Skip to main content

Advertisement

Log in

Distribution and Y397 phosphorylation of focal adhesion kinase on follicular development in the mouse ovary

  • Regular Article
  • Published:
Cell and Tissue Research Aims and scope Submit manuscript

Abstract

Several protein tyrosine kinases (PTKs) are identified as follicle survival factors that suppress apoptosis in granulosa cells. Focal adhesion kinase (FAK/PTK2) interacts with numerous signaling partners and is important for cell adhesion, survival and other vital processes in which FAK autophosphorylation at Y397 (pY397 FAK) is critical for activating signaling pathways. Despite its important roles in apoptosis, the expression and function of FAK in the ovaries remain unknown. Here, we describe FAK expression, including pY397 FAK, in normal healthy mouse ovaries and its association with follicular development and/or atresia. Normal healthy mouse ovaries were used for western blot (n > 60) and immunohistochemical (n > 180) analyses. Western blot results in immature and mature mice revealed that total FAK and pY397 FAK were highly expressed in the ovary and immunohistochemistry results in 3-week-old mice showed they were localized to granulosa cells of ovarian follicles, especially preantral follicles. In 3-week-old mice treated with 5 IU pregnant mare serum gonadotropin (for obtaining homogenous populations of growing or atretic follicles), western blotting revealed that follicular atresia progression involved decreased phosphorylation of Y397 at 72 and 96 h after treatment, particularly in granulosa cells of atretic follicles, as shown by immunohistochemistry results at 72 h after treatment. Moreover, immunostaining patterns of FAK and cleaved caspase-3 were negatively correlated in serial sections of 3-week-old mouse ovaries. These results suggest that FAK is most active in ovarian follicle granulosa cells and that its phosphorylation at Y397 is histologically meaningful in follicular development in normal healthy ovaries.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Adashi EY (1992) The potential relevance of cytokines to ovarian physiology. J Steroid Biochem Mol Biol 43:439–444

    Article  PubMed  CAS  Google Scholar 

  • Adashi EY, Resnick CE, D'Ercole AJ, Svoboda ME, Van Wyk JJ (1985) Insulin-like growth factors as intraovarian regulators of granulosa cell growth and function. Endocr Rev 6:400–420

    Article  PubMed  CAS  Google Scholar 

  • Adashi EY, Resnick CE, Vera A, Hernandez ER (1991) In vivo regulation of granulosa cell type I insulin-like growth factor receptors: evidence for an inhibitory role for the putative endogenous ligand(s) of the ovarian gonadotropin-releasing hormone receptor. Endocrinology 128:3130–3137

    Article  PubMed  CAS  Google Scholar 

  • Alonso-Pozos I, Rosales-Torres AM, Avalos-Rodriguez A, Vergara-Onofre M, Rosado-Garcia A (2003) Mechanism of granulosa cell death during follicular atresia depends on follicular size. Theriogenology 60:1071–1081

    Article  PubMed  CAS  Google Scholar 

  • Beviglia L, Golubovskaya V, Xu L, Yang X, Craven RJ, Cance WG (2003) Focal adhesion kinase N-terminus in breast carcinoma cells induces rounding, detachment and apoptosis. Biochem J 373:201–210

    Article  PubMed  CAS  Google Scholar 

  • Calalb MB, Polte TR, Hanks SK (1995) Tyrosine phosphorylation of focal adhesion kinase at sites in the catalytic domain regulates kinase activity: a role for Src family kinases. Mol Cell Biol 15:954–963

    PubMed  CAS  Google Scholar 

  • Cance WG, Harris JE, Iacocca MV, Roche E, Yang X, Chang J, Simkins S, Xu L (2000) Immunohistochemical analyses of focal adhesion kinase expression in benign and malignant human breast and colon tissues: correlation with preinvasive and invasive phenotypes. Clin Cancer Res 6:2417–2423

    PubMed  CAS  Google Scholar 

  • Cary LA, Guan JL (1999) Focal adhesion kinase in integrin-mediated signaling. Front Biosci 4:D102–113

    Article  PubMed  CAS  Google Scholar 

  • Cary LA, Klinghoffer RA, Sachsenmaier C, Cooper JA (2002) SRC catalytic but not scaffolding function is needed for integrin-regulated tyrosine phosphorylation, cell migration, and cell spreading. Mol Cell Biol 22:2427–2440

