scholarly journals Brain gray matter astroglia-specific connexin 43 ablation attenuates spinal cord inflammatory demyelination

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Hayato Une ◽  
Ryo Yamasaki ◽  
Satoshi Nagata ◽  
Hiroo Yamaguchi ◽  
Yuko Nakamuta ◽  
...  
Keyword(s):  
Gene Expression ◽  
Spinal Cord ◽  
Cerebrospinal Fluid ◽  
Gray Matter ◽  
Connexin 43 ◽  
Critical Role ◽  
Interferon Γ ◽  
Anti Inflammatory ◽  
Onset Phase ◽  
Peak Phase

Abstract Background Brain astroglia are activated preceding the onset of experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis (MS). We characterized the effects of brain astroglia on spinal cord inflammation, focusing on astroglial connexin (Cx)43, because we recently reported that Cx43 has a critical role in regulating neuroinflammation. Methods Because glutamate aspartate transporter (GLAST)+ astroglia are enriched in the brain gray matter, we generated Cx43fl/fl;GLAST-CreERT2/+ mice that were brain gray matter astroglia-specific Cx43 conditional knockouts (Cx43 icKO). EAE was induced by immunization with myelin oligodendroglia glycoprotein (MOG) 35–55 peptide 10 days after tamoxifen injection. Cx43fl/fl mice were used as controls. Results Acute and chronic EAE signs were significantly milder in Cx43 icKO mice than in controls whereas splenocyte MOG-specific responses were unaltered. Histologically, Cx43 icKO mice showed significantly less demyelination and fewer CD45+ infiltrating immunocytes, including F4/80+ macrophages, and Iba1+ microglia in the spinal cord than controls. Microarray analysis of the whole cerebellum revealed marked upregulation of anti-inflammatory A2-specific astroglia gene sets in the pre-immunized phase and decreased proinflammatory A1-specific and pan-reactive astroglial gene expression in the onset phase in Cx43 icKO mice compared with controls. Astroglia expressing C3, a representative A1 marker, were significantly decreased in the cerebrum, cerebellum, and spinal cord of Cx43 icKO mice compared with controls in the peak phase. Isolated Cx43 icKO spinal microglia showed more anti-inflammatory and less proinflammatory gene expression than control microglia in the pre-immunized phase. In particular, microglial expression of Ccl2, Ccl5, Ccl7, and Ccl8 in the pre-immunized phase and of Cxcl9 at the peak phase was lower in Cx43 icKO than in controls. Spinal microglia circularity was significantly lower in Cx43 icKO than in controls in the peak phase. Significantly lower interleukin (IL)-6, interferon-γ, and IL-10 levels were present in cerebrospinal fluid from Cx43 icKO mice in the onset phase compared with controls. Conclusions The ablation of Cx43 in brain gray matter astroglia attenuates EAE by promoting astroglia toward an anti-inflammatory phenotype and suppressing proinflammatory activation of spinal microglia partly through depressed cerebrospinal fluid proinflammatory cytokine/chemokine levels. Brain astroglial Cx43 might be a novel therapeutic target for MS.

Screening anti-inflammatory compounds in injured spinal cord with microarrays: a comparison of bioinformatics analysis approaches

2004 ◽  
Vol 17 (2) ◽  
pp. 201-214 ◽  
Author(s):  
Jonathan Z. Pan ◽  
Rebecka Jörnsten ◽  
Ronald P. Hart
Keyword(s):  
Gene Expression ◽  
Spinal Cord ◽  
Inflammatory Responses ◽  
Expression Profiles ◽  
Slice Culture ◽  
Data Set ◽  
Cord Injury ◽  
In Vivo Testing ◽  
Anti Inflammatory

Inflammatory responses contribute to secondary tissue damage following spinal cord injury (SCI). A potent anti-inflammatory glucocorticoid, methylprednisolone (MP), is the only currently accepted therapy for acute SCI but its efficacy has been questioned. To search for additional anti-inflammatory compounds, we combined microarray analysis with an explanted spinal cord slice culture injury model. We compared gene expression profiles after treatment with MP, acetaminophen, indomethacin, NS398, and combined cytokine inhibitors (IL-1ra and soluble TNFR). Multiple gene filtering methods and statistical clustering analyses were applied to the multi-dimensional data set and results were compared. Our analysis showed a consistent and unique gene expression profile associated with NS398, the selective cyclooxygenase-2 (COX-2) inhibitor, in which the overall effect of these upregulated genes could be interpreted as neuroprotective. In vivo testing demonstrated that NS398 reduced lesion volumes, unlike MP or acetaminophen, consistent with a predicted physiological effect in spinal cord. Combining explanted spinal cultures, microarrays, and flexible clustering algorithms allows us to accelerate selection of compounds for in vivo testing.

