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Neurogenic and neurotrophic effects of BDNF peptides in mouse hippocampal primary neuronal cell cultures.

Cardenas-Aguayo Mdel C, Kazim SF, Grundke-Iqbal I, Iqbal K - PLoS ONE (2013)

Bottom Line: Peptide B-3, when combined with BDNF, potentiated the survival effect of this neurotrophin on H(2)O(2)-treated E18 hippocampal cells.Peptides B-3 and B-5 were found to work as partial agonists and as partial antagonists competing with BDNF to activate the TrkB receptor in a dose-dependent manner.Taken together, these results suggest that the described BDNF tetra peptides are neurotrophic, can modulate BDNF signaling in a partial agonist/antagonist way, and offer a novel therapeutic approach to neural pathologies where BDNF levels are dysregulated.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurochemistry, New York State Institute for Basic Research in Developmental Disabilities, Staten Island, New York, United States of America.

ABSTRACT
The level of brain-derived neurotrophic factor (BDNF), a member of the neurotrophin family, is down regulated in Alzheimer's disease (AD), Parkinson's disease (PD), depression, stress, and anxiety; conversely the level of this neurotrophin is increased in autism spectrum disorders. Thus, modulating the level of BDNF can be a potential therapeutic approach for nervous system pathologies. In the present study, we designed five different tetra peptides (peptides B-1 to B-5) corresponding to different active regions of BDNF. These tetra peptides were found to be non-toxic, and they induced the expression of neuronal markers in mouse embryonic day 18 (E18) primary hippocampal neuronal cultures. Additionally, peptide B-5 induced the expression of BDNF and its receptor, TrkB, suggesting a positive feedback mechanism. The BDNF peptides induced only a moderate activation (phosphorylation at Tyr 706) of the TrkB receptor, which could be blocked by the Trk's inhibitor, K252a. Peptide B-3, when combined with BDNF, potentiated the survival effect of this neurotrophin on H(2)O(2)-treated E18 hippocampal cells. Peptides B-3 and B-5 were found to work as partial agonists and as partial antagonists competing with BDNF to activate the TrkB receptor in a dose-dependent manner. Taken together, these results suggest that the described BDNF tetra peptides are neurotrophic, can modulate BDNF signaling in a partial agonist/antagonist way, and offer a novel therapeutic approach to neural pathologies where BDNF levels are dysregulated.

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BDNF peptides potentiate the effect of BDNF in rescuing H2O2-induced neurotoxicity.(A) LDH cytotoxicity assay showing the percentage of cell death in hippocampal cells treated with increasing concentrations of H2O2 i.e. 0, 60, 80, and 100 µM for 6 h, and then after changing the medium, exposed to B-5, B-3, BDNF or BDNF+B-3 for 24 h. Peptide B-5 significantly reduced cell death caused by 80 µM H2O2 and Peptide B-3 significantly potentiated the neuroprotective effect of BDNF. (B) Percent viability of hippocampal cells by LDH assay. The cells were treated in the same way as in A. Peptide B-3 significantly increased the viability when combined with BDNF in cells not treated or treated with 60 or 80 µM of H2O2. Data were normalized to control (vehicle treated cells). * p<0.05, **p<0.01. ANOVA and/or Student’s t test, n = 3.
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pone-0053596-g004: BDNF peptides potentiate the effect of BDNF in rescuing H2O2-induced neurotoxicity.(A) LDH cytotoxicity assay showing the percentage of cell death in hippocampal cells treated with increasing concentrations of H2O2 i.e. 0, 60, 80, and 100 µM for 6 h, and then after changing the medium, exposed to B-5, B-3, BDNF or BDNF+B-3 for 24 h. Peptide B-5 significantly reduced cell death caused by 80 µM H2O2 and Peptide B-3 significantly potentiated the neuroprotective effect of BDNF. (B) Percent viability of hippocampal cells by LDH assay. The cells were treated in the same way as in A. Peptide B-3 significantly increased the viability when combined with BDNF in cells not treated or treated with 60 or 80 µM of H2O2. Data were normalized to control (vehicle treated cells). * p<0.05, **p<0.01. ANOVA and/or Student’s t test, n = 3.

