A pilot study on searching for peri-nuclear NeuN-positive cells

Yu, Yun; Wu, Meiyu; Zhang, Nan; Yin, Hua; Shu, Bin; Duan, Weigang

doi:10.7717/peerj.8254

Cited by 6 publications

(5 citation statements)

References 36 publications

(10 reference statements)

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“…We observed NeuN expression in both cell lines, but in traditional PC12 cells, expression was localised to cell nuclei, and in adherent cells, it occurred for the whole cell, both in the nucleus and the cytoplasm. For many years, NeuN was considered a specific marker that allows the recognition of adult neurons [38]. Recent studies, however, have shown that quantitative assessment of this protein does not give reliable information, because its expression occurs in many tissues as well as many cell lines that are not-neuronal cells [38,53,54].…”

Section: Discussionmentioning

confidence: 99%

“…For many years, NeuN was considered a specific marker that allows the recognition of adult neurons [38]. Recent studies, however, have shown that quantitative assessment of this protein does not give reliable information, because its expression occurs in many tissues as well as many cell lines that are not-neuronal cells [38,53,54]. Additional assessment is necessary, whether NeuN is expressed in or around the nucleus or in the cytoplasm.…”

Section: Discussionmentioning

confidence: 99%

“…In contrast, in adherent cells, expression was observed in the entire cell-both in the nucleus and the cytoplasm. Expression of NeuN outside the cell nucleus cannot be considered a neuronal marker because it has been reported for cells derived from many different organs (other than the brain), as well as many cell lines known to be non-neuronal cells [38]. The quantitative comparison was not performed due to the expression of the evaluated biomarkers in various cellular structures (the quantitative assessment would be unreliable).…”

Section: Expression Of Neuronal Biomarkersmentioning

confidence: 99%

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PC12 Cell Line: Cell Types, Coating of Culture Vessels, Differentiation and Other Culture Conditions

Wiatrak

Kubis-Kubiak

Piwowar

et al. 2020

Cells

208

126

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The PC12 cell line is one of the most commonly used in neuroscience research, including studies on neurotoxicity, neuroprotection, neurosecretion, neuroinflammation, and synaptogenesis. Two types of this line are available in the ATCC collection: traditional PC12 cells grown in suspension and well-attached adherent phenotype. PC12 cells grown in suspension tend to aggregate and adhere poorly to non-coated surfaces. Therefore, it is necessary to modify the surface of culture vessels. This paper aims to characterise the use of two distinct variants of PC12 cells as well as describe their differentiation and neuronal outgrowth with diverse NGF concentrations (rat or human origin) on various surfaces. In our study, we evaluated cell morphology, neurite length, density and outgrowth (measured spectrofluorimetrically), and expression of neuronal biomarkers (doublecortin and NeuN). We found that the collagen coating was the most versatile method of surface modification for both cell lines. For adherent cells, the coating was definitely less important, and the poly-d-lysine surface was as good as collagen. We also demonstrated that the concentration of NGF is of great importance for the degree of differentiation of cells. For suspension cells, we achieved the best neuronal characteristics (length and density of neurites) after 14 days of incubation with 100 ng/mL NGF (change every 48 h), while for adherent cells after 3–5 days, after which they began to proliferate. In the PC12 cell line, doublecortin (DCX) expression in the cytoplasm and NeuN in the cell nucleus were found. In turn, in the PC12 Adh line, DCX was not expressed, and NeuN expression was located in the entire cell (both in the nucleus and cytoplasm). Only the traditional PC12 line grown in suspension after differentiation with NGF should be used for neurobiological studies, especially until the role of the NeuN protein, whose expression has also been noted in the cytoplasm of adherent cells, is well understood.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Expression Of Neuronal Biomarkersmentioning

confidence: 99%

See 1 more Smart Citation

PC12 Cell Line: Cell Types, Coating of Culture Vessels, Differentiation and Other Culture Conditions

Wiatrak

Kubis-Kubiak

Piwowar

et al. 2020

Cells

208

126

View full text Add to dashboard Cite

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“…Tubb3 is a neuron-specific class III β-tubulin that mainly localizes in neural axons [38], thus revealing important processes of neurite development in PK 11195-treated cells, as were observed microscopically. Although NeuN is commonly localized in neuronal soma, recent studies have reported its cytoplasmic localization in several peripheral tissues-such as heart, liver, lung, and kidneys-where NeuN has functions other than neuronal differentiation, specifically those related to RNA splicing [39]. Expression of NeuN in undifferentiated P19 stem cells supports these non-neural functions of NeuN.…”

