Increased reactivity of cultured chicken blastodermal cells to anti-stage-specific embryonic antigen-1 antibody after exposure to bone morphogenetic proteins.
We evaluated whether bone morphogenetic proteins (BMPs) increased the reactivity of chicken stage X blastodermal cells to the germ cell marker, anti-stage-specific embryonic antigen (SSEA)-1 antibody. In Experiment 1, blastodermal cells cultured on a feeder layer of SIM mouse embryo-derived thioguanine and ouabain resistant (STO) cells were treated with different doses of BMP-2 and/or BMP-4, and the anti-SSEA-1 antibody reactivity of cultured cells was examined 48 h later. A significant (P < 0.05) increase in the number of anti-SSEA-1 antibody-positive cells was detected after the addition of 75 or 100 ng/ml BMP-2. Neither 0-20 ng/ml BMP-4 nor the combined addition of 75 ng/ml BMP-2 with either 10 or 15 ng/ml BMP-4 increased reactivity more than that induced by 75 ng/ml BMP-2 alone. Results of the qualification and quantification of BMP receptor kinase (BRK)-1, BRK-2, and BRK-3 using RT-PCR and real-time PCR showed that all three receptors were detected in blastodermal cells treated with BMPs, intact stage X embryos and 5.5-day-old embryonic gonads, but no expression was detected in STO feeder cells. In Experiment 2, the treatment of stage X embryos with different doses of BMP-2 (0.15-3 ng/embryo) or BMP-4 (0.02-0.4 ng/embryo) did not affect the reactivity of 5.5-day-old embryonic gonadal cells to the anti-SSEA-1 antibody. BRK-1 expression was selectively increased in stage X embryos after the infusion of 3ng BMP-2 than after no infusion, but no changes in other BRKs' expression were detected. In conclusion, the addition of BMP-2 to culture medium in the presence of STO feeder cells promoted the reactivity of blastodermal cells to anti-SSEA-1 antibody, which might contribute to the generation of chicken primordial germ cell precursor or germ cell-like cells. The relationship between BMP action and BRK expression was further discussed.
Kim
Duk Kyung
DK
Department of Food and Animal Biotechnology, Division of Animal Genetic Engineering, Seoul National University, Seoul 151-921, Republic of Korea.
Song
Ki Duk
KD
Kim
Jin Nam
JN
Park
Tae Sub
TS
Lim
Jeong Mook
JM
Han
Jae Yong
JY
D000818:Animals; D001756:Blastoderm / Q000187:drug effects*; D019485:Bone Morphogenetic Proteins / Q000494:pharmacology*; D002478:Cells, Cultured; D002642:Chick Embryo / Q000166:cytology*; D004306:Dose-Response Relationship, Immunologic; D016256:Lewis X Antigen / Q000276:immunology*; D010770:Phosphotransferases / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary
D019485:Bone Morphogenetic Proteins; D016256:Lewis X Antigen; D010770:Phosphotransferases