Novel method for collecting hippocampal interstitial fluid extracellular vesicles (EV<sup>ISF</sup>) reveals sex‐dependent changes in microglial EV proteome in response to Aβ pathology

Pait, Morgan C.; Kaye, Sarah D.; Su, Yixin; Kumar, Ashish; Singh, Sangeeta; Gironda, Stephen C.; Vincent, Samantha; Anwar, Maria; Carroll, Caitlin M.; Snipes, James Andy; Lee, Jingyun; Furdui, Cristina M.; Deep, Gagan; Macauley, Shannon L.

doi:10.1002/jev2.12398

Cited by 6 publications

(1 citation statement)

References 135 publications

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“…Images were imported into Fiji (ImageJ). Image analysis included counting IBA 1 + microglia and brain macrophages with healthy and dystrophy-related phenotypes (Shahidehpour et al, 2021), and collecting CD 68 area fraction (Pait et al, 2024).…”

Section: Image Acquisition and Analysismentioning

confidence: 99%

Early life psychosocial stress increases binge-like ethanol consumption and CSF1R inhibition prevents stress-induced alterations in microglia and brain macrophage population density

Gironda,

Centanni,

Weiner

2024

Preprint

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As key regulators of brain homeostasis, microglia and brain macrophages are vital to development. However, early life stress (ELS) experiences, like bullying, can have lasting consequences on microglial function. During ELS, microglia and brain macrophages alter their engagement with cells and synapses. These alterations can result in lasting adaptations in circuit function that could lead to an increased risk to develop maladaptive drinking behaviors and alcohol use disorder (AUD) in adulthood. Whether microglia and brain macrophages truly mediate the relationship between ELS and AUD remains elusive. Here, we report: 1) a novel early life psychosocial stress exposure, PSS, increases binge-like ethanol consumption in early adulthood. 2) This increase in ethanol drinking is associated with alterations in microglia and brain macrophage populations in the ventral hippocampus and across other brain regions. 3) Inhibiting microglia and brain macrophages using a CSF1-r inhibitor, GW2580, results in a trend towards a reduction in binge-like ethanol consumption and normalizes microglia and brain macrophage populations. Therefore, our study suggests that acutely inhibiting microglia and brain macrophage function during periods of early life psychosocial stress may help reduce the risk to develop risky drinking behaviors and AUD.HighlightsAn early life psychosocial stress (PSS) exposure increases ethanol consumptionPSS and ethanol consumption alter microglia function in the ventral hippocampusPSS and ethanol consumption have further consequences in other brain regionsMicroglial inhibition during PSS trends towards reducing ethanol consumptionMicroglial inhibition prevents microglia population changes in some brain regions

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Section: Image Acquisition and Analysismentioning

confidence: 99%

Early life psychosocial stress increases binge-like ethanol consumption and CSF1R inhibition prevents stress-induced alterations in microglia and brain macrophage population density

Gironda,

Centanni,

Weiner

2024

Preprint

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Exploring microtubule dynamics in Alzheimer's disease: Longitudinal assessment using [¹¹C]MPC‐6827 PET imaging in rodent models of Alzheimer's‐related pathology

Damuka,

Irmen,

Krizan

et al. 2024

Alzheimer's & Dementia

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INTRODUCTIONMicrotubule (MT) stability is crucial for proper neuronal function. Understanding MT dysregulation is critical for connecting amyloid beta (Aβ) and tau‐based degenerative events and early changes in presymptomatic Alzheimer's disease (AD). Herein we present positron emission tomography (PET) imaging properties of our MT‐PET radiotracer, [11C]MPC‐6827, in multiple established AD mouse models.METHODSLongitudinal PET, biodistribution, autoradiography, immunohistochemistry, and behavioral studies were conducted at multiple time points in APPswe/PSEN1dE9 (APP/PS1), P301S‐PS19 (P301S), 5xFAD, and age‐matched control mice.RESULTSLongitudinal [11C]MPC‐6827 brain imaging showed significant increases in APP/PS1, P301S, and 5xFAD mice compared to controls. Longitudinal MT‐PET correlated positively with biodistribution, autoradiography, and immunohistochemistry results and negatively with behavior data.DISCUSSIONOur study demonstrated significant longitudinal [11C]MPC‐6827 PET increases in multiple AD mouse models for the first time. Strong correlations between PET and biomarker data underscored the interplay of MT destabilization, amyloid, and tau pathology in AD. These results suggest [11C]MPC‐6827 PET as a promising tool for monitoring MT dysregulation early in AD progression.Highlights Longitudinal positron emission tomography (PET) imaging studies using [11C]MPC‐6827 in multiple established Alzheimer's disease (AD) mouse models revealed an early onset of microtubule dysregulation, with significant changes in brain radiotracer uptake evident from 2 to 4 months of age. Intra‐group analysis showed a progressive increase in microtubule dysregulation with increasing AD burden, supported by significant correlations between PET imaging data and biodistribution, autoradiography, and molecular pathological markers. [11C]MPC‐6827 PET imaging demonstrated its efficacy in detecting early microtubule alterations preceding observable behavioral changes in AD mouse models, suggesting its potential for early AD imaging. The inclusion of the 5xFAD mouse model further elucidated the impact of amyloid beta (Aβ) toxicity on inducing tau hyperphosphorylation‐mediated microtubule dysregulation, highlighting the versatility of [11C]MPC‐6827 in delineating various aspects of AD pathology. Our study provides immediate clarity on high uptake of the microtubule‐based radiotracer in AD brains in a longitudinal setting, which directly informs clinical utility in Aβ/tau‐based studies.

