Limits...
Transgenic Mouse Expressing Optical MicroRNA Reporter for Monitoring MicroRNA-124 Action during Development.

Choi Y, Hwang do W, Kim MY, Kim JY, Sun W, Lee DS - Front Mol Neurosci (2016)

Bottom Line: A method to monitor the action of miRNAs in vivo shall help understand their dynamic behavior during development.Bioluminescence dramatically decreased in the brain between embryonic day 13 and 16 as endogenous miR-124 expression increased, which sustained into adulthood.The inverse relationship of miR-124 expression was observed with luciferase bioluminescence and activity ex vivo as well as in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Nuclear Medicine, College of Medicine, Seoul National University Seoul, South Korea.

ABSTRACT
MicroRNAs (miRNAs) fine-tune target protein synthesis by suppressing gene expression, temporally changing along development and possibly in pathological conditions. A method to monitor the action of miRNAs in vivo shall help understand their dynamic behavior during development. In this study, we established a transgenic mouse harboring miR-124 responsive element in their luciferase-eGFP reporter transgenes which enabled monitoring the action of miR-124 in the brain and other organs in vivo by the bioluminescence imaging. The mouse model was produced and verified by imaging ex vivo so that luminescence by luciferase shone and then reduced during development with miR-124 expression. Bioluminescence dramatically decreased in the brain between embryonic day 13 and 16 as endogenous miR-124 expression increased, which sustained into adulthood. The inverse relationship of miR-124 expression was observed with luciferase bioluminescence and activity ex vivo as well as in vivo. Taken together, one can use this microRNA-transgenic mouse to investigate the temporal changes of microRNA action in vivo in the brain as well as in other organs.

No MeSH data available.


Related in: MedlinePlus

Developmental stage-dependent miR-124 expression regulates luciferase signal. (A)Ex vivo bioluminescence images of brain and liver of a transgenic mouse along the bodily development in utero and after birth (E13 n = 6, E16 n = 6, P10 n = 3, young n = 3, and old n = 4); 3∼5 month old (young), 16∼24 month old (old). (B) Quantified results of these ex vivo images. Average counts of the brain and the liver was shown as photons/sec/cm2. The brain luciferase was maximal at E13 and decreased to the minimum at young adult period and the liver luciferase was maximal at E16 and minimum at young adult period. (C) Luciferase activity (RLU/mg) of brain lysate in each developmental stage. (D) Quantitative real-time RT-PCR results of mir-124 over U6 of the brain lysates in each developmental stage. Data are shown as means ± SD. RLU/mg means relative light units per mg of protein of the lysate. (E) Relationship between miR-124 expression and luciferase activity of brain lysates which showed inverse relation. After log-log transform, the relation was inverse-proportional (inset). Whisker of each point represents S.D.’s of miR-124 expression (abscissa) and luciferase activity (ordinate).
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4940420&req=5

Figure 4: Developmental stage-dependent miR-124 expression regulates luciferase signal. (A)Ex vivo bioluminescence images of brain and liver of a transgenic mouse along the bodily development in utero and after birth (E13 n = 6, E16 n = 6, P10 n = 3, young n = 3, and old n = 4); 3∼5 month old (young), 16∼24 month old (old). (B) Quantified results of these ex vivo images. Average counts of the brain and the liver was shown as photons/sec/cm2. The brain luciferase was maximal at E13 and decreased to the minimum at young adult period and the liver luciferase was maximal at E16 and minimum at young adult period. (C) Luciferase activity (RLU/mg) of brain lysate in each developmental stage. (D) Quantitative real-time RT-PCR results of mir-124 over U6 of the brain lysates in each developmental stage. Data are shown as means ± SD. RLU/mg means relative light units per mg of protein of the lysate. (E) Relationship between miR-124 expression and luciferase activity of brain lysates which showed inverse relation. After log-log transform, the relation was inverse-proportional (inset). Whisker of each point represents S.D.’s of miR-124 expression (abscissa) and luciferase activity (ordinate).

Mentions: The pattern of miR-124 expression and luciferase reporter bioluminescence was compared using brain tissues in each period of E13, E16, P10, young and old adult with ex vivo luminescence imaging. At E13 period, the bioluminescence of the brain was higher than that of the liver on luciferase imaging but it decreased at E16, further at P10 and it was the lowest in the young adult period (Figures 4A,B). In contrast, bioluminescence of the liver was the highest at E16 and decreased at P10 and it was the lowest in the young adult period (Figures 4A,B).


