Specific interaction of proteins with 5 S RNA and tRNA in the 42 S storage particle of Xenopus oocytes

During early oogenesis in amphibia, most of the 5 S RNA and tRNA is stored in a ribonucleoprotein particle that sediments at 42 S. In Xenopus laevis the 42 S particle contains two major proteins: of M_r 48 000 (P48) and 43 000 (P43). It is shown that heterogeneity in composition of the 42 S particle reflects a changing situation whereby initially, both 5 S RNA and tRNA are complexed with P48 (1 molecule 5 S RNA: 1 molecule P48; 2 or 3 molecules tRNA: 1 molecule P48), but later, tRNA becomes increasingly associated with P43 (in a 1:1 ratio) although 5 S RNA remains complexed with a cleavage product of P48. These changes relate to the eventual utilization of the excess 5 S RNA and tRNA in ribosome assembly and protein synthesis.     

利用腺病毒DNA与爪蟾卵提取物在非细胞体系中重构细胞核(简报)

1983年,Lohka和Masui首先报道,用精子染色质与卵提取物一起温育,可以组装成具有典型结构的细胞核。这种核不仅具有合成DNA的能力,而且还能进入M期。同年,     

Multiple roles of the mitogen-activated protein kinase kinase/mitogen-activated protein kinase cascade in Xenopus laevis

Mitogen-activated protein kinase (MAPK) was originally identified as a serine/threonine protein kinase that is rapidly activated in response to various growth factors and tumor promoters in mammalian cultured cells. The kinase cascade including MAPK and its direct activator, MAPK kinase (MAPKK), is now believed to transmit various extracellular signals into their intracellular targets in eukaryotic cells. It has been reported that activation of MAPKK and MAPK occurs during the meiotic maturation of oocytes in several species, including Xenopus laevis . Studies with neutralizing antibodies against MAPKK, MAPK phosphatases and constitutively active MAPKK or MAPK have revealed a crucial role of the MAPKK/MAPK cascade in a number of developmental processes in Xenopus oocytes and embryos.     

A Drosophila receptor-type tyrosine kinase (DFR1) acts as a fibroblast growth factor receptor in Xenopus embryos

Members of the fibroblast growth factor (FGF) family play important roles in various developmental processes in vertebrates. Since two genes closely related to the vertebrate FGF receptor (FGFR) genes DFR1 and DFR2/breathless have already been reported in Drosophila , the existence of a Drosophila FGF has been predicted. In the present study, we examined whether DFR1 is functionally interchangeable with a vertebrate FGFR in the Xenopus system. First, we found that the expression of DFR1 promoted Ca²⁺ efflux in response to human basic (b)FGF in Xenopus oocytes, whereas the coexpression of a dominant negative form of DFR1 (ΔDFR1) with a chick FGFR1/cek1 inhibited promotion of Ca²⁺ efflux induced by the expression of cek1 in the oocyte. Second, the expression of ΔDFR1 was observed to induce a defect in the posterior structure of the Xenopus embryo at stage 30, as observed with a dominant negative form of cek1 (Δcek1). Third, we found that the expression of ΔDFR1 inhibited the expression of FGF-regulated genes such as Xbra, Xnot , and Xshh in Xenopus embryos at stage 11, while the coexpression of DFR1 with ΔDFR1 could rescue the inhibited expression of FGF-regulated genes. These results indicate that DFR1 acts as an FGFR in Xenopus embryos and that an FGF is likely to exist in Drosophila .     

