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Functional imaging of Rel expression in inflammatory processes using bioluminescence imaging system in transgenic mice.

Yang X, Jing H, Zhao K, Sun R, Liu Z, Ying Y, Ci L, Kuang Y, Huang F, Wang Z, Fei J - PLoS ONE (2013)

Bottom Line: Revealing the dynamic expression of c-Rel in disease processes in vivo is critical for understanding c-Rel functions and for developing anti-inflammatory drugs.Luciferase expression could be tracked in living mice by the method of bioluminescence imaging in a variety of inflammatory processes, including LPS induced sepsis and EAE disease model.These results indicate that the B6-Tg(c-Rel-luc)(Mlit) mouse is a valuable animal model to study Rel expression in physiological and pathological processes, and the effects of various drug treatments in vivo.

View Article: PubMed Central - PubMed

Affiliation: School of Life Science and Technology, Tongji University, Shanghai, China.

ABSTRACT
c-Rel plays important roles in many inflammatory diseases. Revealing the dynamic expression of c-Rel in disease processes in vivo is critical for understanding c-Rel functions and for developing anti-inflammatory drugs. In this paper, a transgenic mouse line, B6-Tg(c-Rel-luc)(Mlit), which incorporated the transgene firefly luciferase driven by a 14.5-kb fragment containing mouse c-Rel gene Rel promoter, was generated to monitor Rel expression in vivo. Luciferase expression could be tracked in living mice by the method of bioluminescence imaging in a variety of inflammatory processes, including LPS induced sepsis and EAE disease model. The luciferase expression in transgenic mice was comparable to the endogenous Rel expression and could be suppressed by administration of anti-inflammatory drug dexamethasone or aspirin. These results indicate that the B6-Tg(c-Rel-luc)(Mlit) mouse is a valuable animal model to study Rel expression in physiological and pathological processes, and the effects of various drug treatments in vivo.

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Comparison of luciferase acitivity profile and endogenous Rel mRNA expression pattern in B6-Tg(c-Rel-luc)8Mlit mice following LPS treament.(A) The Rel expression were analyzed by RT-PCR in the B6-Tg(c-Rel-luc)8Mlit mice (TG) and wild type littermates (WT) without LPS treatment, n = 3 in each group. (B)The luciferase activity in selected organs was measured in male B6-Tg(c-Rel-luc)8Mlit mice at 3 hour after either LPS (3 mg/kg) or saline (NS) treatment. n = 3 in each group. (C) The ratio of LPS induced luciferase activity in each group to that of saline treated control (NS), n = 3. (D) The Rel gene expression in selected organs was measured by RT-PCR in the male transgenic mice at 3 hour after LPS or saline treatment (NS). The Rel mRNA level in saline treated kidney was set as 1, in each group, n = 3. (E) The ratio of LPS induced endogenous Rel expression in each group to that of saline treated control (NS). n = 3, *p<0.05; **p<0.01; ***p<0.001.
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pone-0057632-g003: Comparison of luciferase acitivity profile and endogenous Rel mRNA expression pattern in B6-Tg(c-Rel-luc)8Mlit mice following LPS treament.(A) The Rel expression were analyzed by RT-PCR in the B6-Tg(c-Rel-luc)8Mlit mice (TG) and wild type littermates (WT) without LPS treatment, n = 3 in each group. (B)The luciferase activity in selected organs was measured in male B6-Tg(c-Rel-luc)8Mlit mice at 3 hour after either LPS (3 mg/kg) or saline (NS) treatment. n = 3 in each group. (C) The ratio of LPS induced luciferase activity in each group to that of saline treated control (NS), n = 3. (D) The Rel gene expression in selected organs was measured by RT-PCR in the male transgenic mice at 3 hour after LPS or saline treatment (NS). The Rel mRNA level in saline treated kidney was set as 1, in each group, n = 3. (E) The ratio of LPS induced endogenous Rel expression in each group to that of saline treated control (NS). n = 3, *p<0.05; **p<0.01; ***p<0.001.

Mentions: The Rel expression in B6-Tg(c-Rel-luc)8Mlit mice was comparable with that of wild type littermates (Fig. 3A). This result indicated that the transgene and the process of transgenic processing did not modify the endogenous Rel expression in mouse. The ex vivo measurement of LPS induced luciferase activity and endogenous Rel mRNA expression in B6-Tg(c-Rel-luc)8Mlit mice were performed. The mouse organs were dissected and homogenated at the time point of 3 h after LPS injection. The luciferase activity was high in the heart, liver, spleen, intestine and stomach. Compared with the saline treated mice, luciferase activity in LPS treated group could be increased by 6.5 fold in the liver, 6 fold in the spleen, 5.2 fold in the intestine, 5 fold in the stomach, 4 fold in the heart, 3.8 fold in the lung, 3 fold in the kidney, 2.3 fold in thymus. In addition, the luciferase activity reached to 3.2 fold in the macrophages (Fig. 3B, C). Meanwhile, the endogenous Rel expression was quantified by real time PCR (Fig. 3D, E). Compared with the saline group, LPS stimulated the endogenous Rel expression by 2.1 fold in the liver, 1.8 fold in the spleen, 2.3 fold in the intestine, 2.6 fold in the stomach, 4.1. fold in the heart, 3.3 fold in the lung, 4.4 fold in the kidney, 2.3 fold in the thymus and 4.4 fold in macrophages (Fig. 3D, E). These results showed that the luciferase activity profile and its response to LPS treatment were in parallel with that of endogenous Rel mRNA expression in B6-Tg(c-Rel-luc)8Mlit mice.


