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Article ; Online: The Comprehensive Steroidome in Complete TSPO/PBR Knockout Mice under Basal Conditions.

Liere, Philippe / Liu, Guo-Jun / Pianos, Antoine / Middleton, Ryan J / Banati, Richard B / Akwa, Yvette

International journal of molecular sciences

2023 Volume 24, Issue 3

Abstract: The 18 kDa translocator protein (TSPO/PBR) is a multifunctional evolutionary highly conserved outer mitochondrial membrane protein. Decades of research has reported an obligatory role of TSPO/PBR in both mitochondrial cholesterol transport and, thus, ... ...

Abstract	The 18 kDa translocator protein (TSPO/PBR) is a multifunctional evolutionary highly conserved outer mitochondrial membrane protein. Decades of research has reported an obligatory role of TSPO/PBR in both mitochondrial cholesterol transport and, thus, steroid production. However, the strict dependency of steroidogenesis on TSPO/PBR has remained controversial. The aim of this study was to provide insight into the steroid profile in complete C57BL/6-
MeSH term(s)	Male ; Mice ; Animals ; Receptors, GABA/genetics ; Receptors, GABA/metabolism ; Mice, Knockout ; Tandem Mass Spectrometry ; Mice, Inbred C57BL ; Steroids ; Carrier Proteins ; Pregnenolone
Chemical Substances	Receptors, GABA ; Steroids ; Carrier Proteins ; Pregnenolone (73R90F7MQ8)
Language	English
Publishing date	2023-01-27
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms24032474
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Article ; Online: The Translocator Protein (TSPO) in Mitochondrial Bioenergetics and Immune Processes.

Betlazar, Calina / Middleton, Ryan J / Banati, Richard / Liu, Guo-Jun

Cells

2020 Volume 9, Issue 2

Abstract: The translocator protein (TSPO) is an outer mitochondrial membrane protein that is widely used as a biomarker of neuroinflammation, being markedly upregulated in activated microglia in a range of brain pathologies. Despite its extensive use as a target ... ...

Abstract	The translocator protein (TSPO) is an outer mitochondrial membrane protein that is widely used as a biomarker of neuroinflammation, being markedly upregulated in activated microglia in a range of brain pathologies. Despite its extensive use as a target in molecular imaging studies, the exact cellular functions of this protein remain in question. The long-held view that TSPO plays a fundamental role in the translocation of cholesterol through the mitochondrial membranes, and thus, steroidogenesis, has been disputed by several groups with the advent of TSPO knockout mouse models. Instead, much evidence is emerging that TSPO plays a fundamental role in cellular bioenergetics and associated mitochondrial functions, also part of a greater role in the innate immune processes of microglia. In this review, we examine the more direct experimental literature surrounding the immunomodulatory effects of TSPO. We also review studies which highlight a more central role for TSPO in mitochondrial processes, from energy metabolism, to the propagation of inflammatory responses through reactive oxygen species (ROS) modulation. In this way, we highlight a paradigm shift in approaches to TSPO functioning.
MeSH term(s)	Animals ; Disease Models, Animal ; Energy Metabolism/physiology ; Humans ; Immunity/physiology ; Mice ; Mitochondria/metabolism ; Receptors, GABA/metabolism
Chemical Substances	Receptors, GABA ; TSPO protein, human
Language	English
Publishing date	2020-02-24
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	2661518-6
ISSN	2073-4409 ; 2073-4409
ISSN (online)	2073-4409
ISSN	2073-4409
DOI	10.3390/cells9020512
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Article ; Online: Control of Neuroinflammation through Radiation-Induced Microglial Changes.

Boyd, Alexandra / Byrne, Sarah / Middleton, Ryan J / Banati, Richard B / Liu, Guo-Jun

Cells

2021 Volume 10, Issue 9

Abstract: Microglia, the innate immune cells of the central nervous system, play a pivotal role in the modulation of neuroinflammation. Neuroinflammation has been implicated in many diseases of the CNS, including Alzheimer's disease and Parkinson's disease. It is ... ...

