Inhibition of Sox2 Expression in the Adult Neural Stem Cell Niche In Vivo by Monocationic-based siRNA Delivery

Remaud, Sylvie; López-Juárez, Alejandra; Bolcato-Bellemin, Anne-Laure; Neuberg, Patrick; Stock, Fabrice; Bonnet, Marie-Elise; Ghaddab, Rym; Clerget-Froidevaux, Marie-Stéphanie; Pierre-Simons, Jacqueline; Erbacher, Patrick; Demeneix, Barbara; Morvan-Dubois, Ghislaine

doi:10.1038/mtna.2013.8

Cited by 9 publications

(12 citation statements)

References 31 publications

(46 reference statements)

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“…siRNA (24 pmol per animal) was diluted in glucose (5%) and mixed with monocationic lipid (IC10, Polyplus-transfection) at a ratio of 15 monocationic lipid nitrogens per RNA phosphate as described before [36], [37]. Complexes prepared at room temperature are stable for two hours after preparation.…”

Section: Methodsmentioning

confidence: 99%

“…siRNA against pGL2 (for control group) and NgR1 (Rtn4r, 4 different sequences, Cat no: SI02722888, SI02699186, SI02748333, SI02678249) were purchased from Qiagen (validated siRNA). siRNA (24 pmol per animal) was diluted in glucose (5%) and mixed with monocationic lipid (IC10, Polyplus-transfection) at a ratio of 15 monocationic lipid nitrogens per RNA phosphate as described before [36] , [37] . Complexes prepared at room temperature are stable for two hours after preparation.…”

Section: Methodsmentioning

confidence: 99%

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Nogo Receptor Inhibition Enhances Functional Recovery following Lysolecithin-Induced Demyelination in Mouse Optic Chiasm

et al. 2014

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BackgroundInhibitory factors have been implicated in the failure of remyelination in demyelinating diseases. Myelin associated inhibitors act through a common receptor called Nogo receptor (NgR) that plays critical inhibitory roles in CNS plasticity. Here we investigated the effects of abrogating NgR inhibition in a non-immune model of focal demyelination in adult mouse optic chiasm.Methodology/Principal FindingsA focal area of demyelination was induced in adult mouse optic chiasm by microinjection of lysolecithin. To knock down NgR levels, siRNAs against NgR were intracerebroventricularly administered via a permanent cannula over 14 days, Functional changes were monitored by electrophysiological recording of latency of visual evoked potentials (VEPs). Histological analysis was carried out 3, 7 and 14 days post demyelination lesion. To assess the effect of NgR inhibition on precursor cell repopulation, BrdU was administered to the animals prior to the demyelination induction. Inhibition of NgR significantly restored VEPs responses following optic chiasm demyelination. These findings were confirmed histologically by myelin specific staining. siNgR application resulted in a smaller lesion size compared to control. NgR inhibition significantly increased the numbers of BrdU+/Olig2+ progenitor cells in the lesioned area and in the neurogenic zone of the third ventricle. These progenitor cells (Olig2+ or GFAP+) migrated away from this area as a function of time.Conclusions/SignificanceOur results show that inhibition of NgR facilitate myelin repair in the demyelinated chiasm, with enhanced recruitment of proliferating cells to the lesion site. Thus, antagonizing NgR function could have therapeutic potential for demyelinating disorders such as Multiple Sclerosis.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Nogo Receptor Inhibition Enhances Functional Recovery following Lysolecithin-Induced Demyelination in Mouse Optic Chiasm

et al. 2014

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“…Previous studies have demonstrated the expression of scleraxis [33][34][35], Nanog [36], and Sox 2 [37] in the cytoplasm. Although these transcription factors function in the cell nucleus, stimulatory signal is likely required to induce their translocation to the cell nucleus.…”

Section: Role Of Lrcs During Tendon Repairmentioning

confidence: 99%

In Vivo Identity of Tendon Stem Cells and the Roles of Stem Cells in Tendon Healing