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Efficient enzyme‐free isolation of brain‐derived extracellular vesicles

Matamoros‐Angles,

Karadjuzovic,

Mohammadi

et al. 2024

J of Extracellular Vesicle

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Extracellular vesicles (EVs) have gained significant attention as pathology mediators and potential diagnostic tools for neurodegenerative diseases. However, isolation of brain‐derived EVs (BDEVs) from tissue remains challenging, often involving enzymatic digestion steps that may compromise the integrity of EV proteins and overall functionality. Here, we describe that collagenase digestion, commonly used for BDEV isolation, produces undesired protein cleavage of EV‐associated proteins in brain tissue homogenates and cell‐derived EVs. In order to avoid this effect, we studied the possibility of isolating BDEVs with a reduced amount of collagenase or without any protease. Characterization of the isolated BDEVs from mouse and human samples (both female and male) revealed their characteristic morphology and size distribution with both approaches. However, we show that even minor enzymatic digestion induces ‘artificial’ proteolytic processing in key BDEV markers, such as Flotillin‐1, CD81, and the cellular prion protein (PrPC), whereas avoiding enzymatic treatment completely preserves their integrity. We found no major differences in mRNA and protein content between non‐enzymatically and enzymatically isolated BDEVs, suggesting that the same BDEV populations are purified with both approaches. Intriguingly, the lack of Golgi marker GM130 signal, often referred to as contamination indicator (or negative marker) in EV preparations, seems to result from enzymatic digestion rather than from its actual absence in BDEV samples. Overall, we show that non‐enzymatic isolation of EVs from brain tissue is possible and avoids artificial pruning of proteins while achieving an overall high BDEV yield and purity. This protocol will help to understand the functions of BDEV and their associated proteins in a near‐physiological setting, thus opening new research approaches.

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Novel method for collecting hippocampal interstitial fluid extracellular vesicles (EV^ISF) reveals sex‐dependent changes in microglial EV proteome in response to Aβ pathology

Cited by 6 publications

References 135 publications

Early life psychosocial stress increases binge-like ethanol consumption and CSF1R inhibition prevents stress-induced alterations in microglia and brain macrophage population density

Early life psychosocial stress increases binge-like ethanol consumption and CSF1R inhibition prevents stress-induced alterations in microglia and brain macrophage population density

Exploring microtubule dynamics in Alzheimer's disease: Longitudinal assessment using [¹¹C]MPC‐6827 PET imaging in rodent models of Alzheimer's‐related pathology

Efficient enzyme‐free isolation of brain‐derived extracellular vesicles

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Novel method for collecting hippocampal interstitial fluid extracellular vesicles (EVISF) reveals sex‐dependent changes in microglial EV proteome in response to Aβ pathology

Cited by 6 publications

References 135 publications

Early life psychosocial stress increases binge-like ethanol consumption and CSF1R inhibition prevents stress-induced alterations in microglia and brain macrophage population density

Early life psychosocial stress increases binge-like ethanol consumption and CSF1R inhibition prevents stress-induced alterations in microglia and brain macrophage population density

Exploring microtubule dynamics in Alzheimer's disease: Longitudinal assessment using [11C]MPC‐6827 PET imaging in rodent models of Alzheimer's‐related pathology

Efficient enzyme‐free isolation of brain‐derived extracellular vesicles

Contact Info

Product

Resources

About

Novel method for collecting hippocampal interstitial fluid extracellular vesicles (EV^ISF) reveals sex‐dependent changes in microglial EV proteome in response to Aβ pathology

Exploring microtubule dynamics in Alzheimer's disease: Longitudinal assessment using [¹¹C]MPC‐6827 PET imaging in rodent models of Alzheimer's‐related pathology