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一种含N-连接碳水化合物的细胞外基质决定因素在海胆原肠胚形成中起关键作用。

An N-linked carbohydrate-containing extracellular matrix determinant plays a key role in sea urchin gastrulation.

作者信息

Ingersoll E P, Ettensohn C A

机构信息

Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, Pennsylvania 15213.

出版信息

Dev Biol. 1994 Jun;163(2):351-66. doi: 10.1006/dbio.1994.1154.

Abstract

During the process of gastrulation in the sea urchin embryo, the vegetal plate invaginates to form the archenteron, a long narrow tube that extends across the blastocoel. Rearrangement of cells within the archenteron is thought to be a key component of the process of archenteron elongation. While these cell rearrangements have been well described, the mechanism of the rearrangements and the coordination of the movements of individual cells that results in the elongation of the archenteron are not well understood. We have identified a monoclonal antibody, called ECM 1, that recognizes an N-linked carbohydrate-containing epitope on several high-molecular-weight basal lamina glycoproteins. In an attempt to block the function of this determinant in development, the ECM 1 antibody was injected into the blastocoel of living embryos and the effects on morphogenesis were determined. Injection of intact ECM 1 IgG or monovalent Fab fragments blocks cell rearrangements during secondary invagination and cell movements during segmentation of the gut. Glycopeptides derived from the glycoproteins recognized by ECM 1 also inhibit cell rearrangements during secondary invagination when injected into the blastocoel. The inhibitory activity of these peptides is eliminated by digestion with N-glycosidase F or pronase, enzymes that also disrupt the ECM 1 determinant. ECM 1 recognizes a nonuniformly distributed determinant in the basal lamina and blastocoel matrix. This determinant is stored in cytoplasmic granules in the unfertilized egg and is deposited into the basal lamina by the blastula stage. The determinant becomes concentrated in the basal lamina in the vegetal region of the embryo early in gastrulation. At the prism stage, the determinant accumulates in the basal lamina and blastocoel matrix in the ventral region of the embryo. These data indicate that the vegetally localized, N-linked carbohydrate-containing determinant recognized by ECM 1 plays an important role in cell movements during archenteron morphogenesis in the sea urchin embryo.

摘要

在海胆胚胎原肠胚形成过程中,植物极板内陷形成原肠,即一根横跨囊胚腔的狭长管道。原肠内细胞的重排被认为是原肠伸长过程的关键组成部分。虽然这些细胞重排已得到充分描述,但重排机制以及导致原肠伸长的单个细胞运动的协调过程仍未得到很好的理解。我们鉴定出一种名为ECM 1的单克隆抗体,它能识别几种高分子量基膜糖蛋白上含N - 连接碳水化合物的表位。为了在发育过程中阻断该决定簇的功能,将ECM 1抗体注入活胚胎的囊胚腔,并确定其对形态发生的影响。注入完整的ECM 1 IgG或单价Fab片段会阻断二次内陷过程中的细胞重排以及肠道分割过程中的细胞运动。当注入囊胚腔时,源自ECM 1识别的糖蛋白的糖肽也会抑制二次内陷过程中的细胞重排。用N - 糖苷酶F或链霉蛋白酶消化这些肽可消除其抑制活性,这两种酶也会破坏ECM 1决定簇。ECM 1识别基膜和囊胚腔基质中分布不均匀的决定簇。该决定簇存储在未受精卵的细胞质颗粒中,并在囊胚期沉积到基膜中。在原肠胚形成早期,该决定簇在胚胎植物区域的基膜中富集。在棱柱期,该决定簇在胚胎腹侧区域的基膜和囊胚腔基质中积累。这些数据表明,ECM 1识别的植物区域定位的、含N - 连接碳水化合物的决定簇在海胆胚胎原肠形态发生过程中的细胞运动中起重要作用。

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