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  1. AU="Freeman, Willard M"
  2. AU="Lussier, A M"
  3. AU="J.Castaneda, "
  4. AU="Izquierdo, Ledys"
  5. AU="Werner, F"
  6. AU="Boddington, Marie E"
  7. AU="N Siddaiah"

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  1. Artikel ; Online: Heterochronic Plasma Transfer: Experimental Design, Considerations, and Technical Challenges.

    Ansere, Victor A / Bubak, Matthew P / Miller, Benjamin F / Freeman, Willard M

    Rejuvenation research

    2023  Band 26, Heft 5, Seite(n) 171–179

    Abstract: Experimental approaches such as Heterochronic Plasma Transfer (HPT) provide insights into the aging process and help identify the factors that impact aging, with the aim of developing anti-aging therapies. HPT involves the transfer of plasma from an ... ...

    Abstract Experimental approaches such as Heterochronic Plasma Transfer (HPT) provide insights into the aging process and help identify the factors that impact aging, with the aim of developing anti-aging therapies. HPT involves the transfer of plasma from an animal of one age to an animal of a different age and highlights the effects of the systemic environment on aging. Despite its importance as an aging research tool, HPT is not without limitations and HPT experiments across various studies differ in key experimental designs considerations, presenting a challenge in obtaining comparable outcomes. In this review, we examine the caveats and experimental design considerations of HPT as a research tool. We provide insights into plasma preparation procedures, route of administration, dosing regimen, and appropriate controls to assist investigators in achieving their experimental goals.
    Mesh-Begriff(e) Animals ; Research Design ; Parabiosis ; Aging ; Rejuvenation
    Sprache Englisch
    Erscheinungsdatum 2023-09-25
    Erscheinungsland United States
    Dokumenttyp Journal Article ; Review
    ZDB-ID 2150779-X
    ISSN 1557-8577 ; 1549-1684
    ISSN (online) 1557-8577
    ISSN 1549-1684
    DOI 10.1089/rej.2023.0035
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  2. Artikel ; Online: Exercising your mind.

    Ansere, Victor A / Freeman, Willard M

    Science (New York, N.Y.)

    2020  Band 369, Heft 6500, Seite(n) 144–145

    Mesh-Begriff(e) Brain ; Cognition ; Exercise ; Neurogenesis
    Schlagwörter covid19
    Sprache Englisch
    Erscheinungsdatum 2020-07-06
    Erscheinungsland United States
    Dokumenttyp Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Comment
    ZDB-ID 128410-1
    ISSN 1095-9203 ; 0036-8075
    ISSN (online) 1095-9203
    ISSN 0036-8075
    DOI 10.1126/science.abc8830
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  3. Artikel ; Online: Age- and sex- divergent translatomic responses of the mouse retinal pigmented epithelium.

    Chucair-Elliott, Ana J / Ocañas, Sarah R / Pham, Kevin / Machalinski, Adeline / Plafker, Scott / Stout, Michael B / Elliott, Michael H / Freeman, Willard M

    Neurobiology of aging

    2024  Band 140, Seite(n) 41–59

    Abstract: Aging is the main risk factor for age-related macular degeneration (AMD), a retinal neurodegenerative disease that leads to irreversible blindness, particularly in people over 60 years old. Retinal pigmented epithelium (RPE) atrophy is an AMD hallmark. ... ...

    Abstract Aging is the main risk factor for age-related macular degeneration (AMD), a retinal neurodegenerative disease that leads to irreversible blindness, particularly in people over 60 years old. Retinal pigmented epithelium (RPE) atrophy is an AMD hallmark. Genome-wide chromatin accessibility, DNA methylation, and gene expression studies of AMD and control RPE demonstrate epigenomic/transcriptomic changes occur during AMD onset and progression. However, mechanisms by which molecular alterations of normal aging impair RPE function and contribute to AMD pathogenesis are unclear. Here, we specifically interrogate the RPE translatome with advanced age and across sexes in a novel RPE reporter mouse model. We find differential age- and sex- associated transcript expression with overrepresentation of pathways related to inflammation in the RPE. Concordant with impaired RPE function, the phenotypic changes in the aged translatome suggest that aged RPE becomes immunologically active, in both males and females, with some sex-specific signatures, which supports the need for sex representation for in vivo studies.
    Sprache Englisch
    Erscheinungsdatum 2024-05-03
    Erscheinungsland United States
    Dokumenttyp Journal Article
    ZDB-ID 604505-4
    ISSN 1558-1497 ; 0197-4580
    ISSN (online) 1558-1497
    ISSN 0197-4580
    DOI 10.1016/j.neurobiolaging.2024.04.012
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  4. Artikel: VTA dopamine neurons are hyperexcitable in 3xTg-AD mice due to casein kinase 2-dependent SK channel dysfunction.

