. Die biologische Bulletin. Biologie; Zoologie; Biologie; Meeresbiologie. 30 E.HIROSE UND T. ISHII Materialien und Methoden Tiere die Kolonien von Apliilhtin yainaiii wurden in Nabeta Bay, Shimoda (Präfektur Shizuoka, Japan) gesammelt. Sie waren vorübergehend in Betrieb Meerwasser in der Labo-ratory oder in einer Kultur, in Nabeta Bucht getaucht aufgezogen. A. vamaiii bildet eine relativ flache Kolonie, etwa 2 bis 3 mm Dicke, die sich auf einer flachen Unterlage; seine stabförmigen Zooids sind separat in einem gemeinsamen Tunika (Bild eingebettet. 1). Tunika Rundung assay Kolonien wurden Quer mit einer Rasierklinge in p in Scheiben geschnitten

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. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 30 E. HIROSE AND T. ISHII Materials and Methods Animals The colonies of Apliilhtin yainaiii were collected in Nabeta Bay, Shimoda (Shizuoka Prefecture, Japan). They were temporarily kept in running seawater in the labo- ratory or reared in a culture box immersed in Nabeta Bay. A. vamaiii forms a relatively flat colony, about 2 to 3 mm thick, spreading on a flat substratum; its rod-shaped zooids are embedded separately in a common tunic (Fig. 1). Tunic rounding assay Colonies were transversely sliced with a razor blade into pieces that were 0.5 mm thick or less (e.g.. 7 X 2 X 0.5 mm). The tunic slices (Hiroset'/a/., 1994a) were com- posed of tunic, tunic cells, and small fragments of zooids; many of the zooid fragments were washed out. The fresh tunic slices were placed in a plastic petri dish filled with filtered seawater (FSW) or artificial seawater (ASW), and were incubated overnight at 17° to 20°C. During the in- cubation, each specimen shrank and became a single tunic ball. Video recording of tunic rounding The tunic slices were put in a 100-ml beaker filled with FSW (16°-18°C), and the process of tunic rounding was recorded with a time-lapse videocassette recorder (AG- 6010; National, Osaka, Japan) and a video camera (WV- 1800; National) equipped with a 55-mm macro lens (Mi- cro-Nikkor; Nikon, Tokyo). Recording was performed at about 1/60 of the actual speed. The time course of trans- formation was analyzed by hourly measurements of the length of the longest diagonal line that could be drawn within the profile of the rounding specimens. Tunic rounding assay under experimental conditions Some of the slices were frozen at -20°C. thawed at room temperature, and then incubated in ASW. Live slices were incubated in three kinds of experi- mental media: Ca2+-Mg24-free artificial seawater (CMF- ASW), various concentrations of colchicine-ASW. and various concentrations of cytochalasin B-ASW. Beca

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