity, the LH dose biphasic [Ca2+]i response arises via the recruitment of a larger number of responding gonadal cells rather than by increased [Ca2+]i signal amplitude. Finally, the percentage of individual LH (but not endothelin-1)-responding granulosa cells increases with follicular maturation. Collectively, these data highlight the potential importance of the LH-stimulatable, PLC-transduced [Ca2+]i signaling mechanism in the later stages of granulosa cell differentiation.




This article has been cited by other articles:


Home page
 J. Cell Sci.Home page
D. Tong, T. Y. Li, K. E. Naus, D. Bai, and G. M. Kidder
In vivo analysis of undocked connexin43 gap junction hemichannels in ovarian granulosa cells
J. Cell Sci., November 15, 2007; 120(22): 4016 - 4024.
[Abstract] [Full Text] [PDF]

Home page
 Endocr. Rev.Home page
C. Stocco, C. Telleria, and G. Gibori
The Molecular Control of Corpus Luteum Formation, Function, and Regression
Endocr. Rev., February 1, 2007; 28(1): 117 - 149.
[Abstract] [Full Text] [PDF]

Home page
 EndocrinologyHome page
P. E. Micevych, V. Chaban, J. Ogi, P. Dewing, J. K. H. Lu, and K. Sinchak
Estradiol Stimulates Progesterone Synthesis in Hypothalamic Astrocyte Cultures
Endocrinology, February 1, 2007; 148(2): 782 - 789.
[Abstract] [Full Text] [PDF]

Home page
 Biol. Reprod.Home page
M. N. Diouf, K. Sayasith, R. Lefebvre, D. W. Silversides, J. Sirois, and J. G. Lussier
Expression of Phospholipase A2 Group IVA (PLA2G4A) Is Upregulated by Human Chorionic Gonadotropin in Bovine Granulosa Cells of Ovulatory Follicles
Biol Reprod, June 1, 2006; 74(6): 1096 - 1103.
[Abstract] [Full Text] [PDF]

Home page
 ReproductionHome page
J. Banerjee and C. M Komar
Effects of luteinizing hormone on peroxisome proliferator-activated receptor {gamma} in the rat ovary before and after the gonadotropin surge
Reproduction, January 1, 2006; 131(1): 93 - 101.
[Abstract] [Full Text] [PDF]

Home page
 Biol. Reprod.Home page
A. Sen, E. Choudhary, E. K. Inskeep, and J. A. Flores
Effects of Selective Protein Kinase C Isozymes in Prostaglandin2{alpha}-Induced Ca2+ Signaling and Luteinizing Hormone-Induced Progesterone Accumulation in the Mid-Phase Bovine Corpus Luteum
Biol Reprod, April 1, 2005; 72(4): 976 - 984.
[Abstract] [Full Text] [PDF]

Home page
 EndocrinologyHome page
R. C. Seals, R. J. Urban, N. Sekar, and J. D. Veldhuis
Up-Regulation of Basal Transcriptional Activity of the Cytochrome P450 Cholesterol Side-Chain Cleavage (CYP11A) Gene by Isoform-Specific Calcium-Calmodulin-Dependent Protein Kinase in Primary Cultures of Ovarian Granulosa Cells
Endocrinology, December 1, 2004; 145(12): 5616 - 5622.
[Abstract] [Full Text] [PDF]

Home page
 EndocrinologyHome page
G. Zhang and J. D. Veldhuis
Requirement for Proximal Putative Sp1 and AP-2 cis-Deoxyribonucleic Acid Elements in Mediating Basal and Luteinizing Hormone- and Insulin-Dependent in Vitro Transcriptional Activation of the CYP17 Gene in Porcine Theca Cells
Endocrinology, June 1, 2004; 145(6): 2760 - 2766.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Endocrinol. Metab.Home page
E. K. Asem, W. Qin, and S. G. Rane
Effect of basal lamina of ovarian follicle on T- and L-type Ca2+ currents in differentiated granulosa cells
Am J Physiol Endocrinol Metab, January 1, 2002; 282(1): E184 - E196.
[Abstract] [Full Text] [PDF]

Home page
 EndocrinologyHome page
G. Zhang, J. C. Garmey, and J. D. Veldhuis
Interactive Stimulation by Luteinizing Hormone and Insulin of the Steroidogenic Acute Regulatory (StAR) Protein and 17{alpha}-Hydroxylase/17, 20-Lyase (CYP17) Genes in Porcine Theca Cells
Endocrinology, August 1, 2000; 141(8): 2735 - 2742.
[Abstract] [Full Text] [PDF]

Home page
 EndocrinologyHome page
F. C. L. Jayes, R. N. Day, J. C. Garmey, R. J. Urban, G. Zhang, and J. D. Veldhuis
Calcium Ions Positively Modulate Follicle-Stimulating Hormone- and Exogenous Cyclic 3',5'-Adenosine Monophosphate-Driven Transcription of the P450scc Gene in Porcine Granulosa Cells
Endocrinology, July 1, 2000; 141(7): 2377 - 2384.
[Abstract] [Full Text] [PDF]

Home page
 EndocrinologyHome page
C. Aguirre, F. C. L. Jayes, and J. D. Veldhuis
Luteinizing Hormone (LH) Drives Diverse Intracellular Calcium Second Messenger Signals in Isolated Porcine Ovarian Thecal Cells: Preferential Recruitment of Intracellular Ca2+ Oscillatory Cells by Higher Concentrations of LH
Endocrinology, June 1, 2000; 141(6): 2220 - 2228.
[Abstract] [Full Text] [PDF]

Home page
 EndocrinologyHome page
R. Mamluk, N. Defer, J. Hanoune, and R. Meidan
Molecular Identification of Adenylyl Cyclase 3 in Bovine Corpus Luteum and Its Regulation by Prostaglandin F2{alpha}-Induced Signaling Pathways
Endocrinology, October 1, 1999; 140(10): 4601 - 4608.
[Abstract] [Full Text]

Home page
 Pharmacol. Rev.Home page
G. G. Nussdorfer, G. P. Rossi, L. K. Malendowicz, and G. Mazzocchi
Autocrine-Paracrine Endothelin System in the Physiology and Pathology of Steroid-Secreting Tissues
Pharmacol. Rev., September 1, 1999; 51(3): 403 - 438.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Endocrinology Endocrine Reviews J. Clin. End. & Metab.
Molecular Endocrinology Recent Prog. Horm. Res. All Endocrine Journals
Copyright © 1998 by The Endocrine Society