Knocking down of heat-shock protein 27 directs differentiation of functional glutamatergic neurons from placenta-derived multipotent cells

Cheng, Yu‐Che; Huang, Chun-Ta; Lee, Yih-Jing; Tien, Lu-Tai; Ku, Wei-Chi; Chien, Raymond; Lee, Fa‐Kung; Chien, Chih‐Cheng

doi:10.1038/srep30314

Cited by 17 publications

(19 citation statements)

References 48 publications

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“…The individual provided informed consent. The stem cells (PDMCs) were isolated as described previously (7,27). Prior to any differentiation, a minimal immunophenotyping was performed as previously described, with minor modifications of the antibody titers as described, to validate the stemness of the PDMCs using a FACSCalibur flow cyto�eter and CellQuest software (v.3.3; both BD Biosciences, Franklin Lakes, NJ, USA) (29,30).…”

Section: Methodsmentioning

confidence: 99%

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Dynamic expression of SMAD3 is critical in�osteoblast differentiation of PDMCs

et al. 2018

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Human pluripotent stem cells have the potential assist in the identification of genes involved in mammalian development. The human placenta is considered a repository of stem cells, termed placenta-derived multipotent cells (PDMCs), which are able to differentiate into cells with an osteoblastic phenotype. This plasticity of PDMCs maybe applied clinically to the understanding of osteogenesis and osteoporosis. In the presentstudy, osteoblasts were generated by culturing PDMCs in osteogenic medium. Reverse transcription quantitative polymerase chain reactionand the degree of osteoblast calcification were used to evaluate the efficacy of osteogenesis. The results suggestedthat the expression of mothers against decapentaplegic homolog 3 (SMAD3) increased in the initial stages of osteogenic differentiation but decreased in the later stages. However, osteogenesis was inhibitedwhen the PDMCs overexpressed SMAD3 throughout the differentiation period. In addition, the rate of osteogenic differentiation was decreased when SMAD3 signaling was impaired. In conclusion, SMAD3 serves an important role in osteoblast differentiation and bone formation in a time-dependent manner. The data from the present study indicate that arapid increase in SMAD3 expression is crucial for osteogenesis and suggest a role for PDMCs in the treatment of patients with osteoporosis.

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Section: Methodsmentioning

confidence: 99%

“…A number of growth factors, cytokines, drugs and gene products are critical for the differentiation of stem cells, including osteoblast differentiation (7)(8)(9). Indeed, one of the pivotal factors in the health and maintenance of bone density is the coordinated activity of osteoblasts and osteoclasts (10).…”

Section: Introductionmentioning

confidence: 99%

Dynamic expression of SMAD3 is critical in�osteoblast differentiation of PDMCs

et al. 2018

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“…21 However, in our previous study, knocking down of HSP27 guided the differentiation of glutamatergic neurons from a cultured mesenchymal cell system. 22 This finding suggests that HSP27 may play different roles in different situations. Because Müller cells are highly associated with the secretion of neurotrophic factors, this study concluded that HSP27 might be involved in the developmental and neuroprotective effects on the retina.…”

mentioning

confidence: 76%

“…Our previous study suggested that suppressing HSP27 might induce glutamatergic neuron differentiation in a cultured mesenchymal cell system. 22 It is highly possible that the suppression of HSP27 after light exposure would protect photoreceptors, which are also glutamatergic, and preserve retinal function against damage induced by long-term light exposure.…”

Section: Discussionmentioning

confidence: 99%

Suppression of HSP27 Restores Retinal Function and Protects Photoreceptors From Apoptosis in a Light-Induced Retinal Degeneration Animal Model

Chien

Huang

Tien

et al. 2017

Invest. Ophthalmol. Vis. Sci.

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PURPOSE. We used a light-induced retinal degeneration animal model to investigate possible roles of heat shock protein 27 (HSP27) in retinal/photoreceptor protection.METHODS. Sprague-Dawley rats were used for the light-induced retinal degeneration animal model. The histology of eye sections was observed for morphologic changes in the retina. Cell apoptosis was examined in each group using the terminal deoxynucleotidyl transferase dUTP nick-end labeling assay, and electroretinography was used to evaluate retinal function. Protein and mRNA expression levels of different retinal cell markers were also detected through immunofluorescence staining, Western blotting, and real-time PCR.RESULTS. The thickness of the outer nuclear layer significantly decreased after 7-day light exposure. Moreover, we injected a viral vector for silencing HSP27 expression into the eyes and observed that photoreceptors were better preserved in the HSP27-suppressed (sHSP27) retina 2 weeks after injection. HSP27 suppression also reduced retinal cell apoptosis caused by light exposure. In addition, the loss of retinal function caused by light exposure was reversed on suppressing HSP27 expression. We subsequently found that the expression of the Rho gene and immunofluorescence staining of rhodopsin and arrestin (cell markers for photoreceptors) increased in sHSP27-treated retinas. HSP27 suppression did not affect the survival of ganglion and amacrine cells. CONCLUSIONS.Retinal cell apoptosis and functional loss were observed after 7-day light exposure. However, in the following 2 weeks after light exposure, HSP27 suppression may initiate a protective effect for retinal cells, particularly photoreceptors, from light-induced retinal degeneration.

