Erratum
for
DABBENI-SALA et al., FASEB J. 15 (10) 1786-1788.
(The FASEB Journal. 2001;15:2554.)
© 2001 FASEB
Erratum
Kainic acid induces selective mitochondrial oxidative phosphorylation enzyme dysfunction in cerebellar granule neurons: protective effects of melatonin and GSH ethyl ester. Federica Dabbeni-Sala, Maura Floreani, Davide Franceschini, Stephen D. Skaper, and Pietro Giusti (2001) FASEB J. 15, 1786–1788.
The same figure appears as Figure 4 in the version of record (http://www.fasebj.org/cgi/doi/10.1096/fj.00-0427fje).
The loss of sympathetic nerve fibers in the synovial tissue of patients with rheumatoid arthritis is accompanied by increased norepinephrine release from synovial macrophages. L.E. Miller, H.P. Jüsten, J. Schölmerich, R.H. Straub (2000) FASEB J. 14, 2097–2107.
The double staining technique with APAAP and fluorescence microscopy was incorrect in the original article. The mentioned norepinephrine secreting tyrosine-hydroxylase positive cells are not macrophages/monocytes; neither are they B lymphocytes, T lympho-cytes, fibroblasts, or mast cells. This is only relevant in figure 7; no other parts of the paper are influenced by this error.
The authors regret this error and any inconvenience it might have caused. A complete explanation appears below.
The APAAP staining technique (Dako, Hamburg, Germany) applied in the mentioned paper used three different antibodies: 1) One monoclonal mouse antibody was directed against the human target antigen CD163 (activated macrophage, clone BERMAC 3, Dako). 2) One monoclonal mouse antibody was coupled to the enzyme alkaline phosphatase, which yielded the substrate color reaction (red staining; Dako). 3) The third antibody from rabbit linked the two Fc parts of the first and second monoclonal mouse antibodies (bridging antibody, which may also stem from goat and other animal; Dako). In addition, we used a polyclonal antibody from rabbit in order to stain the human tyrosine hydroxylase in the tissue (Chemicon, Temecula, CA) together with an Alexa-conjugated goat anti-rabbit antibody (MoBiTec, Göttingen, Germany). Since we used the wrong APAAP-bridging antibody (rabbit instead of goat), double staining of both structures in identical high power fields – CD163 (under bright conditions) and tyrosine hydroxylase (under fluorescent light condition) – was an inevitable result. After realizing this error in August 2001, we now used an alternative double staining technique. In this protocol, mouse monoclonal antibodies against several human antigens were used (CD3, T lymphocyte, clone UCHT 1, Dako; CD19, B lymphocyte, clone HD 37, Dako; CD68 monocyte/macrophage, clone PG-M1, Dako; CD 163, activated macrophage, clone BERMAC 3, Dako; chymase, mast cell, Chemicon, Temecula, CA; fibroblast marker, clone AS 02, Dianova, Hamburg, Germany). A secondary antibody from goat (Dianova) against the Fc part of the mouse monoclonal antibody linked to alkaline phosphatase was used in order to visualize the complex (BCIP/NBT substrate 
purple/black staining). As mentioned above, the polyclonal rabbit antibody against tyrosine hydroxylase (Chemicon) was used in order to stainthe tyrosine hydroxylase-positive cells together with an Alexa-conjugated goat anti-rabbit antibody (MoBiTec). As in the mentioned paper, double staining was visualized under fluorescent light and under bright light conditions in 400x high power fields (Fig. 1)
.It is obvious that identified structures in the respective figures are notidentical (arrows, Fig. 1
). Thus, the new tyrosine hydroxylase-positive cell in the synovial tissue is not a monocyte/macrophage or a B lymphocyte, a T lymphocyte, a mast cell, or a fibroblast. One may speculate as to whether its origin is a sympathoadrenal chromaffin cell, which has to be subject of future studies.
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Figure 1. Micrographs of identical fields of view in a section of double-stained synovial tissue with antibodies against different cellular markers (bright light conditions, left side of panel) and tyrosine hydroxylase (fluorescent light conditions, right side of panel). A) CD163, activated macrophage; B) CD3, T lymphocyte; C) CD19, B lymphocyte; D) CD68, monocyte/macrophage; E) chymase, mast cell; F) fibroblast marker, fibroblast. The tissue was photographed at 400x magnification. Arrow identifies the respective structure under bright light conditions; arrowhead identifies tyrosine hydroxylase cells under fluorescent light conditions.
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Figure 2. KA reduced granule neuron content of the catalytic portion of complex II. Cerebellar granule neuron mitochondrial extracts were subjected to Western blot analysis using antibodies against complex II (C II) and complex V (C V). Control (lane 1); 0.5 mM KA (lane 2); 0.5 mM KA + 50 µM DNQX (lane 3).
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