. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 190 M. SHIRAE ET AL. Marine Research Center. University of Tsukuba. Colonies that grew well were used for the experiments. Fusion experiments to test fusibility (compatibility) be- tween two colonies were routinely carried out as follows. A small piece was cut from the periphery of each colony with a razor blade. Two allogeneic or syngeneic colony pieces of the same size were brought into contact at their growing edges on a glass slide and were allowed to adhere to the slide in a moisture chamber for 30 min. These colony pie
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 190 M. SHIRAE ET AL. Marine Research Center. University of Tsukuba. Colonies that grew well were used for the experiments. Fusion experiments to test fusibility (compatibility) be- tween two colonies were routinely carried out as follows. A small piece was cut from the periphery of each colony with a razor blade. Two allogeneic or syngeneic colony pieces of the same size were brought into contact at their growing edges on a glass slide and were allowed to adhere to the slide in a moisture chamber for 30 min. These colony pieces were subsequently reared in a running seawater aquarium and were observed periodically under a binocular stereomi- croscope. Specimens undergoing fusion or rejection were fixed for 2 h on ice in a glutaraldehyde solution containing M sucrose buffered with M sodium cacodylate at pH The fixed specimens were rinsed in the same buffer and were then postfixed with 1% osmium tetroxide in the same buffer without sucrose for h. After dehydration through an ethanol series, the specimens were cleared with //-butyl glycidyl ether and embedded in low-viscosity epoxy resins. Thick sections were stained with toluidine blue and examined under a light microscope. Thin sections were double-stained with uranyl acetate and lead citrate and then examined with a Hitachi HS-9 transmission electron micro- scope at 75 kV. Results Allorejection reaction in Botryllus scalaris In B. scalaris, as in other botryllids. autogeneic or syn- geneic colony pairs fuse and form a single mass. The fusion process was the same as that of other botryllids (Katow and Watanabe. 1980; Saito and Watanabe. 1982). First, the tunics of two colonies fused. Second, the ampullae (termini of blood vessels) penetrated into the opposite colony and the ampullar tips came into contact with the sides of opposite ampullae (tip-to-side contact); at these contact points the colonies fused with each other
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