Characterization of adenylyl cyclases in cultured human granulosa cells

Asboth, G

doi:10.1530/reprod/121.2.217

Cited by 3 publications

(4 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…An elevation of intracellular Ca 2ϩ could lead to such a cAMP increase through the activation of calmodulin-sensitive adenylate cyclase isoforms I and VIII (Taussig and Zimmermann, 1998). These adenylate cyclases have never been reported in mouse cumulus cells, but they are expressed in human granulosa cells (Asboth et al, 2001) and could link an ␣-methyl-5-HT-induced Ca 2ϩ increase through a 5-HT 2 receptor to increased cAMP. In addition, part of the 5-HT-evoked cAMP elevation in cumulus cells could be mediated by this Ca 2ϩ increment because BAPTA-pretreated cells showed a smaller increase in cAMP after 5-HT addition.…”

Section: Discussionmentioning

confidence: 99%

Intracellular cAMP and Calcium Signaling by Serotonin in Mouse Cumulus-Oocyte Complexes

Amireault

Dubé²

2005

Mol Pharmacol

View full text Add to dashboard Cite

cAMP and intracellular Ca 2ϩ are important second messengers involved in mammalian follicular growth and oocyte meiotic maturation. We investigated the capacity of the neurohormone serotonin (5-hydroxytryptamine, 5-HT) to regulate intracellular cAMP and Ca 2ϩ in mouse oocytes and surrounding cumulus cells. On the basis of a reverse transcription-polymerase chain reaction study, 5-HT 7 receptor mRNA is expressed in cumulus cells, oocytes, and embryos up to the four-cell stage, and 5-HT 2A and 5-HT 2B receptor mRNAs are expressed in cumulus cells only, whereas 5-HT 2C , 5-HT 4 , and 5-HT 6 receptors are expressed in neither oocytes nor cumulus cells. The addition of 5-HT (10 nM to 10 M) to isolated metaphase II oocytes had no effect on their internal cAMP or Ca 2ϩ levels, whereas it caused dose-dependent cAMP and Ca 2ϩ increases in cumulus cells. This cAMP increase in cumulus cells could be mimicked by 5-HT agonists with the following order of potency: 5-HT Ͼ 8-hydroxy-2-(di-n-propylamino) tetralin ϭ ␣-methyl-5-HT ϭ 5-carboxamidotryptamine maleate Ͼ 2-[1-(4-piperonyl)piperazinyl]benzo-triazole, thereby supporting a preferential involvement of 5-HT 7 receptors. As measured with cumulus cells preloaded with fura-2/acetoxymethyl ester (AM), the addition of 5-HT also caused dose-dependent Ca 2ϩ increases, which were probably linked to detected 5-HT 2A and 5-HT 2B receptors.

show abstract

Section: Discussionmentioning

confidence: 99%

Intracellular cAMP and Calcium Signaling by Serotonin in Mouse Cumulus-Oocyte Complexes

Amireault

Dubé²

2005

Mol Pharmacol

View full text Add to dashboard Cite

show abstract

“…However, forskolin was able to elicit increases in cAMP generation after 10 min on each day of culture. This response could be explained by the fact that forskolin acts in a receptor-independent manner by stimulating adenylate cyclase in order to raise intracellular cAMP (Jammes et al 1988, Asboth et al 2001). …”

Section: Discussionmentioning

confidence: 99%

Changes in cAMP-dependent protein kinase (PKA) and progesterone secretion in luteinizing human granulosa cells

