. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 158 J. STOLLBERG AND S. E. ERASER croscopy to extend the accuracy and resolution of data describing the distribution of AChRs after exposure to electric fields. The use of these techniques permits rigor- ous comparison to theoretical predictions based on elec- tromigration and diffusion. Experiments presented here document and dissect the mechanisms underlying an in- triguing finding—AChRs, accumulated at the cathode- facing cell pole in an electric field, continue to aggregate there after termination of the field. Materials
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 158 J. STOLLBERG AND S. E. ERASER croscopy to extend the accuracy and resolution of data describing the distribution of AChRs after exposure to electric fields. The use of these techniques permits rigor- ous comparison to theoretical predictions based on elec- tromigration and diffusion. Experiments presented here document and dissect the mechanisms underlying an in- triguing finding—AChRs, accumulated at the cathode- facing cell pole in an electric field, continue to aggregate there after termination of the field. Materials and Methods Reagents, culture system Reagents and culture preparation were as described previously (Stollberg and Fraser, 1988). In brief, myoto- mal cells were dissected from stage 18-20 Xenopus laevis embryos (Nieuwkoop and Faber, 1962) in collagenase/ Steinberg's solution, dissociated in Ca++/Mg++-free Steinberg's solution, and maintained on sterile coverslips in drops of culture medium for 1 day at 24°C. Experimental manipulations Neuraminidase incubations were carried out in 50 /jl of Steinberg's solution, pH In the event of subse- quent trypsin digestion, the neuraminidase solution was removed and replaced with 50 n\ of trypsin in Steinberg's solution, pH Following pre-field treat- ments (where indicated), the cultures were returned to culture medium and assembled into chambers with in- ternal dimensions of 6 X 1 X cm. Following the elec- tric fields, cultures were given the indicated post-field pe- riods, chilled, and labeled with 300 nM tetra-methyl-rho- damine labeled alpha-Bungarotoxin (TMR-«-Bgt), 25 Mg/ml tluorescein labeled con A (FLR-con A) in culture medium. The cultures were then rinsed with 1 ml of cul- ture medium, and kept on ice until video images were acquired. Data analysis Data analysis techniques were as previously described (Stollberg and Fraser, 1988). In brief, fluorescent images were viewed through a Zeiss Universal microscop
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Keywords: ., bookauthorlilliefrankrat, booksubjectbiology, booksubjectzoology