Mentions: In order to evaluate the potential neuroprotective effect of BDNF peptides, we challenged the primary E18 hippocampal cells with 0, 60, 80 and 100 µM H2O2 for 6 h, and then cells were washed with culture medium and fresh culture medium was added containing the peptides B-5 or B-3 or BDNF or BDNF plus B-3 (Fig. 4 A, B). After 24 h, the cell viability was assayed by the LDH method. Cells treated with H2O2 showed a significant increase in cell death (ANOVA, p<0.0001) (Fig. 4A) with a concomitant reduction in cell viability (Fig. 4B) (ANOVA, p<0.001). The cells treated with BDNF after being exposed to H2O2 showed some reduction in cell death when compared to control medium treated cells, however, this was not statistically significant for most except for BDNF with 60 µM H2O2 treatment (0 µM H2O2, ANOVA, p = 0.9377, Student’s t test, p = 0.2700; 60 µM H2O2, ANOVA, p = 0.0304,Student’s t test, p = 0.1076; 80 µM H2O2, ANOVA, p = 0.3699, Student’s t test, p = 0.3094; 100 µM H2O2, ANOVA, p = 0.3764, Student’s t test, p = 0.9519). Conversely, BDNF increased cell viability significantly at 0 and 60 µM H2O2 but was not effective at high H2O2 concentrations (80 µM and 100 µM) (0 µM H2O2, ANOVA, p = 0.3167, Student’s t test, p = 0.0124; 60 µM H2O2, ANOVA, p = 0.0675, Student’s t test, p = 0.1072; 80 µM H2O2, ANOVA, p = 0.5762, Student’s t test, p = 0.7943; 100 µM H2O2, ANOVA, p = 0.4162, Student’s t test, p = 0.9508). These effects were enhanced by the combination of BDNF with Peptide B-3 in cells treated with 0, 60, and 80 µM but not 100 µM H2O2 (Cell death, 0 µM H2O2, ANOVA, p = 0.7534, Student’s t test, p = 0.7876; 60 µM H2O2, ANOVA, p = 0.0247, Student’s t test, p = 0.0256; 80 µM H2O2, ANOVA, p = 0.4880, Student’s t test, p = 0.0189; 100 µM H2O2, ANOVA, p = 0.1603, Student’s t test, p = 0.5894; Cell viability, 0 µM H2O2, ANOVA, p = 0.6739, Student’s t test, p = 0.0074; 60 µM H2O2, ANOVA, p = 0.0129, Student’s t test, p = 0.0790; 80 µM H2O2, ANOVA, p = 0.1956, Student’s t test, p = 0.0573; 100 µM H2O2, ANOVA, p = 0.2682, Student’s t test, p = 0.4745). These results suggest that peptide B-3 potentiates the neuroprotective effect of BDNF but alone is not sufficient to exert a significant effect. Peptide B-5 alone had a moderate effect in reducing the percentage of cell death when the hippocampal cells were treated with 80 µM of H2O2 (0 µM H2O2, ANOVA, p = 0.1119, Student’s t test, p = 0.1911; 60 µM H2O2, ANOVA, p = 0.2057, Student’s t test, p = 0.7086; 80 µM H2O2, ANOVA, p = 0.4007, Student’s t test, p = 0.051; 100 µM H2O2, ANOVA, p = 0.4124, Student’s t test, p = 0.6888); also it significantly increased the cell viability with 60 and 80 µM of H2O2 (0 µM H2O2, ANOVA, p = 0.6015, Student’s t test, p = 0.2478; 60 µM H2O2, ANOVA, p = 0.3191, Student’s t test, p = 0.084; 80 µM H2O2, ANOVA, p = 0.7923, Student’s t test, p = 0.0229; 100 µM H2O2, ANOVA, p = 0.7709, Student’s t test, p = 0.0717). We did not observe any potentiation of the neuroprotective effect of BDNF when used in combination with peptide B-5 (data not shown).


