Biolistic transfection and morphological analysis of cultured sympathetic neurons

Klimaschewski, Lars; Nindl, W; Pimpl, Michael; Waltinger, Peter; Pfaller, Kristian

doi:10.1016/s0165-0270(01)00473-3

Cited by 37 publications

(26 citation statements)

References 13 publications

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“…8). Over 98% of cells expressing eYFP also expressed the cotransfected plasmid (data not shown), consistent with results showing that over 90% of neurons in slices transfected with two plasmids express both proteins (Klimaschewski et al, 2002;Wirth et al, 2003).…”

Section: Developmental Expression and Localization Of Endogenous Centsupporting

confidence: 90%

The neuronal Arf GAP centaurin α1 modulates dendritic differentiation

Moore

Thacker

Larimore

et al. 2007

Journal of Cell Science

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ReferencesAggensteiner, M. and Reiser, G. (2003). Expression of the brain-specific membrane adapter protein p42 IP4 /centaurin a, a Ins(1,3,4,5)P 4 /PtdIns(3,4,5)P 3 binding protein, in developing rat brain. Brain Res. Dev. Brain Res. 142,[77][78][79][80][81][82][83][84][85][86][87]. Albertinazzi, C., Za, L., Paris, S. and de Curtis, I. (2003). ADP-ribosylation factor 6 and a functional PIX/p95-APP1 complex are required for Rac1B-mediated neurite outgrowth. Mol. Biol. Cell 14, 1295-1307. Allen, P., Ouimet, C. and Greengard, P. (1997 (1996). Identification and cloning of centaurin alpha: a novel phosphatidylinositol (3,4,5)-trisphosphate binding protein from rat brain.

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Section: Developmental Expression and Localization Of Endogenous Centsupporting

confidence: 90%

The neuronal Arf GAP centaurin α1 modulates dendritic differentiation

Moore

Thacker

Larimore

et al. 2007

Journal of Cell Science

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“…Fluorescent images of f100 cells from multiple fields per sample were viewed under a 20Â objective using a FITC filter in a Confocal Microscope (Nikon, PCM2000, with a Spot Camera) and quantitated using Metamorph software (Universal Imaging Corp., Downingtown, PA; refs. 32,33).…”

Section: Methodsmentioning

confidence: 99%

MYC Can Induce DNA Breaks In vivo and In vitro Independent of Reactive Oxygen Species

Ray¹,

Atkuri²,

Deb-Basu³

et al. 2006

Cancer Research

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MYC overexpression is thought to initiate tumorigenesis by inducing cellular proliferation and growth and to be restrained from causing tumorigenesis by inducing cell cycle arrest, cellular senescence, and/or apoptosis. Here we show that MYC can induce DNA breaks both in vitro and in vivo independent of increased production of reactive oxygen species (ROS). We provide an insight into the specific circumstances under which MYC generates ROS in vitro and propose a possible mechanism. We found that MYC induces DNA double-strand breaks (DSBs) independent of ROS production in murine lymphocytes in vivo as well as in normal human foreskin fibroblasts (NHFs) in vitro in normal (10%) serum, as measured by ;H2AX staining. However, NHFs cultured in vitro in low serum (0.05%) and/or ambient oxygen saturation resulted in ROS-associated oxidative damage and DNA single-strand breaks (SSBs), as measured by Ape-1 staining. In NHFs cultured in low versus normal serum, MYC induced increased expression of CYP2C9, a gene product well known to be associated with ROS production. Specific inhibition of CYP2C9 by small interfering RNA was shown to partially inhibit MYC-induced ROS production. Hence, MYC overexpression can induce ROS and SSBs under some conditions, but generally induces widespread DSBs in vivo and in vitro independent of ROS production. (Cancer Res 2006; 66(13): 6598-605)

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“…Morphometric analyses were performed to quantify number and area of eosinophils using MetaMorph software (version 5; Universal Imaging Corp.). Five to 6 images from 5 different levels per lung (n = 3 animals per group) were obtained and reassembled using the MetaMorph software montage stage stitching algorithm (78). The integrated morphometric analysis function was used to transform total pixel area of the fluorescent signal from the eosinophils into µm 2 units after calibration (79).…”

Section: Rwe Endotoxin-free Rwe (Lot Xp56-d18-1320) Was Purchased Frmentioning

confidence: 99%

ROS generated by pollen NADPH oxidase provide a signal that augments antigen-induced allergic airway inflammation

Boldogh¹

2005

Journal of Clinical Investigation

329

366

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Pollen exposure induces allergic airway inflammation in sensitized subjects. The role of antigenic pollen proteins in the induction of allergic airway inflammation is well characterized, but the contribution of other constituents in pollen grains to this process is unknown. Here we show that pollen grains and their extracts contain intrinsic NADPH oxidases. The pollen NADPH oxidases rapidly increased the levels of ROS in lung epithelium as well as the amount of oxidized glutathione (GSSG) and 4-hydroxynonenal (4-HNE) in airwaylining fluid. These oxidases, as well as products of oxidative stress (such as GSSG and 4-HNE) generated by these enzymes, induced neutrophil recruitment to the airways independent of the adaptive immune response. Removal of pollen NADPH oxidase activity from the challenge material reduced antigen-induced allergic airway inflammation, the number of mucin-containing cells in airway epithelium, and antigen-specific IgE levels in sensitized mice. Furthermore, challenge with Amb a 1, the major antigen in ragweed pollen extract that does not possess NADPH oxidase activity, induced low-grade allergic airway inflammation. Addition of GSSG or 4-HNE to Amb a 1 challenge material boosted allergic airway inflammation. We propose that oxidative stress generated by pollen NADPH oxidases (signal 1) augments allergic airway inflammation induced by pollen antigen (signal 2).

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Biolistic transfection and morphological analysis of cultured sympathetic neurons

Cited by 37 publications

References 13 publications

The neuronal Arf GAP centaurin α1 modulates dendritic differentiation

The neuronal Arf GAP centaurin α1 modulates dendritic differentiation

MYC Can Induce DNA Breaks In vivo and In vitro Independent of Reactive Oxygen Species

ROS generated by pollen NADPH oxidase provide a signal that augments antigen-induced allergic airway inflammation

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