TGFβ1 downregulates neurite outgrowth, expression of Ca2+ transporters, and mitochondrial dynamics of in vitro cerebellar granule cells

Jašková, Katarína; Pavlovicova, Michaela; Cagalinec, Michal; Lacinová, Ľubica; Jurkovičová, Dana

doi:10.1097/wnr.0000000000000106

Cited by 11 publications

(13 citation statements)

References 24 publications

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“…In cell-free assays, LY364947 inhibits TGFβRI (ALK5) with an IC 50 of 0.58 μM [42], but nonspecific effects on other kinases have been demonstrated [43]. RIKI has been used at concentrations ranging from 0.4 to 10 μM in cell-based assays [8], [44], [45]. Because drug bioavailability deep within explants cannot be predicted by previous in vitro assays, we tested two concentrations of RIKI (0.4 μM and 2 μM; 24-hour treatment) in six explants using immunofluorescence analysis for nuclear p-Smad2.…”

Section: Resultsmentioning

confidence: 99%

Patient-Specific Screening Using High-Grade Glioma Explants to Determine Potential Radiosensitization by a TGF-β Small Molecule Inhibitor

Bayın¹,

Thomas

et al. 2016

Neoplasia

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High-grade glioma (HGG), a deadly primary brain malignancy, manifests radioresistance mediated by cell-intrinsic and microenvironmental mechanisms. High levels of the cytokine transforming growth factor-β (TGF-β) in HGG promote radioresistance by enforcing an effective DNA damage response and supporting glioma stem cell self-renewal. Our analysis of HGG TCGA data and immunohistochemical staining of phosphorylated Smad2, which is the main transducer of canonical TGF-β signaling, indicated variable levels of TGF-β pathway activation across HGG tumors. These data suggest that evaluating the putative benefit of inhibiting TGF-β during radiotherapy requires personalized screening. Thus, we used explant cultures of seven HGG specimens as a rapid, patient-specific ex vivo platform to test the hypothesis that LY364947, a small molecule inhibitor of the TGF-β type I receptor, acts as a radiosensitizer in HGG. Immunofluorescence detection and image analysis of γ-H2AX foci, a marker of cellular recognition of radiation-induced DNA damage, and Sox2, a stem cell marker that increases post-radiation, indicated that LY364947 blocked these radiation responses in five of seven specimens. Collectively, our findings suggest that TGF-β signaling increases radioresistance in most, but not all, HGGs. We propose that short-term culture of HGG explants provides a flexible and rapid platform for screening context-dependent efficacy of radiosensitizing agents in patient-specific fashion. This time- and cost-effective approach could be used to personalize treatment plans in HGG patients.

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Section: Resultsmentioning

confidence: 99%

Patient-Specific Screening Using High-Grade Glioma Explants to Determine Potential Radiosensitization by a TGF-β Small Molecule Inhibitor

Bayın¹,

Thomas

et al. 2016

Neoplasia

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“…Neurite length was analysed using ImageJ software, as described previously (Jaskova et al ., ; Ondacova et al ., ). The length of the ruler was calibrated in micrometers using the scale tool of the microscope camera and statistically evaluated by GraphPad InStat 3 software.…”

Section: Methodsmentioning

confidence: 96%

“…Conversely, TGF‐β1‐neutralizing antibodies significantly attenuated the deposition of fibrous scar tissue and the formation of a limiting glial membrane that borders the lesion (Logan et al ., ). Effects of TGF‐β1 itself may underlie some pathophysiological processes that neurons undergo in a CNS injury (Jaskova et al ., ; Ondacova et al ., ).…”

Section: Introductionmentioning

confidence: 96%

“…Recently, our group compared effects of a complex environment created by a fibrotic scar model (Kimura‐Kuroda et al ., ) with effects of simple addition of TGF‐β1 on characteristics of rat cerebellar granule cells (CGCs) (Ondacova et al ., ). Both interventions inhibited the neurite outgrowth to a similar extent (Jaskova et al ., ; Ondacova et al ., ). Current density of voltage‐dependent sodium and potassium currents decreased when CGCs were cultured on the fibrotic scar model, and this decrease was prevented when ChABC was applied to the cell culture (Ondacova et al ., ).…”