Chin

Harris

Abayasekara³

2004

Journal of Endocrinology

View full text Add to dashboard Cite

Luteinization of follicular granulosa cells leads to an increase in progesterone secretion that is regulated by luteinizing hormone (LH). LH acts mainly by elevating intracellular cyclic 3 ,5 -adenosine monophosphate (cAMP) and activating cAMP-dependent protein kinase (PKA). In this study, we have examined the role of PKA in relation to progesterone output by luteinizing human granulosa cells. Human granulosa cells were obtained by percoll gradient centrifugation of follicular aspirates of patients undergoing oocyte retrieval for assisted conception. Cells were cultured in serum-supplemented medium for up to 3 days in the presence and/or absence of human (h)LH and other cAMP-elevating agents. Spent medium was assayed for cAMP and progesterone content by specific RIA. Cell lysates were collected and assessed for PKA regulatory (R)II /catalytic (C) expression by Western blotting. Although basal progesterone secretion increased progressively throughout culture, cAMP levels remained unchanged. Under basal conditions, PKA RII /C expression appeared to increase throughout the 3-day culture period. In the presence of hLH and other cAMP-elevating agents, progesterone secretion increased in a dose-dependent manner coincident with an increase in cAMP. However, despite the increase in both progesterone secretion and cAMP accumulation, there was a dose-dependent decrease in both PKA RII and C expression. Thus, data presented in this study show that increases in progesterone secretion in luteinizing human granulosa cells can be dissociated from increases in PKA expression. This notion implies that progesterone secretion may be regulated by PKA-dependent as well as PKAindependent mechanisms.

show abstract

“…Follicular fluid aspirated is an excellent source of granulosa-lutein cells that retain functional Gs-adenylyl cyclase and Gq-phospholipase C coupling (Asboth et al, 2001, Carrasco et al, 1997. To obtain a sufficient number of GCs Serum FSH (IU/L) levels were similar in the two groups: 6.5±1.6 versus 6±1.3 for normal and PCOS, respectively.…”

Section: Gcs Isolation and Culturementioning

confidence: 99%

“…The GCs resuspended in 10ml Dulbecco's modified Eagle's medium (DMEM) and over layered onto 12.5ml sterile 35% Percoll in PBS-BSA and centrifuged for 30min at 400g to obtain GCs as the layer at the interface of medium and percoll (Asboth et al, 2001). GCs were then washed three times with PBS-BSA and re-suspended in DMEM containing 2mM glutamine, 100U/ml penicillin, 0.1mg/ml streptomycin and 10% fetal calf serum (complete medium) and determined cell number and viability by trypan blue (0.4%) staining.…”

Section: Gcs Isolation and Culturementioning

confidence: 99%

Luteinizing hormone/chorionic gonadotrophin receptor overexpressed in granulosa cells from polycystic ovary syndrome ovaries is functionally active

Kanamarlapudi

Gordon

Bernal

2016

Reproductive BioMedicine Online

View full text Add to dashboard Cite

General rightsThis document is made available in accordance with publisher policies. Please cite only the published version using the reference above. AbstractPolycystic ovarian syndrome (PCOS) is associated with anovulatory infertility. Luteinizing hormone/chorionic gonadotropin receptor (LHCGR), which is critical for ovulation, has been suggested to be expressed prematurely in the ovarian follicles of women with PCOS. The objective of this study was to analyse the expression and activity of LHCGR in ovarian granulosa cells from PCOS patients and the involvement of ARF6 small GTPase in LHCGR internalisation. Granulosa cells (GCs) isolated from follicular fluid collected during oocyte retrieval from normal women (n=19) and women with PCOS (n=17) were used to study differences in LHCGR protein expression and activity between normal and PCOS patients.LHCGR expression is up-regulated in GCs from PCOS women. LHCGR in PCOS GCs is functionally active as evidenced by increased cAMP production upon human gonadotrophin (HCG)-stimulation. Moreover, ARF6 is highly expressed in GCs from PCOS patients and HCG-stimulation increases the levels of active ARF6. The inhibition of ARF6 activation attenuates HCG-induced LHCGR internalisation in both normal and PCOS GCs, indicating that there are no alterations in LHCGR internalisation in GCs from PCOS. In conclusion, the expression and activation of LHCGR and ARF6 are up-regulated in GCs from PCOS women but the mechanism of agonist-induced LHCGR internalisation is unaltered.

show abstract

Characterization of adenylyl cyclases in cultured human granulosa cells

Cited by 3 publications

References 37 publications

Intracellular cAMP and Calcium Signaling by Serotonin in Mouse Cumulus-Oocyte Complexes

Intracellular cAMP and Calcium Signaling by Serotonin in Mouse Cumulus-Oocyte Complexes

Changes in cAMP-dependent protein kinase (PKA) and progesterone secretion in luteinizing human granulosa cells

Luteinizing hormone/chorionic gonadotrophin receptor overexpressed in granulosa cells from polycystic ovary syndrome ovaries is functionally active

Contact Info

Product

Resources

About