摘要:

AbstractThis study was undertaken to characterize the expression of a chimeric growth factor receptor composed of the extracellular and transmembrane domains of the platelet-derived growth factor (PDGF)β-receptor (PDGFR-β) fused to the intracellular domain of the fibroblast growth factor receptor-1 (FGFR-1) and to assess its effect on the growth potential of pancreatic islet cells. For this purpose rat pancreatic islets or monolayers of pancreatic islet cells were transfected with recombinant DNA constructs coding for the PDGF B-chain, the PDGFR-β, the FGFR-1 and the chimera between PDGFR-βand FGFR-1. DNA synthesis, monitored as the percentage of labelled nuclei and [3H]thymidine incorporation, was stimulated in pancreatic islet cells cotransfected with the constructs coding for the PDGF B-chain and the PDGFR-βor the chimeric PDGFR-β/FGFR-1 as compared with that determined after transfection with control plasmid. PDGF-BB stimulated DNA synthesis when islet cells had been transfected with PDGFR-βor PDGFR-β/FGFR-1. Cotransfection of the PDGFR-βand the chimeric PDGFR-β/FGFR-1 constructs attenuated the stimulation of DNA synthesis in response to PDGF-BB. Receptor binding studies showed binding with a Kdof 0.7 nM to the chimeric receptor. The present findings show that when the chimeric PDGFR-β/FGFR-1 construct is expressed inβ-cells it is efficient in increasing DNA synthesis when stimulated with ligand.

ISSN:0897-7194    

DOI:10.3109/08977199209011013

出版商:Taylor&Francis    

年代:1992

数据来源: Taylor

摘要:

AbstractThe structure of the murine interleukin-4 receptor (mIL-4R) gene has been determined. The gene spans approximately 25 kilobases (kb) of DNA and is composed of 12 exons interrupted by 11 introns. The gene contains sequences accounting for all the sequences present in the functional mIL-4R cDNAs, including several exons which can encode DNA inserts found in recently cloned IL-4R cDNA variants. Thus expression of the gene may be regulated, at least in part, by alternative splicing. A combination of SI nuclease protection and primer extension assays was used to localize the 5′end of the gene and demonstrated the use of multiple transcription initiation sites within this region. We have found that the overall intron-exon organization of the murine IL-4R gene is markedly similar to that of the murine erythropoietin receptor (m-epoR) and the human interleukin-2 receptor beta chain (ML-2Rβ), as well as the human growth hormone receptor (hGhR). This is consistent with the recent grouping of these receptors, on the basis of protein sequence homology, into the hematopoietin receptor gene superfamily. Such homology at the levels of both protein and gene structure suggest a divergent evolution of the receptors from a single primordial gene.

ISSN:0897-7194    

DOI:10.3109/08977199209011014

出版商:Taylor&Francis    

年代:1992

数据来源: Taylor

摘要:

AbstractMurine receptors for granulocyte-macrophage colony-stimulating factor (GM-CSF) and Multi-CSF (interleukin-3) can exist in both high- and low-affinity forms and demonstrate trans-modulation by several different ligands. In contrast the recently cloned human GM-CSF receptor and murine interleukin-3 (IL-3) receptor display only low-affinity binding. to begin to understand the molecular basis of the formation of high- and low-affinity receptors and their trans-modulation we have developed methods for the solubilization and assay of GM-CSF and interleukin-3 receptors so that their binding characteristics can be studied in cell-free solution. Both receptors displayed a single class of high-affinity binding on intact FDC-P1 cells and IL-3 receptors had unaltered binding characteristics in cells, membranes and in detergent solution. However, GM-CSF receptors were converted to a single class of low-affinity binding in detergent solution while both high- and low-affinity forms were evident in membranes. The basis of affinity conversion of GM-CSF receptors was exclusively a change in the kinetic dissociation rate of ligand. Cross-linking experiments suggested that high-affinity receptors for GM-CSF and IL-3 might consist of two different protein species and, if this is so, the data suggest that this association is more stable for IL-3 than for GM-CSF receptors.

