Infection of Fetal Feline Brain Cells in Culture with Bartonella henselae

ABSTRACT Bartonella henselae is known to cause central nervous system (CNS) disease in humans, and neurological signs have been observed in experimentally infected cats. However, the pathogenesis of CNS disease remains unclear. This study was undertaken to determine whether B. henselae infects feline fetal brain cells in vitro. Microglial-cell- and astrocyte-enriched cultures were inoculated with B. henselae. Giménez staining identified bacterial organisms within microglial cells by day 7 postinoculation. The viability of the intracellular bacteria was demonstrated by incubating cultures with gentamicin and plating cell lysate on agar. Electron microscopy identified intracellular organisms with characteristic Bartonella morphology but identified no ultrastructural abnormalities within infected microglial cells. No evidence of infection was seen in Bartonella-inoculated astrocyte cultures. These findings suggest a role for microglia in the pathogenesis of B. henselae-associated neurological disease.

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Effect of memantine, an anti-Alzheimer’s drug, on rodent microglial cells in vitro

Scientific Reports ◽

10.1038/s41598-021-85625-4 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Toru Murakawa-Hirachi ◽

Yoshito Mizoguchi ◽

Masahiro Ohgidani ◽

Yoshinori Haraguchi ◽

Akira Monji

Keyword(s):

Neuronal Death ◽

Phagocytic Activity ◽

Microglial Cell ◽

Cell Function ◽

Microglial Cells ◽

Phagocytosis Assay ◽

Intracellular Ca ◽

Β Amyloid ◽

The Brain

AbstractThe pathophysiology of Alzheimer’s disease (AD) is related to neuroinflammatory responses mediated by microglia. Memantine, an antagonist of N-methyl-d-aspartate (NMDA) receptors used as an anti-Alzheimer’s drug, protects from neuronal death accompanied by suppression of proliferation and activation of microglial cells in animal models of AD. However, it remains to be tested whether memantine can directly affect microglial cell function. In this study, we examined whether pretreatment with memantine affects intracellular NO and Ca2+ mobilization using DAF-2 and Fura-2 imaging, respectively, and tested the effects of memantine on phagocytic activity by human β-Amyloid (1–42) phagocytosis assay in rodent microglial cells. Pretreatment with memantine did not affect production of NO or intracellular Ca2+ elevation induced by TNF in rodent microglial cells. Pretreatment with memantine also did not affect the mRNA expression of pro-inflammatory (TNF, IL-1β, IL-6 and CD45) or anti-inflammatory (IL-10, TGF-β and arginase) phenotypes in rodent microglial cells. In addition, pretreatment with memantine did not affect the amount of human β-Amyloid (1–42) phagocytosed by rodent microglial cells. Moreover, we observed that pretreatment with memantine did not affect 11 major proteins, which mainly function in the phagocytosis and degradation of β-Amyloid (1–42), including TREM2, DAP12 and neprilysin in rodent microglial cells. To the best of our knowledge, this is the first report to suggest that memantine does not directly modulate intracellular NO and Ca2+ mobilization or phagocytic activity in rodent microglial cells. Considering the neuroinflammation hypothesis of AD, the results might be important to understand the effect of memantine in the brain.

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An in vitro study of the effects of lovastatin on human fetal brain cells

Neurotoxicology and Teratology ◽

10.1016/0892-0362(95)91641-w ◽

1995 ◽

Vol 17 (1) ◽

pp. 31-39 ◽

Cited By ~ 22

Author(s):

O.V. Pavlov ◽

Yu.V. Bobryshev ◽

Yu.V. Balabanov ◽

K. Ashwell

Keyword(s):

Fetal Brain ◽

In Vitro Study ◽

Vitro Study ◽

Brain Cells ◽

Human Fetal Brain

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Functional maturation of human neural stem cells in a 3D bioengineered brain model enriched with fetal brain-derived matrix

10.1101/691907 ◽

2019 ◽

Cited By ~ 1

Author(s):

Disha Sood ◽

Dana M. Cairns ◽

Jayanth M. Dabbi ◽

Charu Ramakrishnan ◽

Karl Deisseroth ◽

...

