Abstract

Receptors for the glycoprotein hormone follicle stimulating hormone (follitropin; FSH) are present in the Sertoli cells of testis and granulosa cells of ovaries. FSH is essential for normal high quality gametogenesis in males and females. Glycoprotein hormones which function in fertility and normal thyroid function are of general medical significance. Function of the FSH receptor (FSHR) beyond the initial binding event is regulated by cytoplasmic proteins which bind to receptor intracellular loops (iL) and regulate receptor function and act to organize receptors. Adapter proteins that have been identified which interact with human FSHR (hFSHR) are the focus of these studies.
Aim one is to study the role of newly discovered adapter proteins APPL 1 and 2, that bind to FSHR-iL1. Using siRNA to inhibit their synthesis in primary cultures of rat granulosa cells, the role of APPL1 and APPL2 regulation in FSH driven granulosa cell functions will be determined. In addition, amino acids in FSHR-IL1 which are essential for interaction with APPL proteins will be determined so that binding defective mutants can be studied for trafficking defects.
Aim two will study the adapter protein 14-3-3 tau which binds to FSHR-iL2. It will be determined how inhibition of 14-3-3 tau affects FSHR signal transduction pathways in primary cultures of rat granulosa cells. Mutants of hFSHR-il-2 defective in binding to 14-3-3 tau will be prepared and FSH activation of the Erk pathway will be studied in rat granulosa cell lines expressing these FSHR mutants.
The third aim will determine the functional significance of the protein kinase CK2 (PKCK2) consensus site of the hFSHR-iL3. Consequences of inhibition of PKCK2 on steroidogenesis in primary cultures of human granulosa cells will be studied. It will be determined if PKCK2 interacts with hFSHR-iL3, and if consensus site mutants of the receptor are ubiquitination defective. Phosphorylation of hFSHR by PKCK2 will also be studied. The overall goal of the studies is to provide new information to better our understanding of regulation of gonadal function by gonadotropins and provide new avenues for fertility.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD018407-18
Application #
7166815
Study Section
Cellular, Molecular and Integrative Reproduction Study Section (CMIR)
Program Officer
Yoshinaga, Koji
Project Start
1997-04-01
Project End
2010-12-31
Budget Start
2007-01-01
Budget End
2007-12-31
Support Year
18
Fiscal Year
2007
Total Cost
$240,937
Indirect Cost

  Institution

Name
Wadsworth Center
Department
Type
DUNS #
153695478
City
Menands
State
NY
Country
United States
Zip Code
12204

  Publications

Mazurkiewicz, Joseph E; Herrick-Davis, Katharine; Barroso, Margarida et al. (2015) Single-molecule analyses of fully functional fluorescent protein-tagged follitropin receptor reveal homodimerization and specific heterodimerization with lutropin receptor. Biol Reprod 92:100
Dias, James A; Campo, Brice; Weaver, Barbara A et al. (2014) Inhibition of follicle-stimulating hormone-induced preovulatory follicles in rats treated with a nonsteroidal negative allosteric modulator of follicle-stimulating hormone receptor. Biol Reprod 90:19
Ulloa-Aguirre, Alfredo; Zariñán, Teresa; Dias, James A et al. (2014) Mutations in G protein-coupled receptors that impact receptor trafficking and reproductive function. Mol Cell Endocrinol 382:411-423
Ulloa-Aguirre, Alfredo; Dias, James A; Bousfield, George et al. (2013) Trafficking of the follitropin receptor. Methods Enzymol 521:17-45
Casas-Gonzalez, Patricia; Scaglia, Hugo E; Perez-Solis, Marco A et al. (2012) Normal testicular function without detectable follicle-stimulating hormone. A novel mutation in the follicle-stimulating hormone receptor gene leading to apparent constitutive activity and impaired agonist-induced desensitization and internalization. Mol Cell Endocrinol 364:71-82
Bousfield, George R; Dias, James A (2011) Synthesis and secretion of gonadotropins including structure-function correlates. Rev Endocr Metab Disord 12:289-302
Kluetzman, Kerri S; Thomas, Richard M; Nechamen, Cheryl A et al. (2011) Decreased degradation of internalized follicle-stimulating hormone caused by mutation of aspartic acid 6.30(550) in a protein kinase-CK2 consensus sequence in the third intracellular loop of human follicle-stimulating hormone receptor. Biol Reprod 84:1154-63
Thomas, Richard M; Nechamen, Cheryl A; Mazurkiewicz, Joseph E et al. (2011) The adapter protein APPL1 links FSH receptor to inositol 1,4,5-trisphosphate production and is implicated in intracellular Ca(2+) mobilization. Endocrinology 152:1691-701
Dias, James A; Bonnet, Béatrice; Weaver, Barbara A et al. (2011) A negative allosteric modulator demonstrates biased antagonism of the follicle stimulating hormone receptor. Mol Cell Endocrinol 333:143-50
Zarinan, Teresa; Perez-Solis, Marco A; Maya-Nunez, Guadalupe et al. (2010) Dominant negative effects of human follicle-stimulating hormone receptor expression-deficient mutants on wild-type receptor cell surface expression. Rescue of oligomerization-dependent defective receptor expression by using cognate decoys. Mol Cell Endocrinol 321:112-22

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