    Blankenship, Harris E / Carter, Kelsey A / Cassidy, Nina T / Markiewicz, Andrea N / Thellmann, Michael I / Sharpe, Amanda L / Freeman, Willard M / Beckstead, Michael J

    bioRxiv : the preprint server for biology

    2023  

    Abstract: Alzheimer's disease (AD) patients exhibit neuropsychiatric symptoms that extend beyond classical cognitive deficits, suggesting involvement of subcortical areas. Here, we investigated the role of midbrain dopamine (DA) neurons in AD using the amyloid + ... ...

    Abstract Alzheimer's disease (AD) patients exhibit neuropsychiatric symptoms that extend beyond classical cognitive deficits, suggesting involvement of subcortical areas. Here, we investigated the role of midbrain dopamine (DA) neurons in AD using the amyloid + tau-driven 3xTg-AD mouse model. We found deficits in reward-based operant learning in AD mice, suggesting possible VTA DA neuron dysregulation. Physiological assessment revealed hyperexcitability and disrupted firing in DA neurons caused by reduced activity of small-conductance calcium-activated potassium (SK) channels. RNA sequencing from contents of single patch-clamped DA neurons (Patch-seq) identified up-regulation of the SK channel modulator casein kinase 2 (CK2). Pharmacological inhibition of CK2 restored SK channel activity and normal firing patterns in 3xTg-AD mice. These findings shed light on a complex interplay between neuropsychiatric symptoms and subcortical circuits in AD, paving the way for novel treatment strategies.
    Sprache Englisch
    Erscheinungsdatum 2023-11-17
    Erscheinungsland United States
    Dokumenttyp Preprint
    DOI 10.1101/2023.11.16.567486
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  5. Artikel: Differential usage of DNA modifications in neurons, astrocytes, and microglia.

    Tooley, Kyla B / Chucair-Elliott, Ana J / Ocañas, Sarah R / Machalinski, Adeline H / Pham, Kevin D / Stanford, David R / Freeman, Willard M

    bioRxiv : the preprint server for biology

    2023  

    Abstract: Background: Cellular identity is determined partly by cell type-specific epigenomic profiles that regulate gene expression. In neuroscience, there is a pressing need to isolate and characterize the epigenomes of specific CNS cell types in health and ... ...

    Abstract Background: Cellular identity is determined partly by cell type-specific epigenomic profiles that regulate gene expression. In neuroscience, there is a pressing need to isolate and characterize the epigenomes of specific CNS cell types in health and disease. This is especially true as for DNA modifications where most data are derived from bisulfite sequencing that cannot differentiate between DNA methylation and hydroxymethylation. In this study, we developed an
    Results: After validating the cell-specificity of the Camk2a-NuTRAP model, we performed TRAP-RNA-Seq and INTACT whole genome oxidative bisulfite sequencing to assess the neuronal translatome and epigenome in the hippocampus of young mice (3 months old). These data were then compared to microglial and astrocytic data from NuTRAP models. When comparing the different cell types, microglia had the highest global mCG levels followed by astrocytes and then neurons, with the opposite pattern observed for hmCG and mCH. Differentially modified regions between cell types were predominantly found within gene bodies and distal intergenic regions, with limited differences occurring within proximal promoters. Across cell types there was a negative correlation between DNA modifications (mCG, mCH, hmCG) and gene expression at proximal promoters. In contrast, a negative correlation of mCG with gene expression within the gene body while a positive relationship between distal promoter and gene body hmCG and gene expression was observed. Furthermore, we identified a neuron-specific inverse relationship between mCH and gene expression across promoter and gene body regions.
    Conclusions: In this study, we identified differential usage of DNA modifications across CNS cell types, and assessed the relationship between DNA modifications and gene expression in neurons and glia. Despite having different global levels, the general modification-gene expression relationship was conserved across cell types. The enrichment of differential modifications in gene bodies and distal regulatory elements, but not proximal promoters, across cell types highlights epigenomic patterning in these regions as potentially greater determinants of cell identity.
    Sprache Englisch
    Erscheinungsdatum 2023-06-07
    Erscheinungsland United States
    Dokumenttyp Preprint
    DOI 10.1101/2023.06.05.543497
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  6. Artikel ; Online: Chromosomal and gonadal factors regulate microglial sex effects in the aging brain.