Neurogenic and neurotrophic effects of BDNF peptides in mouse hippocampal primary neuronal cell cultures.

Cardenas-Aguayo Mdel C, Kazim SF, Grundke-Iqbal I, Iqbal K - PLoS ONE (2013)

BDNF peptides potentiate the effect of BDNF in rescuing H2O2-induced neurotoxicity.(A) LDH cytotoxicity assay showing the percentage of cell death in hippocampal cells treated with increasing concentrations of H2O2 i.e. 0, 60, 80, and 100 µM for 6 h, and then after changing the medium, exposed to B-5, B-3, BDNF or BDNF+B-3 for 24 h. Peptide B-5 significantly reduced cell death caused by 80 µM H2O2 and Peptide B-3 significantly potentiated the neuroprotective effect of BDNF. (B) Percent viability of hippocampal cells by LDH assay. The cells were treated in the same way as in A. Peptide B-3 significantly increased the viability when combined with BDNF in cells not treated or treated with 60 or 80 µM of H2O2. Data were normalized to control (vehicle treated cells). * p<0.05, **p<0.01. ANOVA and/or Student’s t test, n = 3.
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pone-0053596-g004: BDNF peptides potentiate the effect of BDNF in rescuing H2O2-induced neurotoxicity.(A) LDH cytotoxicity assay showing the percentage of cell death in hippocampal cells treated with increasing concentrations of H2O2 i.e. 0, 60, 80, and 100 µM for 6 h, and then after changing the medium, exposed to B-5, B-3, BDNF or BDNF+B-3 for 24 h. Peptide B-5 significantly reduced cell death caused by 80 µM H2O2 and Peptide B-3 significantly potentiated the neuroprotective effect of BDNF. (B) Percent viability of hippocampal cells by LDH assay. The cells were treated in the same way as in A. Peptide B-3 significantly increased the viability when combined with BDNF in cells not treated or treated with 60 or 80 µM of H2O2. Data were normalized to control (vehicle treated cells). * p<0.05, **p<0.01. ANOVA and/or Student’s t test, n = 3.
Mentions: In order to evaluate the potential neuroprotective effect of BDNF peptides, we challenged the primary E18 hippocampal cells with 0, 60, 80 and 100 µM H2O2 for 6 h, and then cells were washed with culture medium and fresh culture medium was added containing the peptides B-5 or B-3 or BDNF or BDNF plus B-3 (Fig. 4 A, B). After 24 h, the cell viability was assayed by the LDH method. Cells treated with H2O2 showed a significant increase in cell death (ANOVA, p<0.0001) (Fig. 4A) with a concomitant reduction in cell viability (Fig. 4B) (ANOVA, p<0.001). The cells treated with BDNF after being exposed to H2O2 showed some reduction in cell death when compared to control medium treated cells, however, this was not statistically significant for most except for BDNF with 60 µM H2O2 treatment (0 µM H2O2, ANOVA, p = 0.9377, Student’s t test, p = 0.2700; 60 µM H2O2, ANOVA, p = 0.0304,Student’s t test, p = 0.1076; 80 µM H2O2, ANOVA, p = 0.3699, Student’s t test, p = 0.3094; 100 µM H2O2, ANOVA, p = 0.3764, Student’s t test, p = 0.9519). Conversely, BDNF increased cell viability significantly at 0 and 60 µM H2O2 but was not effective at high H2O2 concentrations (80 µM and 100 µM) (0 µM H2O2, ANOVA, p = 0.3167, Student’s t test, p = 0.0124; 60 µM H2O2, ANOVA, p = 0.0675, Student’s t test, p = 0.1072; 80 µM H2O2, ANOVA, p = 0.5762, Student’s t test, p = 0.7943; 100 µM H2O2, ANOVA, p = 0.4162, Student’s t test, p = 0.9508). These effects were enhanced by the combination of BDNF with Peptide B-3 in cells treated with 0, 60, and 80 µM but not 100 µM H2O2 (Cell death, 0 µM H2O2, ANOVA, p = 0.7534, Student’s t test, p = 0.7876; 60 µM H2O2, ANOVA, p = 0.0247, Student’s t test, p = 0.0256; 80 µM H2O2, ANOVA, p = 0.4880, Student’s t test, p = 0.0189; 100 µM H2O2, ANOVA, p = 0.1603, Student’s t test, p = 0.5894; Cell viability, 0 µM H2O2, ANOVA, p = 0.6739, Student’s t test, p = 0.0074; 60 µM H2O2, ANOVA, p = 0.0129, Student’s t test, p = 0.0790; 80 µM H2O2, ANOVA, p = 0.1956, Student’s t test, p = 0.0573; 100 µM H2O2, ANOVA, p = 0.2682, Student’s t test, p = 0.4745). These results suggest that peptide B-3 potentiates the neuroprotective effect of BDNF but alone is not sufficient to exert a significant effect. Peptide B-5 alone had a moderate effect in reducing the percentage of cell death when the hippocampal cells were treated with 80 µM of H2O2 (0 µM H2O2, ANOVA, p = 0.1119, Student’s t test, p = 0.1911; 60 µM H2O2, ANOVA, p = 0.2057, Student’s t test, p = 0.7086; 80 µM H2O2, ANOVA, p = 0.4007, Student’s t test, p = 0.051; 100 µM H2O2, ANOVA, p = 0.4124, Student’s t test, p = 0.6888); also it significantly increased the cell viability with 60 and 80 µM of H2O2 (0 µM H2O2, ANOVA, p = 0.6015, Student’s t test, p = 0.2478; 60 µM H2O2, ANOVA, p = 0.3191, Student’s t test, p = 0.084; 80 µM H2O2, ANOVA, p = 0.7923, Student’s t test, p = 0.0229; 100 µM H2O2, ANOVA, p = 0.7709, Student’s t test, p = 0.0717). We did not observe any potentiation of the neuroprotective effect of BDNF when used in combination with peptide B-5 (data not shown).