Molecular signature of late-stage human ALS revealed by expression profiling of postmortem spinal cord gray matter

2004 ◽  
Vol 16 (2) ◽  
pp. 229-239 ◽  
Author(s):  
Fernando Dangond ◽  
Daehee Hwang ◽  
Sandra Camelo ◽  
Piera Pasinelli ◽  
Matthew P. Frosch ◽  
...  
Keyword(s):  
Gene Expression ◽  
Spinal Cord ◽  
Gray Matter ◽  
Expression Profiles ◽  
Expression Patterns ◽  
Gene Expression Profiles ◽  
Terminal State ◽  
Molecular Signature ◽  
Fisher Discriminant Analysis ◽  
Sporadic Als

Little is known about global gene expression patterns in the human neurodegenerative disease amyotrophic lateral sclerosis (ALS). To address this, we used high-density oligonucleotide microarray technology to compare expression levels of ∼6,800 genes in postmortem spinal cord gray matter obtained from individuals with ALS as well as normal individuals. Using Fisher discriminant analysis (FDA) and leave-one-out cross-validation (LOOCV), we discerned an ALS-specific signature. Moreover, it was possible to distinguish familial ALS (FALS) from sporadic ALS (SALS) gene expression profiles. Characterization of the specific genes significantly altered in ALS uncovered a pro-inflammatory terminal state. Moreover, we found alterations in genes involved in mitochondrial function, oxidative stress, excitotoxicity, apoptosis, cytoskeletal architecture, RNA transcription and translation, proteasomal function, and growth and signaling. It is apparent from this study that DNA microarray analysis and appropriate bioinformatics can reveal distinct phenotypic changes that underlie the terminal stages of neurodegeneration in ALS.

scholarly journals Nanoparticle Estrogen in Rat Spinal Cord Injury Elicits Rapid Anti-Inflammatory Effects in Plasma, Cerebrospinal Fluid, and Tissue

2015 ◽  
Vol 32 (18) ◽  
pp. 1413-1421 ◽  
Author(s):  
April Cox ◽  
Abhay Varma ◽  
John Barry ◽  
Alexey Vertegel ◽  
Naren Banik
Keyword(s):  
Spinal Cord ◽  
Spinal Cord Injury ◽  
Cerebrospinal Fluid ◽  
Rat Spinal Cord ◽  
Cord Injury ◽  
Anti Inflammatory

scholarly journals The Study of Cerebrospinal Fluid microRNAs in Spinal Cord Injury and Neurodegenerative Diseases: Methodological Problems and Possible Solutions

2021 ◽  
Vol 23 (1) ◽  
pp. 114
Author(s):  
Irina Baichurina ◽  
Victor Valiullin ◽  
Victoria James ◽  
Albert Rizvanov ◽  
Yana Mukhamedshina
Keyword(s):  
Gene Expression ◽  
Spinal Cord ◽  
Spinal Cord Injury ◽  
Cerebrospinal Fluid ◽  
Neurodegenerative Diseases ◽  
Neurological Disorders ◽  
Molecular Mechanisms ◽  
Current Knowledge ◽  
Rna Isolation ◽  
Cord Injury

Despite extensive research on neurological disorders, unanswered questions remain regarding the molecular mechanisms underpinning the course of these diseases, and the search continues for effective biomarkers for early diagnosis, prognosis, or therapeutic intervention. These questions are especially acute in the study of spinal cord injury (SCI) and neurodegenerative diseases. It is believed that the changes in gene expression associated with processes triggered by neurological disorders are the result of post-transcriptional gene regulation. microRNAs (miRNAs) are key regulators of post-transcriptional gene expression and, as such, are often looked to in the search for effective biomarkers. We propose that cerebrospinal fluid (CSF) is potentially a source of biomarkers since it is in direct contact with the central nervous system and therefore may contain biomarkers indicating neurodegeneration or damage to the brain and spinal cord. However, since the abundance of miRNAs in CSF is low, their isolation and detection is technically difficult. In this review, we evaluate the findings of recent studies of CSF miRNAs as biomarkers of spinal cord injury (SCI) and neurodegenerative diseases. We also summarize the current knowledge concerning the methods of studying miRNA in CSF, including RNA isolation and normalization of the data, highlighting the caveats of these approaches and possible solutions.