Transgenic Mouse Expressing Optical MicroRNA Reporter for Monitoring MicroRNA-124 Action during Development.

Choi Y, Hwang do W, Kim MY, Kim JY, Sun W, Lee DS - Front Mol Neurosci (2016)

Developmental stage-dependent miR-124 expression regulates luciferase signal. (A)Ex vivo bioluminescence images of brain and liver of a transgenic mouse along the bodily development in utero and after birth (E13 n = 6, E16 n = 6, P10 n = 3, young n = 3, and old n = 4); 3∼5 month old (young), 16∼24 month old (old). (B) Quantified results of these ex vivo images. Average counts of the brain and the liver was shown as photons/sec/cm2. The brain luciferase was maximal at E13 and decreased to the minimum at young adult period and the liver luciferase was maximal at E16 and minimum at young adult period. (C) Luciferase activity (RLU/mg) of brain lysate in each developmental stage. (D) Quantitative real-time RT-PCR results of mir-124 over U6 of the brain lysates in each developmental stage. Data are shown as means ± SD. RLU/mg means relative light units per mg of protein of the lysate. (E) Relationship between miR-124 expression and luciferase activity of brain lysates which showed inverse relation. After log-log transform, the relation was inverse-proportional (inset). Whisker of each point represents S.D.’s of miR-124 expression (abscissa) and luciferase activity (ordinate).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4940420&req=5

Figure 4: Developmental stage-dependent miR-124 expression regulates luciferase signal. (A)Ex vivo bioluminescence images of brain and liver of a transgenic mouse along the bodily development in utero and after birth (E13 n = 6, E16 n = 6, P10 n = 3, young n = 3, and old n = 4); 3∼5 month old (young), 16∼24 month old (old). (B) Quantified results of these ex vivo images. Average counts of the brain and the liver was shown as photons/sec/cm2. The brain luciferase was maximal at E13 and decreased to the minimum at young adult period and the liver luciferase was maximal at E16 and minimum at young adult period. (C) Luciferase activity (RLU/mg) of brain lysate in each developmental stage. (D) Quantitative real-time RT-PCR results of mir-124 over U6 of the brain lysates in each developmental stage. Data are shown as means ± SD. RLU/mg means relative light units per mg of protein of the lysate. (E) Relationship between miR-124 expression and luciferase activity of brain lysates which showed inverse relation. After log-log transform, the relation was inverse-proportional (inset). Whisker of each point represents S.D.’s of miR-124 expression (abscissa) and luciferase activity (ordinate).
Mentions: The pattern of miR-124 expression and luciferase reporter bioluminescence was compared using brain tissues in each period of E13, E16, P10, young and old adult with ex vivo luminescence imaging. At E13 period, the bioluminescence of the brain was higher than that of the liver on luciferase imaging but it decreased at E16, further at P10 and it was the lowest in the young adult period (Figures 4A,B). In contrast, bioluminescence of the liver was the highest at E16 and decreased at P10 and it was the lowest in the young adult period (Figures 4A,B).

Bottom Line: A method to monitor the action of miRNAs in vivo shall help understand their dynamic behavior during development.Bioluminescence dramatically decreased in the brain between embryonic day 13 and 16 as endogenous miR-124 expression increased, which sustained into adulthood.The inverse relationship of miR-124 expression was observed with luciferase bioluminescence and activity ex vivo as well as in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Nuclear Medicine, College of Medicine, Seoul National University Seoul, South Korea.

ABSTRACT
MicroRNAs (miRNAs) fine-tune target protein synthesis by suppressing gene expression, temporally changing along development and possibly in pathological conditions. A method to monitor the action of miRNAs in vivo shall help understand their dynamic behavior during development. In this study, we established a transgenic mouse harboring miR-124 responsive element in their luciferase-eGFP reporter transgenes which enabled monitoring the action of miR-124 in the brain and other organs in vivo by the bioluminescence imaging. The mouse model was produced and verified by imaging ex vivo so that luminescence by luciferase shone and then reduced during development with miR-124 expression. Bioluminescence dramatically decreased in the brain between embryonic day 13 and 16 as endogenous miR-124 expression increased, which sustained into adulthood. The inverse relationship of miR-124 expression was observed with luciferase bioluminescence and activity ex vivo as well as in vivo. Taken together, one can use this microRNA-transgenic mouse to investigate the temporal changes of microRNA action in vivo in the brain as well as in other organs.

No MeSH data available.


Related in: MedlinePlus