Abstract	Microglia, the innate immune cells of the central nervous system, play a pivotal role in the modulation of neuroinflammation. Neuroinflammation has been implicated in many diseases of the CNS, including Alzheimer's disease and Parkinson's disease. It is well documented that microglial activation, initiated by a variety of stressors, can trigger a potentially destructive neuroinflammatory response via the release of pro-inflammatory molecules, and reactive oxygen and nitrogen species. However, the potential anti-inflammatory and neuroprotective effects that microglia are also thought to exhibit have been under-investigated. The application of ionising radiation at different doses and dose schedules may reveal novel methods for the control of microglial response to stressors, potentially highlighting avenues for treatment of neuroinflammation associated CNS disorders, such as Alzheimer's disease and Parkinson's disease. There remains a need to characterise the response of microglia to radiation, particularly low dose ionising radiation.
MeSH term(s)	Animals ; Dose-Response Relationship, Radiation ; Humans ; Immunity, Innate/radiation effects ; Inflammation Mediators/metabolism ; Microglia/immunology ; Microglia/metabolism ; Microglia/pathology ; Microglia/radiation effects ; Neurodegenerative Diseases/immunology ; Neurodegenerative Diseases/metabolism ; Neurodegenerative Diseases/pathology ; Neurodegenerative Diseases/radiotherapy ; Neuroimmunomodulation/radiation effects ; Nitrosative Stress/radiation effects ; Oxidative Stress/radiation effects ; Phenotype ; Reactive Nitrogen Species/metabolism ; Reactive Oxygen Species/metabolism ; Receptors, GABA/metabolism
Chemical Substances	Inflammation Mediators ; Reactive Nitrogen Species ; Reactive Oxygen Species ; Receptors, GABA ; TSPO protein, human
Language	English
Publishing date	2021-09-10
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2661518-6
ISSN	2073-4409 ; 2073-4409
ISSN (online)	2073-4409
ISSN	2073-4409
DOI	10.3390/cells10092381
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Article ; Online: Subcellular distribution of the 18kDa translocator protein and transcript variant PBR-S in human cells.

Liu, Guo-Jun / Middleton, Ryan J / Banati, Richard B

Gene

2017 Volume 613, Page(s) 45–56

Abstract: Despite continued interest in the 18kDa translocator protein (PBR/TSPO) as a biomarker and a therapeutic target for a range of diseases, its functional role, such as in the steroid synthesis pathway and energy metabolism has either become contentious or ... ...

Abstract	Despite continued interest in the 18kDa translocator protein (PBR/TSPO) as a biomarker and a therapeutic target for a range of diseases, its functional role, such as in the steroid synthesis pathway and energy metabolism has either become contentious or remains to be described more precisely. The PBR/TSPO gene consists of four exons, while a shorter isoform termed PBR-S lacks exon 2. The PBR-S 102-codon open reading frame differs to that of PBR/TSPO, resulting in a protein that is completely unrelated to PBR/TSPO. To our knowledge, PBR-S protein has never been described and has no known or proposed function. To obtain possible clues on the role of this uncharacterised protein, we compared the subcellular distribution of PBR-S to that of PBR/TSPO. By expressing fluorescently tagged PBR/TSPO, we confirmed that full-length PBR/TSPO co-localises with mitochondria in HeLa, HEK-293, MDA-MB-231, BJ and U87-MG human cell lines. Unlike the strictly mitochondrial localisation of PBR/TSPO, PBR-S has a punctate distribution throughout the cytosol that co-localises with lysosomes in HeLa, HEK-293, MDA-MB-231, BJ and U87-MG cells. In summary, within the cell lines examined we confirm mitochondria rather than occasionally reported other localisations, such as the cell nucleus, to be the only site where PBR/TSPO resides. Due to the lack of any shared protein sequences and the different subcellular locations, we suggest that the previously uncharacterised PBR-S protein variant of the PBR/TSPO gene is likely to serve a different yet to be discovered function compared to PBR/TSPO.
MeSH term(s)	Cell Line ; Cell Line, Tumor ; Humans ; Lysosomes/chemistry ; Mitochondria/chemistry ; Receptors, GABA/analysis ; Receptors, GABA/genetics ; Transfection
Chemical Substances	Receptors, GABA ; TSPO protein, human
Language	English
Publishing date	2017-05-20
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	391792-7
ISSN	1879-0038 ; 0378-1119
ISSN (online)	1879-0038
ISSN	0378-1119
DOI	10.1016/j.gene.2017.02.035
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Article: Mitochondrial Translocator Protein (TSPO) Expression in the Brain After Whole Body Gamma Irradiation.

Betlazar, Calina / Middleton, Ryan J / Howell, Nicholas / Storer, Ben / Davis, Emma / Davies, Justin / Banati, Richard / Liu, Guo-Jun

Frontiers in cell and developmental biology

2021 Volume 9, Page(s) 715444

Abstract: The brain's early response to low dose ionizing radiation, as may be encountered during diagnostic procedures and space exploration, is not yet fully characterized. In the brain parenchyma, the mitochondrial translocator protein (TSPO) is constitutively ... ...