ISSN:0897-7194    

DOI:10.3109/08977199209011015

出版商:Taylor&Francis    

年代:1992

数据来源: Taylor

摘要:

AbstractTransgenic mice aberrantly expressing the granulocyte-macrophage colony stimulating factor (GM-CSF) gene develop an unusual syndrome of blindness, tissue damage and wasting which is associated with accumulations of hemopoietic cells. In order to further characterize this disease state, we have used messenger RNA detection techniques to show that the genes for rumor necrosis factor (TNFα), interleukin-1α(IL-1α) and basic fibroblast growth factor (bFGF) are expressed at abnormally high levels in both macrophages and granulocytes in transgenic mice. Furthermore, since these cell types also express the GM-CSF transgene, it is likely that they are autocrine stimulated by GM-CSF. These observations raise the possibilities that, first, the expression of tumor necrosis factorα, interleukin-1αand basic fibroblast growth factor in hemopoietic cells is a direct consequence of their autostimulation by GM-CSF, and second, that these cytokines may be responsible for some aspects of the transgenic mouse disease.

ISSN:0897-7194    

DOI:10.3109/08977199209011016

出版商:Taylor&Francis    

年代:1992

数据来源: Taylor

摘要:

AbstractThe localization of acidic fibroblast growth factor (aFGF) in the male mouse brain was studied with biochemical and immunocytochemical techniques. Using two peptide-based aFGF antisera directed against independent epitopes, Western gel analysis of dissected brain demonstrated significant levels of aFGF immunoreactivity in the pons-medulla, hypothalamus and cerebellum. The cortex contained much less immunoreactivity. Consistent with the biochemical data, immunocytochemical analysis with the same two antisera demonstrated that aFGF immunoreactivity is localized in neuronal cell bodies in these regions. Numerous immunoreactive neurons were observed in the reticular formation of the pons and medulla, as well as in several other brainstem nuclei and areas. Immunoreactive neurons were also present in the lateral and medial hypothalamus, and some thalamic, subthalamic and epithalamic nuclei. In the basal ganglia, immunoreactive neurons were present in the amygdala and septum. Few intensely stained immunoreactive neurons were observed in the striatum, pallidum and neocortex. Limbic cortices contained more numerous immunoreactive neurons than neocortex. These results support the concept that aFGF is present in the brain, where it is heterogeneously distributed in neuronal cell bodies in regions involved in sensory, extrapyramidal motor, limbic and autonomic functions. The results are consistent with various neurotrophic, autogenic, and neuromodulatory functions associated with aFGF in the mammalian central nervous system.

ISSN:0897-7194    

DOI:10.3109/08977199209011017

出版商:Taylor&Francis    

年代:1992

数据来源: Taylor

摘要:

AbstractPreviously we reported that,in vitro, lens cells proliferate, migrate or differentiate in response to low, medium and high concentrations of FGF respectively. to examine further the role of FGF in lens development we used immunohistochemistry to study the distribution of aFGF and bFGF in the eye of the 20 day rat foetus. Strong aFGF-like reactivity was localised in a band of cells near the lens equator which included the germinative zone where most cell proliferation occurs and the transitional zone where epithelial cells differentiate into fibres. The closely apposed inner epithelial layer of the ciliary and iridial retina also reacted strongly. Reactivity for aFGF was also found in the epidermis and in the corneal and conjunctival epithelia. In the neural retina, reactivity was found in the nerve fibre layer and in isolated cells of the inner plexiform layer. bFGF-like reactivity was found in the retinal ganglion cell layer, extra-ocular muscles and associated with endothelial cells of the hyaloid, lenticular and choroid vasculatures. Pre-digestion of sections with hyaluronidase caused loss of cell-associated reactivity but revealed strong bFGF-like reactivity in ocular basement membranes, in particular, the lens capsule. The sensitivity of this capsular bFGF localisation to heparinase indicates that bFGF in the extracellular matrix is complexed with heparan sulphate proteoglycans. The results of this study are consistent with the hypothesis that FGF plays an important role in lens development via both autocrine and paracrine mechanisms.

ISSN:0897-7194    

DOI:10.3109/08977199209011018

出版商:Taylor&Francis    

年代:1992

数据来源: Taylor