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Article ; Online: The importance of controlling in vitro oxygen tension to accurately model in vivo neurophysiology.

Bordt, Evan A

2017 Volume 66, Page(s) 213–220

Abstract: The majority of in vitro studies modeling in vivo conditions are performed on the lab bench in atmospheric air. However, the oxygen tension ( ... ...

Abstract	The majority of in vitro studies modeling in vivo conditions are performed on the lab bench in atmospheric air. However, the oxygen tension (pO
MeSH term(s)	Animals ; Atmospheric Pressure ; Brain/metabolism ; Cell Differentiation ; Humans ; Hypoxia-Inducible Factor 1/metabolism ; Mitochondria/metabolism ; Models, Neurological ; Neuroglia/metabolism ; Oxygen/metabolism ; Reactive Oxygen Species ; Research Design ; Sex Characteristics
Chemical Substances	Hypoxia-Inducible Factor 1 ; Reactive Oxygen Species ; Oxygen (S88TT14065)
Language	English
Publishing date	2017-11-01
Publishing country	Netherlands
Document type	Journal Article ; Review
ZDB-ID	800820-6
ISSN	1872-9711 ; 0161-813X
ISSN (online)	1872-9711
ISSN	0161-813X
DOI	10.1016/j.neuro.2017.10.008
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Article ; Online: Stressed-Out T Cells Fragment the Mind.

Bordt, Evan A / Bilbo, Staci D

Trends in immunology

2020 Volume 41, Issue 2, Page(s) 94–97

Abstract: The immune system is increasingly recognized to play an integral role in regulating stress responses. In a recent article in Cell, Fan et al. demonstrate a novel mechanism through which stress drives mitochondrial fragmentation-induced xanthine ... ...

Abstract	The immune system is increasingly recognized to play an integral role in regulating stress responses. In a recent article in Cell, Fan et al. demonstrate a novel mechanism through which stress drives mitochondrial fragmentation-induced xanthine accumulation in mouse CD4
MeSH term(s)	ATP Citrate (pro-S)-Lyase ; Acetylation ; Adenosine Triphosphate ; Animals ; Histones ; Humans ; Macrophages ; Mice ; Multienzyme Complexes ; Oxo-Acid-Lyases ; T-Lymphocytes ; Toll-Like Receptors
Chemical Substances	Histones ; Multienzyme Complexes ; Toll-Like Receptors ; Adenosine Triphosphate (8L70Q75FXE) ; ATP Citrate (pro-S)-Lyase (EC 2.3.3.8) ; Oxo-Acid-Lyases (EC 4.1.3.-) ; citrate (pro-3S)-lyase (EC 4.1.3.6)
Language	English
Publishing date	2020-01-06
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Comment
ZDB-ID	2036831-8
ISSN	1471-4981 ; 1471-4906
ISSN (online)	1471-4981
ISSN	1471-4906
DOI	10.1016/j.it.2019.12.008
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Zs.A 1601: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (1.OG) ab Jg. 2022: Lesesaal (EG)
Zs.MG 2: Show issues

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Article: The Non-Specific Drp1 Inhibitor Mdivi-1 Has Modest Biochemical Antioxidant Activity.

Bordt, Evan A / Zhang, Naibo / Waddell, Jaylyn / Polster, Brian M

Antioxidants (Basel, Switzerland)

2022 Volume 11, Issue 3

Abstract: Mitochondrial division inhibitor-1 (mdivi-1), a non-specific inhibitor of Drp1-dependent mitochondrial fission, is neuroprotective in numerous preclinical disease models. These include rodent models of Alzheimer's disease and ischemic or traumatic brain ... ...

