The objective of this research proposal is to gain an understanding of the mechanisms underlying cell-type specific actions of activin on key targets of pituitary gonadotropes. Activins are pleiotropic regulators of diverse tissues and cellular functions, often acting through autocrine/paracrine mechanisms. They play a critical role in the pituitary to locally regulate gonadotrope function and promote the differential expression and secretion of the two gonadotropins, FSH and LH. Follistatins are secreted glycoproteins that function extracellularly to bind and modulate the local bioavailability of activin and other related ligands of the TGF-? family through a feedback loop. In the pituitary, as in other tissues, the precise control of the activin/follistatin network helps maintain the functional integrity of target cells and prevents the development of pathologies. The dynamic control of activin/follistatin tone is achieved by the reciprocal actions of activin and follistatin and the self- modulating action of activin that exerts control on follistatin expression. Genetic models have substantiated the importance of follistatin by demonstrating that mice null for follistatin exhibit many embryonic defects and die shortly after birth while follistatin over-expression is associated with varying degrees of infertility. The follistatin gene is a transcriptional target of activin. In gonadotropes, this effect is mediated through the coordinated actions of the activin mediator, Smad3, and a member of the forkhead family of transcription factors, FoxL2. Functional and biochemical experiments have illustrated that FoxL2 is an obligatory partner of Smad3 for activin-dependent transcription of the follistatin gene in gonadotrope-derived cell types. FoxL2 is expressed in a limited number of tissues and human patients with FoxL2 deficiency display the Blepherophimosis-Ptosis- Epicanthus Inversus syndrome (BPES) associated with eyelid defects and premature ovarian failure in a sub- set of affected women. FoxL2 knockout mice exhibit a similar phenotype. FoxL2 is expressed in the pituitary of adult male and female mice and co-localizes with a-glycoprotein and FSH? subunits. FoxL2 protein is also readily detectable in gonadotrope-derived cell lines and siRNA-mediated knockdown strategies have demonstrated that FoxL2 is a critical mediator of activin actions on key transcriptional targets that have thus far been identified such as follistatin, FSH? and GnRH-R. By utilizing complimentary in vitro and in vivo models, the proposed studies of this application aim to provide a better understanding of the central role of FoxL2 in coordinating the Smad3-dependent activin program of gonadotropes, elucidate the mechanism underlying the partnership between FoxL2 and Smad3 and determine how FoxL2 function contributes to the local control of the activin/follistatin network of the pituitary, mechanisms that might prove to be relevant to other FoxL2-expressing tissues. Understanding the mechanism underlying FoxL2 action could illuminate strategies for the differential control of Smad3 targets and identify novel molecular targets with relevance to reproductive and other endocrine disorders.

Public Health Relevance

Activins are ubiquitous growth factors known to regulate many cellular processes, including differential FSH expression and secretion from pituitary gonadotropes. Activins control their own bioactivity in part by exerting positive effects on the expression of activin-binding proteins known as follistatins, which are also ubiquitous and often co-localize with activins. Through a better understanding of how activin regulates the expression of follistatin and other key targets in gonadotropes, these studies will facilitate the identification of novel targets that have the potential to selectively target the tissue-specific actions of activin relevant to reproductive and/or other endocrine pathologies.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD046941-09
Application #
8433979
Study Section
Integrative and Clinical Endocrinology and Reproduction Study Section (ICER)
Program Officer
Lamar, Charisee A
Project Start
2004-04-01
Project End
2015-02-28
Budget Start
2013-03-01
Budget End
2014-02-28
Support Year
9
Fiscal Year
2013
Total Cost
$347,106
Indirect Cost
$164,898
Name
Salk Institute for Biological Studies
Department
Type
DUNS #
078731668
City
La Jolla
State
CA
Country
United States
Zip Code
92037
Bilezikjian, Louise M; Justice, Nicholas J; Blackler, Alissa N et al. (2012) Cell-type specific modulation of pituitary cells by activin, inhibin and follistatin. Mol Cell Endocrinol 359:43-52
Bilezikjian, Louise M; Vale, Wylie W (2011) The Local Control of the Pituitary by Activin Signaling and Modulation. Open Neuroendocrinol J 4:90-101
Justice, Nicholas J; Blount, Amy L; Pelosi, Emanuele et al. (2011) Impaired FSHbeta expression in the pituitaries of Foxl2 mutant animals. Mol Endocrinol 25:1404-15
Blount, Amy L; Schmidt, Karsten; Justice, Nicholas J et al. (2009) FoxL2 and Smad3 coordinately regulate follistatin gene transcription. J Biol Chem 284:7631-45
Blount, Amy L; Vaughan, Joan M; Vale, Wylie W et al. (2008) A Smad-binding element in intron 1 participates in activin-dependent regulation of the follistatin gene. J Biol Chem 283:7016-26
Bilezikjian, Louise M; Blount, Amy L; Donaldson, Cindy J et al. (2006) Pituitary actions of ligands of the TGF-beta family: activins and inhibins. Reproduction 132:207-15