. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. I'ROTAMINES IN A HOLOCEPHALAN FISH 189 •"•"' '.:-- •'.« - : ' V * -. ' B V •' s" & £ - 3 -^ft- D '**$* lx A v<. Rl tf R2 « R3 -H4 -SP3-5 -SP6 -P A B C D E Figure 1. Left: Midsagittal sections of Hydrolagus collici testis showing follicles containing different stages of spermiogenesis. Feulgen staining, zones A-D; hematoxylin-eosin staining. Zone E. Zones A and B, sperniatids beginning the process of nuclear elongation; Zone C, mixture of spermatids undergoing nuclear elongation and spermatids beginning
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. I'ROTAMINES IN A HOLOCEPHALAN FISH 189 •"•"' '.:-- •'.« - : ' V * -. ' B V •' s" & £ - 3 -^ft- D '**$* lx A v<. Rl tf R2 « R3 -H4 -SP3-5 -SP6 -P A B C D E Figure 1. Left: Midsagittal sections of Hydrolagus collici testis showing follicles containing different stages of spermiogenesis. Feulgen staining, zones A-D; hematoxylin-eosin staining. Zone E. Zones A and B, sperniatids beginning the process of nuclear elongation; Zone C, mixture of spermatids undergoing nuclear elongation and spermatids beginning the process of spiralization; Zone D, spermatids undergoing spiralization; Zone E, sperm organized into tightly packed cysts. Scale (in A) denotes 10 /jm for each zone. Right: Elec- trophoretic profiles of basic proteins extracted with N HC1 (without reduction) from dissected testicular zones. Lane 1, //. collici whole testis; lanes 2-6, //. colliei testicular zones A-E. respectively; lane 7, standards of frog (XeniipM /i/cv/.v) sperm basic proteins (histones and proteins SP 3-5, SP 6) and herring protamine (P). H4 = evolutionary conservative histone H4. Electrophoresis in this and subsequent figures is from top (+) to bottom (-). that sperm-specific basic protamines of //. collici consist of three major (Rl, R2, and R3) and four minor (mO, ml, m2, and m3) proteins. Only two of them (R3 and mO) can be extracted from sperm nuclei without previous reduction. In addition, Rl and R2 reproducibly have a slightly lower mobility when extracted without reduction from testis (Fig. 2, lane f) than when extracted with re- duction from epididymal sperm (Fig. 2, lane g). Purification ami analysis of proteins We used sperm cells from epididymis, as well as anterior and posterior parts of the ampulla ductus deferentis, for the purification of ratfish nuclear sperm proteins. Nuclei were extracted directly with N HC1, cleared by cen- trifugation; and reduced, alkylated, and
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Keywords: ., bookauthorlilliefrankrat, booksubjectbiology, booksubjectzoology