Section: Discussionmentioning

confidence: 90%

Neurogenic Potential of the 18-kDa Mitochondrial Translocator Protein (TSPO) in Pluripotent P19 Stem Cells

González-Blanco

Bermejo-Millo

Oliveira

et al. 2021

Cells

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The 18-kDa translocator protein (TSPO) is a key mitochondrial target by which different TSPO ligands exert neuroprotective effects. We assayed the neurogenic potential of TSPO to induce the neuronal differentiation of pluripotent P19 stem cells in vitro. We studied changes in cell morphology, cell proliferation, cell death, the cell cycle, mitochondrial functionality, and the levels of pluripotency and neurogenesis of P19 stem cells treated with the TSPO ligand, PK 11195, in comparison to differentiation induced by retinoid acid (RA) and undifferentiated P19 stem cells. We observed that PK 11195 was able to activate the differentiation of P19 stem cells by promoting the development of embryoid bodies. PK 11195 also induced changes in the cell cycle, decreased cell proliferation, and activated cell death. Mitochondrial metabolism was also enhanced by PK 11195, thus increasing the levels of reactive oxygen species, Ca2+, and ATP as well as the mitochondrial membrane potential. Markers of pluripotency and neurogenesis were also altered during the cell differentiation process, as PK 11195 induced the differentiation of P19 stem cells with a high predisposition toward a neuronal linage, compared to cell differentiation induced by RA. Thus, we suggest a relevant neurogenic potential of TSPO along with broad therapeutic implications.

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“…Some of the speci c proteins expressed by neural cells can be used as their markers. In particular, NeuN was widely accepted as the "speci c" cell marker for neurons [37].In the rats after MCAO/R surgery, the number of neurons decreased, treatment with DHA increased the number of NeuN + cells in the cerebral ischemia penumbra, GSK0660 redud the protective effect of DHA on NeuN + cells. In combination with Hematoxylin-Eosin Staining, the ischemic cortical neurons showed atrophic morphologcey, accompanied by nuclear pyknosis, fragmentation and dissolution.…”

Section: Discussionmentioning

confidence: 99%

Activation of PPARβ/δ by DHA induces VEGF-A expression to promote angiogenesis in the cerebral ischemia penumbra

Zhao

Jia

et al. 2022

Preprint

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The ischemia penumbra is an area of brain tissue where is damaged but not yet dead after ischemic stroke,it has the potential for functional recovery provided if local blood flow can be reestablished.Docosahexaenoic acid (DHA) is protective after experimental ischemic stroke.To explore DHA promote angiogenesis underlying mechanisms of angiogenesis in the cerebral ischemia penumbra, Male Sprague-Dawley rats underwent 2h of middle cerebral artery occlusion (MCAo) and treated with DHA (5mg/kg, IV) 1h after.Neuro-behavioral assessments and 2, 3, 5-Triphenyltetrazolium chloride (TTC) staining was used to assess the cerebral infarction volume on hours 6, 12, 24,48 and 72, PPARβ/δ、VEGF-A、VEGFR2 and P-VEGFR2 expression were measured by Western blotting、brain tissue expressions of TNF-α, IL-1β and NF-κB were assessed using ELISA, NeuN、CD31、α-sma cells in the ischemia penumbra were analyzed using immunofluorescenceon hour 72 after onset of stroke. Post-treatment with DHA ameliorated neurological defects, diminished cerebral infarction volume, enhanced neuronal survival, downregulated inflammation、promoted the expression of angiogenesis related proteins and protected ischemic penumbra by increasing CD31、α-sma cells and vessels .On the contrary, this promote angiogenesis in the cerebral ischemia penumbraprotective effect was reversed after DHA cotreatment with GSK0660, a selective antagonist of the PPARβ/δ.Finally, our experiment clearly demonstrating DHA protect the rat cerebral ischemic injury (MCAO) model, more importantly, DHA was first demonstrated to promote ischemic penumbra angiogenesis through the PPARβ/δ pathway.

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A pilot study on searching for peri-nuclear NeuN-positive cells

Cited by 6 publications

References 36 publications

PC12 Cell Line: Cell Types, Coating of Culture Vessels, Differentiation and Other Culture Conditions

PC12 Cell Line: Cell Types, Coating of Culture Vessels, Differentiation and Other Culture Conditions

Neurogenic Potential of the 18-kDa Mitochondrial Translocator Protein (TSPO) in Pluripotent P19 Stem Cells

Activation of PPARβ/δ by DHA induces VEGF-A expression to promote angiogenesis in the cerebral ischemia penumbra

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