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* Department of Biological Chemistry and Israel Center for Neuronal Computation, The Hebrew University of Jerusalem, Jerusalem, Israel;
School of Chemistry, The Raymond and Beverly Sackler Faculty of Exact Sciences, Tel-Aviv University, Ramat Aviv, Israel; and
Tel Aviv Sourasky Medical Center, Human Brain Imaging Unit, The Wohl Institute for Advanced Imaging, Tel Aviv, Israel
1Correspondence: Department of Biological Chemistry and Israel Center for Neuronal Computation, The Hebrew University of Jerusalem, Jerusalem, Israel. E-mail: soreq{at}cc.huji.ac.il
In his letter, J. Badaut points out a possible AQP4 immunostaining abnormality in Fig. 7 of our manuscript (Meshorer et al., 2005). The abnormality involved staining of whole cell bodies instead of astrocyte endfeet. J. Badaut suggested this discrepancy might be due to the fixation method used in our study. He demonstrated the correct staining pattern of AQP4 in the mouse brain using two different antibodies, including the monoclonal antibody we used in our study (Serotec) and an additional polyclonal antibody that we did not use (Chemicon). To address this, we repeated the immunostaining experiments presented in Fig. 7 using our same original fixation method and immunolabeling protocol with the polyclonal anti-AQP4 antibody from Chemicon, kindly provided to us by J. Badaut. Using this antibody, we observed selective perivascular staining as described by J. Badaut and others in both control (Fig. 1A
, Ct) and transgenic (Fig. 1B
, TgS) animals. Nevertheless, signal quantification using the MetaMorph software in five different brain sectionsfrom two different animals showed an increase in AQP4 intensity in TgS animals (Fig. 1C
), in agreement with microarray data, RT-PCR analysis, and Western blots presented in our original paper. We conclude that the discrepancy was not a result of the perfusion or the distinct fixation methods used, but rather that our original immunolabeling experiments were likely performed with a defective batch of AQP4 antibodies. We are grateful to J. Badaut for correcting this error. Apart from the AQP4 staining abnormality, which has now been corrected (Fig. 1)
, all results and conclusions presented in our original paper remain sound.
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FOOTNOTES
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