FISH mapping of the porcine vWF gene to chromosome 5q21 extends synteny homology with human Chromosome 12

Sjöberg, Anita Herrström; Wt, Seaman; Da, Bellinger; Tr, Griggs; Tc, Nichols; Bp, Chowdhary

doi:10.1111/j.1601-5223.1996.00199.x

Cited by 9 publications

(2 citation statements)

References 19 publications

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“…The polymerase chain reaction (PCR) was performed using GeneAmp (PCR system 9700, Perkin-Elmer, Norwalk, CT, USA) with rat intercellular adhesion molecule-1 (ICAM-1), PDGF-Rβ, angiopoietin-1 (Ang-1), osteopontin (OPN), and von Willebrand factor (vWF) specific primers through 35-40 cycles of 94°C for 0.5 min, 60°C for 1 min and 72°C for 1 min. The sequences of the specific primers are follows: the sense sequence (241-260) was 5'-TCG AGT GGA CAC AAC TGG AA-3' and the antisense sequence (678-659) was 5'-AGT CGG AAG ATC GAA AGT CC-3' for ICAM-1 (Genebank accession number D00913): the sense sequence (10-29) was 5'-TGG GCC AGA GTT CGT CCT CA-3' and the antisense sequence (501-482) was 5'-TCG GAG TCC ACT TCC CTG TC-3' for PDGF-Rβ (Genebank accession number Z14119): the sense sequence (2-21) was 5'-AAA ATT ATA CTC AGT GGC TG-3' and the antisense sequence (330-311) was 5'-TTC TAG GAT TTT ATG CTC TA-3' for Ang-1 (Genebank accession number AF030376): the sense sequence (116-135) was 5'-ATG AGA CTG GCA GTG GTT TG-3' and the antisense sequence (622-603) was 5'-GGC ATC GGG ATA CTG TTC AT-3' for OPN (Genebank accession number M99252): the sense sequence (7635-7657) was 5'-CCC TGC CTC ATC AAT GAG TGT GT-3' and the antisense sequence (8221-8199) was 5'-GGC TCC TCA CAT GTG TCA CAG CA-3' for vWF (Genebank accession number L76227) (Mancuso et al, 1989;Sjöberg et al, 1996). The PCR products were sequenced by electrophoresis on an agarose gel in the presence of ethidium bromide and visualized with an imager (Epipro 7000, Aisin, Kariya, Japan).…”

Section: Reverse Transcription-polymerase Chain Reaction (Rt-pcr) Anamentioning

confidence: 99%

Establishment of Conditionally Immortalized Rat Retinal Pericyte Cell Lines (TR-rPCT) and Their Application in a Co-culture System Using Retinal Capillary Endothelial Cell Line (TR-iBRB2)

Kondo

Hosoya

Hori

et al. 2003

Cell Struct. Funct.

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ABSTRACT. The purpose of this study was to establish and characterize a retinal pericyte cell line from retinal capillaries of transgenic rats harboring the temperature-sensitive simian virus 40 large T-antigen gene (tsA58 Tg rat), and to apply this to the co-culture with a retinal capillary endothelial cell line. The conditionally immortalized rat retinal pericyte cell lines (TR-rPCTs), which express a temperature-sensitive large T-antigen, were obtained from two tsA58 Tg rats. These cell lines had a multicellular nodule morphology and reacted positively with von Kossa staining, a marker of calcification. TR-rPCTs cells expressed mRNA of pericyte markers such as rat intercellular adhesion molecule-1, platelet-derived growth factor-receptor β, angiopoietin-1, and osteopontin. Western blot analysis indicated that α-smooth muscle actin (α-SMA) was expressed in TR-rPCT3 and 4 cells. In contrast, α-SMA was induced by transforming growth factor-β1 and its enhancement was reduced by basic fibroblast growth factor in TR-rPCT1 and 2 cells. When TR-rPCT1 cells were cultured with a rat retinal endothelial cell line (TR-iBRB2) in a contact co-culture system, the number of TR-iBRB2 cells were significantly reduced in comparison with that of a single culture of TR-iBRB2 cells, suggesting that physical contact between pericytes and retinal endothelial cells is important for the growth of retinal endothelial cells. In conclusion, conditionally immortalized retinal pericyte cell lines were established from tsA58 Tg rats. These cell lines exhibited the properties of retinal pericytes and can be applied in co-culture systems with a retinal capillary endothelial cell line.Key words: retinal pericyte cell line/SV 40 large T-antigen/co-culture/retinal endothelial cell growth/α-smooth muscle actinRetinal pericytes, the cells that surround retinal capillary endothelial cells which make up the inner blood-retinal barrier (inner BRB), have been proposed to play a role in regulating endothelial cell proliferation (Orlidge and D 'Amore, 1987;Yamagishi et al., 1993). Platelet-derived growth factor-B (PDGF-B), which is one of the ligands of PDGFreceptor β (PDGF-Rβ), and PDGF-Rβ play an important role in microvessel assembly (Lindahl et al., 1997;Hellström et al., 1999). This suggests that PDGF-Rβ in pericytes and pericytes per se are important for forming the inner BRB.To investigate physiological roles of pericytes, primary *To whom correspondence should be addressed: Professor Tetsuya Terasaki, Ph.D., Department of Molecular Biopharmacy and Genetics, Graduate School of Pharmaceutical Sciences, Tohoku University, Aoba, Aramaki, Aoba-ku, Sendai 980-8578, Japan. Tel: +81-22-217-6831, Fax: +81-22-217-6886 E-mail: terasaki@mail.pharm.tohoku.ac.jp Abbreviations: Ac-LDL, acetylated low density lipoprotein; Ang-1, angiopoietin-1; bFGF, basic fibroblast growth factor; BRB, blood-retinal barrier; BSA, bovine serum albumin; DMEM, Dulbecco's modified Eagle's medium; DiI, 1,[1][2][3]3,3',3'-tetramethyliodo-carbocyanine perchlorate; FBS, fetal bovine ...