    Ocañas, Sarah R / Ansere, Victor A / Kellogg, Collyn M / Isola, Jose V V / Chucair-Elliott, Ana J / Freeman, Willard M

    Brain research bulletin

    2023  Band 195, Seite(n) 157–171

    Abstract: Biological sex contributes to phenotypic sex effects through genetic (sex chromosomal) and hormonal (gonadal) mechanisms. There are profound sex differences in the prevalence and progression of age-related brain diseases, including neurodegenerative ... ...

    Abstract Biological sex contributes to phenotypic sex effects through genetic (sex chromosomal) and hormonal (gonadal) mechanisms. There are profound sex differences in the prevalence and progression of age-related brain diseases, including neurodegenerative diseases. Inflammation of neural tissue is one of the most consistent age-related phenotypes seen with healthy aging and disease. The pro-inflammatory environment of the aging brain has primarily been attributed to microglial reactivity and adoption of heterogeneous reactive states dependent upon intrinsic (i.e., sex) and extrinsic (i.e., age, disease state) factors. Here, we review sex effects in microglia across the lifespan, explore potential genetic and hormonal molecular mechanisms of microglial sex effects, and discuss currently available models and methods to study sex effects in the aging brain. Despite recent attention to this area, significant further research is needed to mechanistically understand the regulation of microglial sex effects across the lifespan, which may open new avenues for sex informed prevention and treatment strategies.
    Mesh-Begriff(e) Male ; Female ; Humans ; Microglia/physiology ; Brain ; Brain Diseases ; Inflammation
    Sprache Englisch
    Erscheinungsdatum 2023-02-15
    Erscheinungsland United States
    Dokumenttyp Journal Article ; Review ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
    ZDB-ID 197620-5
    ISSN 1873-2747 ; 0361-9230
    ISSN (online) 1873-2747
    ISSN 0361-9230
    DOI 10.1016/j.brainresbull.2023.02.008
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  7. Artikel ; Online: Isolation of Neuronal Synaptic Membranes by Sucrose Gradient Centrifugation.

    Hopiavuori, Blake R / Masser, Dustin R / Wilkerson, Joseph L / Brush, Richard S / Mandal, Nawajes A / Anderson, Robert E / Freeman, Willard M

    Methods in molecular biology (Clifton, N.J.)

    2023  Band 2625, Seite(n) 7–15

    Abstract: Sucrose gradient centrifugation is a very useful technique for isolating specific membrane types based on their size and density. This is especially useful for detecting fatty acids and lipid molecules that are targeted to specialized membranes. Without ... ...

    Abstract Sucrose gradient centrifugation is a very useful technique for isolating specific membrane types based on their size and density. This is especially useful for detecting fatty acids and lipid molecules that are targeted to specialized membranes. Without fractionation, these types of molecules could be below the levels of detection after being diluted out by the more abundant lipid molecules with a more ubiquitous distribution throughout the various cell membranes. Isolation of specific membrane types where these lipids are concentrated allows for their detection and analysis. We describe herein our synaptic membrane isolation protocol that produces excellent yield and clear resolution of five major membrane fractions from a starting neural tissue homogenate: P1 (nuclear), P2 (cytoskeletal), P3 (neurosynaptosomal), PSD (post-synaptic densities), and SV (synaptic vesicle).
    Mesh-Begriff(e) Synaptic Membranes/metabolism ; Sucrose/metabolism ; Centrifugation, Density Gradient/methods ; Cell Membrane ; Centrifugation ; Lipids ; Cell Fractionation/methods
    Chemische Substanzen Sucrose (57-50-1) ; Lipids
    Sprache Englisch
    Erscheinungsdatum 2023-01-18
    Erscheinungsland United States
    Dokumenttyp Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ISSN 1940-6029
    ISSN (online) 1940-6029
    DOI 10.1007/978-1-0716-2966-6_2
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  8. Artikel ; Online: Characterization of novel mouse models to study the role of necroptosis in aging and age-related diseases.

    Selvarani, Ramasamy / Van Michelle Nguyen, Hoang / Thadathil, Nidheesh / Wolf, Roman F / Freeman, Willard M / Wiley, Christopher D / Deepa, Sathyaseelan S / Richardson, Arlan

    GeroScience

    2023  Band 45, Heft 6, Seite(n) 3241–3256

    Abstract: To study the impact of necroptosis-induced chronic inflammation on age-related diseases and aging, two knockin mouse models (Ripk3-KI and Mlkl-KI) were generated that overexpress two genes involved in necroptosis (Ripk3 or Mlkl) when crossed to Cre ... ...

