Desensitisation of cultured glial cells to epidermal growth factor by receptor down-regulation.

Epidermal growth factor (EGF), which can be purified from the mouse submaxillary gland or from pregnant human urine, is a potent multiplication-stimulating factor for several types of cultured cells, including human fibroblasts and glial cells. The molecule binds with high affinity and saturation kinetics to a cell-surface receptor, is subsequently internalised and finally degraded. The binding event is accompanied by a reduction in the number of EGF receptors. This phenomenon--'receptor down-regulation'--has been demonstrated with several hormones and may be a general principle for the modulation of binding groups on the outer cell surface. Further, it has been proposed that receptor loss acts to regulate the cellular response to the binding ligand. The present study provides direct experimental support for this hypothesis. It demonstrates that down-regulation of EGF receptors on glial cells causes desensitisation of the mitogenic response of these cells to subsequent stimulation with EGF.     

A RhoGDP dissociation inhibitor spatially regulates growth in root hair cells

Root hairs are cellular protuberances extending from the root surface into the soil; there they provide access to immobile inorganic ions such as phosphate, which are essential for growth. Their cylindrical shape results from a polarized mechanism of cell expansion called tip growth in which elongation is restricted to a small area at the surface of the hair-forming cell (trichoblast) tip. Here we identify proteins that spatially control the sites at which cell growth occurs by isolating Arabidopsis mutants (scn1) that develop ectopic sites of growth on trichoblasts. We cloned SCN1 and showed that SCN1 is a RhoGTPase GDP dissociation inhibitor (RhoGDI) that spatially restricts the sites of growth to a single point on the trichoblast. We showed previously that localized production of reactive oxygen species by RHD2/AtrbohC NADPH oxidase is required for hair growth; here we show that SCN1/AtrhoGDI1 is a component of the mechanism that focuses RHD2/AtrbohC-catalysed production of reactive oxygen species to hair tips during wild-type development. We propose that the spatial organization of growth in plant cells requires the local RhoGDI-regulated activation of the RHD2/AtrbohC NADPH oxidase.     

Nerve growth factor is a mitogen for cultured chromaffin cells

Nerve growth factor (NGF) is essential for the survival and differentiation of a number of neural crest derivatives, including sympathetic and sensory neurones. While early studies suggested that NGF might also have a mitogenic effect on these neurones, subsequent work has favoured the interpretation that NGF promotes cell survival or differentiation rather than proliferation. We have addressed the issue of a mitogenic effect of NGF using adrenal chromaffin cells, which are endocrine cells derived from the neural crest, and are closely related to sympathetic neurones. Adrenal chromaffin cells respond to NGF in vitro by expressing neuronal traits. We now report that NGF elicits a mitotic response in cultured chromaffin cells from young rats, and that this response is blocked by an antiserum to 2.5S NGF. The chromaffin cells that divided in response to NGF can subsequently become neuronal in the continued presence of NGF.     

Induction of a novel epidermal growth factor-secreting cell lineage by mucosal ulceration in human gastrointestinal stem cells

Epidermal growth factor, and its human homologue urogastrone (EGF/URO), are secreted by the gut-associated salivary and Brunner's glands. Recombinant EGF/URO is a powerful stimulator of cell proliferation and differentiation in the rodent and neonatal human intestine. But EGF/URO is not absorbed from the adult gut and has no action when given through the gut lumen; thus the role of secreted EGF/URO is unknown. We now report that ulceration of the epithelium anywhere in the human gastrointestinal tract induces the development of a novel cell lineage from gastrointestinal stem cells. This lineage initially appears as a bud from the base of intestinal crypts, adjacent to the ulcer, and grows locally as a tubule, ramifying to form a new small gland, and ultimately emerges onto the mucosal surface. The lineage produces neutral mucin, shows a unique lectin-binding profile and immunophenotype, is nonproliferative, and contains and secretes abundant immunoreactive EGF/URO. We propose that all gastrointestinal stem cells can produce this cell lineage after mucosal ulceration, secreting EGF/URO to stimulate cell proliferation, regeneration and ulcer healing. This cell lineage is very commonly associated with gastrointestinal mucosal ulceration, and we conclude that a principal in vivo role for EGF/URO is to stimulate ulcer healing throughout the gut through induction of this cell lineage in the adjacent mucosa.     

Involvement of the epidermal growth factor receptor in the invasion of cultured mammalian cells by Salmonella typhimurium.