scholarly journals Fosl1 Targets AMPK to Inhibit Autophagy-Mediated Anti-Apoptotic and Anti-Inflammatory Effects in Spinal Cord Injury

2020 ◽  
Author(s):  
Lin Zhong ◽  
Sheng Fang ◽  
An-Quan Wang ◽  
Tao Wang ◽  
Wei Huang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Spinal Cord ◽  
Spinal Cord Injury ◽  
Animal Model ◽  
Inflammatory Factor ◽  
Mrna Levels ◽  
Ampk Activation ◽  
Cord Injury ◽  
Anti Inflammatory

Abstract Background: The objective of this study was to delineate the role of Fosl1 in regulating inflammation and apoptosis following spinal cord injury.Methods: GSE45006 datasets from Gene Expression Omnibus (GEO) were explored to analyze Fosl1 gene expression. Next, we established an animal model to assess Fosl1 and AMPK by western blotting, real-time PCR, and immunohistochemical staining and used immunofluorescence staining to check Fosl1 expression in neurons. Fosl1 silencing was used to assess the effect on AMPK, cell viability, autophagy, inflammation and apoptosis. Subsequently, an AMPK activator and inhibitor were added to PC-12 cells with H2O2-induced injury subjected to si-Fosl1 treatment to examine the change in the above indexes and to determine whether the benefits from Fosl1 silencing occurred via AMPK. Moreover, we employed chloroquine (CQ) and rapamycin (RAP) to activate and inhibit autophagy, respectively, and revealed the effects of the upregulation and downregulation of autophagy following AMPK interference. Finally, an animal model was used to identify the effect of si-Fosl1 in vivo.Results: Based on the analysis of the GSE45006 datasets, Fosl1 was found to be highly expressed and was also found to be significantly enhanced in our animal model. Fosl1 knockdown upregulated AMPK at the protein and mRNA levels, promoted autophagic proteins (LC3 II/I, Beclin1) and inhibited inflammatory factors (IL-1β, IL-6, TNF-α) and apoptosis markers (caspase3, Bax). However, Fosl1 decreased the negatively related autophagic protein p62, the anti-inflammatory factor IL-10 and the anti-apoptotic marker Bcl-2. By utilizing compound C (com, an AMPK inhibitor), we learned that AMPK inhibition exhibited unfavorable effects on autophagy but promoted inflammation and apoptosis following Fosl1 silencing. AMPK activation showed contrasting effects. Moreover, we used CQ (an autophagic inhibitor), which indicated that CQ reversed the benefits of AMPK activation on inflammation and apoptosis. The autophagic activator RAP attenuated the negative effects after com treatment. In vivo, si-Fosl1 increased BBB scores at 7 d and 14 d and motor neurons, meanwhile, it decreased the number of apoptotic cells, and inflammatory cytokine expression at 14 d postoperation. Conclusion: Fosl1 can suppress AMPK to promote inflammation and apoptosis through autophagy in SCI.

scholarly journals Sensory neurons contacting the cerebrospinal fluid require the Reissner fiber to detect spinal curvature in vivo

2019 ◽  
Author(s):  
Adeline Orts-Del’Immagine ◽  
Yasmine Cantaut-Belarif ◽  
Olivier Thouvenin ◽  
Julian Roussel ◽  
Asha Baskaran ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Cerebrospinal Fluid ◽  
Sensory Neurons ◽  
Critical Role ◽  
Central Canal ◽  
Spinal Curvature ◽  
List Type ◽  
Close Vicinity

