- Title
-
Characterization of Zebrafish Abcc4 as an Efflux Transporter of Organochlorine Pesticides
- Authors
- Lu, X., Long, Y., Lin, L., Sun, R., Zhong, S., Cui, Z.
- Source
- Full text @ PLoS One
Transcriptional expression of zebrafish abcc4 gene. (A) Spatiotemporal expression of abcc4 gene in developing embryos at indicated stages was detected with whole-mount in situ hybridization (WISH). (a) 1 hpf, lateral view; (b) 6 hpf, lateral view with dorsal to the right; (c) 12 hpf, lateral view with anterior to the top and dorsal to the right; (d) 24 hpf, lateral view with anterior to the left; (e, g, h and j) 48, 72, 96 and 120 hpf, lateral views with dorsal to the right; (f, i and k) 48, 96 and 120 hpf, dorsal views with anterior to the top. ey, eyes; le, lens; pa, pancreas; gi, gills; in, intestine; bl, swim bladder. (B) Transcriptional analysis of zebrafish abcc4 gene in developing embryos. Sixty embryos at 1-hpf stage or thirty embryos at 6- to 120-hpf stage were pooled for total RNA extraction and subjected to qPCR analysis. (C) Tissue-specific expression of abcc4 gene in adult zebrafish. Values are given as mean ± standard deviation, n = 3. |
Effects of DDT and lindane on the hatching and abcc4 expression of zebrafish embryos. (A and B) Hatching rates of 96-hpf embryos exposed to DDT or lindane at indicated concentrations. (C and D) Total RNAs of zebrafish embryos in (A) and (B) were extracted for real-time PCR. (E) Embryos treated with 0.05 µg/L DDT or 0.01 µg/L lindane from 24 to 96 hpf were subjected to WISH analysis of induced abcc4 expression, respectively. CK represents the untreated control. Black arrows point to intestine. All values are expressed as mean ± standard deviation, n = 3. Significant differences are indicated by *p<0.05 and **p<0.01. |
Effects of Abcc4-morpholino on the mortalities of zebrafish embryos treated with DDT or lindane. (A-B) Death rates of embryos injected with Abcc4 morpholino (Abcc4-MO) and exposed to 100 µg/L DTT or lindane. The controls include wild type (ctrl) and ctrl-MO-injected embryos. Values are expressed as mean ± standard deviation (n = 5). Significant differences are indicated by *p<0.05. PHENOTYPE:
|
Zebrafish Abcc4 has functions in transport of MCB out of LLC-PK1 cells. (A) Western blot analysis of Flag-tagged Abcc4 or Abcc4-G1188D in stably transfected LLC-PK1 cells and the control cells (CTRL) transfected with empty vectors. The β-actin was used as a loading control. (B) Green signals under the confocal microscope indicate that foreign Abcc4 or Abcc4-G1188D molecules are localized on plasma membrane of LLC-PK1 cells. (C) Fluorescence intensities in LLC-PK1 cells expressing Abcc4 or Abcc4-G1188D and the control (CTRL) after treatment with MCB for 30 min at indicated concentrations. (D) Fluorescence intensities in LLC-PK1 cells expressing Abcc4 or Abcc4-G1188D and the control (CTRL) after treatment with 25 µM MCB for indicated time periods. (E) Fluorescence intensities in Abcc4-expressing LLC-PK1 cells after treatment with MK571 for indicated time periods. Cells were incubated in medium containing 25 µM MCB and different concentrations of MK571. Data are expressed as means ± standard deviations (n = 3). Significant differences are indicated by *p<0.05 and **p<0.01. |