Abstract	The brain's early response to low dose ionizing radiation, as may be encountered during diagnostic procedures and space exploration, is not yet fully characterized. In the brain parenchyma, the mitochondrial translocator protein (TSPO) is constitutively expressed at low levels by endothelial cells, and can therefore be used to assess the integrity of the brain's vasculature. At the same time, the inducible expression of TSPO in activated microglia, the brain's intrinsic immune cells, is a regularly observed early indicator of subtle or incipient brain pathology. Here, we explored the use of TSPO as a biomarker of brain tissue injury following whole body irradiation. Post-radiation responses were measured in C57BL/6 wild type (
Language	English
Publishing date	2021-10-25
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2737824-X
ISSN	2296-634X
ISSN	2296-634X
DOI	10.3389/fcell.2021.715444
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Article ; Online: Neutron Capture Enhances Dose and Reduces Cancer Cell Viability in and out of Beam During Helium and Carbon Ion Therapy.

International journal of radiation oncology, biology, physics

2024

Abstract: Purpose: Neutron capture enhanced particle therapy (NCEPT) is a proposed augmentation of charged particle therapy that exploits thermal neutrons generated internally, within the treatment volume via nuclear fragmentation, to deliver a biochemically ... ...

Abstract	Purpose: Neutron capture enhanced particle therapy (NCEPT) is a proposed augmentation of charged particle therapy that exploits thermal neutrons generated internally, within the treatment volume via nuclear fragmentation, to deliver a biochemically targeted radiation dose to cancer cells. This work is the first experimental demonstration of NCEPT, performed using both carbon and helium ion beams with 2 different targeted neutron capture agents (NCAs). Methods and materials: Human glioblastoma cells (T98G) were irradiated by carbon and helium ion beams in the presence of NCAs [ Results: Cells placed inside the treatment volume reached 10% survival by 2 Gy of carbon or 2 to 3 Gy of helium in the presence of NCAs compared with 5 Gy of carbon and 7 Gy of helium with no NCA. Cells placed adjacent to the treatment volume showed a dose-dependent decrease in cell growth when treated with NCAs, reaching 10% survival by 6 Gy of carbon or helium (to the treatment volume), compared with no detectable effect on cells without NCA. The mean thermal neutron fluence at the center of the SOBP was approximately 2.2 × 10 Conclusions: The addition of NCAs to cancer cells during carbon and helium beam irradiation has a measurable effect on cell survival and growth in vitro. Through the capture of internally generated neutrons, NCEPT introduces the concept of a biochemically targeted radiation dose to charged particle therapy. NCEPT enables the established pharmaceuticals and concepts of neutron capture therapy to be applied to a wider range of deeply situated and diffuse tumors, by targeting this dose to microinfiltrates and cells outside of defined treatment regions. These results also demonstrate the potential for NCEPT to provide an increased dose to tumor tissue within the treatment volume, with a reduction in radiation doses to off-target tissue.
Language	English
Publishing date	2024-03-11
Publishing country	United States
Document type	Journal Article
ZDB-ID	197614-x
ISSN	1879-355X ; 0360-3016
ISSN (online)	1879-355X
ISSN	0360-3016
DOI	10.1016/j.ijrobp.2024.02.052
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Zs.A 1218: Show issues

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Article ; Online: Guwiyang Wurra--'Fire Mouse': a global gene knockout model for TSPO/PBR drug development, loss-of-function and mechanisms of compensation studies.

Middleton, Ryan J / Liu, Guo-Jun / Banati, Richard B

Biochemical Society transactions

2015 Volume 43, Issue 4, Page(s) 553–558

Abstract: The highly conserved 18-kDa translocator protein (TSPO) or peripheral benzodiazepine receptor (PBR), is being investigated as a diagnostic and therapeutic target for disease conditions ranging from inflammation to neurodegeneration and behavioural ... ...