Abstract	Mitochondrial division inhibitor-1 (mdivi-1), a non-specific inhibitor of Drp1-dependent mitochondrial fission, is neuroprotective in numerous preclinical disease models. These include rodent models of Alzheimer's disease and ischemic or traumatic brain injury. Among its Drp1-independent actions, the compound was found to suppress mitochondrial Complex I-dependent respiration but with less resultant mitochondrial reactive oxygen species (ROS) emission compared with the classical Complex I inhibitor rotenone. We employed two different methods of quantifying Trolox-equivalent antioxidant capacity (TEAC) to test the prediction that mdivi-1 can directly scavenge free radicals. Mdivi-1 exhibited moderate antioxidant activity in the 2,2'-azinobis (3-ethylbenzothiazoline 6-sulfonate) (ABTS) assay. Half-maximal ABTS radical depletion was observed at ~25 μM mdivi-1, equivalent to that achieved by ~12.5 μM Trolox. Mdivi-1 also showed antioxidant activity in the α, α-diphenyl-β-picrylhydrazyl (DPPH) assay. However, mdivi-1 exhibited a reduced capacity to deplete the DPPH radical, which has a more sterically hindered radical site compared with ABTS, with 25 μM mdivi-1 displaying only 0.8 μM Trolox equivalency. Both assays indicate that mdivi-1 possesses biochemical antioxidant activity but with modest potency relative to the vitamin E analog Trolox. Future studies are needed to evaluate whether the ability of mdivi-1 to directly scavenge free radicals contributes to its mechanisms of neuroprotection.
Language	English
Publishing date	2022-02-24
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2704216-9
ISSN	2076-3921
ISSN	2076-3921
DOI	10.3390/antiox11030450
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Article: The Non-Specific Drp1 Inhibitor Mdivi-1 Has Modest Biochemical Antioxidant Activity

Bordt, Evan A. / Zhang, Naibo / Waddell, Jaylyn / Polster, Brian M.

Antioxidants. 2022 Feb. 24, v. 11, no. 3

2022

Abstract	Mitochondrial division inhibitor-1 (mdivi-1), a non-specific inhibitor of Drp1-dependent mitochondrial fission, is neuroprotective in numerous preclinical disease models. These include rodent models of Alzheimer’s disease and ischemic or traumatic brain injury. Among its Drp1-independent actions, the compound was found to suppress mitochondrial Complex I-dependent respiration but with less resultant mitochondrial reactive oxygen species (ROS) emission compared with the classical Complex I inhibitor rotenone. We employed two different methods of quantifying Trolox-equivalent antioxidant capacity (TEAC) to test the prediction that mdivi-1 can directly scavenge free radicals. Mdivi-1 exhibited moderate antioxidant activity in the 2,2′-azinobis (3-ethylbenzothiazoline 6-sulfonate) (ABTS) assay. Half-maximal ABTS radical depletion was observed at ~25 μM mdivi-1, equivalent to that achieved by ~12.5 μM Trolox. Mdivi-1 also showed antioxidant activity in the α, α-diphenyl-β-picrylhydrazyl (DPPH) assay. However, mdivi-1 exhibited a reduced capacity to deplete the DPPH radical, which has a more sterically hindered radical site compared with ABTS, with 25 μM mdivi-1 displaying only 0.8 μM Trolox equivalency. Both assays indicate that mdivi-1 possesses biochemical antioxidant activity but with modest potency relative to the vitamin E analog Trolox. Future studies are needed to evaluate whether the ability of mdivi-1 to directly scavenge free radicals contributes to its mechanisms of neuroprotection.
Keywords	antioxidant activity ; brain damage ; mitochondria ; neuroprotective effect ; prediction ; reactive oxygen species ; rodents ; rotenone ; vitamin E
Language	English
Dates of publication	2022-0224
Publishing place	Multidisciplinary Digital Publishing Institute
Document type	Article
ZDB-ID	2704216-9
ISSN	2076-3921
ISSN	2076-3921
DOI	10.3390/antiox11030450
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Microglia and sexual differentiation of the developing brain: A focus on ontogeny and intrinsic factors.

Bordt, Evan A / Ceasrine, Alexis M / Bilbo, Staci D

Glia

2019 Volume 68, Issue 6, Page(s) 1085–1099

Abstract: Sexual differentiation of the brain during early development likely underlies the strong sex biases prevalent in many neurological conditions. Mounting evidence indicates that microglia, the innate immune cells of the central nervous system, are ... ...

