Cochaperone immunophilin FKBP52 is critical to uterine receptivity for embryo implantation

Susanne Tranguch

Joyce Cheung-Flynn

Takiko Daikoku

Viravan Prapapanich

Departments of ^Pediatrics,^{Cell and Developmental Biology,}^{Cancer Biology, and}^{Pharmacology, Vanderbilt University Medical Center, Nashville, TN 37232; and}^{Department of Biochemistry and Molecular Biology, Mayo Clinic, Scottsdale, AZ 85259}

^{To whom correspondence should be addressed. E-mail:}ude.tlibrednav@yed.ks.

^{S.T. and J.C.-F. contributed equally to this work.}

Edited by R. Michael Roberts, University of Missouri, Columbia, MO, and approved August 22, 2005

Received 2005 Jul 9

Abstract

Embryo implantation in the uterus is a critical step in mammalian reproduction, requiring preparation of the uterus receptive to blastocyst implantation. Uterine receptivity, also known as the window of implantation, lasts for a limited period, and it is during this period blastocysts normally implant. Ovarian steroid hormones estrogen and progesterone (P₄) are the primary regulators of this process. The immunophilin FKBP52 serves as a cochaperone for steroid hormone nuclear receptors to govern appropriate hormone action in target tissues. Here we show a critical role for FKBP52 in mouse implantation. This immunophilin has unique spatiotemporal expression in the uterus during implantation, and females missing the Fkbp52 gene have complete implantation failure due to lack of attainment of uterine receptivity. The overlapping uterine expression of FKBP52 with nuclear progesterone receptor (PR) in wild-type mice together with reduced P₄ binding to PR, attenuated PR transcriptional activity and down-regulation of several P₄-regulated genes in uteri of Fkbp52^{mice, establishes this cochaperone as a critical regulator of uterine P}₄ function. Interestingly, ovulation, another P₄-mediated event, remains normal. Collectively, the present investigation provides evidence for an in vivo role for this cochaperone in regulating tissue-specific hormone action and its critical role in uterine receptivity for implantation.

Keywords: mouse, uterus, ovulation, blastocyst, progesterone receptor

Abstract

Progesterone (P₄) is essential for implantation and pregnancy maintenance in all mammalian species studied. In mice, P₄ priming of the uterus is obligatory for estrogen to prepare the uterus to the receptive state conducive to blastocyst implantation. P₄ acting through the nuclear P₄ receptor (PR) modulates uterine physiology and expression of various genes that are required for implantation (1, 2). Numerous defects in mice lacking the Pgr gene that encodes PR include failure in ovulation, mammary gland development, and sexual behavior along with uterine hyperplasia and inflammation, reflecting the critical role of P₄ in female reproduction (3). Appropriate functioning of nuclear steroid hormone receptors depends on interactions with the molecular chaperone machinery to maintain a functional state competent for hormone binding and subsequent transcriptional activation. Functionally mature steroid receptor complexes consist of a receptor monomer, a 90-kDa heat shock protein (Hsp90) dimer, the cochaperone p23, and one of four cochaperones that contain a tetratricopeptide repeat (TPR) domain. The TPR cochaperones include two members of the FK506 binding family of immunophilins, FKBP52/FKBP4 and FKBP51/FKBP5, a member of the cyclosporin-binding immunophilin cyclophilin 40 (CyP40) or the protein phosphatase PP5. FKBP52 and FKBP51 are similar to other FKBP family members in that both contain an active peptidylprolyl cis/trans isomerase domain that catalyzes conformational changes in protein substrates (4, 5).

Roles for Hsp90 and p23 in initiating and maintaining receptor competency for hormone binding are well established (6), but the contribution of cochaperones in receptor complexes are not clearly understood. Although FKBP51 and FKBP52 share 60% sequence identity, have similar x-ray crystallographic structures (7, 8), and display similar peptidylprolyl cis/trans isomerase and Hsp90-binding activities, there are clear functional distinctions between these cochaperones in steroid receptor complexes. There is evidence based on in vitro cellular assays that, whereas FKBP52 potentiates the function of glucocorticoid receptors (GR) (9), androgen receptors (10), and PR (5), FKBP51 antagonizes GR and PR functions (11, 12). However, physiological roles of FKBP52 and FKBP51 in an in vivo context have not been examined. More specifically, a physiological role for FKBP52 and FKBP51 in P₄-dependent processes including ovarian and uterine functions remains unknown. Here we show that female mice missing the Fkbp52 gene have compromised PR functions leading to total failure of the uterus to support blastocyst implantation.

+/+, +/−, and −/− females were mated with wild-type fertile males and the number of implantation sites (IS) was examined on days 5 and 6 of pregnancy by the blue dye method. Uteri without IS were flushed with saline to recover any unimplanted embryos.

Day 4 wild-type (+/+) blastocysts were transferred into uteri of +/+, +/−, or −/− recipients on day 4 of pseudopregnancy. The recipients were examined for implantation sites (IS) on day 5 by the blue dye method. Uteri without IS were flushed with saline to recover any unimplanted blastocysts.

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Acknowledgments

This work was supported in parts by the National Institutes of Health and the Mayo Foundation. S. K. Dey is recipient of Method to Extend Research in Time Awards from the National Institute of Child Health and Human Development and the National Institute on Drug Abuse. H.W. is recipient of Solvay/Mortola Research Award from the Society for Gynecologic Investigation. S.T. and M.B.C. are supported by National Institutes of Health Training Grants 5 T 32 DK07563 and 1 F32 DK068983, respectively.

Acknowledgments

Notes

Author contributions: S.T., J.C.-F., T.D., H.W., D.F.S., and S. K. Dey designed research; S.T., J.C.-F., T.D., V.P., M.B.C., H.X., H.W., and S. K. Das performed research; S.T., J.C.-F., T.D., V.P., M.B.C., H.W., S. K. Das, D.F.S., and S. K. Dey analyzed data; and S.T., D.F.S., and S. K. Dey wrote the paper.

This paper was submitted directly (Track II) to the PNAS office.

Abbreviations: P₄, progesterone; PR, P₄ receptor.

Notes

This paper was submitted directly (Track II) to the PNAS office.

Abbreviations: P₄, progesterone; PR, P₄ receptor.

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