Keyword(s):

Stem Cells ◽

Neural Stem Cells ◽

Brain Tissue ◽

Fetal Brain ◽

Functional Differentiation ◽

Adult Brain ◽

Human Neural Stem Cells ◽

Functional Maturation ◽

Enriched Cultures

AbstractBrain extracellular matrix (ECM) is often overlooked in vitro brain tissue models, despite its instructive roles during development. Using developmental stage-sourced brain ECM in reproducible 3D bioengineered culture systems, we demonstrate enhanced functional differentiation of human induced neural stem cells (hiNSCs) into healthy neurons and astrocytes. Particularly, fetal brain tissue-derived ECM supported long-term maintenance of differentiated neurons, demonstrated by morphology, gene expression and secretome profiling. Astrocytes were evident within the second month of differentiation, and reactive astrogliosis was inhibited in brain ECM-enriched cultures when compared to unsupplemented cultures. Functional maturation of the differentiated hiNSCs within fetal ECM-enriched cultures was confirmed by calcium signaling and unsupervised cluster analysis. Additionally, the study identified native biochemical cues in decellularized ECM with notable comparisons between fetal and adult brain-derived ECMs. The development of novel brain-specific biomaterials for generating mature in vitro brain models provides an important path forward for interrogation of neuron-glia interactions.

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In Vitro Transformation of Fetal Brain Cells From CDF Rats Exposed In Utero to N-Ethyl-N-nitrosourea: Morphologic and Immunologic Studies234

JNCI Journal of the National Cancer Institute ◽

10.1093/jnci/64.5.1231 ◽

1980 ◽

Keyword(s):

Fetal Brain ◽

In Utero ◽

Brain Cells ◽

In Vitro Transformation

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Microglia induce neurogenesis by stimulating PI3K/AKT intracellular signaling in vitro

10.21203/rs.2.11343/v3 ◽

2020 ◽

Author(s):

Kristi Lorenzen ◽

Nicholas W. Mathy ◽

Erin R. Whiteford ◽

Alex Eischeid ◽

Jing Chen ◽

...

Keyword(s):

Cell Viability ◽

Intracellular Signaling ◽

Microglial Cell ◽

Cortical Cell ◽

Model System ◽

Microglial Cells ◽

Cortical Cells ◽

Akt Phosphorylation ◽

Cortical Neurogenesis

Abstract Background: Emerging evidence suggests that microglia can support neuronal survival, synapse development, and neurogenesis in classic neurogenic niches. Little is known about the ability of microglia to regulate the cortical environment and stimulate cortical neurogenesis outside classic neurogenic niches. We used an in vitro co-culture model system to investigate the hypothesis that microglia respond to soluble signals from cortical cells, particularly following injury, by altering the cortical environment to promote cortical cell proliferation, differentiation, and survival. Results: Analyses of cell proliferation, apoptosis, protein expression, and intracellular signaling were performed on uninjured and injured cortical cells in co-culture with an EOC2 microglial cell line. Microglia soluble cues enhanced cortical cell viability and proliferation of uninjured and injured cortical cells. Co-culture of injured cortical cells with microglial cells significantly reduced cortical cell apoptosis. Microglial significantly increased Nestin+ and a-internexin+ cells within and outside the injury site. NeuN+ cells increased in injured cortical cultures with microglia. Multiplex ELISA assays showed decreased levels of inflammatory cytokines in conditioned media collected from injured cortical cell and microglial co-culture. RTPCR analysis of microglial mRNA was performed. AKT phosphorylation in uninjured, and particularly injured cortical cells, significantly increased when co-cultured with EOC2 microglia. Inhibition of AKT phosphorylation in cortical cells blocked the microglial-enhanced cortical cell viability and expression of neurogenic markers in vitro . Conclusion: This in vitro model system allows for assessment of the effect of microglial-derived soluble signals on cortical cell viability, proliferation, and stages of differentiation during homeostasis or following injury. These data suggest that EOC2 microglia downregulate inflammatory cytokine production following activation by acute cortical injury to enhance proliferation of new cells capable of neurogenesis. Inhibition of AKT signaling in cortical cells blocks the microglial-derived enhanced proliferation and expression of neurogenic markers in injured cortical cultures. This in vitro system is useful for continued studies with other microglial cell lines and primary microglial cells. Increasing our understanding of the mechanisms that drive cortical neurogenesis stimulated by microglial cells during homeostasis and following injury will provide insight into the potential mechanisms of the neuroprotective role of immune activity in the central nervous system (CNS).

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Bartonella henselae Invasion of Feline Erythrocytes In Vitro

Infection and Immunity ◽

10.1128/iai.66.7.3462-3466.1998 ◽

1998 ◽

Vol 66 (7) ◽

pp. 3462-3466 ◽

Cited By ~ 46

Author(s):

Jane R. Mehock ◽

Craig E. Greene ◽

Frank C. Gherardini ◽

Tae-Wook Hahn ◽

Duncan C. Krause

Keyword(s):

Confocal Microscopy ◽

Bartonella Henselae ◽

Intracellular Bacteria ◽

Cat Scratch Disease ◽

Slow Process ◽

Laser Confocal Microscopy ◽

Bacterial Surface ◽

Selection For

ABSTRACT Bartonella henselae, the causative agent of cat scratch disease, establishes long-term bacteremia in cats, in which it attaches to and invades feline erythrocytes (RBC). Feline RBC invasion was assessed in vitro, based on gentamicin selection for intracellular bacteria or by laser confocal microscopy and digital sectioning. Invasion rates ranged from 2 to 20% of the inoculum, corresponding to infection of less than 1% of the RBC. Invasion was a slow process, requiring >8 h before significant numbers of intracellular bacteria were detected. Pretreatment of the bacteria with trypsin, or of the RBC with trypsin or neuraminidase, had no effect, but pronase pretreatment of RBC resulted in a slight increase in invasion frequency. The ability to model B. henselae invasion of feline RBC in vitro should permit identification of bacterial surface components involved in this process and elucidate the significance of RBC invasion to transmission and infection in cats.

