. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 4cS6 LUTZ WIESE AND CHARLES B. METZ types were mixed and immediately 50 ml of the mixed suspension were added to each of the following samples: A. 50 nil Tris-buffer + 25 mg trypsin + ing MgSO4 B. 50 ml Tris-buffer + 25 mg trypsin + mg MgSO4 -f- 50 mg trypsin inhibitor C. 50 ml Tris-buffer + mg MgSO4 D. 50 ml Tris-buffer + mg MgSO4 + 50 mg trypsin inhibitor E. 50 ml Tris-buffer + 25 mg trypsin + mg MgSO4. 50 mg trypsin inhibitor (dissolved in additional 10 ml buffer) were added to this sample after 1
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 4cS6 LUTZ WIESE AND CHARLES B. METZ types were mixed and immediately 50 ml of the mixed suspension were added to each of the following samples: A. 50 nil Tris-buffer + 25 mg trypsin + ing MgSO4 B. 50 ml Tris-buffer + 25 mg trypsin + mg MgSO4 -f- 50 mg trypsin inhibitor C. 50 ml Tris-buffer + mg MgSO4 D. 50 ml Tris-buffer + mg MgSO4 + 50 mg trypsin inhibitor E. 50 ml Tris-buffer + 25 mg trypsin + mg MgSO4. 50 mg trypsin inhibitor (dissolved in additional 10 ml buffer) were added to this sample after 15 min. 80 60 40 ^. 20 =3 80 60 40 20 PRETREATED mixing of the\sexes f^l-di it- A it- it i> UNTREATED mixing // of the i sexes I. mm 15 10 5 0 5 10 15 20 25 30 35 time FIGURE 2. Trypsin inhibition of pair formation in C. mocit'itsii. Different application of trypsin (at mixing of the sexes or as pre-incubation) in presence or absence or trypsin inhibitor. A and B: Gametes pretreated with trypsin 15 min. before mixing of the sexes. A. Trypsin inhibitor () added at mixing; trypsin content adjusted to final concentration. B. Without inhibitor. C, D and E: Same material not pretreated. Added at mixing of the sexes were C. trypsin, D. trypsin and inhibitor, and E. inhibitor. Microscopic examination of the samples showed that during the entire course of the experiment the trypsin-treated gametes were not impaired with respect to flagellation, motility, and agglutinability. However, the trypsin-treated samples showed markedly reduced pair formation. In a representative experiment (Fig. 1) the control gamete mixture yielded over 80% pairing (curve C , curve D ) whereas the trypsin-treated sample (curve A) was practically devoid of pairs. Trypsin-inhibitor, added simultaneously with trypsin at the mixing of the gametes, yielded full pairing (curve B, ). Addition of trypsin-inhibitor 15 minutes after trypsin application still
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Keywords: ., bookauthorlilliefrankrat, booksubjectbiology, booksubjectzoology