Functional imaging of Rel expression in inflammatory processes using bioluminescence imaging system in transgenic mice.

Yang X, Jing H, Zhao K, Sun R, Liu Z, Ying Y, Ci L, Kuang Y, Huang F, Wang Z, Fei J - PLoS ONE (2013)

Comparison of luciferase acitivity profile and endogenous Rel mRNA expression pattern in B6-Tg(c-Rel-luc)8Mlit mice following LPS treament.(A) The Rel expression were analyzed by RT-PCR in the B6-Tg(c-Rel-luc)8Mlit mice (TG) and wild type littermates (WT) without LPS treatment, n = 3 in each group. (B)The luciferase activity in selected organs was measured in male B6-Tg(c-Rel-luc)8Mlit mice at 3 hour after either LPS (3 mg/kg) or saline (NS) treatment. n = 3 in each group. (C) The ratio of LPS induced luciferase activity in each group to that of saline treated control (NS), n = 3. (D) The Rel gene expression in selected organs was measured by RT-PCR in the male transgenic mice at 3 hour after LPS or saline treatment (NS). The Rel mRNA level in saline treated kidney was set as 1, in each group, n = 3. (E) The ratio of LPS induced endogenous Rel expression in each group to that of saline treated control (NS). n = 3, *p<0.05; **p<0.01; ***p<0.001.
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pone-0057632-g003: Comparison of luciferase acitivity profile and endogenous Rel mRNA expression pattern in B6-Tg(c-Rel-luc)8Mlit mice following LPS treament.(A) The Rel expression were analyzed by RT-PCR in the B6-Tg(c-Rel-luc)8Mlit mice (TG) and wild type littermates (WT) without LPS treatment, n = 3 in each group. (B)The luciferase activity in selected organs was measured in male B6-Tg(c-Rel-luc)8Mlit mice at 3 hour after either LPS (3 mg/kg) or saline (NS) treatment. n = 3 in each group. (C) The ratio of LPS induced luciferase activity in each group to that of saline treated control (NS), n = 3. (D) The Rel gene expression in selected organs was measured by RT-PCR in the male transgenic mice at 3 hour after LPS or saline treatment (NS). The Rel mRNA level in saline treated kidney was set as 1, in each group, n = 3. (E) The ratio of LPS induced endogenous Rel expression in each group to that of saline treated control (NS). n = 3, *p<0.05; **p<0.01; ***p<0.001.
Mentions: The Rel expression in B6-Tg(c-Rel-luc)8Mlit mice was comparable with that of wild type littermates (Fig. 3A). This result indicated that the transgene and the process of transgenic processing did not modify the endogenous Rel expression in mouse. The ex vivo measurement of LPS induced luciferase activity and endogenous Rel mRNA expression in B6-Tg(c-Rel-luc)8Mlit mice were performed. The mouse organs were dissected and homogenated at the time point of 3 h after LPS injection. The luciferase activity was high in the heart, liver, spleen, intestine and stomach. Compared with the saline treated mice, luciferase activity in LPS treated group could be increased by 6.5 fold in the liver, 6 fold in the spleen, 5.2 fold in the intestine, 5 fold in the stomach, 4 fold in the heart, 3.8 fold in the lung, 3 fold in the kidney, 2.3 fold in thymus. In addition, the luciferase activity reached to 3.2 fold in the macrophages (Fig. 3B, C). Meanwhile, the endogenous Rel expression was quantified by real time PCR (Fig. 3D, E). Compared with the saline group, LPS stimulated the endogenous Rel expression by 2.1 fold in the liver, 1.8 fold in the spleen, 2.3 fold in the intestine, 2.6 fold in the stomach, 4.1. fold in the heart, 3.3 fold in the lung, 4.4 fold in the kidney, 2.3 fold in the thymus and 4.4 fold in macrophages (Fig. 3D, E). These results showed that the luciferase activity profile and its response to LPS treatment were in parallel with that of endogenous Rel mRNA expression in B6-Tg(c-Rel-luc)8Mlit mice.

Bottom Line: Revealing the dynamic expression of c-Rel in disease processes in vivo is critical for understanding c-Rel functions and for developing anti-inflammatory drugs.Luciferase expression could be tracked in living mice by the method of bioluminescence imaging in a variety of inflammatory processes, including LPS induced sepsis and EAE disease model.These results indicate that the B6-Tg(c-Rel-luc)(Mlit) mouse is a valuable animal model to study Rel expression in physiological and pathological processes, and the effects of various drug treatments in vivo.

View Article: PubMed Central - PubMed

Affiliation: School of Life Science and Technology, Tongji University, Shanghai, China.

ABSTRACT
c-Rel plays important roles in many inflammatory diseases. Revealing the dynamic expression of c-Rel in disease processes in vivo is critical for understanding c-Rel functions and for developing anti-inflammatory drugs. In this paper, a transgenic mouse line, B6-Tg(c-Rel-luc)(Mlit), which incorporated the transgene firefly luciferase driven by a 14.5-kb fragment containing mouse c-Rel gene Rel promoter, was generated to monitor Rel expression in vivo. Luciferase expression could be tracked in living mice by the method of bioluminescence imaging in a variety of inflammatory processes, including LPS induced sepsis and EAE disease model. The luciferase expression in transgenic mice was comparable to the endogenous Rel expression and could be suppressed by administration of anti-inflammatory drug dexamethasone or aspirin. These results indicate that the B6-Tg(c-Rel-luc)(Mlit) mouse is a valuable animal model to study Rel expression in physiological and pathological processes, and the effects of various drug treatments in vivo.

Show MeSH
Related in: MedlinePlus