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Lipid Rafts play a role in human normal follicle stimlating hormone receptor (hFSHR) Signaling
Author(s) -
Libous Jennifer Lauren,
Dias James A,
Cohen Brian D
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.645.15
Subject(s) - lipid raft , microbiology and biotechnology , signal transduction , phosphorylation , chemistry , receptor , stimulation , hormone , protein kinase a , endocrinology , kinase , medicine , biology , biochemistry
The hFSHR is a member of the family of g protein‐coupled receptors that has been shown to signal through both g protein linked and MAP kinase pathways. Little is known about the importance of the membrane microenvironment in normal signaling. To study the role of lipid rafts in hFSHR signaling, cholesterol depletion experiments were carried out with methyl‐â‐cyclodextrin (MBCD) in HEK293 cells expressing hFSHR. Cells were treated with MBCD prior to stimulation with FSH for 0, 15, and 30 minutes. Cells were harvested and activation of MAP kinase enzymes (p38 and p44/42) were measured by western blot. MBCD treatment resulted in constant phosphorylation for p38 and a decrease in phosphorylation for p44/42 in response to FSH. In parallel experiments, cells were treated with MBCD prior to FSH stimulation to measure cAMP accumulation. cAMP production in MBCD treated cells was almost completely abolished, although hormone binding remained normal. One interpretation of the data suggests that the disruption of lipid rafts by depleting cholesterol disturbs the membrane microevironment sufficiently to disturb hFSHR signaling. Currently, a cholesterol add‐back experiment is being carried out to follow up on the cholesterol depletion to see if signaling is restored.