Abstract	The highly conserved 18-kDa translocator protein (TSPO) or peripheral benzodiazepine receptor (PBR), is being investigated as a diagnostic and therapeutic target for disease conditions ranging from inflammation to neurodegeneration and behavioural illnesses. Many functions have been attributed to TSPO/PBR including a role in the mitochondrial permeability transition pore (MPTP), steroidogenesis and energy metabolism. In this review, we detail the recent developments in determining the physiological role of TSPO/PBR, specifically based on data obtained from the recently generated Tspo knockout mouse models. In addition to defining the role of TSPO/PBR, we also describe the value of Tspo knockout mice in determining the selectivity, specificity and presence of any off-target effects of TSPO/PBR ligands.
MeSH term(s)	Animals ; Bridged Bicyclo Compounds, Heterocyclic/pharmacology ; Bridged Bicyclo Compounds, Heterocyclic/therapeutic use ; Disease Models, Animal ; Energy Metabolism ; Inflammation/drug therapy ; Inflammation/genetics ; Inflammation/metabolism ; Isoquinolines/pharmacology ; Isoquinolines/therapeutic use ; Mice ; Mice, Knockout ; Mitochondria/metabolism ; Mitochondrial Membrane Transport Proteins/metabolism ; Mutation ; Neurodegenerative Diseases/drug therapy ; Neurodegenerative Diseases/genetics ; Neurodegenerative Diseases/metabolism ; Receptors, GABA/genetics ; Receptors, GABA/metabolism ; Steroids/biosynthesis
Chemical Substances	6-chloro-2-(4'-iodophenyl)-3-(N,N-diethyl)imidazo(1,2-a)pyridine-3-acetamide ; Bridged Bicyclo Compounds, Heterocyclic ; Bzrp protein, mouse ; Isoquinolines ; Mitochondrial Membrane Transport Proteins ; Receptors, GABA ; Steroids ; mitochondrial permeability transition pore ; PK 11195 (YNF83VN1RL)
Language	English
Publishing date	2015-08
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	184237-7
ISSN	1470-8752 ; 0300-5127
ISSN (online)	1470-8752
ISSN	0300-5127
DOI	10.1042/BST20150039
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Article ; Online: The water chemistry and microbiome of household wells in Medawachchiya, Sri Lanka, an area with high prevalence of chronic kidney disease of unknown origin (CKDu).

McDonough, Liza K / Meredith, Karina T / Nikagolla, Chandima / Middleton, Ryan J / Tan, Jian K / Ranasinghe, Asanga V / Sierro, Frederic / Banati, Richard B

Scientific reports

2020 Volume 10, Issue 1, Page(s) 18295

Abstract: Chronic kidney disease (CKD) of unknown etiology (CKDu) mostly affects agricultural communities in Central America, South Asia, Africa, but likely also in North America and Australia. One such area with increased CKDu prevalence is the Medawachchiya ... ...

Abstract	Chronic kidney disease (CKD) of unknown etiology (CKDu) mostly affects agricultural communities in Central America, South Asia, Africa, but likely also in North America and Australia. One such area with increased CKDu prevalence is the Medawachchiya District Secretariat Division of the Anuradhapura District in the North Central Province of Sri Lanka. Recent research has focused on the presence of various microbial pathogens in drinking water as potential causal or contributing factors to CKDu, yet no study to date has performed a more comprehensive microbial and water chemistry assessment of household wells used for domestic water supply in areas of high CKDu prevalence. In this study, we describe the chemical composition and total microbial content in 30 domestic household wells in the Medawachchiya District Secretariat Division. While the chemical composition in the tested wells mostly lies within standard drinking water limits, except for high levels of fluoride (F), magnesium (Mg), sodium (Na), chloride (Cl) and calcium (Ca) in some samples, we find a frequent presence of cyanotoxin-producing Microcystis, confirming earlier studies in Sri Lanka. Since the total microbial content of drinking water also directly influences the composition of the human gut microbiome, it can be considered an important determinant of health. Several bacterial phyla were previously reported in the gut microbiome of patients with CKD. Using these bacteria phyla to define operational taxonomic units, we found that these bacteria also occur in the microbiome of the sampled well water. Based on available environmental data, our study demonstrates associations between the abundances of these bacteria with geographical distribution, well water temperature and likely fertilizer use in the local surface water catchment area of the individual household wells. Our results reinforce the recommendation that household wells with stagnant or infrequently used water should be purged prior to use for drinking water, bathing and irrigation. The latter is suggested because of the reported potential accumulation of bacterial toxins by agricultural crops. The observation that bacteria previously found in chronic kidney disease patients are also present in household wells requires a more detailed systematic study of both the human gut and drinking water microbiomes in CKDu patients, in relation to disease prevalence and progression.
MeSH term(s)	Bacteria/classification ; Bacteria/isolation & purification ; Disease Progression ; Drinking Water/analysis ; Drinking Water/chemistry ; Drinking Water/microbiology ; Gastrointestinal Microbiome ; Humans ; Phylogeny ; Prevalence ; Renal Insufficiency, Chronic/epidemiology ; Renal Insufficiency, Chronic/etiology ; Sri Lanka/epidemiology ; Water Microbiology ; Water Pollutants, Chemical/analysis ; Water Wells
Chemical Substances	Drinking Water ; Water Pollutants, Chemical
Language	English
Publishing date	2020-10-26
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-020-75336-7
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Article ; Online: Cellular Sources and Regional Variations in the Expression of the Neuroinflammatory Marker Translocator Protein (TSPO) in the Normal Brain.

