Lin S, Long W, Chen J, Hopkins N
Biology Department, Massachusetts Institute of Technology, Cambridge 02139.
Proc Natl Acad Sci U S A. 1992 May 15;89(10):4519-23. doi: 10.1073/pnas.89.10.4519.
To determine whether embryonic cells transplanted from one zebrafish embryo to another can contribute to the germ line of the recipient, and to determine whether pigmentation can be used as a dominant visible marker to monitor cell transplants, we introduced cells from genetically pigmented (donor) embryos to albino recipients at midblastula stage. By 48 hr many of the resulting chimeras expressed dark pigment in their eyes and bodies, characteristics of donor but not albino embryos. By 4-6 weeks of age pigmentation was observed on the body of 23 of 70 chimeras. In contrast to fully pigmented wild-type fish, pigmentation in chimeras appeared within transverse bands running from dorsal to ventral. Pigmentation patterns differed from one fish to another and in almost every case were different on each side of a single fish. At 2-3 months of age chimeras were mated to albino fish to determine whether pigmented donor cells had contributed to the germ line. Of 28 chimeric fish that have yielded at least 50 offspring each, 5 have given rise to pigmented progeny at frequencies of 1-40%. The donor cells for some chimeras were derived from embryos that, in addition to being pigmented, were transgenic for a lacZ plasmid. Pigmented offspring of some germ-line chimeras inherited the transgene, confirming that they descended from transplanted donor cells. Our ability to make germ-line chimeras suggests that it is possible to introduce genetically engineered cells into zebrafish embryos and to identify the offspring of these cells by pigmentation at 2 days of age.
为了确定从一个斑马鱼胚胎移植到另一个胚胎的胚胎细胞是否能对受体的生殖系有贡献，以及确定色素沉着是否可用作监测细胞移植的显性可见标记，我们在囊胚中期将来自基因色素沉着（供体）胚胎的细胞引入白化受体。到48小时时，许多产生的嵌合体在其眼睛和身体中表达深色色素，这是供体胚胎而非白化胚胎的特征。到4至6周龄时，在70个嵌合体中的23个的身体上观察到色素沉着。与完全色素沉着的野生型鱼不同，嵌合体中的色素沉着出现在从背侧到腹侧的横向带内。色素沉着模式因鱼而异，并且在几乎每种情况下，单个鱼的两侧都不同。在2至3个月大时，将嵌合体与白化鱼交配，以确定色素沉着的供体细胞是否对生殖系有贡献。在28条每条至少产生50个后代的嵌合体鱼中，有5条产生了色素沉着的后代，频率为1%至40%。一些嵌合体的供体细胞来自除了色素沉着外还对lacZ质粒进行转基因的胚胎。一些生殖系嵌合体的色素沉着后代继承了转基因，证实它们来自移植的供体细胞。我们制造生殖系嵌合体的能力表明，有可能将基因工程细胞引入斑马鱼胚胎，并在2日龄时通过色素沉着识别这些细胞的后代。
