Shi, Xinyu
Impact of microrna-29a-3p on the neural stem cell differentiation process into neurons - Vol.85(4), Jul-Aug - Mumbai Indian Journal of Pharmaceutical Science 2023 - 1025-1032p.
To explore how microRNA-29a-3p influences the differentiation of neural stem cells into neurons and to
identify any potential mechanisms involved. The program TargetScan, available online was utilized to
examine and make predictions about the microRNA-29a-3p located upstream that regulate doublecortin
X messenger ribonucleic acid. Ultimately, microRNA-29a-3p was preferred as the main central point of the
investigation. The targeting of doublecortin X gene by microRNA-29a-3p was validated by employing a
dual-luciferase reporter gene system. Before subjecting to adherent differentiation, the neural stem cells were
transfected with microRNA-29a-3p inhibitor, microRNA-29a-3p mimic and normal control. MicroRNA and
doublecortin X messenger ribonucleic acid levels were measured through quantitative reverse transcription-
polymerase chain reaction and doublecortin X protein levels were determined through Western blot assays.
The differentiation of neuronal precursor cells was detected by doublecortin X immunofluorescence.
Neuronal differentiation was detected by microtubule-associated protein 2 immunofluorescence. According
to the dual-luciferase reporter gene system, miR-29a-3p targeted the doublecortin X gene; microRNA-29a-
3p significantly decreased in differentiated cells for 3 d was observed, along with a significant reduction in
levels of doublecortin X protein and messenger ribonucleic acid. On the other hand, a significant raise in
microRNA-29a-3p in differentiated cells of microRNA-29a-3p mimic group for a period of 3 d was noticed
with a significant drop in protein levels, doublecortin X messenger ribonucleic acid and doublecortin X
immunofluorescence-positive neuronal precursor cells. From the microtubule-associated protein 2
immunofluorescence, it was confirmed that microRNA-29a-3p inhibitor group significantly elevated the total
number of neurons, while microRNA-29a-3p mimic reduced the neurons on 7 d. Therefore, low expression
of microRNA-29a-3p promote differentiation of neural stem cells into neurons via targeting doublecortin X.
PHARMACEUTICS
Impact of microrna-29a-3p on the neural stem cell differentiation process into neurons - Vol.85(4), Jul-Aug - Mumbai Indian Journal of Pharmaceutical Science 2023 - 1025-1032p.
To explore how microRNA-29a-3p influences the differentiation of neural stem cells into neurons and to
identify any potential mechanisms involved. The program TargetScan, available online was utilized to
examine and make predictions about the microRNA-29a-3p located upstream that regulate doublecortin
X messenger ribonucleic acid. Ultimately, microRNA-29a-3p was preferred as the main central point of the
investigation. The targeting of doublecortin X gene by microRNA-29a-3p was validated by employing a
dual-luciferase reporter gene system. Before subjecting to adherent differentiation, the neural stem cells were
transfected with microRNA-29a-3p inhibitor, microRNA-29a-3p mimic and normal control. MicroRNA and
doublecortin X messenger ribonucleic acid levels were measured through quantitative reverse transcription-
polymerase chain reaction and doublecortin X protein levels were determined through Western blot assays.
The differentiation of neuronal precursor cells was detected by doublecortin X immunofluorescence.
Neuronal differentiation was detected by microtubule-associated protein 2 immunofluorescence. According
to the dual-luciferase reporter gene system, miR-29a-3p targeted the doublecortin X gene; microRNA-29a-
3p significantly decreased in differentiated cells for 3 d was observed, along with a significant reduction in
levels of doublecortin X protein and messenger ribonucleic acid. On the other hand, a significant raise in
microRNA-29a-3p in differentiated cells of microRNA-29a-3p mimic group for a period of 3 d was noticed
with a significant drop in protein levels, doublecortin X messenger ribonucleic acid and doublecortin X
immunofluorescence-positive neuronal precursor cells. From the microtubule-associated protein 2
immunofluorescence, it was confirmed that microRNA-29a-3p inhibitor group significantly elevated the total
number of neurons, while microRNA-29a-3p mimic reduced the neurons on 7 d. Therefore, low expression
of microRNA-29a-3p promote differentiation of neural stem cells into neurons via targeting doublecortin X.
PHARMACEUTICS