Abstract	Sexual differentiation of the brain during early development likely underlies the strong sex biases prevalent in many neurological conditions. Mounting evidence indicates that microglia, the innate immune cells of the central nervous system, are intricately involved in these sex-specific processes of differentiation. In this review, we synthesize literature demonstrating sex differences in microglial number, morphology, transcriptional state, and functionality throughout spatiotemporal development as well as highlight current literature regarding ontogeny of microglia. Along with vanRyzin et al. in this issue, we explore the idea that differences in microglia imparted by chromosomal or ontogeny-related programming can influence microglial-driven sexual differentiation of the brain, as well as the idea that extrinsic differences in the male and female brain microenvironment may in turn impart sex differences in microglia.
MeSH term(s)	Animals ; Brain/cytology ; Brain/growth & development ; Humans ; Male ; Microglia/physiology ; Nervous System Diseases/pathology ; Neurons/cytology ; Sex Characteristics ; Sex Differentiation/physiology
Language	English
Publishing date	2019-11-19
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Review
ZDB-ID	639414-0
ISSN	1098-1136 ; 0894-1491
ISSN (online)	1098-1136
ISSN	0894-1491
DOI	10.1002/glia.23753
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Zs.A 2345: Show issues

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Article ; Online: NADPH oxidase- and mitochondria-derived reactive oxygen species in proinflammatory microglial activation: a bipartisan affair?

Bordt, Evan A / Polster, Brian M

Free radical biology & medicine

2014 Volume 76, Page(s) 34–46

Abstract: Microglia are the resident immune cells of the brain and play major roles in central nervous system development, maintenance, and disease. Brain insults cause microglia to proliferate, migrate, and transform into one or more activated states. Classical ... ...

Abstract	Microglia are the resident immune cells of the brain and play major roles in central nervous system development, maintenance, and disease. Brain insults cause microglia to proliferate, migrate, and transform into one or more activated states. Classical M1 activation triggers the production of proinflammatory factors such as tumor necrosis factor-α, interleukin-1β (IL-1β), nitric oxide, and reactive oxygen species (ROS), which, in excess, can exacerbate brain injury. The mechanisms underlying microglial activation are not fully understood, yet reactive oxygen species are increasingly implicated as mediators of microglial activation. In this review, we highlight studies linking reactive oxygen species, in particular hydrogen peroxide derived from NADPH oxidase-generated superoxide, to the classical activation of microglia. In addition, we critically evaluate controversial evidence suggesting a specific role for mitochondrial reactive oxygen species in the activation of the NLRP3 inflammasome, a multiprotein complex that mediates the production of IL-1β and IL-18. Finally, the limitations of common techniques used to implicate mitochondrial ROS in microglial and inflammasome activation, such as the use of the mitochondrially targeted ROS indicator MitoSOX and the mitochondrially targeted antioxidant MitoTEMPO, are also discussed.
MeSH term(s)	Animals ; Humans ; Inflammation Mediators/metabolism ; Microglia/immunology ; Microglia/metabolism ; Microglia/pathology ; Mitochondria/metabolism ; NADPH Oxidases/metabolism ; Reactive Oxygen Species/metabolism
Chemical Substances	Inflammation Mediators ; Reactive Oxygen Species ; NADPH Oxidases (EC 1.6.3.-)
Language	English
Publishing date	2014-08-01
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Review
ZDB-ID	807032-5
ISSN	1873-4596 ; 0891-5849
ISSN (online)	1873-4596
ISSN	0891-5849
DOI	10.1016/j.freeradbiomed.2014.07.033
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Zs.A 2109: Show issues

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ab Jg. 2022: Lesesaal (EG)

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Article: Hofbauer cells and fetal brain microglia share transcriptional profiles and responses to maternal diet-induced obesity.

Batorsky, Rebecca / Ceasrine, Alexis M / Shook, Lydia L / Kislal, Sezen / Bordt, Evan A / Devlin, Benjamin A / Perlis, Roy H / Slonim, Donna K / Bilbo, Staci D / Edlow, Andrea G

bioRxiv : the preprint server for biology

2023

Abstract: Maternal immune activation is associated with adverse offspring neurodevelopmental outcomes, many mediated by in utero microglial programming. As microglia remain inaccessible throughout development, identification of noninvasive biomarkers reflecting ... ...

