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First published online September 19, 2008
Journal of Experimental Biology 211, 3077-3084 (2008)
Published by The Company of Biologists 2008
doi: 10.1242/jeb.019950

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Effects of hypothermia on gene expression in zebrafish gills: upregulation in differentiation and function of ionocytes as compensatory responses

Ming-Yi Chou^1,2, Chung-Der Hsiao^1,3, Shyh-Chi Chen¹, I-Wen Chen¹, Sian-Tai Liu¹ and Pung-Pung Hwang^1,2,*

¹ Institute of Cellular and Organismic Biology, Academia Sinica, Taipei, 115 Taiwan
² Institute of Fishery Science, National Taiwan University, Taipei, 10617 Taiwan
³ Department of Bioscience Technology, Chung Yuan Christian University, Chung Li, 32023 Taiwan

View larger version (6K): [in this window] [in a new window]   Fig. 1. Expression of cirbp and hmgb1 mRNA in zebrafish gills. After cold treatment for 1 day or 30 days, expression of cirbp and hmgb1 mRNAs was strongly induced. The values were normalized to β-actin. Values are means ± s.d. (N=4 or 5). Different letters above the bars indicate significant differences (one-way ANOVA, Tukey's pair-wise comparison).

View larger version (16K): [in this window] [in a new window]   Fig. 2. Validation of the microarray data by real-time RT–PCR. Nine selected genes were subjected to real-time RT–PCR, and their relative expression levels in the control, 1 day and 30 days cold-acclimated groups were compared. Basically, the expression levels detected by real-time RT–PCR were consistent with the microarray data. The values were normalized to β-actin. Values are means ± s.d. (N=4 or 5). Different letters above the bars indicate significant differences (one-way ANOVA, Tukey's pair-wise comparison).

View larger version (69K): [in this window] [in a new window]   Fig. 3. In situ hybridization of trpv6 and ca2 in zebrafish gills. Expression of trpv6 and ca2 mRNA increased significantly in 30 day cold-acclimated gills. trpv6- and ca2-expressing cells were found in the gill filaments and the lamellae in the cold-acclimated group, whereas in the control group these cells only appeared in gill filaments. Scale bar, 50µm.

View larger version (7K): [in this window] [in a new window]   Fig. 4. The effects of cold treatments on Ca2+ influx in zebrafish. Ca2+ influx decreased after 9 h exposure to 12°C but recovered after acclimation at 12°C for 30 days. Values are means ± s.d. (N=5). Different letters above the bars indicate significant differences (one-way ANOVA, Tukey's pair-wise comparison).

View larger version (71K): [in this window] [in a new window]   Fig. 5. Cell proliferation and apoptosis in zebrafish gills. (A) Immunohistochemistry of mitotic cells in normal and cold-acclimated gills. (B) Western blot analysis of phosphohistone H3 (pH3) expression in normal and cold-acclimated gills. (C) TUNEL assay for apoptotic cells in normal and cold-acclimated gills. Cold acclimation retarded both cell proliferation and apoptosis.

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