SummaryRecent evidence indicate active roles for the cerebrospinal fluid (CSF) on body axis development and morphogenesis of the spine implying CSF-contacting neurons (CSF-cNs) in the spinal cord. CSF-cNs project a ciliated apical extension into the central canal that is enriched in the channel PKD2L1 and enables the detection of spinal curvature in a directional manner. Dorsolateral CSF-cNs ipsilaterally respond to lateral bending while ventral CSF-cNs respond to longitudinal bending. Historically, the implication of the Reissner fiber (RF), a long extracellular thread in the CSF, to CSF-cN sensory functions has remained a subject of debate. Here, we reveal using electron microscopy in zebrafish larvae that the RF is in close vicinity with cilia and microvilli of ventral and dorsolateral CSF-cNs. We investigate in vivo the role of cilia and the Reissner fiber in the mechanosensory functions of CSF-cNs by combining calcium imaging with patch-clamp recordings. We show that disruption of cilia motility affects CSF-cN sensory responses to passive and active curvature of the spinal cord without affecting the Pkd2l1 channel activity. Since ciliary defects alter the formation of the Reissner fiber, we investigated whether the Reissner fiber contributes to CSF-cN mechanosensitivity in vivo. Using a hypomorphic mutation in the scospondin gene that forbids the aggregation of SCO-spondin into a fiber, we demonstrate in vivo that the Reissner fiber per se is critical for CSF-cN mechanosensory function. Our study uncovers that neurons contacting the cerebrospinal fluid functionally interact with the Reissner fiber to detect spinal curvature in the vertebrate spinal cord.Abstract FigureeToCThe role of the Reissner fiber, a long extracellular thread running in the cerebrospinal fluid (CSF), has been since its discovery in 1860 a subject of debate. Orts-Del’Immagine et al. report that the Reissner fiber plays a critical role in the detection of spinal curvature by sensory neurons contacting the CSF.HighlightsSince its discovery, the role of the Reissner fiber has long been a subject of debateMechanoreception in CSF-contacting neurons (CSF-cNs) in vivo requires the Reissner fiberCSF-cN apical extension is in close vicinity of the Reissner fiberCSF-cNs and the Reissner fiber form in vivo a sensory organ detecting spinal curvature

scholarly journals Critical Role of Connexin 43 in Secondary Expansion of Traumatic Spinal Cord Injury

2012 ◽  
Vol 32 (10) ◽  
pp. 3333-3338 ◽  
Author(s):  
C. Huang ◽  
X. Han ◽  
X. Li ◽  
E. Lam ◽  
W. Peng ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Spinal Cord Injury ◽  
Connexin 43 ◽  
Critical Role ◽  
Traumatic Spinal Cord Injury ◽  
Cord Injury

scholarly journals Activation of Neuroprotective Microglia and Astrocytes at the Lesion Site and in the Adjacent Segments Is Crucial for Spontaneous Locomotor Recovery after Spinal Cord Injury

Cells ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 1943
Author(s):  
Alexandra Kisucká ◽  
Katarína Bimbová ◽  
Mária Bačová ◽  
Ján Gálik ◽  
Nadežda Lukáčová
Keyword(s):  
Gene Expression ◽  
Spinal Cord ◽  
Lesion Site ◽  
Post Injury ◽  
Reactive Microglia ◽  
Locomotor Recovery ◽  
Cord Injury ◽  
Polarization States ◽  
Anti Inflammatory ◽  
Adjacent Segments

Microglia and astrocytes play an important role in the regulation of immune responses under various pathological conditions. To detect environmental cues associated with the transformation of reactive microglia (M1) and astrocytes (A1) into their polarization states (anti-inflammatory M2 and A2 phenotypes), we studied time-dependent gene expression in naive and injured spinal cord. The relationship between astrocytes and microglia and their polarization states were studied in a rat model after Th9 compression (40g/15 min) in acute and subacute stages at the lesion site, and both cranially and caudally. The gene expression of microglia/macrophages and M1 microglia was strongly up-regulated at the lesion site and caudally one week after SCI, and attenuated after two weeks post-SCI. GFAP and S100B, and A1 astrocytes were profoundly expressed predominantly two weeks post-SCI at lesion site and cranially. Gene expression of anti-inflammatory M2a microglia (CD206, CHICHI, IL1rn, Arg-1), M2c microglia (TGF-β, SOCS3, IL4R α) and A2 astrocytes (Tgm1, Ptx3, CD109) was greatly activated at the lesion site one week post-SCI. In addition, we observed positive correlation between neurological outcome and expression of M2a, M2c, and A2 markers. Our findings indicate that the first week post‑injury is critical for modulation of reactive microglia/astrocytes into their neuroprotective phenotypes.

