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W Risau

Publications and source records attributed to W Risau.

133 records · Page 8Linked to original sources
PubMed1986

Differentiation-dependent expression of proteins in brain endothelium during development of the blood-brain barrier.

The blood-brain barrier is a specific property of differentiated brain endothelium. To study the differentiation of blood vessels in the brain, we have correlated the expression of a number of proteins in brain endothelial cells with the development of the blood-brain barrier in mouse, quail, and chick embryos. Using histochemical methods, alkaline phosphatase activity was found to be present in all species and appeared around embryonic Days 17 (mouse), 14 (quail), and 12 (chick). Butyrylcholinesterase activity was found in the mouse and quail but not the chick brain vasculature, and appeared around Days 17 (mouse) and 15 (quail). gamma-Glutamyltranspeptidase activity was demonstrated histochemically in mouse but not in chick and quail brain capillaries, beginning at Day 15. Transferrin receptor was localized on brain endothelium in all species by immunofluorescence methods using monoclonal antibodies. It appeared at Days 15 and 11 in mouse and chick embryonic brain, respectively. The staining of all markers in embryonic brain was compared with adult brain endothelium and the leptomeningeal blood vessels. The expression of these proteins was correlated with the development of the blood-brain barrier by studying the permeability of brain endothelium for the protein horseradish peroxidase during mouse embryogenesis. Vessels in the telencephalon were found to become impermeable around Day 16 of development. Taken together the results of previous investigations and those presented here, we conclude that a number of proteins are sequentially expressed in brain endothelial cells correlating in time with the formation of the blood-brain barrier in different species.

Alkaline Phosphatase↗
PubMed1986

Developing brain produces an angiogenesis factor.

Embryonic chicken brain was analyzed for the presence of growth factors for capillary endothelial cells. A growth factor was found whose total and specific activities began to increase around the 12th day of development and reached a plateau around the 14th-16th day. A high specific activity was observed early in embryonic development at days 3 and 4. The growth factor was partially purified from extracts of 18-day-old embryonic chicken brain by heparin-Sepharose affinity chromatography where it eluted as a single peak of activity at 1.2-1.4 M sodium chloride. In chromatography on Sephacryl S200 gels, the activity eluted at a molecular size of 16- to 18-kDa. The growth factor is mitogenic for endothelial cells but not for smooth muscle or glial cells. In vivo assays using rabbit cornea and chicken chorioallantoic membrane demonstrated the angiogenic capacity of the chromatographed growth factor. The relationship between this early factor, the subsequent invasion of blood vessels into neural tissue, and the formation of the blood-brain barrier is discussed.

Age Factors↗
PubMed1986

Production of a heparin-binding angiogenesis factor by the embryonic kidney.

Embryonic mouse kidneys induce angiogenesis when transplanted on the quail chorioallantoic membrane (Ekblom, P., H. Sariola, M. Karkinen, and L. Saxén, 1982, Cell Differ., 11:35-39). In these experiments all blood vessels were derived from the quail host, suggesting that kidney endothelium is derived from outside blood vessels. We have now analyzed whether kidney angiogenesis is regulated by kidney-derived soluble factors that stimulate the growth of new blood vessels. In the rabbit cornea, 11-d embryonic kidneys induced angiogenesis, whereas uninduced 11-d kidney mesenchymes did not. To characterize and purify this activity from an embryonic organ, we dissected between 600 and 1,000 14-17-d-old embryonic mouse kidneys for each purification experiment. Growth factor activity for capillary endothelial cells was found to bind to heparin-Sepharose and eluted at 0.9-1.1 M sodium chloride. Gel filtration revealed a molecular weight of 16,000-20,000 of this factor. A major 18,000-mol-wt band was seen after gel electrophoresis and silver staining of partially purified growth factor material. The chromatographed factor is mitogenic for endothelial cells but not for smooth muscle cells and stimulates angiogenesis in vivo in the rabbit cornea. Adult kidneys contained two heparin-binding endothelial cell growth factors. The differentiation-dependent production of an angiogenesis factor by the embryonic kidney suggests an important role of angiogenesis in organogenesis.

Angiogenesis Inducing Agents↗
PubMed1983

Nonpackaging and packaging proteins of hnRNA in Drosophila melanogaster.

Monoclonal antibodies have previously been raised against chromosomal proteins of Drosophila. Using a biochemical fractionation method for the isolation of large hnRNA-containing structures (hnRNP) of Drosophila tissue culture cells, we show that seven of these antibodies recognize different antigens, and that these antigens are associated with RNA. Analysis of the sedimentation behavior of antigen-containing structures in sucrose gradients reveals that the antigens are differentially distributed with respect both to one another and to pulse-labeled RNA. We demonstrate that the antigens are minor components of hnRNP and are different from the major Drosophila hnRNP packaging proteins, which we have also identified. The antigens are probably involved in the processing of hnRNA in the nucleus.

Animals↗
PubMedDate not supplied

Retrovirus producer cells encoding antisense VEGF prolong survival of rats with intracranial GS9L gliomas.

With increasing size tumors are continually dependent on a functional blood vessel system to guarantee the supply with oxygen and nutrients. Vascular endothelial growth factor (VEGF) is a key mediator not only of developmental but also of hypoxia-mediated and tumor-induced angiogenesis. Gene therapy using antisense VEGF with the aim to inhibit tumor angiogenesis may be a successful strategy for the treatment of highly vascular and invasive malignant gliomas. We investigated whether retrovirus producer cells encoding antisense VEGF can be used for in vivo gene transfer. The full length mouse VEGF164 cDNA was cloned in a sense and antisense direction into the retroviral expression vector pLEN. pLEN-VEGF (sense) and pLEN-FGEV (antisense) expression vectors were used to transfect the packaging cell line GP + E86 and to establish ecotropic virus producer cell lines. GP + E86:LEN-FGEV (#5) cells showed high expression of antisense VEGF mRNA, whereas GP+ E86:LEN-VEGF (#8) showed high expression of sense VEGF mRNA and active VEGF protein. Co-implantation of GS-9L cells with retrovirus producing cells containing the antisense VEGF construct into the brains of syngeneic rats showed a statistically significant inhibition of tumor growth and prolongation of survival time, while co-implantation of retrovirus producer cells containing the sense VEGF expression vector resulted in an increasing tumor growth and reduced survival time of the rats compared to control animals. Histological analysis of the tumors co-implanted with GP + E86:LEN-FGEV (#5) cells showed the suppression of angiogenesis, high degree of necrosis and no evidence of a significant immune response. Expression of antisense VEGF mRNA in these tumors was confirmed by in situ hybridization analysis. This is the first report demonstrating the potential utility of virus producer cells as in vivo gene transfer vehicles for antisense VEGF gene therapy of malignant gliomas.

Animals↗