    Article  PubMed  CAS  Google Scholar 

  • Chan PC, Lai JF, Cheng CH, Tang MJ, Chiu CC, Chen HC (1999) Suppression of ultraviolet irradiation-induced apoptosis by overexpression of focal adhesion kinase in Madin-Darby canine kidney cells. J Biol Chem 274:26901–26906

    Article  PubMed  CAS  Google Scholar 

  • Chen HC, Appeddu PA, Isoda H, Guan JL (1996) Phosphorylation of tyrosine 397 in focal adhesion kinase is required for binding phosphatidylinositol 3-kinase. J Biol Chem 271:26329–26334

    Article  PubMed  CAS  Google Scholar 

  • Chun SY, Billig H, Tilly JL, Furuta I, Tsafriri A, Hsueh AJ (1994) Gonadotropin suppression of apoptosis in cultured preovulatory follicles: mediatory role of endogenous insulin-like growth factor I. Endocrinology 135:1845–1853

    Article  PubMed  CAS  Google Scholar 

  • Chun SY, Eisenhauer KM, Minami S, Billig H, Perlas E, Hsueh AJ (1996) Hormonal regulation of apoptosis in early antral follicles: follicle-stimulating hormone as a major survival factor. Endocrinology 137:1447–1456

    Article  PubMed  CAS  Google Scholar 

  • Cobb BS, Schaller MD, Leu TH, Parsons JT (1994) Stable association of pp 60src and pp59fyn with the focal adhesion-associated protein tyrosine kinase, pp125FAK. Mol Cell Biol 14:147–155

    PubMed  CAS  Google Scholar 

  • Crouch DH, Fincham VJ, Frame MC (1996) Targeted proteolysis of the focal adhesion kinase pp 125 FAK during c-MYC-induced apoptosis is suppressed by integrin signalling. Oncogene 12:2689–2696

    PubMed  CAS  Google Scholar 

  • Dhanasekaran N, Moudgal NR (1989) Biochemical and histological validation of a model to study follicular atresia in rats. Endocrinol Exp 23:155–166

    PubMed  CAS  Google Scholar 

  • Dissen GA, Romero C, Hirshfield AN, Ojeda SR (2001) Nerve growth factor is required for early follicular development in the mammalian ovary. Endocrinology 142:2078–2086

    Article  PubMed  CAS  Google Scholar 

  • Erickson GF, Shimasaki S (2000) The role of the oocyte in folliculogenesis. Trends Endocrinol Metab 11:193–198

    Article  PubMed  CAS  Google Scholar 

  • Erickson GF, Garzo VG, Magoffin DA (1991) Progesterone production by human granulosa cells cultured in serum free medium: effects of gonadotrophins and insulin-like growth factor I (IGF-I). Hum Reprod 6:1074–1081

    PubMed  CAS  Google Scholar 

  • Gilchrist RB, Ritter LJ, Armstrong DT (2004) Oocyte-somatic cell interactions during follicle development in mammals. Anim Reprod Sci 82–83:431–446

    Article  PubMed  Google Scholar 

  • Guthrie HD, Garrett WM, Cooper BS (1998) Follicle-stimulating hormone and insulin-like growth factor-I attenuate apoptosis in cultured porcine granulosa cells. Biol Reprod 58:390–396

    Article  PubMed  CAS  Google Scholar 

  • Halder J, Kamat AA, Landen CN Jr, Han LY, Lutgendorf SK, Lin YG, Merritt WM, Jennings NB, Chavez-Reyes A, Coleman RL, Gershenson DM, Schmandt R, Cole SW, Lopez-Berestein G, Sood AK (2006) Focal adhesion kinase targeting using in vivo short interfering RNA delivery in neutral liposomes for ovarian carcinoma therapy. Clin Cancer Res 12:4916–4924

    Article  PubMed  CAS  Google Scholar 

  • Hanks SK, Polte TR (1997) Signaling through focal adhesion kinase. Bioessays 19:137–145

