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An Aminopropyl Carbazole Derivative Induces Neurogenesis by Increasing Final Cell Division in Neural Stem Cells.

Shin JY, Kong SY, Yoon HJ, Ann J, Lee J, Kim HJ - Biomol Ther (Seoul) (2015)

Bottom Line: Although P7C3 is known to increase neurogenesis by protecting newborn neurons, it is not known whether its derivatives also have protective effects to increase neurogenesis.In the current study, we examined how 1 induces neurogenesis.Compound 1 did not induce astrocytogenesis during NSC differentiation. 5-Bromo-2'-deoxyuridine (BrdU) pulsing experiments showed that 1 significantly enhanced BrdU-positive neurons.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular and Stem Cell Pharmacology, College of Pharmacy, Chung-Ang University, Seoul 156-756.

ABSTRACT
P7C3 and its derivatives, 1-(3,6-dibromo-9H-carbazol-9-yl)-3-(p-tolylamino)propan-2-ol (1) and N-(3-(3,6-dibromo-9H-carbazol-9-yl)-2-hydroxypropyl)-N-(3-methoxyphenyl)-4-methylbenzenesulfonamide (2), were previously reported to increase neurogenesis in rat neural stem cells (NSCs). Although P7C3 is known to increase neurogenesis by protecting newborn neurons, it is not known whether its derivatives also have protective effects to increase neurogenesis. In the current study, we examined how 1 induces neurogenesis. The treatment of 1 in NSCs increased numbers of cells in the absence of epidermal growth factor (EGF) and fibroblast growth factor 2 (FGF2), while not affecting those in the presence of growth factors. Compound 1 did not induce astrocytogenesis during NSC differentiation. 5-Bromo-2'-deoxyuridine (BrdU) pulsing experiments showed that 1 significantly enhanced BrdU-positive neurons. Taken together, our data suggest that 1 promotes neurogenesis by the induction of final cell division during NSC differentiation.

No MeSH data available.


Related in: MedlinePlus

1 does not affect NSC proliferation in the presence of mitogens. (A) Individual neurospheres were treated with either 5.0 μM compound 1 or DMSO for 4 days in the presence of EGF and FGF2. Digital images of neurospheres on day 0–3 are shown. Scale bar, 100 μm. (B) Volumes of neurospheres were calculated using the measured diameters. The values are the mean ± S.D. (n≥4, for each treatment). (C) Passaged NSCs were dissociated, plated as single cells at clonal density (1 cell/μl), and grown as neurospheres for 7 days in the presence of growth factors added with 5.0 μM compound 1 or DMSO. Scale bar, 100 μm. (D) The number of spheres was counted from 3 independent randomly chosen fields. The values are the mean ± S.D. (n=3). Statistical analysis of all data was performed using Student’s t-test.
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f5-bt-23-313: 1 does not affect NSC proliferation in the presence of mitogens. (A) Individual neurospheres were treated with either 5.0 μM compound 1 or DMSO for 4 days in the presence of EGF and FGF2. Digital images of neurospheres on day 0–3 are shown. Scale bar, 100 μm. (B) Volumes of neurospheres were calculated using the measured diameters. The values are the mean ± S.D. (n≥4, for each treatment). (C) Passaged NSCs were dissociated, plated as single cells at clonal density (1 cell/μl), and grown as neurospheres for 7 days in the presence of growth factors added with 5.0 μM compound 1 or DMSO. Scale bar, 100 μm. (D) The number of spheres was counted from 3 independent randomly chosen fields. The values are the mean ± S.D. (n=3). Statistical analysis of all data was performed using Student’s t-test.

Mentions: We next examined whether 1 affected NSC proliferation in the presence of EGF and FGF2. Firstly, we performed a neurosphere growth assay. The sizes of the neurospheres treated with 5.0 μM compound 1 or DMSO in the presence of growth factors were measured daily for 4 days (day 0–3). As shown in Fig. 5A, the sizes of the neurospheres became bigger in a time-dependent manner. To evaluate the effect of 1 on NSC proliferation, we measured the diameters of neurospheres and calculated the volumes of spheres using the equation V=4/3πr3, where r=1/2 diameter. Neurosphere volume on each day was divided by the volume on day 0 and then multiplied by 100 to obtain the percentage increase in sphere volume. However, there was no significant difference in the volume increase of the neurospheres between the two groups (Fig. 5B). Furthermore, when we dissociated, plated NSCs at clonal density (1 cell/μl), and cultured for 7 days in the presence of mitogens with 5.0 μM compound 1 or DMSO to determine neurosphere formation ability, we observed no significant difference between the two groups (Fig. 5C, 5D). Taken together, these data suggest that 1 does not change the proliferation or neurosphere formation ability of NSCs in the presence of growth factors.