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Receptor-Independent Spread of a Highly Neurotropic Murine Coronavirus JHMV Strain from Initially Infected Microglial Cells in Mixed Neural Cultures

Journal of Virology ◽

10.1128/jvi.79.10.6102-6110.2005 ◽

2005 ◽

Vol 79 (10) ◽

pp. 6102-6110 ◽

Cited By ~ 32

Author(s):

Keiko Nakagaki ◽

Kazuhide Nakagaki ◽

Fumihiro Taguchi

Keyword(s):

Cell Cultures ◽

Hepatitis Virus ◽

Resistant Mutant ◽

Neural Cell ◽

Microglial Cells ◽

Brain Cells ◽

Neural Cells ◽

Virus Attachment ◽

Enriched Cultures ◽

The Brain

ABSTRACT Although neurovirulent mouse hepatitis virus (MHV) strain JHMV multiplies in a variety of brain cells, expression of its receptor carcinoembryonic antigen cell adhesion molecule 1 (CEACAM 1) (MHVR) is restricted only in microglia. The present study was undertaken to clarify the mechanism of an extensive JHMV infection in the brain by using neural cells isolated from mouse brain. In contrast to wild-type (wt) JHMV, a soluble-receptor-resistant mutant (srr7) infects and spreads solely in an MHVR-dependent fashion (F. Taguchi and S. Matsuyama, J. Virol. 76:950-958, 2002). In mixed neural cell cultures, srr7 infected a limited number of cells and infection did not spread, although wt JHMV induced syncytia in most of the cells. srr7-infected cells were positive for GS-lectin, a microglia marker. Fluorescence-activated cell sorter analysis showed that about 80% of the brain cells stained with anti-MHVR antibody (CC1) were also positive for GS-lectin. Pretreatment of those cells with CC1 prevented virus attachment to the cell surface and also blocked virus infection. These results show that microglia express functional MHVR that mediates JHMV infection. As expected, in microglial cell-enriched cultures, both srr7and wt JHMV produced syncytia in a majority of cells. Treatment with CC1 of mixed neural cell cultures and microglia cultures previously infected with wt virus failed to block the spread of infection, indicating that wt infection spreads in an MHVR-independent fashion. Thus, the present study indicates that microglial cells are the major population of the initial target for MHV infection and that the wt spreads from initially infected microglia to a variety of cells in an MHVR-independent fashion.

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Functional maturation of human neural stem cells in a 3D bioengineered brain model enriched with fetal brain-derived matrix

Scientific Reports ◽

10.1038/s41598-019-54248-1 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 4

Author(s):

Disha Sood ◽

Dana M. Cairns ◽

Jayanth M. Dabbi ◽

Charu Ramakrishnan ◽

Karl Deisseroth ◽

...

Keyword(s):

Stem Cells ◽

Neural Stem Cells ◽

Brain Tissue ◽

Fetal Brain ◽

Functional Differentiation ◽

Adult Brain ◽

Human Neural Stem Cells ◽

Functional Maturation ◽

Enriched Cultures

AbstractBrain extracellular matrix (ECM) is often overlooked in vitro brain tissue models, despite its instructive roles during development. Using developmental stage-sourced brain ECM in reproducible 3D bioengineered culture systems, we demonstrate enhanced functional differentiation of human induced neural stem cells (hiNSCs) into healthy neurons and astrocytes. Particularly, fetal brain tissue-derived ECM supported long-term maintenance of differentiated neurons, demonstrated by morphology, gene expression and secretome profiling. Astrocytes were evident within the second month of differentiation, and reactive astrogliosis was inhibited in brain ECM-enriched cultures when compared to unsupplemented cultures. Functional maturation of the differentiated hiNSCs within fetal ECM-enriched cultures was confirmed by calcium signaling and spectral/cluster analysis. Additionally, the study identified native biochemical cues in decellularized ECM with notable comparisons between fetal and adult brain-derived ECMs. The development of novel brain-specific biomaterials for generating mature in vitro brain models provides an important path forward for interrogation of neuron-glia interactions.

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