Bottom Line: Peptide B-3, when combined with BDNF, potentiated the survival effect of this neurotrophin on H(2)O(2)-treated E18 hippocampal cells.Peptides B-3 and B-5 were found to work as partial agonists and as partial antagonists competing with BDNF to activate the TrkB receptor in a dose-dependent manner.Taken together, these results suggest that the described BDNF tetra peptides are neurotrophic, can modulate BDNF signaling in a partial agonist/antagonist way, and offer a novel therapeutic approach to neural pathologies where BDNF levels are dysregulated.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurochemistry, New York State Institute for Basic Research in Developmental Disabilities, Staten Island, New York, United States of America.

ABSTRACT
The level of brain-derived neurotrophic factor (BDNF), a member of the neurotrophin family, is down regulated in Alzheimer's disease (AD), Parkinson's disease (PD), depression, stress, and anxiety; conversely the level of this neurotrophin is increased in autism spectrum disorders. Thus, modulating the level of BDNF can be a potential therapeutic approach for nervous system pathologies. In the present study, we designed five different tetra peptides (peptides B-1 to B-5) corresponding to different active regions of BDNF. These tetra peptides were found to be non-toxic, and they induced the expression of neuronal markers in mouse embryonic day 18 (E18) primary hippocampal neuronal cultures. Additionally, peptide B-5 induced the expression of BDNF and its receptor, TrkB, suggesting a positive feedback mechanism. The BDNF peptides induced only a moderate activation (phosphorylation at Tyr 706) of the TrkB receptor, which could be blocked by the Trk's inhibitor, K252a. Peptide B-3, when combined with BDNF, potentiated the survival effect of this neurotrophin on H(2)O(2)-treated E18 hippocampal cells. Peptides B-3 and B-5 were found to work as partial agonists and as partial antagonists competing with BDNF to activate the TrkB receptor in a dose-dependent manner. Taken together, these results suggest that the described BDNF tetra peptides are neurotrophic, can modulate BDNF signaling in a partial agonist/antagonist way, and offer a novel therapeutic approach to neural pathologies where BDNF levels are dysregulated.

Show MeSH
Related in: MedlinePlus