. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 15 C 5 10 15 TIME ( min ) 20 Figure 3. Aggregation of hemocytes of Halocynthia rnrci:i in the presence of 10% plasma at 5°C and at 15°C. In the absence of plasma, no aggregation occurred even at 15°C. The extent of light scattering is shown on the ordmate in arbitrary units. inducing effects. Withinti shaking, however. Mg2+ had lit- tle inducing effect in contrast to the case of plasma: the inducing effects of plasma were detectable irrespective of shaking. Effects of various preparations ol P/CIMHU on liemocyte aggreg
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 15 C 5 10 15 TIME ( min ) 20 Figure 3. Aggregation of hemocytes of Halocynthia rnrci:i in the presence of 10% plasma at 5°C and at 15°C. In the absence of plasma, no aggregation occurred even at 15°C. The extent of light scattering is shown on the ordmate in arbitrary units. inducing effects. Withinti shaking, however. Mg2+ had lit- tle inducing effect in contrast to the case of plasma: the inducing effects of plasma were detectable irrespective of shaking. Effects of various preparations ol P/CIMHU on liemocyte aggregation We developed method A to analyze the aggregation of H. roretii hemocytes. This method is useful for tracing the progress of the reaction, but is too time-consuming for investigating the effects of various substances on ag- gregation. Therefore, we developed another quantitative method (B) in which an ELISA reader is used to measure the increased transmittance that should occur as hemocyte aggregates form. In fact, the transmission was increased at a rate consistent with the formation of aggregates de- tected microscopically. By this method, we confirmed that both plasma and Mg:+ induced the aggregation of//. n>r- etii hemocytes. To characterize the plasma factor that can induce the aggregation of //. roretii hemocytes, we used method B to examine the effects of various preparations of plasma (10%) on the aggregation. The plasma factor was stable under acidic and alkaline conditions and resistant to heat treatment; its molecular weight was less than 10,000 be- cause dialyzed plasma had little inducing activity (Fig. 5a). In fact, the filtrate (containing low molecular weight substances) obtained from intact plasma by filtration through a Centricut filter (10,000 molecular weight cut- off") had inducing activity (control experiment in Fig. 5b), whereas that obtained from dialyzed plasma had little ac- tivity (second bar in Fig. 5b). The filtrate from trypsin- tre
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Keywords: ., bookauthorlilliefrankrat, booksubjectbiology, booksubjectzoology