鲫鱼视网膜谷氨酸受体和GABA受体在爪蟾卵母细胞中的表达

我们以两栖类卵母细胞为功能表达系统,通过注射鲫鱼（Ｃａｒａｓｓｉｕｓｃａｒａｓｓｉｕｓ）视网膜ｍＲＮＡ,利用电压箝及药物灌流手段,系统地研究了鲫鱼视网膜内氨基酸受体的类型和特征,结果如下：（１）Ｇｌｕ受体：ＫＡ可以诱发明显的去极化电流,而且Ｄｉａｚｏｘｉｄｅ能增强ＫＡ诱导的反应,这提示鲫鱼视网膜内某些Ｃｌｕ受体是ＡＭＰＡ选择性亚型（ＡＭＰＡ－ｐｒｅｆｅｒｒｉｎｇｓｕｂｔｙｐｅ）。（２）ＣＡＢＡ受体：ＧＡＢＡ能诱发一个快速、光滑的内向电流,绝大部分对ＧＡＢＡ的反应可被ｂｉｃｕｃｕｌｌｉｎｅ所压抑,而ＧＡＢＡ＿Ｂ受体的激动剂ｂａｃｌｏｆｅｎ则无任何作用,这提示,鲫鱼视网膜内大部分是ＧＡＢＡ＿Ａ受体。     

反义寡核苷酸片段可抑制鲫鱼视网膜电压依赖性钾离子通道的表达

爪蟾卵母细胞经注射鲫鱼视网总ＲＮＡ后,可表达大量电压依赖性钾离子通道。在此基础上,我们进一步发现,一个特定序列的寡核苷酸能专一地抑制该通道的表达。由于我们设计与合成的该片段与果蝇、小鼠中已克隆的钾通道中编码Ｎ端的一个多肽的ｍＲＮＡ完全互补,因此推测：鲫鱼视网膜中的Ｋ这个区域与其它物种的Ｋ通道有高度同源性,这为克隆该基因和研究它的功能提供了重要资料。     

Identification of members of the HSP30 small heat shock protein family and characterization of their developmental regulation in heat-shocked xenopus laevis embryos

In the present study we have characterized the synthesis of members of the HSP30 family during Xenopus laevis development using a polyclonal antipeptide antibody derived from the carboxyl end of HSP30C. Two-dimensional PAGE/immunoblot analysis was unable to detect any heat-inducible small HSPs in cleavage, blastula, gastrula, or neurula stage embryos. However, heat-inducible accumulation of a single protein was first detectable in early tailbud embryos with an additional 5 HSPs at the late tailbud stage and a total of 13 small HSPs at the early tadpole stage. In the Xenopus A6 kidney epithelial cell line, a total of eight heat-inducible small HSPs were detected by this antibody. Comparison of the pattern of protein synthesis in embryos and somatic cells revealed a number of common and unique heat inducible proteins in Xenopus embryos and cultured kidney epithelial cells. To specifically identify the protein product of the HSP30C gene, we made a chimeric gene construct with the Xenopus HSP30C coding sequence under the control of a constitutive promoter. This construct was microinjected into fertilized eggs and resulted in the premature and constitutive synthesis of the HSP30C protein in gastrula stage embryos. Through a series of mixing experiments, we were able to specifically identify the protein encoded by the HSP30C gene in embryos and somatic cells and to conclude that HSP30C synthesis was first heat-inducible at the early tailbud stage of development. The differential pattern of heat-inducible accumulation of members of the HSP30 family during Xenopus development suggests that these proteins may have distinct functions at specific embryonic stages during a stress response.     

爪蟾脊索在决定过程中和相邻细胞的关系

已经知道,对预定脊索的决定起重要作用的是位于它两侧的预定肌节。电子显微镜的观察指出,预定脊索和肌节细胞相互靠得很近,或者相隔一定距离,以突起相连形成腔隙。有被小窝和小泡在两类细胞的外缘常被观察到。最引人注意的是在肌节细胞近腔隙的部位或者附近,球状体的出现。它们大小不等,内含物主要是颗粒,有的松散分布,有的致密地充满整个球状体。这些颗粒的大小和电子染色与这时期胚胎细胞中的核糖体很相似。在预定脊索细胞中以及附近,未见上述结构,但是,观察到它们伸出突起包吞腔隙中物质的现象。讨论了这些球状体的出现与脊索决定之间可能存在的关系。