The giant panda skeletal muscle cells, uterus epithelial cells and mammary gland cells from an adult individual were cultured and used as nucleus donor for the construction of interspecies embryos by transferring them...The giant panda skeletal muscle cells, uterus epithelial cells and mammary gland cells from an adult individual were cultured and used as nucleus donor for the construction of interspecies embryos by transferring them into enucleated rabbit eggs. All the three kinds of somatic cells were able to reprogram in rabbit ooplasm and support early embryo development, of which mammary gland cells were proven to be the best, followed by uterus epithelial cells and skeletal muscle cells. The experiments showed that direct injection of mammary gland cell into enucleated rabbit ooplasm, combined with in vivo development in ligated rabbit oviduct, achieved higher blastocyst development than in vitro culture after the somatic cell was injected into the perivitelline space and fused with the enucleated egg by electrical stimulation. The chromosome analysis demonstrated that the genetic materials in reconstructed blastocyst cells were the same as that in panda somatic cells. In addition, giant panda mitochondrial DNA (mtDNA) was shown to exist in the interspecies reconstructed blastocyst. The data suggest that (i) the ability of ooplasm to dedifferentiate somatic cells is not species-specific; (ii) there is compatibility between interspecies somatic nucleus and ooplasm during early development of the reconstructed egg.展开更多
The ultracytochemical localization of ATPase in the secondary xylem cells during their differentiation and dedifferentiation in the girdled Eucommia ulmoides Oliv. was carried out using a lead phosphate precipitation ...The ultracytochemical localization of ATPase in the secondary xylem cells during their differentiation and dedifferentiation in the girdled Eucommia ulmoides Oliv. was carried out using a lead phosphate precipitation technique. Throughout the differentiation, which is a typical programmed cell death (PCD) process, ATPase deposits increased in the nucleus but decreased and progressively disappeared in the cell organelles. At the same time, the distribution of ATPase increased in the inner face of the cell wall and pits with cytoplasmic degeneration. The results demonstrated that the PCD was an energy dependent active process and was controlled by nuclear genes. On the other hand, the distribution of ATPase in the intercellular spaces increased with the formation of the new cambium resulted from the dedifferentiation of the secondary xylem cells after girdling. However, ATPase was not found in the nucleus of the dividing cells, suggesting that nutrients were transported through protoplast during differentiation, and through both protoplast and apoplast during dedifferentiation. Thus, the energy required in cell division was provided mainly by intercellular spaces. These findings indicate that the dynamic distribution of ATPase reflected which cell component was actively taking part in the cell metabolism at various stages of the plant development, and its distribution was associated with the physiological state of the cell. Based on the characteristic distributions of ATPase, the critical stage of cell differentiation and the relationship between the critical stage and dedifferentiation were discussed.展开更多
地钱(Marchantia polymorpha L.)的胞芽和配子体先在 MS 培养基上补加1mg/l 2,4-D和3%蔗糖,经过启动部分脱分化后,再移入1/2KNOP 培养基补加4—8mg/l 2,4-D,0.25～0.5mg/l BA 与 MS 的铁盐,20g 蔗糖,此后愈伤组织肉眼可见,但仍伴有假根...地钱(Marchantia polymorpha L.)的胞芽和配子体先在 MS 培养基上补加1mg/l 2,4-D和3%蔗糖,经过启动部分脱分化后,再移入1/2KNOP 培养基补加4—8mg/l 2,4-D,0.25～0.5mg/l BA 与 MS 的铁盐,20g 蔗糖,此后愈伤组织肉眼可见,但仍伴有假根。最后移入 White 培养基添加丙酮酸、延胡索酸与柠檬酸三者混合物(5mmol/l)及1mg/l 2,4-D与4%葡萄糖后,始呈现彻底的脱分化状态,愈伤组织才能正常生长。整个脱分化时间长达10个月。而再分化成配子体却比高等植物容易,甚至移入不含激素的 MS 基本培养基即可形成正常的配子体。展开更多
基金Project supported by the 1998 Special Fund of the Chinese Academy of Sciences (KY95-J1-318)a 1999 Special Fund from the Ministry of ScienceTechnology.Instruments were dominated by Mr. Shum Yam Wa, Heal Force Development Ltd
文摘The giant panda skeletal muscle cells, uterus epithelial cells and mammary gland cells from an adult individual were cultured and used as nucleus donor for the construction of interspecies embryos by transferring them into enucleated rabbit eggs. All the three kinds of somatic cells were able to reprogram in rabbit ooplasm and support early embryo development, of which mammary gland cells were proven to be the best, followed by uterus epithelial cells and skeletal muscle cells. The experiments showed that direct injection of mammary gland cell into enucleated rabbit ooplasm, combined with in vivo development in ligated rabbit oviduct, achieved higher blastocyst development than in vitro culture after the somatic cell was injected into the perivitelline space and fused with the enucleated egg by electrical stimulation. The chromosome analysis demonstrated that the genetic materials in reconstructed blastocyst cells were the same as that in panda somatic cells. In addition, giant panda mitochondrial DNA (mtDNA) was shown to exist in the interspecies reconstructed blastocyst. The data suggest that (i) the ability of ooplasm to dedifferentiate somatic cells is not species-specific; (ii) there is compatibility between interspecies somatic nucleus and ooplasm during early development of the reconstructed egg.
文摘The ultracytochemical localization of ATPase in the secondary xylem cells during their differentiation and dedifferentiation in the girdled Eucommia ulmoides Oliv. was carried out using a lead phosphate precipitation technique. Throughout the differentiation, which is a typical programmed cell death (PCD) process, ATPase deposits increased in the nucleus but decreased and progressively disappeared in the cell organelles. At the same time, the distribution of ATPase increased in the inner face of the cell wall and pits with cytoplasmic degeneration. The results demonstrated that the PCD was an energy dependent active process and was controlled by nuclear genes. On the other hand, the distribution of ATPase in the intercellular spaces increased with the formation of the new cambium resulted from the dedifferentiation of the secondary xylem cells after girdling. However, ATPase was not found in the nucleus of the dividing cells, suggesting that nutrients were transported through protoplast during differentiation, and through both protoplast and apoplast during dedifferentiation. Thus, the energy required in cell division was provided mainly by intercellular spaces. These findings indicate that the dynamic distribution of ATPase reflected which cell component was actively taking part in the cell metabolism at various stages of the plant development, and its distribution was associated with the physiological state of the cell. Based on the characteristic distributions of ATPase, the critical stage of cell differentiation and the relationship between the critical stage and dedifferentiation were discussed.
文摘地钱(Marchantia polymorpha L.)的胞芽和配子体先在 MS 培养基上补加1mg/l 2,4-D和3%蔗糖,经过启动部分脱分化后,再移入1/2KNOP 培养基补加4—8mg/l 2,4-D,0.25～0.5mg/l BA 与 MS 的铁盐,20g 蔗糖,此后愈伤组织肉眼可见,但仍伴有假根。最后移入 White 培养基添加丙酮酸、延胡索酸与柠檬酸三者混合物(5mmol/l)及1mg/l 2,4-D与4%葡萄糖后,始呈现彻底的脱分化状态,愈伤组织才能正常生长。整个脱分化时间长达10个月。而再分化成配子体却比高等植物容易,甚至移入不含激素的 MS 基本培养基即可形成正常的配子体。
