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Effects of Progranulin on Blastocyst Hatching and Subsequent Adhesion and Outgrowth in the Mouse¹

Laboratory of Animal Breeding,³ Graduate School of Agriculture and Life Sciences, University of Tokyo, Tokyo 113-8657, Japan Department of Obstetrics and Gynecology,⁴ University of Pennsylvania Medical Center, University of Pennsylvania, Philadelphia, Pennsylvania 19104 Research Unit in Reproductive Medicine,⁵ Hospital de Ginecobstetricia "Dr. Luis Castelazo Ayala," Instituto Mexicano del Seguro Social, Mexico D.F. 10101, Mexico Korea Research Institute of Bioscience and Biotechnology,⁶ Taejon, 305-333, Korea

Using cDNA microarray methodology, we have shown previously that transcripts of progranulin gene (Grn, also known as acrogranin), a recently identified autocrine growth factor, were upregulated in mouse blastocysts adhered to the filter membrane in an in vitro-culture system. In the present study, we investigated the expression and effects of progranulin on blastocyst hatching, adhesion, and embryo outgrowth during the peri-implantation period in the mouse. During this period, substantial amounts of Grn mRNA were present in both inner cell mass (ICM) and trophectoderm. Progranulin was localized exclusively to the surface of the trophectoderm in early and pre- and postadhesion blastocysts as well as in trophoblast cells and ICM of outgrowth embryos, being secreted as a single, 88-kDa form into the surrounding medium. NIH3T3 cells that had been transfected with a progranulin expression construct secreted the 88-kDa form of the protein, from which a 68-kDa form could be generated by deglycosylation. In vitro treatment of blastocysts with recombinant progranulin promoted blastocyst hatching, adhesion, and outgrowth, whereas rabbit anti-mouse progranulin immunoglobulin G reduced the incidence of blastocyst hatching, adhesion, and outgrowth. Studies of bromodeoxyuridine incorporation and immunodissection of the ICM revealed that progranulin was effective on the trophectoderm but not on the ICM. These results indicate that progranulin is an important factor for the processes of blastocyst hatching, adhesion, and outgrowth, and they suggest that the effects of progranulin on blastocyst adhesion and outgrowth may have been triggered by the previous action of progranulin to induce hatching of the blastocysts.

cytokines, early development, embryo, growth factors implantation

² Correspondence: Kazuhiko Imakawa, Laboratory of Animal Breeding, Graduate School of Agriculture and Life Sciences, University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan. FAX: 81 3 5841 8180; akaz{at}mail.ecc.u-tokyo.ac.jp

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