Regulatory role of endoplasmic reticulum resident chaperone protein ERp29 in anti-murine β-coronavirus host cell response

Bose, Abhishek; Kasle, Grishma; Jana, Rishika; Maulik, Mahua; Thomas, Deepthi; Mulchandani, Vaishali; Mukherjee, Priyanka; Koval, Michael; Sarma, Jayasri Das

doi:10.1016/j.jbc.2022.102836

Cited by 4 publications

(2 citation statements)

References 67 publications

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“…MHV-A59 infection in primary astrocytes increases Endoplasmic reticulum (ER) stress along with downregulation of the ER-resident chaperone protein, Erp29. Additionally, it has been demonstrated that 4-PBA treatment reduces ER stress by enhancing the expression of the chaperone Erp29 [refer to Figures 1 , 2 , and 3 ( Bose et al, 2023 )] [38]. This suggests that the current isolation procedure can also be used to comprehend neuro-glial-cells-specific stress responses [38].…”

Section: Validation Of Protocolmentioning

confidence: 99%

Isolation and Enrichment of Major Primary Neuroglial Cells from Neonatal Mouse Brain

Kumar Samal,

Sharma,

Das Sarma

2024

BIO-PROTOCOL

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The central nervous system (CNS) relies on the complex interaction of neuroglial cells to carry out vital physiological functions. To comprehensively understand the structural and functional interplay between these neuroglial cells, it is essential to establish an appropriate in vitro system that can be utilized for thorough investigation. Traditional protocols for establishing primary neuronal and mixed glial cultures from prenatal mice or neural stem cells require sacrificing pregnant mice and have the drawback of yielding only specific types of cells. Our current protocol overcomes these drawbacks by utilizing the brain from day-0 pups to isolate CNS resident neuroglial cells including astrocytes, microglia, oligodendrocytes [oligodendrocyte precursor cells (OPCs) and differentiated oligodendrocytes], and meningeal fibroblasts, as well as hippocampal neurons, avoiding sacrificing pregnant mice, which makes this procedure efficient and cost effective. Furthermore, through this protocol, we aim to provide step-by-step instructions for isolating and establishing different primary neuroglial cells and their characterization using cell-specific markers. This study presents an opportunity to isolate, culture, and establish all major CNS resident cells individually. These cells can be utilized in various cell-based and biochemical assays to comprehensively investigate the cell-specific roles and behaviors of brain resident cells in a reductionist approach. Key features • Efficient isolation of major neuroglial cells like meningeal fibroblasts, neurons, astrocytes, oligodendrocytes, and microglia from a single day-0 neonatal mouse pup’s brain. • Circumvents the sacrifice of pregnant female mice. • Acts as a bridging experimental method between secondary cell lines and in vivo systems. • Isolated cells can be used for performing various cell-based and biochemical assays.

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Section: Validation Of Protocolmentioning

confidence: 99%

Isolation and Enrichment of Major Primary Neuroglial Cells from Neonatal Mouse Brain

Kumar Samal,

Sharma,

Das Sarma

2024

BIO-PROTOCOL

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“…The expression of gap junction proteins, particularly connexin, has been reported to be modulated following virus infections [4]- [6]. Connexin 43 (Cx43), the most ubiquitously expressed and widely studied among all connexins, has been extensively studied in response to murine β-coronavirus infections [7]- [10]. The regulation in Cx43 following virus infection is attributed to either viral burden on the endoplasmic reticulum (ER) causing cellular stress or hijacking microtubule-mediated transport for trafficking of viral proteins within the cells.…”

Section: Introductionmentioning

confidence: 99%

Human coronavirus OC43 infection remodels Connexin 43 mediated gap junction intercellular communicationin vitro

Karmakar,

Sarma

2023

Preprint

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Abstractβ-coronaviruses cause acute infection in the upper respiratory tract, resulting in various symptoms and clinical manifestations. OC43 is a human β-coronavirus that induces mild clinical symptoms and can be safely studied in the BSL2 laboratory. Due to its low risk, OC43 can be a valuable and accessible model for understanding β-coronavirus pathogenesis. One potential target for limiting virus infectivity could be gap junction-mediated communication. This study aims to unveil the status of cell-to-cell communications through gap junctions in human β-coronavirus infection. Infection with OC43 leads to reduced expression of Cx43 in A549, a lung epithelial carcinoma cell line. Infection with this virus also showed a significant ER and oxidative stress increase. Internal localization of Cx43 is observed post OC43 infection in the ERGIC region, which impairs the gap junction communication between two adjacent cells, confirmed by Lucifer yellow dye transfer assay. It also affects hemichannel formation, as depicted by the EtBr uptake assay. Altogether, these results suggest that several physiological changes accompany OC43 infection in A549 cells and can be considered an appropriate model system for understanding the differences in gap junction communication post-viral infections. This model system can provide valuable insights for developing therapies against human β-coronavirus infections.ImportanceThe enduring impact of the recent SARS-CoV-2 pandemic underscores the importance of studying human β-coronaviruses, advancing our preparedness for future coronavirus infections. Due to SARS-CoV-2 being highly infectious, another human β-coronavirus OC43 can be considered as an experimental model. One of the crucial pathways that can be considered is gap junction communication, as it is vital for cellular homeostasis. Our study seeks to understand the change in Cx43-mediated cell-to-cell communication during human β-coronavirus OC43 infection.In vitrostudies showed the downregulation of the gap junction protein Cx43 and the upregulation of endoplasmic reticulum and oxidative stress markers post-OC43 infection. Furthermore, OC43 infection causes impairment of functional hemichannel and gap junction formation. Overall, this current study infers that OC43 infection reshapes intercellular communication, suggesting that this pathway may be a promising target for designing highly effective therapeutics against human coronaviruses by regulating Cx43 expression.

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Differential expression of cellular prion protein (PrPC) in mouse hepatitis virus induced neuroinflammation

Ghosh,

Jana,

Jana

et al. 2024

J. Neurovirol.

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Regulatory role of endoplasmic reticulum resident chaperone protein ERp29 in anti-murine β-coronavirus host cell response

Cited by 4 publications

References 67 publications

Isolation and Enrichment of Major Primary Neuroglial Cells from Neonatal Mouse Brain

Isolation and Enrichment of Major Primary Neuroglial Cells from Neonatal Mouse Brain

Human coronavirus OC43 infection remodels Connexin 43 mediated gap junction intercellular communicationin vitro

Differential expression of cellular prion protein (PrPC) in mouse hepatitis virus induced neuroinflammation

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