Betlazar, Calina / Harrison-Brown, Meredith / Middleton, Ryan J / Banati, Richard / Liu, Guo-Jun

International journal of molecular sciences

2018 Volume 19, Issue 9

Abstract: The inducible expression of the mitochondrial translocator protein 18 kDa (TSPO) by activated microglia is a prominent, regular feature of acute and chronic-progressive brain pathology. This expression is also the rationale for the continual development ... ...

Abstract	The inducible expression of the mitochondrial translocator protein 18 kDa (TSPO) by activated microglia is a prominent, regular feature of acute and chronic-progressive brain pathology. This expression is also the rationale for the continual development of new TSPO binding molecules for the diagnosis of "neuroinflammation" by molecular imaging. However, there is in the normal brain an ill-defined, low-level constitutive expression of TSPO. Taking advantage of healthy TSPO knockout mouse brain tissue to validate TSPO antibody specificity, this study uses immunohistochemistry to determine the regional distribution and cellular sources of TSPO in the normal mouse brain. Fluorescence microscopy revealed punctate TSPO immunostaining in vascular endothelial cells throughout the brain. In the olfactory nerve layers and glomeruli of the olfactory bulb, choroid plexus and ependymal layers, we confirm constitutive TSPO expression levels similar to peripheral organs, while some low TSPO expression is present in regions of known neurogenesis, as well as cerebellar Purkinje cells. The distributed-sparse expression of TSPO in endothelial mitochondria throughout the normal brain can be expected to give rise to a low baseline signal in TSPO molecular imaging studies. Finally, our study emphasises the need for valid and methodologically robust verification of the selectivity of TSPO ligands through the use of TSPO knockout tissues.
MeSH term(s)	Animals ; Brain/cytology ; Brain/ultrastructure ; Brain Chemistry ; Immunohistochemistry/methods ; Mice, Inbred C57BL ; Mice, Knockout ; Microscopy, Fluorescence/methods ; Positron-Emission Tomography ; Receptors, GABA/analysis ; Receptors, GABA/genetics
Chemical Substances	Bzrp protein, mouse ; Receptors, GABA
Language	English
Publishing date	2018-09-11
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms19092707
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Article ; Online: The impact of high and low dose ionising radiation on the central nervous system.

Betlazar, Calina / Middleton, Ryan J / Banati, Richard B / Liu, Guo-Jun

Redox biology

2016 Volume 9, Page(s) 144–156

Abstract: Responses of the central nervous system (CNS) to stressors and injuries, such as ionising radiation, are modulated by the concomitant responses of the brains innate immune effector cells, microglia. Exposure to high doses of ionising radiation in brain ... ...

Abstract	Responses of the central nervous system (CNS) to stressors and injuries, such as ionising radiation, are modulated by the concomitant responses of the brains innate immune effector cells, microglia. Exposure to high doses of ionising radiation in brain tissue leads to the expression and release of biochemical mediators of 'neuroinflammation', such as pro-inflammatory cytokines and reactive oxygen species (ROS), leading to tissue destruction. Contrastingly, low dose ionising radiation may reduce vulnerability to subsequent exposure of ionising radiation, largely through the stimulation of adaptive responses, such as antioxidant defences. These disparate responses may be reflective of non-linear differential microglial activation at low and high doses, manifesting as an anti-inflammatory or pro-inflammatory functional state. Biomarkers of pathology in the brain, such as the mitochondrial Translocator Protein 18kDa (TSPO), have facilitated in vivo characterisation of microglial activation and 'neuroinflammation' in many pathological states of the CNS, though the exact function of TSPO in these responses remains elusive. Based on the known responsiveness of TSPO expression to a wide range of noxious stimuli, we discuss TSPO as a potential biomarker of radiation-induced effects.
Language	English
Publishing date	2016-10
Publishing country	Netherlands
Document type	Review ; Journal Article
ISSN	2213-2317
ISSN (online)	2213-2317
DOI	10.1016/j.redox.2016.08.002
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