    Abstract To study the impact of necroptosis-induced chronic inflammation on age-related diseases and aging, two knockin mouse models (Ripk3-KI and Mlkl-KI) were generated that overexpress two genes involved in necroptosis (Ripk3 or Mlkl) when crossed to Cre transgenic mice. Crossing Ripk3-KI or Mlkl-KI mice to albumin-Cre transgenic mice produced hepatocyte specific hRipk3-KI or hMlkl-KI mice, which express the two transgenes only in the liver. Ripk3 and Mlkl proteins were overexpressed 10- and fourfold, respectively, in the livers of the hRipk3-KI or hMlkl-KI mice. Treating young (2-month) hRipk3-KI or hMlkl-KI mice with carbon tetrachloride (CCl
    Mesh-Begriff(e) Mice ; Animals ; Protein Kinases/genetics ; Necroptosis ; Inflammation ; Aging ; Mice, Transgenic
    Chemische Substanzen Protein Kinases (EC 2.7.-)
    Sprache Englisch
    Erscheinungsdatum 2023-10-04
    Erscheinungsland Switzerland
    Dokumenttyp Journal Article
    ZDB-ID 2886586-8
    ISSN 2509-2723 ; 2509-2715
    ISSN (online) 2509-2723
    ISSN 2509-2715
    DOI 10.1007/s11357-023-00955-7
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  9. Artikel ; Online: A single-cell atlas of the aging mouse ovary.

    Isola, José V V / Ocañas, Sarah R / Hubbart, Chase R / Ko, Sunghwan / Mondal, Samim Ali / Hense, Jessica D / Carter, Hannah N C / Schneider, Augusto / Kovats, Susan / Alberola-Ila, José / Freeman, Willard M / Stout, Michael B

    Nature aging

    2024  Band 4, Heft 1, Seite(n) 145–162

    Abstract: Ovarian aging leads to diminished fertility, dysregulated endocrine signaling and increased chronic disease burden. These effects begin to emerge long before follicular exhaustion. Female humans experience a sharp decline in fertility around 35 years of ... ...

    Abstract Ovarian aging leads to diminished fertility, dysregulated endocrine signaling and increased chronic disease burden. These effects begin to emerge long before follicular exhaustion. Female humans experience a sharp decline in fertility around 35 years of age, which corresponds to declines in oocyte quality. Despite a growing body of work, the field lacks a comprehensive cellular map of the transcriptomic changes in the aging mouse ovary to identify early drivers of ovarian decline. To fill this gap we performed single-cell RNA sequencing on ovarian tissue from young (3-month-old) and reproductively aged (9-month-old) mice. Our analysis revealed a doubling of immune cells in the aged ovary, with lymphocyte proportions increasing the most, which was confirmed by flow cytometry. We also found an age-related downregulation of collagenase pathways in stromal fibroblasts, which corresponds to rises in ovarian fibrosis. Follicular cells displayed stress-response, immunogenic and fibrotic signaling pathway inductions with aging. This report provides critical insights into mechanisms responsible for ovarian aging phenotypes. The data can be explored interactively via a Shiny-based web application.
    Mesh-Begriff(e) Humans ; Female ; Mice ; Animals ; Ovary/metabolism ; Aging/genetics ; Oocytes/metabolism ; Fertility/genetics ; Signal Transduction
    Sprache Englisch
    Erscheinungsdatum 2024-01-10
    Erscheinungsland United States
    Dokumenttyp Journal Article
    ISSN 2662-8465
    ISSN (online) 2662-8465
    DOI 10.1038/s43587-023-00552-5
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  10. Artikel ; Online: Minimizing the

    Ocañas, Sarah R / Pham, Kevin D / Blankenship, Harris E / Machalinski, Adeline H / Chucair-Elliott, Ana J / Freeman, Willard M

    eNeuro

    2022  Band 9, Heft 2

    Abstract: Modern molecular and biochemical neuroscience studies require analysis of specific cellular populations derived from brain tissue samples to disambiguate cell type-specific events. This is particularly true in the analysis of minority glial populations ... ...

    Abstract Modern molecular and biochemical neuroscience studies require analysis of specific cellular populations derived from brain tissue samples to disambiguate cell type-specific events. This is particularly true in the analysis of minority glial populations in the brain, such as microglia, which may be obscured in whole tissue analyses. Microglia have central functions in development, aging, and neurodegeneration and are a current focus of neuroscience research. A long-standing concern for glial biologists using
    Mesh-Begriff(e) Animals ; Cell Separation/methods ; Flow Cytometry/methods ; Mice ; Microglia/physiology ; Neuroglia ; Transcriptome
    Sprache Englisch
    Erscheinungsdatum 2022-03-28
    Erscheinungsland United States
    Dokumenttyp Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S.
    ZDB-ID 2800598-3
    ISSN 2373-2822 ; 2373-2822
    ISSN (online) 2373-2822
    ISSN 2373-2822
    DOI 10.1523/ENEURO.0348-21.2022
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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