Regulation of embryo outgrowth by a morphogenic factor, epimorphin, in the mouse.
Conceptus implantation to the uterine endometrium represents a complex series of events, including synchronized development of conceptus and uterus through up- and/or down-regulation of numerous gene products. In a previous study using the DNA microarray technique, we had discovered evidence that increase in a transcript for mesenchymal morphogen, epimorphin, was noted as the conceptus attached to the matrix in vitro (Qin et al., 2003). In the present study, the expression and potential function of epimorphin in developing conceptuses was investigated through the use of reverse transcription-polymerase chain reaction (RT-PCR), whole mount in situ hybridization/immunohistochemistry, and in vitro blastocyst culture. RT-PCR and in situ hybridization analysis revealed that epimorphin mRNA was expressed weakly in murine conceptuses during early developmental stages (1 cell to post-adhesion blastocyst stages) and higher levels of epimorphin transcripts were observed in both inner cell mass (ICM) and trophectoderm of outgrowing blastocysts. Immunohistochemical analysis confirmed that epimorphin was localized in outgrowing trophoblast cells and ICM. Treating blastocysts in culture with a 115 kDa form of recombinant epimorphin promoted trophoblast outgrowth (P < 0.05), but a 34 kDa form of recombinant epimorphin had no effect. Treatment with a function inhibitor, rat anti-mouse epimorphin IgM, reduced the number of embryos progressing to blastocyst outgrowth to the levels similar to those observed with plain culture medium. Reverse transcription-polymerase chain reaction (RT-PCR) analysis also revealed that epimorphin increased the expression of a trophoblast cell differentiation marker, placental lactogen-1 (PL-1), mRNA (P < 0.01). These results suggest that epimorphin is involved in trophoblast outgrowth, a process required for conceptus implantation into the endometrium.
Qin
Junwen
J
Laboratory of Animal Breeding, Graduate School of Agriculture and Life Sciences, The University of Tokyo, Yayoi, Tokyo, Japan.
Takahashi
Yuji
Y
Isuzugawa
Kazuto
K
Imai
Misa
M
Yamamoto
Shinya
S
Hirai
Yohei
Y
Imakawa
Kazuhiko
K
D000818:Animals; D001755:Blastocyst / Q000378:metabolism*; D005260:Female; D015151:Immunoblotting; D008297:Male; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008813:Mice, Inbred ICR; D016376:Oligonucleotides, Antisense; D010928:Placental Lactogen / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism
C504582:Epim protein, mouse; D008562:Membrane Glycoproteins; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; C085699:placental lactogen I, rat; D010928:Placental Lactogen