Abstract	Maternal immune activation is associated with adverse offspring neurodevelopmental outcomes, many mediated by in utero microglial programming. As microglia remain inaccessible throughout development, identification of noninvasive biomarkers reflecting fetal brain microglial programming could permit screening and intervention. We used lineage tracing to demonstrate the shared ontogeny between fetal brain macrophages (microglia) and fetal placental macrophages (Hofbauer cells) in a mouse model of maternal diet-induced obesity, and single-cell RNA-seq to demonstrate shared transcriptional programs. Comparison with human datasets demonstrated conservation of placental resident macrophage signatures between mice and humans. Single-cell RNA-seq identified common alterations in fetal microglial and Hofbauer cell gene expression induced by maternal obesity, as well as sex differences in these alterations. We propose that Hofbauer cells, which are easily accessible at birth, provide novel insights into fetal brain microglial programs, and may facilitate the early identification of offspring vulnerable to neurodevelopmental disorders in the setting of maternal exposures.
Language	English
Publishing date	2023-12-19
Publishing country	United States
Document type	Preprint
DOI	10.1101/2023.12.16.571680
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Article ; Online: Gonadal hormones impart male-biased behavioral vulnerabilities to immune activation via microglial mitochondrial function.

Bordt, Evan A / Moya, Haley A / Jo, Young Chan / Ravichandran, Caitlin T / Bankowski, Izabella M / Ceasrine, Alexis M / McDougle, Christopher J / Carlezon, William A / Bilbo, Staci D

Brain, behavior, and immunity

2023 Volume 115, Page(s) 680–695

Abstract: There is a strong male bias in the prevalence of many neurodevelopmental disorders such as autism spectrum disorder. However, the mechanisms underlying this sex bias remain elusive. Infection during the perinatal period is associated with an increased ... ...

Abstract	There is a strong male bias in the prevalence of many neurodevelopmental disorders such as autism spectrum disorder. However, the mechanisms underlying this sex bias remain elusive. Infection during the perinatal period is associated with an increased risk of neurodevelopmental disorder development. Here, we used a mouse model of early-life immune activation that reliably induces deficits in social behaviors only in males. We demonstrate that male-biased alterations in social behavior are dependent upon microglial immune signaling and are coupled to alterations in mitochondrial morphology, gene expression, and function specifically within microglia, the innate immune cells of the brain. Additionally, we show that this behavioral and microglial mitochondrial vulnerability to early-life immune activation is programmed by the male-typical perinatal gonadal hormone surge. These findings demonstrate that social behavior in males over the lifespan are regulated by microglia-specific mechanisms that are shaped by events that occur in early development.
MeSH term(s)	Animals ; Mice ; Pregnancy ; Female ; Male ; Microglia/metabolism ; Autism Spectrum Disorder ; Brain/metabolism ; Gonadal Hormones/metabolism ; Mitochondria/metabolism
Chemical Substances	Gonadal Hormones
Language	English
Publishing date	2023-11-14
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural
ZDB-ID	639219-2
ISSN	1090-2139 ; 0889-1591
ISSN (online)	1090-2139
ISSN	0889-1591
DOI	10.1016/j.bbi.2023.11.010
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Zs.A 2276: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (2.OG) ab Jg. 2022: Lesesaal (EG)
Zs.MO 327: Show issues

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Article ; Online: DNAJB1-PRKACA in HEK293T cells induces LINC00473 overexpression that depends on PKA signaling.

Kim, Stephanie S / Kycia, Ina / Karski, Michael / Ma, Rosanna K / Bordt, Evan A / Kwan, Julian / Karki, Anju / Winter, Elle / Aktas, Ranan G / Wu, Yuxuan / Emili, Andrew / Bauer, Daniel E / Sethupathy, Praveen / Vakili, Khashayar

PloS one

2022 Volume 17, Issue 2, Page(s) e0263829

Abstract: Fibrolamellar carcinoma (FLC) is a primary liver cancer that most commonly arises in adolescents and young adults in a background of normal liver tissue and has a poor prognosis due to lack of effective chemotherapeutic agents. The DNAJB1-PRKACA gene ... ...

