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Secreted miR-34a in astrocytic shedding vesicles enhanced the vulnerability of dopaminergic neurons to neurotoxins by targeting Bcl-2.

Mao S, Sun Q, Xiao H, Zhang C, Li L - Protein Cell (2015)

Bottom Line: To elucidate the potential role of glial MVs in disease, we evaluated the effects of secreted astrocytic MVs on stress condition.Further investigation showed that increased astrocytic miR-34a in SVs was involved in this progress via targeting anti-apoptotic protein Bcl-2 in dopaminergic neurons.These data revealed a novel mechanism underlying astrocyte-neuron interaction in disease.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Pharmaceutical Biotechnology, Nanjing Advanced Institute for Life Sciences (NAILS), Nanjing University School of Life Sciences, Nanjing, 210093, China.

ABSTRACT
MicroRNAs (miRNAs) are a class of noncoding RNAs that regulates target gene expression at posttranscriptional level, leading to further biological functions. We have demonstrated that microvesicles (MVs) can deliver miRNAs into target cells as a novel way of intercellular communication. It is reported that in central nervous system, glial cells release MVs, which modulate neuronal function in normal condition. To elucidate the potential role of glial MVs in disease, we evaluated the effects of secreted astrocytic MVs on stress condition. Our results demonstrated that after Lipopolysaccharide (LPS) stimulation, astrocytes released shedding vesicles (SVs) that enhanced vulnerability of dopaminergic neurons to neurotoxin. Further investigation showed that increased astrocytic miR-34a in SVs was involved in this progress via targeting anti-apoptotic protein Bcl-2 in dopaminergic neurons. We also found that inhibition of astrocytic miR-34a after LPS stimulation can postpone dopaminergic neuron loss under neurotoxin stress. These data revealed a novel mechanism underlying astrocyte-neuron interaction in disease.

No MeSH data available.


Related in: MedlinePlus

SVs derived from LPS-stimulated U-87 MG cells contain increased miR-34a and reduce SH-SY5Y cell viability to neurotoxins. (A) Cell viability assessed by CCK-8 showed that pretreatment with LPS SVs increased the vulnerability of SH-SY5Y cells to 0.2 mmol/L MPP+ treatment, *P < 0.05; (B) Cell viability assessed by CCK-8 showed that pretreatment with LPS SVs increased the vulnerability of SH-SY5Y cells to 10 μmol/L 6-OHDA treatment, *P < 0.05. Control SV: SVs derived from PBS-stimulated U-87 MG cells; LPS SVs: SVs derived from LPS-stimulated U-87 MG cells; Control Exo: Exosomes derived from control U-87 MG cells; LPS Exo: Exosomes derived from LPS-stimulated U-87 MG cells; (C) Relative miRNA levels between control SV and LPS SVs, *P < 0.05; (D) Fluorescent images showed that labeled SVs derived from U-87 MG cells can enter SH-SY5Y cells after co-incubation; red dots are labeled shedding vesicles, Scale bar = 10 μm; (E) FACS analysis showed that more than 80% of SH-SY5Y cells carry red fluorescence after co-incubation with labeled SVs; the left histogram represents the control group; the right histogram represents SH-SY5Y cells co-cultured with the fluorescence-labeled SVs
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Fig2: SVs derived from LPS-stimulated U-87 MG cells contain increased miR-34a and reduce SH-SY5Y cell viability to neurotoxins. (A) Cell viability assessed by CCK-8 showed that pretreatment with LPS SVs increased the vulnerability of SH-SY5Y cells to 0.2 mmol/L MPP+ treatment, *P < 0.05; (B) Cell viability assessed by CCK-8 showed that pretreatment with LPS SVs increased the vulnerability of SH-SY5Y cells to 10 μmol/L 6-OHDA treatment, *P < 0.05. Control SV: SVs derived from PBS-stimulated U-87 MG cells; LPS SVs: SVs derived from LPS-stimulated U-87 MG cells; Control Exo: Exosomes derived from control U-87 MG cells; LPS Exo: Exosomes derived from LPS-stimulated U-87 MG cells; (C) Relative miRNA levels between control SV and LPS SVs, *P < 0.05; (D) Fluorescent images showed that labeled SVs derived from U-87 MG cells can enter SH-SY5Y cells after co-incubation; red dots are labeled shedding vesicles, Scale bar = 10 μm; (E) FACS analysis showed that more than 80% of SH-SY5Y cells carry red fluorescence after co-incubation with labeled SVs; the left histogram represents the control group; the right histogram represents SH-SY5Y cells co-cultured with the fluorescence-labeled SVs

Mentions: Next, we investigated the impact of astrocyte-derived microvesicles under stress conditions on cell survival. We found that neither SVs nor exosomes derived from the LPS-stimulated U87-MG astroglial cell line had any effects on SH-SY5Y cell viability under normal conditions (Fig. S1). However, further investigation showed that pretreatment of SVs derived from LPS-stimulated U87-MG cells increased the vulnerability of the SH-SY5Y cells to threshold concentrations of neurotoxins, such as 0.2 mmol/L MPP+ or 10 μmol/L 6-OHDA (Figs. 2A, 2B and S1).Figure 2


Secreted miR-34a in astrocytic shedding vesicles enhanced the vulnerability of dopaminergic neurons to neurotoxins by targeting Bcl-2.