Tan

Lui

Lee

2013

Stem Cells and Development

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We investigated the spatial distribution of stem cells in tendons and the roles of stem cells in early tendon repair. The relationship between tendon-derived stem cells (TDSCs) isolated in vitro and tendon stem cells in vivo was also explored. Iododeoxyuridine (IdU) label-retaining method was used for labeling stem cells in rat patellar tendons with and without injury. Co-localization of label-retaining cells (LRCs) with different markers was done by immunofluorescent staining. TDSCs were isolated from patellar tendon mid-substance after IdU pulsing, and the expression of different markers in fresh and expanded cells was done by immunofluorescent staining. More LRCs were found at the peritenon and tendon-bone junction compared with the mid-substance. Some LRCs at the peritenon were located at the perivascular niche. The LRC number and the expression of proliferative, tendon-related, pluripotency, and pericyte-related markers in LRCs in the window wound increased. Most of the freshly isolated TDSCs expressed IdU, and some TDSCs expressed pericyte-related markers, which were lost during expansion. Both freshly isolated and subcultured TDSCs expressed pluripotency markers, which were absent in LRCs in intact tendons. In conclusion, we identified LRCs at the peritenon, mid-substance, and tendon-bone junction. There were both vascular and non-vascular sources of LRCs at the peritenon, while the source of LRCs at the mid-substance was non-vascular. LRCs participated in tendon repair via migration, proliferation, activation for tenogenesis, and increased pluripotency. Some LRCs in the window wound were pericyte like. Most of the mid-substance TDSCs were LRCs. The pluripotency markers and pericyte-related marker in LRCs might be important for function after injury. Recently, stem/progenitor cells have been isolated from tendon tissues of varies species, including human, horse, rabbit, rat, and mouse [3][4][5][6]. These stem/progenitor cells isolated from tendon tissues exhibited self-renewal and multilineage differentiation potential [3][4][5][6]. Since the origin and identity of these cells were not clear, we called them tendon-derived stem cells (TDSCs) to indicate only the tissue from which the cells were isolated [5].Many studies suggested that the wall of capillaries, small vessels, and large vessels harbored stem/progenitor cells [7][8][9][10][11]. Some studies further suggested that mesenchymal stem cells (MSCs) were derived from pericytes [9,12].Pericytes/perivascular cells from a variety of tissues were reported to exhibit characteristics that were strikingly similar to those of MSCs [7][8][9][10][11]. For tendons, there was also evidence that the vasculature of tendon tissue might harbor stem cells [13]. However, tendon mid-substance is hypovascular compared with other tissues and receives its blood supply mainly from the endotenon and paratenon [14]. TDSCs isolated from the tendon mid-substance, while positive for alpha smooth muscle actin [5], were not positive for other pericyte-related markers [3,15]. Tend...

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“…Interestingly, the RT-PCR analysis showed decreased expression of CD133 , a stem cell specific marker. NESTIN and SOX2 expression were increased after treatment, but both markers are also expressed by neural progenitors [ 43 ], suggesting a neuronal commitment of the cells. The mRNA of early neuronal markers Tubulin βIII and DCX were both increased at 60 µmol/L of NFL peptide, validating the more advanced neuronal state.…”

Section: Discussionmentioning

confidence: 99%

The neurofilament derived-peptide NFL-TBS.40-63 enters in-vitro in human neural stem cells and increases their differentiation

2018

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Regenerative medicine is a promising approach to treat neurodegenerative diseases by replacing degenerating cells like neurons or oligodendrocytes. Targeting human neural stem cells directly in the brain is a big challenge in such a strategy. The neurofilament derived NFL-TBS.40-63 peptide has recently been introduced as a novel tool to target neural stem cells. Previous studies showed that this peptide can be internalized by rat neural stem cells in vitro and in vivo, which coincided with lower proliferation and self-renewal capacity and increase of differentiation. In this study, we analyzed the uptake and potential effects of the NFL-TBS.40-63 peptide on human neural stem cells isolated from human fetuses. We showed that the peptide inhibits proliferation and the ability to produce neurospheres in vitro, which is consistent with an increase in cell adhesion and differentiation. These results confirm that the peptide could be a promising molecule to target and manipulate human neural stem cells and thus could serve as a strategic tool for regenerative medicine.

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Inhibition of Sox2 Expression in the Adult Neural Stem Cell Niche In Vivo by Monocationic-based siRNA Delivery

Cited by 9 publications

References 31 publications

Nogo Receptor Inhibition Enhances Functional Recovery following Lysolecithin-Induced Demyelination in Mouse Optic Chiasm

Nogo Receptor Inhibition Enhances Functional Recovery following Lysolecithin-Induced Demyelination in Mouse Optic Chiasm

In Vivo Identity of Tendon Stem Cells and the Roles of Stem Cells in Tendon Healing

The neurofilament derived-peptide NFL-TBS.40-63 enters in-vitro in human neural stem cells and increases their differentiation

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