Abstract	Fibrolamellar carcinoma (FLC) is a primary liver cancer that most commonly arises in adolescents and young adults in a background of normal liver tissue and has a poor prognosis due to lack of effective chemotherapeutic agents. The DNAJB1-PRKACA gene fusion (DP) has been reported in the majority of FLC tumors; however, its oncogenic mechanisms remain unclear. Given the paucity of cellular models, in particular FLC tumor cell lines, we hypothesized that engineering the DP fusion gene in HEK293T cells would provide insight into the cellular effects of the fusion gene. We used CRISPR/Cas9 to engineer HEK293T clones expressing DP fusion gene (HEK-DP) and performed transcriptomic, proteomic, and mitochondrial studies to characterize this cellular model. Proteomic analysis of DP interacting partners identified mitochondrial proteins as well as proteins in other subcellular compartments. HEK-DP cells demonstrated significantly elevated mitochondrial fission, which suggests a role for DP in altering mitochondrial dynamics. Transcriptomic analysis of HEK-DP cells revealed a significant increase in LINC00473 expression, similar to what has been observed in primary FLC samples. LINC00473 overexpression was reversible with siRNA targeting of PRKACA as well as pharmacologic targeting of PKA and Hsp40 in HEK-DP cells. Therefore, our model suggests that LINC00473 is a candidate marker for DP activity.
MeSH term(s)	CRISPR-Cas Systems ; Carcinoma, Hepatocellular/genetics ; Cyclic AMP-Dependent Protein Kinase Catalytic Subunits/genetics ; Cyclic AMP-Dependent Protein Kinase Catalytic Subunits/metabolism ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; HEK293 Cells ; HSP40 Heat-Shock Proteins/genetics ; HSP40 Heat-Shock Proteins/metabolism ; Humans ; Mitochondria/metabolism ; Models, Biological ; Oncogene Proteins, Fusion/genetics ; Proteomics ; RNA, Long Noncoding/genetics ; Up-Regulation
Chemical Substances	DNAJB1 protein, human ; HSP40 Heat-Shock Proteins ; LINC00473 RNA, human ; Oncogene Proteins, Fusion ; RNA, Long Noncoding ; Cyclic AMP-Dependent Protein Kinase Catalytic Subunits (EC 2.7.11.11) ; PRKACA protein, human (EC 2.7.11.11)
Language	English
Publishing date	2022-02-15
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2267670-3
ISSN	1932-6203 ; 1932-6203
ISSN (online)	1932-6203
ISSN	1932-6203
DOI	10.1371/journal.pone.0263829
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Article ; Online: Isolation of Microglia from Mouse or Human Tissue.

Bordt, Evan A / Block, Carina L / Petrozziello, Tiziana / Sadri-Vakili, Ghazaleh / Smith, Caroline J / Edlow, Andrea G / Bilbo, Staci D

STAR protocols

2020 Volume 1, Issue 1

Abstract: Microglia are the innate immune cells of the central nervous system. Although numerous methods have been developed to isolate microglia from the brain, the method of dissociation and isolation can have a profound effect on the function of these highly ... ...

Abstract	Microglia are the innate immune cells of the central nervous system. Although numerous methods have been developed to isolate microglia from the brain, the method of dissociation and isolation can have a profound effect on the function of these highly dynamic cells. Here, we present an optimized protocol to isolate CD11b+ cells (microglia) from mouse or human brain tissue using magnetic bead columns. Isolated microglia can be used to model diseases with neuroinflammatory components for potential therapeutic discoveries. For complete details on the use and execution of this protocol, please refer to Hanamsagar et al. (2017), Rivera et al. (2019), and Edlow et al. (2019).
MeSH term(s)	Animals ; CD11b Antigen/metabolism ; Cell Separation/methods ; Humans ; Mice ; Microglia/metabolism
Chemical Substances	CD11b Antigen
Language	English
Publishing date	2020-06-03
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ISSN	2666-1667
ISSN (online)	2666-1667
DOI	10.1016/j.xpro.2020.100035
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