    Article  PubMed  CAS  Google Scholar 

  • Hirshfield AN (1991) Development of follicles in the mammalian ovary. Int Rev Cytol 124:43–101

    Article  PubMed  CAS  Google Scholar 

  • Hussein TS, Froiland DA, Amato F, Thompson JG, Gilchrist RB (2005) Oocytes prevent cumulus cell apoptosis by maintaining a morphogenic paracrine gradient of bone morphogenetic proteins. J Cell Sci 118:5257–5268

    Article  PubMed  CAS  Google Scholar 

  • Judson PL, He X, Cance WG, Van Le L (1999) Overexpression of focal adhesion kinase, a protein tyrosine kinase, in ovarian carcinoma. Cancer 86:1551–1556

    Article  PubMed  CAS  Google Scholar 

  • Kaipia A, Hsueh AJ (1997) Regulation of ovarian follicle atresia. Annu Rev Physiol 59:349–363

    Article  PubMed  CAS  Google Scholar 

  • Lark AL, Livasy CA, Calvo B, Caskey L, Moore DT, Yang X, Cance WG (2003) Overexpression of focal adhesion kinase in primary colorectal carcinomas and colorectal liver metastases: immunohistochemistry and real-time PCR analyses. Clin Cancer Res 9:215–222

    PubMed  CAS  Google Scholar 

  • Levkau B, Herren B, Koyama H, Ross R, Raines EW (1998) Caspase-mediated cleavage of focal adhesion kinase pp 125FAK and disassembly of focal adhesions in human endothelial cell apoptosis. J Exp Med 187:579–586

    Article  PubMed  CAS  Google Scholar 

  • Manabe N, Imai Y, Ohno H, Takahagi Y, Sugimoto M, Miyamoto H (1996) Apoptosis occurs in granulosa cells but not cumulus cells in the atretic antral follicles in pig ovaries. Experientia 52:647–651

    Article  PubMed  CAS  Google Scholar 

  • Owen JD, Ruest PJ, Fry DW, Hanks SK (1999) Induced focal adhesion kinase (FAK) expression in FAK-null cells enhances cell spreading and migration requiring both auto- and activation loop phosphorylation sites and inhibits adhesion-dependent tyrosine phosphorylation of Pyk2. Mol Cell Biol 19:4806–4818

    PubMed  CAS  Google Scholar 

  • Owens LV, Xu L, Craven RJ, Dent GA, Weiner TM, Kornberg L, Liu ET, Cance WG (1995) Overexpression of the focal adhesion kinase (p125FAK) in invasive human tumors. Cancer Res 55:2752–2755

    PubMed  CAS  Google Scholar 

  • Roy SK, Greenwald GS (1996) Follicular development through preantral stages: signalling via growth factors. J Reprod Fertil Suppl 50:83–94

    PubMed  CAS  Google Scholar 

  • Sakamaki K, Yoshida H, Nishimura Y, Nishikawa S, Manabe N, Yonehara S (1997) Involvement of Fas antigen in ovarian follicular atresia and luteolysis. Mol Reprod Dev 47:11–18

    Article  PubMed  CAS  Google Scholar 

  • Schaller MD (2001) Biochemical signals and biological responses elicited by the focal adhesion kinase. Biochim Biophys Acta 1540:1–21

    Article  PubMed  CAS  Google Scholar 

  • Schaller MD, Borgman CA, Cobb BS, Vines RR, Reynolds AB, Parsons JT (1992) pp 125FAK a structurally distinctive protein-tyrosine kinase associated with focal adhesions. Proc Natl Acad Sci USA 89:5192–5196

    Article  PubMed  CAS  Google Scholar 

  • Schaller MD, Hildebrand JD, Shannon JD, Fox JW, Vines RR, Parsons JT (1994) Autophosphorylation of the focal adhesion kinase, pp 125FAK, directs SH2-dependent binding of pp60src. Mol Cell Biol 14:1680–1688

    PubMed  CAS  Google Scholar 

  • Schlaepfer DD, Hunter T (1996) Evidence for in vivo phosphorylation of the Grb2 SH2-domain binding site on focal adhesion kinase by Src-family protein-tyrosine kinases. Mol Cell Biol 16:5623–5633

