- Title
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Action of Vitamin D and the Receptor, VDRa, in Calcium Handling in Zebrafish (Danio rerio)
- Authors
- Lin, C.H., Su, C.H., Tseng, D.Y., Ding, F.C., and Hwang, P.P.
- Source
- Full text @ PLoS One
Effects of exogenous 1α,25(OH)2D3 (20 µg/l) in 3-d post-fertilization (dpf) zebrafish embryos. Ca2+ influx (A), Ca2+ content (B), mRNA expressions in 3-dpf zebrafish embryos (C). mRNA expressions were analyzed by a qPCR, and values were normalized to β-actin. Values are the mean ± SD (n = 7~10). Student’s t-test, *p<0.05; **p<0.01, ***p<0.001. EXPRESSION / LABELING:
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Zebrafish vdra and vdrb expression profiles. mRNA expression patterns in developing stages (A) and various tissues (B) were determined by an RT-PCR using β-actin as the internal control. (C) vdra and vdrb mRNA expressions in different tissues of adult zebrafish were analyzed by a qPCR using β-actin as the internal control. Values are the mean ± SD (n = 3). E, eye; K, kidney; Sk, skin; I, intestine; G, gill; H, heart; M, muscle; B, brain; L, liver; Ov, ovary; Te, testis. |
Effects of vitamin D receptor (VDR)a and VDRb morpholino oligonucleotides (MOs) in 3-d post-fertilization (dpf) zebrafish embryos. Ca2+ influx (A), Ca2+ content (B), mRNA expression (C), density of ecac-expressing cells (D), and ecac signals (E). mRNA expressions were analyzed by a qPCR using β-actin as the internal control. Different letters indicate a significant difference (p<0.05) using a one-way ANOVA followed by Tukey’s multiple-comparison test. Values are the mean ± SD (n = 6 or 7). Scale bar:100 μm. EXPRESSION / LABELING:
PHENOTYPE:
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Effects of vitamin D receptor (VDR)a and VDRb morpholino oligonucleotides (MOs) on cyp27b1 and cyp24a1 mRNA expressions in 3-d post-fertilization (dpf) zebrafish embryos treated with 1α,25(OH)2D3 (20 µg/l). (A) cyp27b1 mRNA expression. (B) cyp24a1 mRNA expression. mRNA expressions were analyzed by a qPCR using β-actin as the internal control. Different letters indicate a significant difference (p<0.05) using one-way ANOVA followed by Tukey’s multiple-comparison test. Values are the mean ± SD. (n = 5-6). EXPRESSION / LABELING:
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Specificity and effectiveness of vitamin D receptor (VDR)a and VDRb morpholino oligonucleotides (MOs). VDRa and VDRb MOs were respectively injected into 1- or 2-cell embryos. To clarify the MO specificity and effectiveness, Western blotting was used to detect VDRa and VDRb protein expressions in wild-type (WT) and MO-injected embryos at 3 d post-fertilization (dpf). An asterisk (*) indicates the position of VDRa or VDRb expression. |
Effect of vitamin D receptor (VDR)a and VDRb morpholino oligonucleotides (MOs) on ossification of vertebrae of 5-d post-fertilization (dpf) zebrafish embryos. Control (A), VDRa (B), and VDRb (C) MO. Ossification of vertebrae was observed by calcein staining. Scale bar: 100 μm. PHENOTYPE:
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Colocalization of vdra mRNA with Na,K-ATPase rich (NaR) cells in zebrafish gill cryosections. (A) In situ hybridization of vdra mRNA; (B) immunocytochemical staining of Na,K-ATPase. The arrow indicates colocalization of vdra mRNA and Na,K-ATPase protein signals in the same cells. Scale bar: 5 μm. EXPRESSION / LABELING:
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