Mao S, Sun Q, Xiao H, Zhang C, Li L - Protein Cell (2015)

SVs derived from LPS-stimulated U-87 MG cells contain increased miR-34a and reduce SH-SY5Y cell viability to neurotoxins. (A) Cell viability assessed by CCK-8 showed that pretreatment with LPS SVs increased the vulnerability of SH-SY5Y cells to 0.2 mmol/L MPP+ treatment, *P < 0.05; (B) Cell viability assessed by CCK-8 showed that pretreatment with LPS SVs increased the vulnerability of SH-SY5Y cells to 10 μmol/L 6-OHDA treatment, *P < 0.05. Control SV: SVs derived from PBS-stimulated U-87 MG cells; LPS SVs: SVs derived from LPS-stimulated U-87 MG cells; Control Exo: Exosomes derived from control U-87 MG cells; LPS Exo: Exosomes derived from LPS-stimulated U-87 MG cells; (C) Relative miRNA levels between control SV and LPS SVs, *P < 0.05; (D) Fluorescent images showed that labeled SVs derived from U-87 MG cells can enter SH-SY5Y cells after co-incubation; red dots are labeled shedding vesicles, Scale bar = 10 μm; (E) FACS analysis showed that more than 80% of SH-SY5Y cells carry red fluorescence after co-incubation with labeled SVs; the left histogram represents the control group; the right histogram represents SH-SY5Y cells co-cultured with the fluorescence-labeled SVs
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Related In: Results  -  Collection

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Fig2: SVs derived from LPS-stimulated U-87 MG cells contain increased miR-34a and reduce SH-SY5Y cell viability to neurotoxins. (A) Cell viability assessed by CCK-8 showed that pretreatment with LPS SVs increased the vulnerability of SH-SY5Y cells to 0.2 mmol/L MPP+ treatment, *P < 0.05; (B) Cell viability assessed by CCK-8 showed that pretreatment with LPS SVs increased the vulnerability of SH-SY5Y cells to 10 μmol/L 6-OHDA treatment, *P < 0.05. Control SV: SVs derived from PBS-stimulated U-87 MG cells; LPS SVs: SVs derived from LPS-stimulated U-87 MG cells; Control Exo: Exosomes derived from control U-87 MG cells; LPS Exo: Exosomes derived from LPS-stimulated U-87 MG cells; (C) Relative miRNA levels between control SV and LPS SVs, *P < 0.05; (D) Fluorescent images showed that labeled SVs derived from U-87 MG cells can enter SH-SY5Y cells after co-incubation; red dots are labeled shedding vesicles, Scale bar = 10 μm; (E) FACS analysis showed that more than 80% of SH-SY5Y cells carry red fluorescence after co-incubation with labeled SVs; the left histogram represents the control group; the right histogram represents SH-SY5Y cells co-cultured with the fluorescence-labeled SVs
Mentions: Next, we investigated the impact of astrocyte-derived microvesicles under stress conditions on cell survival. We found that neither SVs nor exosomes derived from the LPS-stimulated U87-MG astroglial cell line had any effects on SH-SY5Y cell viability under normal conditions (Fig. S1). However, further investigation showed that pretreatment of SVs derived from LPS-stimulated U87-MG cells increased the vulnerability of the SH-SY5Y cells to threshold concentrations of neurotoxins, such as 0.2 mmol/L MPP+ or 10 μmol/L 6-OHDA (Figs. 2A, 2B and S1).Figure 2

Bottom Line: To elucidate the potential role of glial MVs in disease, we evaluated the effects of secreted astrocytic MVs on stress condition.Further investigation showed that increased astrocytic miR-34a in SVs was involved in this progress via targeting anti-apoptotic protein Bcl-2 in dopaminergic neurons.These data revealed a novel mechanism underlying astrocyte-neuron interaction in disease.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Pharmaceutical Biotechnology, Nanjing Advanced Institute for Life Sciences (NAILS), Nanjing University School of Life Sciences, Nanjing, 210093, China.

ABSTRACT
MicroRNAs (miRNAs) are a class of noncoding RNAs that regulates target gene expression at posttranscriptional level, leading to further biological functions. We have demonstrated that microvesicles (MVs) can deliver miRNAs into target cells as a novel way of intercellular communication. It is reported that in central nervous system, glial cells release MVs, which modulate neuronal function in normal condition. To elucidate the potential role of glial MVs in disease, we evaluated the effects of secreted astrocytic MVs on stress condition. Our results demonstrated that after Lipopolysaccharide (LPS) stimulation, astrocytes released shedding vesicles (SVs) that enhanced vulnerability of dopaminergic neurons to neurotoxin. Further investigation showed that increased astrocytic miR-34a in SVs was involved in this progress via targeting anti-apoptotic protein Bcl-2 in dopaminergic neurons. We also found that inhibition of astrocytic miR-34a after LPS stimulation can postpone dopaminergic neuron loss under neurotoxin stress. These data revealed a novel mechanism underlying astrocyte-neuron interaction in disease.

No MeSH data available.


Related in: MedlinePlus