    PubMed  CAS  Google Scholar 

  • Schlaepfer DD, Hanks SK, Hunter T, van der Geer P (1994) Integrin-mediated signal transduction linked to Ras pathway by GRB2 binding to focal adhesion kinase. Nature 372:786–791

    PubMed  CAS  Google Scholar 

  • Simpson ER, Clyne C, Rubin G, Boon WC, Robertson K, Britt K, Speed C, Jones M (2002) Aromatase–a brief overview. Annu Rev Physiol 64:93–127

    Article  PubMed  CAS  Google Scholar 

  • Singh B, Armstrong DT (1997) Insulin-like growth factor-1, a component of serum that enables porcine cumulus cells to expand in response to follicle-stimulating hormone in vitro. Biol Reprod 56:1370–1375

    Article  PubMed  CAS  Google Scholar 

  • Sonoda Y, Matsumoto Y, Funakoshi M, Yamamoto D, Hanks SK, Kasahara T (2000) Anti-apoptotic role of focal adhesion kinase (FAK). Induction of inhibitor-of-apoptosis proteins and apoptosis suppression by the overexpression of FAK in a human leukemic cell line, HL-60. J Biol Chem 275:16309–16315

    Article  PubMed  CAS  Google Scholar 

  • Sood AK, Coffin JE, Schneider GB, Fletcher MS, DeYoung BR, Gruman LM, Gershenson DM, Schaller MD, Hendrix MJ (2004) Biological significance of focal adhesion kinase in ovarian cancer: role in migration and invasion. Am J Pathol 165:1087–1095

    Article  PubMed  CAS  Google Scholar 

  • Takahashi R, Sonoda Y, Ichikawa D, Yoshida N, Eriko AY, Tadashi K (2007) Focal adhesion kinase determines the fate of death or survival of cells in response to TNFalpha in the presence of actinomycin D. Biochim Biophys Acta 1770:518–526

    Article  PubMed  CAS  Google Scholar 

  • Tilly JL (1996) Apoptosis and ovarian function. Rev Reprod 1:162–172

    Article  PubMed  CAS  Google Scholar 

  • Tilly JL, Billig H, Kowalski KI, Hsueh AJ (1992) Epidermal growth factor and basic fibroblast growth factor suppress the spontaneous onset of apoptosis in cultured rat ovarian granulosa cells and follicles by a tyrosine kinase-dependent mechanism. Mol Endocrinol 6:1942–1950

    Article  PubMed  CAS  Google Scholar 

  • Tsafriri A, Braw RH (1984) Experimental approaches to atresia in mammals. Oxf Rev Reprod Biol 6:226–265

    PubMed  CAS  Google Scholar 

  • Weiner TM, Liu ET, Craven RJ, Cance WG (1993) Expression of focal adhesion kinase gene and invasive cancer. Lancet 342:1024–1025

    Article  PubMed  CAS  Google Scholar 

  • Wen LP, Fahrni JA, Troie S, Guan JL, Orth K, Rosen GD (1997) Cleavage of focal adhesion kinase by caspases during apoptosis. J Biol Chem 272:26056–26061

    Article  PubMed  CAS  Google Scholar 

  • Yoshimura Y, Ando M, Nagamatsu S, Iwashita M, Adachi T, Sueoka K, Miyazaki T, Kuji N, Tanaka M (1996) Effects of insulin-like growth factor-I on follicle growth, oocyte maturation, and ovarian steroidogenesis and plasminogen activator activity in the rabbit. Biol Reprod 55:152–160

    Article  PubMed  CAS  Google Scholar 

  • Zhou J, Chin E, Bondy C (1991) Cellular pattern of insulin-like growth factor-I (IGF-I) and IGF-I receptor gene expression in the developing and mature ovarian follicle. Endocrinology 129:3281–3288

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgement

This work was supported by the Japan Society for the Promotion of Science Grant to E. Sato (No. 21248032).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Masahiro Sakurai.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sakurai, M., Ohtake, J., Ishikawa, T. et al. Distribution and Y397 phosphorylation of focal adhesion kinase on follicular development in the mouse ovary. Cell Tissue Res 347, 457–465 (2012). https://doi.org/10.1007/s00441-011-1307-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00441-011-1307-2

Keywords

Navigation