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Section: Reverse Transcription-polymerase Chain Reaction (Rt-pcr) Anamentioning

confidence: 99%

Establishment of Conditionally Immortalized Rat Retinal Pericyte Cell Lines (TR-rPCT) and Their Application in a Co-culture System Using Retinal Capillary Endothelial Cell Line (TR-iBRB2)

Kondo

Hosoya

Hori

et al. 2003

Cell Struct. Funct.

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“…The porcine VWF gene is on pig chromosome 5q21, a region that is syntenic with human chromosome 12p13.2, the location of the human gene [12, 13]. The human VWF gene is located near the telomere, perhaps in part accounting for the high frequency of mutations resulting in various types of VWD, while the pig VWF gene is more centromeric.…”

Section: Porcine Vwf and Vwdmentioning

confidence: 99%

Porcine and Canine von Willebrand Factor and von Willebrand Disease: Hemostasis, Thrombosis, and Atherosclerosis Studies

et al. 2010

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Use of animal models of inherited and induced von Willebrand factor (VWF) deficiency continues to advance the knowledge of VWF-related diseases: von Willebrand disease (VWD), thrombotic thrombocytopenic purpura (TTP), and coronary artery thrombosis. First, in humans, pigs, and dogs, VWF is essential for normal hemostasis; without VWF bleeding events are severe and can be fatal. Second, the ADAMTS13 cleavage site is preserved in all three species suggesting all use this mechanism for normal VWF multimer processing and that all are susceptible to TTP when ADAMTS13 function is reduced. Third, while the role of VWF in atherogenesis is debated, arterial thrombosis complicating atherosclerosis appears to be VWF-dependent. The differences in the VWF gene and protein between humans, pigs, and dogs are relatively few but important to consider in the design of VWF-focused experiments. These homologies and differences are reviewed in detail and their implications for research projects are discussed. The current status of porcine and canine VWD are also reviewed as well as their potential role in future studies of VWF-related disorders of hemostasis and thrombosis.

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Conditionally Immortalized Retinal Capillary Endothelial Cell Lines (TR-iBRB) Expressing Differentiated Endothelial Cell Functions Derived from a Transgenic Rat

Hosoya

Tomi

Ohtsuki

et al. 2001

Experimental Eye Research

139

133

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FISH mapping of the porcine vWF gene to chromosome 5q21 extends synteny homology with human Chromosome 12

Cited by 9 publications

References 19 publications

Establishment of Conditionally Immortalized Rat Retinal Pericyte Cell Lines (TR-rPCT) and Their Application in a Co-culture System Using Retinal Capillary Endothelial Cell Line (TR-iBRB2)

Establishment of Conditionally Immortalized Rat Retinal Pericyte Cell Lines (TR-rPCT) and Their Application in a Co-culture System Using Retinal Capillary Endothelial Cell Line (TR-iBRB2)

Porcine and Canine von Willebrand Factor and von Willebrand Disease: Hemostasis, Thrombosis, and Atherosclerosis Studies

Conditionally Immortalized Retinal Capillary Endothelial Cell Lines (TR-iBRB) Expressing Differentiated Endothelial Cell Functions Derived from a Transgenic Rat

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