scholarly journals Effects of Triptolide on Cell Viability, Secretion of Inflammatory Cytokines, and Gene Expression of Severe Coronavirus Disease 2019（COVID-19）Pseudovirus Cell Model

2021 ◽  
Author(s):  
junyong han ◽  
Huiqing Que ◽  
Wei Li ◽  
Shijie Xue ◽  
Sui Lin ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Viability ◽  
Inflammatory Cytokines ◽  
Cell Model ◽  
Critical Role ◽  
Membrane Integrity ◽  
Mapk Signaling ◽  
Lung Epithelial Cells ◽  
Normal Lung ◽  
Anti Inflammatory

Abstract Background: Acute respiratory distress syndrome (ARDS), which is caused by severe immune cell response and associated alveolar inflammation, is also a critical complication in hospitalized patients with COVID-19. Triptolide is a powerful anti-inflammatory and immunosuppressive drug and is proved to help relieve the inflammation of ARDS. However, its anti-inflammatory effect on COVID-19 patients with ARDS complications remains uncertain. Methods: In this study, human normal lung epithelial cells (BEAS-2B), the pseudovirus system of syndrome coronavirus 2(SARS-CoV-2) and lipopolysaccharide (LPS) were used to construct as severe COVID-19-pseudovirus cell model to explore the effects of triptolide on cell viability, secretion of inflammatory cytokines, and gene expression. Results: The results showed that triptolide increased cell viability, decreased the secretion levels of cytokines IL-6, TNF-a, and increased the expression of IL-10. Furthermore, transcriptome analysis in this cell models showed that the Differentially expressed genes (DEGs) were related to plasma membrane integrity, metabolic activity and mitochondrial function, and were associated with TNF, FOXO, mTOR and MAPK signaling pathways. Conclusion: Take into consideration previous studies on the functions of triptolide in BEAS-2B cells, the current study indicated that triptolide can play a critical role in protecting against inflammatory damage and maintaining the normal physiological function of BEAS-2B cells in response to pseudovirus and LPS infection.

Selective inhibition of interleukin-4 gene expression in human T cells by aspirin

Blood ◽  
2001 ◽  
Vol 97 (6) ◽  
pp. 1742-1749 ◽  
Author(s):  
Antonella Cianferoni ◽  
John T. Schroeder ◽  
Jean Kim ◽  
John W. Schmidt ◽  
Lawrence M. Lichtenstein ◽  
...  
Keyword(s):  
Gene Expression ◽  
T Cells ◽  
Cd4 T Cells ◽  
Inhibitory Effect ◽  
Interferon Γ ◽  
Interleukin 4 ◽  
Cytokine Gene Expression ◽  
Cytokine Gene ◽  
Activated T Cells ◽  
Anti Inflammatory

Previous studies indicated that aspirin (acetylsalicylic acid [ASA]) can have profound immunomodulatory effects by regulating cytokine gene expression in several types of cells. This study is the first in which concentrations of ASA in the therapeutic range were found to significantly reduce interleukin (IL)-4 secretion and RNA expression in freshly isolated and mitogen-primed human CD4+ T cells. In contrast, ASA did not affect IL-13, interferon-γ, and IL-2 expression. ASA inhibited IL-4, but not IL-2, promoter-driven chloramphenicol acetyltransferase expression in transiently transfected Jurkat T cells. The structurally unrelated nonsteroidal anti-inflammatory drugs indomethacin and flurbiprofen did not affect cytokine gene expression in T cells, whereas the weak cyclo-oxygenase inhibitor salicylic acid was at least as effective as ASA in inhibiting IL-4 expression and promoter activity. The inhibitory effect of ASA on IL-4 transcription was not mediated by decreased nuclear expression of the known salicylate target nuclear factor (NF)–κB and was accompanied by reduced binding of an inducible factor to an IL-4 promoter region upstream of, but not overlapping, the NF of activated T cells– and NF-κB–binding P1 element. It is concluded that anti-inflammatory salicylates, by means of a previously unrecognized mechanism of action, can influence the nature of adaptive immune responses by selectively inhibiting the expression of IL-4, a critical effector of these responses, in CD4+ T cells.

Sign in / Sign up

Export Citation Format

Share Document