An Aminopropyl Carbazole Derivative Induces Neurogenesis by Increasing Final Cell Division in Neural Stem Cells.

Shin JY, Kong SY, Yoon HJ, Ann J, Lee J, Kim HJ - Biomol Ther (Seoul) (2015)

1 does not affect NSC proliferation in the presence of mitogens. (A) Individual neurospheres were treated with either 5.0 μM compound 1 or DMSO for 4 days in the presence of EGF and FGF2. Digital images of neurospheres on day 0–3 are shown. Scale bar, 100 μm. (B) Volumes of neurospheres were calculated using the measured diameters. The values are the mean ± S.D. (n≥4, for each treatment). (C) Passaged NSCs were dissociated, plated as single cells at clonal density (1 cell/μl), and grown as neurospheres for 7 days in the presence of growth factors added with 5.0 μM compound 1 or DMSO. Scale bar, 100 μm. (D) The number of spheres was counted from 3 independent randomly chosen fields. The values are the mean ± S.D. (n=3). Statistical analysis of all data was performed using Student’s t-test.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4489824&req=5

f5-bt-23-313: 1 does not affect NSC proliferation in the presence of mitogens. (A) Individual neurospheres were treated with either 5.0 μM compound 1 or DMSO for 4 days in the presence of EGF and FGF2. Digital images of neurospheres on day 0–3 are shown. Scale bar, 100 μm. (B) Volumes of neurospheres were calculated using the measured diameters. The values are the mean ± S.D. (n≥4, for each treatment). (C) Passaged NSCs were dissociated, plated as single cells at clonal density (1 cell/μl), and grown as neurospheres for 7 days in the presence of growth factors added with 5.0 μM compound 1 or DMSO. Scale bar, 100 μm. (D) The number of spheres was counted from 3 independent randomly chosen fields. The values are the mean ± S.D. (n=3). Statistical analysis of all data was performed using Student’s t-test.
Mentions: We next examined whether 1 affected NSC proliferation in the presence of EGF and FGF2. Firstly, we performed a neurosphere growth assay. The sizes of the neurospheres treated with 5.0 μM compound 1 or DMSO in the presence of growth factors were measured daily for 4 days (day 0–3). As shown in Fig. 5A, the sizes of the neurospheres became bigger in a time-dependent manner. To evaluate the effect of 1 on NSC proliferation, we measured the diameters of neurospheres and calculated the volumes of spheres using the equation V=4/3πr3, where r=1/2 diameter. Neurosphere volume on each day was divided by the volume on day 0 and then multiplied by 100 to obtain the percentage increase in sphere volume. However, there was no significant difference in the volume increase of the neurospheres between the two groups (Fig. 5B). Furthermore, when we dissociated, plated NSCs at clonal density (1 cell/μl), and cultured for 7 days in the presence of mitogens with 5.0 μM compound 1 or DMSO to determine neurosphere formation ability, we observed no significant difference between the two groups (Fig. 5C, 5D). Taken together, these data suggest that 1 does not change the proliferation or neurosphere formation ability of NSCs in the presence of growth factors.

Bottom Line: Although P7C3 is known to increase neurogenesis by protecting newborn neurons, it is not known whether its derivatives also have protective effects to increase neurogenesis.In the current study, we examined how 1 induces neurogenesis.Compound 1 did not induce astrocytogenesis during NSC differentiation. 5-Bromo-2'-deoxyuridine (BrdU) pulsing experiments showed that 1 significantly enhanced BrdU-positive neurons.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular and Stem Cell Pharmacology, College of Pharmacy, Chung-Ang University, Seoul 156-756.

ABSTRACT
P7C3 and its derivatives, 1-(3,6-dibromo-9H-carbazol-9-yl)-3-(p-tolylamino)propan-2-ol (1) and N-(3-(3,6-dibromo-9H-carbazol-9-yl)-2-hydroxypropyl)-N-(3-methoxyphenyl)-4-methylbenzenesulfonamide (2), were previously reported to increase neurogenesis in rat neural stem cells (NSCs). Although P7C3 is known to increase neurogenesis by protecting newborn neurons, it is not known whether its derivatives also have protective effects to increase neurogenesis. In the current study, we examined how 1 induces neurogenesis. The treatment of 1 in NSCs increased numbers of cells in the absence of epidermal growth factor (EGF) and fibroblast growth factor 2 (FGF2), while not affecting those in the presence of growth factors. Compound 1 did not induce astrocytogenesis during NSC differentiation. 5-Bromo-2'-deoxyuridine (BrdU) pulsing experiments showed that 1 significantly enhanced BrdU-positive neurons. Taken together, our data suggest that 1 promotes neurogenesis by the induction of final cell division during NSC differentiation.

No MeSH data available.


Related in: MedlinePlus