Acid Ethanol Fixation and Polyester Wax Embedding Combines Preservation of Antigenic Determinants with Good Morphology and Enables Simultaneous Bromodeoxyuridine (BRDU) Labeling

Roholl, P. J. M.; Dullens, H. F. J.; Kleijne, J.; Dubbink, E. J.; Otter, W. Den

doi:10.3109/10520299109110551

Cited by 4 publications

(2 citation statements)

References 21 publications

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“…Nonradioactive in situ hybridization was performed essentially as described by Coen et al (1990). except that the wax embedding medium was replaced by a low-temperature polyester wax to maintain protein antigenicity in the tissue block (Labacq and Ritter, 1979;Roholl et al, 1991). The temperature for infiltration of polyester wax was 42OC.…”

Section: In Situ Hybridizationmentioning

confidence: 99%

A Model for Seed Transmission of a Plant Virus: Genetic and Structural Analyses of Pea Embryo Invasion by Pea Seed-Borne Mosaic Virus.

Wang

Maule

1994

Plant Cell

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Pea seed-borne mosaic virus (PSbMV), a seed-transmitted virus in pea and other legumes, invades pea embryos early in development. This process is controlled by maternal genes and, in a cultivar that shows no seed transmission, is prevented through the action of multiple host genes segregating as quantitative trait loci. These genes control the ability of PSbMV to spread into and/or multiply in the nonvascular testa tissues, thereby preventing the virus from crossing the boundary between the maternal and progeny tissues. lmmunocytochemical and in situ hybridization studies suggested that the virus uses the embryonic suspensor as the route for the direct invasion of the embryo. The programmed degeneration of the suspensor during embryo development may provide a transient window for embryo invasion by the virus and could explain the inverse relationship between the age of the mother plant for virus infection and the extent of virus seed transmission.

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Section: In Situ Hybridizationmentioning

confidence: 99%

A Model for Seed Transmission of a Plant Virus: Genetic and Structural Analyses of Pea Embryo Invasion by Pea Seed-Borne Mosaic Virus.

Wang

Maule

1994

Plant Cell

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“…Fixed Matrigels were embedded in polyester wax and sectioned at 8 m for immunohistochemical analysis as previously described. 15 Identification of endothelial cells was performed by immunostaining for PECAM-1 using a monoclonal antibody against mouse PECAM-1 at 4°C overnight. 16 In a parallel experiment, recovered Matrigel was homogenized in PBS and digested in Sigma diagnostic hemoglobin reagents (No.…”

Section: In Vivo Assessment Of Angiogenesis Using the Matrigel Plug Amentioning

confidence: 99%

Ephrin-B2 Induces Migration of Endothelial Cells Through the Phosphatidylinositol-3 Kinase Pathway and Promotes Angiogenesis in Adult Vasculature

Maekawa

Oike

Kanda

et al. 2003

ATVB

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Objective-Ephrin-B2 plays a key role in vascular development. The purpose of this study was to elucidate the molecular mechanisms of ephrin-B2 signaling through the EphB receptor in endothelial cells and to determine whether ephrin-B2 contributes to in vivo angiogenesis in adult mice. Methods and Results-A chemotaxis assay on a polycarbonate membrane revealed that ephrin-B2/Fc chimeric protein induced migration of human umbilical vein endothelial cells (HUVECs) at a level 98% greater than control (PϽ0.01).To determine the signaling pathways activated in the HUVECs by Eph stimulation, phosphatidylinositol-3 kinase (PI3 kinase) activity was determined in an immune complex PI3 kinase assay. Serum-starved HUVECs were stimulated with ephrin-B2/Fc and compared with unstimulated cells. PI3 kinase activity in stimulated cells was higher than that seen in unstimulated cells. In a chemotaxis assay, the PI3 kinase-specific inhibitor LY294002 blocked the migratory response of HUVECs induced by addition of ephrin-B2/Fc. Finally, ephrin-B2/Fc promoted angiogenesis in vivo in corneal neovascularization and Matrigel plug assays in adult mice, whereas LY294002 reduced angiogenesis in Matrigel that was induced by ephrin-B2/Fc. Key Words: ephrin-B2 Ⅲ angiogenesis Ⅲ phosphatidylinositol-3 kinase Ⅲ migration Ⅲ endothelial cells E ph receptor tyrosine kinases (RTKs) constitute the largest known family of RTKs identified, consisting of at least 14 receptors and 8 ephrin ligands. 1,2 Unlike most soluble RTK ligands, ephrins are membrane-attached cell surface molecules. 1 It has been shown that the ephrins and Ephs play essential roles in vascular development during embryogenesis. 3 Targeted mutation of ephrin-B2 in mice results in embryonic lethality at embryonic day 10.5 in homozygous nulls. 3 In these mice, vascular development is arrested at the primary plexus stage, and features of angiogenic remodeling are not observed. 3 Mouse embryos lacking EphB4 essentially phenocopy defects observed in ephrin-B2-null mice, suggesting reciprocal functions for the 2 molecules during vascular development in the embryo. 4 By contrast, little is known about the function of ephrin-B2 in the adult vasculature. Expression studies indicate that the endothelium of a subset of new vessels strongly expresses ephrin-B2 in adult angiogenesis as in tumor angiogenesis and in the female reproductive system, 5 suggesting that ephrin-B2 may play a role in adult angiogenesis. Here, by using in vivo model of angiogenesis in adult mice, we ask whether ephrin-B2 has an angiogenic effect. Conclusions-Ephrin-B2/FcSeveral proteins function in the Eph signaling pathway. Among these, phosphatidylinositol-3 (PI3) kinase, Grb2, Grb10, Nck, RasGAP, and Src are implicated in regulating cell morphology, attachment, and motility. 2,6,7 However, the molecular mechanism of angiogenesis induced by Eph activation has not been elucidated. In this study, we demonstrate that ephrin-B2 induces angiogenesis in vivo and that the PI3 kinase signaling pathway contributes to angiogenic ev...

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Immunohistochemistry Using Polyester Wax

Kent

2008

Methods in Molecular Biology™

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Acid Ethanol Fixation and Polyester Wax Embedding Combines Preservation of Antigenic Determinants with Good Morphology and Enables Simultaneous Bromodeoxyuridine (BRDU) Labeling

Cited by 4 publications

References 21 publications

A Model for Seed Transmission of a Plant Virus: Genetic and Structural Analyses of Pea Embryo Invasion by Pea Seed-Borne Mosaic Virus.

A Model for Seed Transmission of a Plant Virus: Genetic and Structural Analyses of Pea Embryo Invasion by Pea Seed-Borne Mosaic Virus.

Ephrin-B2 Induces Migration of Endothelial Cells Through the Phosphatidylinositol-3 Kinase Pathway and Promotes Angiogenesis in Adult Vasculature

Immunohistochemistry Using Polyester Wax

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