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  1. Article ; Online: Comparative evaluation of glomerular morphometric techniques reveals differential technical artifacts between focal segmental glomerulosclerosis and normal glomeruli.

    Reghuvaran, Anand C / Lin, Qisheng / Basgen, John M / Banu, Khadija / Shi, Hongmei / Vashist, Anushree / Pell, John / Perinchery, Sudhir / He, John C / Moledina, Dennis / Wilson, F Perry / Menon, Madhav C

    Physiological reports

    2023  Volume 11, Issue 13, Page(s) e15688

    Abstract: Morphometric estimates of mean or individual glomerular volume (MGV, IGV) have biological implications, over and above qualitative histologic data. However, morphometry is time-consuming and requires expertise limiting its utility in clinical cases. We ... ...

    Abstract Morphometric estimates of mean or individual glomerular volume (MGV, IGV) have biological implications, over and above qualitative histologic data. However, morphometry is time-consuming and requires expertise limiting its utility in clinical cases. We evaluated MGV and IGV using plastic- and paraffin-embedded tissue from 10 control and 10 focal segmental glomerulosclerosis (FSGS) mice (aging and 5/6th nephrectomy models) using the gold standard Cavalieri (Cav) method versus the 2-profile and Weibel-Gomez (WG) methods and a novel 3-profile method. We compared accuracy, bias and precision, and quantified results obtained when sampling differing numbers of glomeruli. In both FSGS and controls, we identified an acceptable precision for MGV of 10-glomerular sampling versus 20-glomerular sampling using the Cav method, while 5-glomerular sampling was less precise. In plastic tissue, 2- or 3-profile MGVs showed greater concordance with MGV when using Cav, versus MGV with WG. IGV comparisons using the same glomeruli reported a consistent underestimation bias with both 2- or 3-profile methods versus the Cav method. FSGS glomeruli showed wider variations in bias estimation than controls. Our 3-profile method offered incremental benefit to the 2-profile method in both IGV and MGV estimation (improved correlation coefficient, Lin's concordance and reduced bias). In our control animals, we quantified a shrinkage artifact of 52% from tissue processed for paraffin-embedded versus plastic-embedded tissue. FSGS glomeruli showed overall reduced shrinkage albeit with variable artifact signifying periglomerular/glomerular fibrosis. A novel 3-profile method offers slightly improved concordance with reduced bias versus 2-profile. Our findings have implications for future studies using glomerular morphometry.
    MeSH term(s) Animals ; Mice ; Glomerulosclerosis, Focal Segmental ; Artifacts ; Kidney Glomerulus/pathology ; Kidney Diseases/pathology ; Nephrectomy
    Language English
    Publishing date 2023-06-13
    Publishing country United States
    Document type Journal Article ; Research Support, U.S. Gov't, P.H.S. ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 2724325-4
    ISSN 2051-817X ; 2051-817X
    ISSN (online) 2051-817X
    ISSN 2051-817X
    DOI 10.14814/phy2.15688
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: AMPK mediates regulation of glomerular volume and podocyte survival.

    Banu, Khadija / Lin, Qisheng / Basgen, John M / Planoutene, Marina / Wei, Chengguo / Reghuvaran, Anand C / Tian, Xuefei / Shi, Hongmei / Garzon, Felipe / Garzia, Aitor / Chun, Nicholas / Cumpelik, Arun / Santeusanio, Andrew D / Zhang, Weijia / Das, Bhaskar / Salem, Fadi / Li, Li / Ishibe, Shuta / Cantley, Lloyd G /
    Kaufman, Lewis / Lemley, Kevin V / Ni, Zhaohui / He, John Cijiang / Murphy, Barbara / Menon, Madhav C

    JCI insight

    2021  Volume 6, Issue 19

    Abstract: Herein, we report that Shroom3 knockdown, via Fyn inhibition, induced albuminuria with foot process effacement (FPE) without focal segmental glomerulosclerosis (FSGS) or podocytopenia. Interestingly, knockdown mice had reduced podocyte volumes. Human ... ...

    Abstract Herein, we report that Shroom3 knockdown, via Fyn inhibition, induced albuminuria with foot process effacement (FPE) without focal segmental glomerulosclerosis (FSGS) or podocytopenia. Interestingly, knockdown mice had reduced podocyte volumes. Human minimal change disease (MCD), where podocyte Fyn inactivation was reported, also showed lower glomerular volumes than FSGS. We hypothesized that lower glomerular volume prevented the progression to podocytopenia. To test this hypothesis, we utilized unilateral and 5/6th nephrectomy models in Shroom3-KD mice. Knockdown mice exhibited less glomerular and podocyte hypertrophy after nephrectomy. FYN-knockdown podocytes had similar reductions in podocyte volume, implying that Fyn was downstream of Shroom3. Using SHROOM3 or FYN knockdown, we confirmed reduced podocyte protein content, along with significantly increased phosphorylated AMPK, a negative regulator of anabolism. AMPK activation resulted from increased cytoplasmic redistribution of LKB1 in podocytes. Inhibition of AMPK abolished the reduction in glomerular volume and induced podocytopenia in mice with FPE, suggesting a protective role for AMPK activation. In agreement with this, treatment of glomerular injury models with AMPK activators restricted glomerular volume, podocytopenia, and progression to FSGS. Glomerular transcriptomes from MCD biopsies also showed significant enrichment of Fyn inactivation and Ampk activation versus FSGS glomeruli. In summary, we demonstrated the important role of AMPK in glomerular volume regulation and podocyte survival. Our data suggest that AMPK activation adaptively regulates glomerular volume to prevent podocytopenia in the context of podocyte injury.
    MeSH term(s) Adenylate Kinase/antagonists & inhibitors ; Adenylate Kinase/metabolism ; Adolescent ; Adult ; Aged ; Albuminuria/genetics ; Animals ; Cell Size ; Cell Survival/genetics ; Child ; Child, Preschool ; Female ; Gene Knockdown Techniques ; Glomerulonephritis, Membranous/genetics ; Glomerulonephritis, Membranous/pathology ; Glomerulosclerosis, Focal Segmental/genetics ; Glomerulosclerosis, Focal Segmental/pathology ; Humans ; Hypertrophy ; Infant ; Kidney Glomerulus/metabolism ; Kidney Glomerulus/pathology ; Male ; Mice ; Microfilament Proteins/genetics ; Middle Aged ; Nephrectomy ; Nephrosis, Lipoid/genetics ; Nephrosis, Lipoid/pathology ; Nephrotic Syndrome/genetics ; Nephrotic Syndrome/pathology ; Podocytes/metabolism ; Podocytes/pathology ; Proportional Hazards Models ; Proto-Oncogene Proteins c-fyn/genetics ; Young Adult
    Chemical Substances Microfilament Proteins ; Shrm protein, mouse ; FYN protein, human (EC 2.7.10.2) ; Fyn protein, mouse (EC 2.7.10.2) ; Proto-Oncogene Proteins c-fyn (EC 2.7.10.2) ; Adenylate Kinase (EC 2.7.4.3)
    Language English
    Publishing date 2021-10-08
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ISSN 2379-3708
    ISSN (online) 2379-3708
    DOI 10.1172/jci.insight.150004
    Database MEDical Literature Analysis and Retrieval System OnLINE

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