Salmonella infection continues to be a major world-wide health problem. One essential pathogenic feature common to all Salmonella is their ability to penetrate the cells of the intestinal epithelium which are normally non-phagocytic. The internalization of Salmonella into mammalian cells is thought to be a receptor-mediated phenomenon and the invasion of cultured epithelial cells depends on several Salmonella genes, but nothing is known about the host determinants participating in this interaction. Protein tyrosine phosphorylation follows stimulation of many cell-surface receptors to initiate signal transduction pathways that stimulate cellular responses. We report here that invasion of cultured Henle-407 cells by Salmonella typhimurium induces the tyrosine phosphorylation of the epidermal growth factor (EGF) receptor. In contrast, an isogenic strain of S. typhimurium that is defective in invasion owing to a mutation in the invA gene is unable to induce such phosphorylation. Addition of EGF to cultured Henle-407 cells allowed the internalization of the invasion-defective S. typhimurium invA mutant although it did not cause the internalization of an adherent, but non-invasive, strain of Escherichia coli. This result indicates that stimulation of the EGF receptor is involved in the invasion of cultured Henle-407 cells by S. typhimurium.     

PSK对杂交鹅掌楸悬浮培养细胞生长的影响

以杂交鹅掌楸悬浮培养胚性细胞为材料,在培养过程中添加不同浓度的新型植物肽类生长调节物质植物磺肽素(PSK),研究PSK对悬浮细胞生理的影响,分析其在细胞培养中的作用。结果表明:PSK能够促进细胞分裂和增殖,0.8 mg/L的PSK处理后能够使杂交鹅掌楸胚性愈伤组织细胞结构致密、胚性细胞小、直径一致性增加、细胞质变浓,细胞内大量积累淀粉等,同时PSK显著提高了细胞的活力,促进了培养细胞低密度的增殖,并且能够促进蛋白质的积累,有利于杂交鹅掌楸体细胞胚胎的发生。     

不同培养基对山羊毛囊体外培养的影响研究

为得到1种简捷而适宜的山羊毛囊体外培养方法,采用3因子3水平的拉丁方正交试验设计(L9(34))方法,对比研究了无血清的Williams E培养基、无血清的DMEM培养基和添加10％FCS(胎牛血清)的DMEM培养基,及其在3种培养基中分别添加0、2和10ng／mL 3种不同质量浓度的血管内皮细胞生长因子(VEGF)和表皮细胞生长因子(EGF)时对毛囊体外培养的影响。将成年青山羊皮肤上的单根毛囊完整地分离出来,并同时平行培养在9种培养基中,借助于倒置显微镜和目镜测微尺随时不断地观察毛囊生长的形态变化和测量毛根的长度。结果发现:山羊游离毛囊在9种培养基中均能很好地生长;但以无血清的培养基和在Williams E培养基的效果最好;分别和同时添加2％～10％的VEGF和EGF时可加快毛囊的生长;添加10％FCS和不添加细胞生长因子时毛囊也能生长,但生长速度较慢。     

Induction of yeast mating pheromone a-factor by alpha cells

Saccharomyces cerevisiae cells of a and alpha mating types constitutively secrete cell-specific peptide mating pheromones. a-Factor is secreted by a cells and acts on alpha cells, while alpha-factor is secreted by alpha cells and acts on a cells. Confirming preliminary studies, we demonstrate here that cultures of a cells contain higher than constitutive levels of a-factor activity when grown with alpha cells or alpha-factor. This induction of a-factor may result from increased synthesis or increased secretion of a-factor, as opposed to modification or stabilization of preexisting a-factor, as part of the a cell response to alpha-factor, as an a ste2 mutant (which cannot respond to alpha-factor) is not induced by alpha-factor. In mixed cultures inoculated with equal numbers of a cells and alpha cells, a cells predominate by stationary phase. Thus, a series of sequential interactions between a and alpha cells may be involved in establishing optimal hormone concentrations and cell ratios for conjugation.     

Mycoplasma infection of cultured cells

Mycoplasma contamination is tough to detect and even more difficult to eradicate. It is best to start over fresh from clean cell stocks, but several elimination options are available.     

水解酪蛋白对烟草愈伤组织和悬浮培养细胞生长的促进作用

报道了水解酪蛋白对烟草愈伤组织和悬浮细胞生长的促进作用，并确定了水解酪蛋白的适宜用量。