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Maechler, P.

Publications and source records attributed to Maechler, P..

2 recordsLinked to original sources

Third Harmonic Generation Microscopy Reveals Structure and Mucus Dynamics in Human Airway Epithelium Models

Airway epithelium plays a major role as the primary interface between human body and the external environment, acting both as a physical and functional barrier. In vitro airway models that reproduce the epithelium architecture are therefore a valuable tool for studying infection, inflammation, and transport processes. In this work, we present a label-free, non-invasive method to visualize and measure mucociliary transport in air-liquid human models using third-harmonic generation (THG) microscopy with an optical parametric amplifier laser source at 1300 nm. By exploiting the intrinsic nonlinear contrast at optical heterogeneities, THG provides high-resolution images of both epithelial structures and of the overlying mucus layer without the need for fluorescence staining or sample processing. Time-lapse THG imaging reveals depth-dependent transport dynamics within the mucus, offering new insights into mucociliary transport mechanism. Our approach offers a physiologically relevant way to assess mucociliary function in vitro and could support studies on respiratory diseases, drug delivery and efficacy, and epithelial remodeling. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=117 SRC="FIGDIR/small/717621v1_ufig1.gif" ALT="Figure 1"> View larger version (52K): org.highwire.dtl.DTLVardef@62e8acorg.highwire.dtl.DTLVardef@199a8b7org.highwire.dtl.DTLVardef@113bb84org.highwire.dtl.DTLVardef@7be3f8_HPS_FORMAT_FIGEXP M_FIG For Table of Contents Only C_FIG

biophysics↗

GLUD1 dictates muscle stem cell differentiation by controlling mitochondrial glutamate levels

Muscle stem cells (MuSCs) enable muscle growth and regeneration after exercise or injury. Upon activation MuSCs metabolically rewire to meet the changing demands of proliferation. Here we describe that primary changes in metabolism itself can dictate MuSC fate decisions to control differentiation and fusion. We found that glutamine anaplerosis into the TCA cycle decreases during MuSC differentiation and coincides with decreased expression of the mitochondrial glutamate deaminase GLUD1. Genetic deletion of Glud1 in proliferating MuSCs resulted in precocious differentiation and imbalanced fusion combined with loss of self-renewal in vitro and in vivo. Mechanistically, deleting Glud1 caused mitochondrial glutamate accumulation in proliferating MuSCs and inhibited the malate-aspartate shuttle (MAS). Restoring MAS activity by supplementation of alanine normalized differentiation. In conclusion, high GLUD1 activity in proliferating MuSCs prevents deleterious mitochondrial glutamate accumulation and inactivation of the MAS. It thereby acts as a compartment specific metabolic brake on MuSC differentiation. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=194 SRC="FIGDIR/small/560525v1_ufig1.gif" ALT="Figure 1"> View larger version (49K): org.highwire.dtl.DTLVardef@e3585dorg.highwire.dtl.DTLVardef@1be4a54org.highwire.dtl.DTLVardef@1f077org.highwire.dtl.DTLVardef@15220f1_HPS_FORMAT_FIGEXP M_FIG C_FIG HighlightsO_LIGlutamine is the major TCA cycle substrate in MuSCs with decreasing contribution upon differentiation. C_LIO_LILoss of Glud1 impairs MuSC self-renewal capacity and causes imbalanced fusion in vitro and in vivo. C_LIO_LIGlud1 deletion leads to mitochondrial glutamate trapping and malate-aspartate shuttle (MAS) dysfunction. C_LIO_LIRestoration of MAS activity in Glud1 deficient MuSCs reverses precocious differentiation and imbalanced fusion. C_LI

cell biology↗