. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 402 C. LEMA-FOLEY ET AL. • B cells+ HB o B cells + HA • B cells + HA(1°C) 10 15 20 time (min) 25 30 Figure 13. Tissue trauma as a possible factor in Hx level. Two prep- arations of cell-free hemolymph were made from clam A: HA(1,C, was obtained via syringe after pre-cooling clam A to 1°C, and HA from the same clam was obtained by cutting muscle after 3 h of rewarming. N-cells (CB) and cell-free hemolymph for control bioassay were obtained from clam B. Three mixtures were made as indicated, with HA(I 0 exhibiting less than ha
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 402 C. LEMA-FOLEY ET AL. • B cells+ HB o B cells + HA • B cells + HA(1°C) 10 15 20 time (min) 25 30 Figure 13. Tissue trauma as a possible factor in Hx level. Two prep- arations of cell-free hemolymph were made from clam A: HA(1,C, was obtained via syringe after pre-cooling clam A to 1°C, and HA from the same clam was obtained by cutting muscle after 3 h of rewarming. N-cells (CB) and cell-free hemolymph for control bioassay were obtained from clam B. Three mixtures were made as indicated, with HA(I 0 exhibiting less than half the activity of HA. Dialysis and initial fractionation of hemolymph Active hemolymph was dialyzed for 12 h against physi- ological medium at 0°C. with a molecular weight cutoff of about 12,000. Control hemolymph was similarly treated except that the dialysis sac was simply kept moist. Bioassay with fresh N-cells showed that both samples exhibited ex- cellent activity, with the experimental comparable to the control except for an extended activation lag (Fig. 14). Centrifugation through filters with different molecular weight cutoffs (Centricon) indicated that the Mr for Hx is greater than 500,000, and chromatogaphy on Sephadex G-200SF confirmed that Hx moved together with the blue dextran marker, indicating the M, to be greater than 250,000. Discussion Shape transformation and recovery The results show that shape transformation occurs in response to an activity (Hx) in the cell-free hemolymph, and that the extent of response is a property of the erythrocyte population. In typical assays, more than 907r of cells were responsive to Hx, but individual cells, as well as cells from certain clams, varied in both responsiveness and rate of recovery. Variability was particularly clear at high hemo- lymph dilutions ( Fig. 5, H/IO), in which only a fraction nl the erythrocytes responded. This was verified by exper- iments using erythrocytes from clams with high vs. low per
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