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“…Briefly, tissues were dewaxed and incubated in 10 mM Citrate buffer, pH6.0 solution for antigen retrieval. Sections were blocked for 1 h (2% normal goat serum, 0.4% BSA, 0.3% Triton X‐100), at room temperature, before overnight incubation in primary antibodies glial fibrillary protein (GFAP, G3893, 1:1000, Sigma; St. Louis, USA; (Poesen et al, 2008)), neuronal nuclei (NeuN, MAB377, 1:500, Millipore‐Chemicon; Darmstadt, Germany; (Tsutsui‐Kimura et al, 2015)), microtubule‐associated protein 2 (MAP2, M9942, 1:20000, Sigma; St. Louis, USA; (Teng et al, 2001)), myelin basic protein (MBP, M9434, 1:1000, Sigma; St. Louis, USA; (Lampe et al, 1983)) or glutamic acid decarboxylase 67 (GAD67, ab26116, 1:1000, Abcam; Cambridge UK; (Cheng et al, 2016)), at room temperature. Primary antibody for GABA Transporter 1 (GAT1, ab426, 1:500, Abcam; Cambridge UK; (Aligny et al, 2014)) was incubated overnight at 4 °C.…”

Section: Methodsmentioning

confidence: 99%

Increased anxiety‐like phenotype in female guinea pigs following reduced neurosteroid exposure in utero

Cumberland¹,

Palliser

Crombie

et al. 2017

Intl J of Devlp Neuroscience

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Neurosteroids are essential for aiding proper fetal neurodevelopment. Pregnancy compromises such as preterm birth, prenatal stress and intrauterine growth restriction are associated with an increased risk of developing behavioural and mood disorders, particularly during adolescence. These pathologies involve the premature loss or alteration of trophic steroid hormones reaching the fetus leading to impaired neurodevelopment. While the specific programming mechanisms are yet to be fully elucidated, in adult life, dysfunctions of allopregnanolone action are prevalent in individuals with depression, post-traumatic stress disorder and anxiety disorders. The objective of this study was to assess if changes in concentrations of the neurosteroid, allopregnanolone, may be a fetal programming factor in priming the brain towards a negative behavioural phenotype during the childhood to adolescent period using a guinea pig model. Pregnant guinea pigs received either vehicle (45% (2-hydroxypropyl)-β-cyclodextrin) or the 5α-reductase inhibitor, finasteride (25mg/kg maternal weight) from gestational age 60 until spontaneous delivery (∼71days gestation). Male and female offspring from vehicle and finasteride treated dams were tested at postnatal day 20 (juvenile-equivalence) in an open field arena, and hippocampus and amygdala subsequently assessed for neurological changes in markers of development and GABA production pathways 24h later. Females with reduced allopregnanolone exposure in utero displayed increased neophobic-like responses to a change in their environment compared to female controls. There were no differences in the neurodevelopmental markers assessed; MAP2, NeuN, MBP, GFAP or GAD67 between intrauterine finasteride or vehicle exposure, in either the hippocampus or amygdala whereas GAT1 staining was decreased. This study indicates that an intrauterine reduction in the supply of allopregnanolone programs vulnerability of female offspring to anxiety-like disorders in juvenility without impacting long term allopregnanolone concentrations.

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Knocking down of heat-shock protein 27 directs differentiation of functional glutamatergic neurons from placenta-derived multipotent cells

Cited by 17 publications

References 48 publications

Dynamic expression of SMAD3 is critical in�osteoblast differentiation of PDMCs

Dynamic expression of SMAD3 is critical in�osteoblast differentiation of PDMCs

Suppression of HSP27 Restores Retinal Function and Protects Photoreceptors From Apoptosis in a Light-Induced Retinal Degeneration Animal Model

Increased anxiety‐like phenotype in female guinea pigs following reduced neurosteroid exposure in utero

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