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Spatiotemporal Expression of C-CAM in the Rat Placenta
Hiroki Sawaa, Hiroyuki Ukitab, Minoru Fukudac, Hajime Kamadab, Isamu Saitoa, and Björn Öbrinkda Department of Neurosurgery, Kyorin University, School of Medicine, Shinkawa, Mitaka, Tokyo, Japan
b Hokuto Hospital, Obihiro, Hokkaido, Japan
c Department of Electron Microscopy, Kyorin University, School of Medicine, Mitaka 181, Tokyo, Japan
d Department of Cell and Molecular Biology, Karolinska Institute, Stockholm, Sweden
Correspondence to: Hiroki Sawa, Dept. of Neurosurgery, Kyorin Univ., School of Medicine, Mitaka, Tokyo 181, Japan.
We investigated the expression of the immunoglobulin superfamily cell adhesion molecule, C-CAM, in developing and mature rat placenta. By immunohistochemical staining at the light microscopic level, no C-CAM-expression was seen before Day 9 of gestation, when it appeared in the trophoblasts of ectoplacental cones. On Day 10.5, spongiotrophoblasts and invasive trophoblasts around the maternal vessels of the decidua basalis were stained positively. On Day 12.5, C-CAM was detected in the spongiotrophoblasts of the junctional layer, but labyrinth trophoblasts and secondary giant trophoblasts were not stained. On Day 17.5, C-CAM was found only in the labyrinth and lacunae of the junctional layer. At this stage, both the labyrinth cytotrophoblasts of the maternal blood vessels and the endothelial cells of the embryonic capillaries were strongly stained. Placental tissues from gestational Days 12.5 and 17.5 were analyzed by immunoelectron microscopy to determine the location of C-CAM at the subcellular level. On Day 12.5, positive staining of the spongiotrophoblasts was observed, mainly on surface membranes and microvilli between loosely associated cells. On Day 17.5, staining was found primarily on the microvilli of the maternal luminal surfaces of the labyrinth cytotrophoblasts, and both on the luminal surface and in the cytoplasm of endothelial cells of the embryonic vessels. RT-PCR analysis and Southern blotting of the PCR products revealed expression of mRNA species for both of the major isoforms, C-CAM1 and C-CAM2. Immunoblotting analysis of C-CAM isolated from 12.5-day and 14.5-day placentae showed that it appeared as a broad band with an apparent molecular mass of 110-170 kD. In summary, C-CAM was strongly expressed in a specific spatiotemporal pattern in trophoblasts actively involved in formation of the placental tissue, suggesting an important role in placental development. In the mature placenta, C-CAM expression was confined to the trophoblastic and endothelial cells lining the maternal and embryonic vessels, respectively, suggesting important functions in placental physiology. (J Histochem Cytochem 45:1021-1034, 1997)
Key Words: C-CAM, rat placenta, immunohistochemistry, RT-PCR, immunoelectron microscopy
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