Section: Introductionmentioning

confidence: 96%

“…Current density of voltage‐dependent sodium and potassium currents decreased when CGCs were cultured on the fibrotic scar model, and this decrease was prevented when ChABC was applied to the cell culture (Ondacova et al ., ). Effects on ion currents were not mimicked by the TGF‐β1 administration suggesting that observed changes in excitability cannot be attributed to the increased expression of this cytokine in the lesion (Jaskova et al ., ; Ondacova et al ., ).…”

Section: Introductionmentioning

confidence: 96%

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Fibrotic scar model and TGF‐β1 differently modulate action potential firing and voltage‐dependent ion currents in hippocampal neurons in primary culture

Ondacova

Moravcikova

Jurkovičová

et al. 2017

Eur J of Neuroscience

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Traumatic injury of the central nervous system is accompanied by various functional and morphological changes. Animal models of traumatic brain injury are commonly used to investigate changes in behaviour, morphology, in the expression of various proteins around the site of the injury, or the expression of diagnostically important biomarkers. Excitability of a single neuron at, or close to, the site of injury was rarely investigated. Several in vitro models were developed which allow such investigation. In the present work, we employed a fibrotic scar model according to Kimura-Kuroda and coauthors to analyse altered excitability of rat hippocampal neurons under the conditions mimicking traumatic brain injury. Hippocampal neurons from newborn rats were cultured either on a fibrotic scar model or in the presence of TGF-β1, a cytokine secreted at a brain injury site that may have both neuroprotective and neurodegenerative function. Fibrotic scar facilitated ability of neonatal hippocampal neurons to fire action potential series by increasing the density of voltage activated sodium and potassium currents. Chondroitin sulphate proteoglycans played substantial role in these effects, as proven by their full reversion after administration of Chondroitinase ABC. In contrast, TGF-β1 did not contribute to them. An application of TGF-β1 itself attenuated generation of action potentials, inhibited sodium current and potentiated potassium currents. Main alteration of electrophysiological parameters of neonatal hippocampal neurons caused by a fibrotic scar model is enhanced excitability. TGF-β1 may have predominantly neuroprotective role in injured rat hippocampus.

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Altered Sodium and Potassium, but not Calcium Currents in Cerebellar Granule Cells in an In Vitro Model of Neuronal Injury

2016

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Acute injury of central nervous system (CNS) starts a cascade of morphological, molecular, and functional changes including formation of a fibrotic scar, expression of transforming growth factor beta 1 (TGF-β1), and expression of extracellular matrix proteins leading to arrested neurite outgrowth and failed regeneration. We assessed alteration of electrophysiological properties of cerebellar granule cells (CGCs) in two in vitro models of neuronal injury: (i) model of fibrotic scar created from coculture of meningeal fibroblasts and cerebral astrocytes with addition of TGF-β1; (ii) a simplified model based on administration of TGF-β1 to CGCs culture. Both models reproduced suppression of neurite outgrowth caused by neuronal injury, which was equally restored by chondroitinase ABC (ChABC), a key disruptor of fibrotic scar formation. Voltage-dependent calcium current was not affected in either injury model. However, intracellular calcium concentration could be altered as an expression of inositol trisphosphate receptor type 1 was suppressed by TGF-β1 and restored by ChABC. Voltage-dependent sodium current was significantly suppressed in CGCs cultured on a model of fibrotic scar and was only partly restored by ChABC. Administration of TGF-β1 significantly shifted current-voltage relation of sodium current toward more positive membrane potential without change to maximal current amplitude. Both transient and sustained potassium currents were significantly suppressed on a fibrotic scar and restored by ChABC to their control amplitudes. In contrast, TGF-β1 itself significantly upregulated transient and did not change sustained potassium current. Observed changes of voltage-dependent ion currents may contribute to known morphological and functional changes in injured CNS.

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TGFβ1 downregulates neurite outgrowth, expression of Ca2+ transporters, and mitochondrial dynamics of in vitro cerebellar granule cells

Cited by 11 publications

References 24 publications

Patient-Specific Screening Using High-Grade Glioma Explants to Determine Potential Radiosensitization by a TGF-β Small Molecule Inhibitor

Patient-Specific Screening Using High-Grade Glioma Explants to Determine Potential Radiosensitization by a TGF-β Small Molecule Inhibitor

Fibrotic scar model and TGF‐β1 differently modulate action potential firing and voltage‐dependent ion currents in hippocampal neurons in primary culture

Altered Sodium and Potassium, but not Calcium Currents in Cerebellar Granule Cells in an In Vitro Model of Neuronal Injury

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