scholarly journals High resolution dynamic mapping of the C. elegans intestinal brush border

2021 ◽  
Author(s):  
Aurelien Bidaud-Meynard ◽  
Flora Demouchy ◽  
Ophelie Nicolle ◽  
Anne Pacquelet ◽  
Gregoire Michaux
Keyword(s):  
High Resolution ◽  
Brush Border ◽  
Membrane Surface ◽  
Super Resolution ◽  
C Elegans ◽  
Mapping Tool ◽  
Nutrient Processing ◽  
Intestinal Brush Border ◽  
Genetic Perturbations

The intestinal brush border is made of an array of microvilli that increases the membrane surface area for nutrient processing, absorption, and host defence. Studies on mammalian cultured epithelial cells uncovered some of the molecular players, structural components and physical constrains required to establish this apical specialized membrane. However, the building and maintenance of a brush border in vivo has not been investigated in detail yet. Here, we combined super-resolution imaging, transmission electron microscopy and genome editing in the developing nematode C. elegans to build a high-resolution and dynamic localization map of known and new markers of the brush border. Notably, we show that microvilli components are dynamically enriched at the apical membrane during microvilli outgrowth and maturation but become highly stable when microvilli are built. This new mapping tool will be instrumental to understand the molecular processes of microvilli growth and maintenance in vivo as well as the effect of genetic perturbations, notably in the context of disorders affecting the brush border integrity.

scholarly journals High resolution dynamic mapping of the C. elegans intestinal brush border

Development ◽  
2021 ◽  
Author(s):  
Aurélien Bidaud-Meynard ◽  
Flora Demouchy ◽  
Ophélie Nicolle ◽  
Anne Pacquelet ◽  
Shashi Kumar Suman ◽  
...  
Keyword(s):  
High Resolution ◽  
Brush Border ◽  
Membrane Surface ◽  
Super Resolution ◽  
C Elegans ◽  
Nutrient Processing ◽  
Intestinal Brush Border ◽  
Genetic Perturbations ◽  
Defense Studies

The intestinal brush border is made of an array of microvilli that increases the membrane surface area for nutrient processing, absorption, and host defense. Studies on mammalian cultured epithelial cells uncovered some of the molecular players and physical constrains required to establish this apical specialized membrane. However, the building and maintenance of a brush border in vivo has not been investigated in detail yet. Here, we combined super-resolution imaging, transmission electron microscopy and genome editing in the developing nematode C. elegans to build a high-resolution and dynamic localization map of known and new brush border markers. Notably, we show that microvilli components are dynamically enriched at the apical membrane during microvilli outgrowth and maturation but become highly stable once microvilli are built. This new toolbox will be instrumental to understand the molecular processes of microvilli growth and maintenance in vivo as well as the effect of genetic perturbations, notably in the context of disorders affecting brush border integrity.

scholarly journals Brush border protocadherin CDHR2 promotes the elongation and maximized packing of microvilli in vivo

2019 ◽  
Vol 30 (1) ◽  
pp. 108-118 ◽  
Author(s):  
Julia A. Pinette ◽  
Suli Mao ◽  
Bryan A. Millis ◽  
Evan S. Krystofiak ◽  
James J. Faust ◽  
...  
Keyword(s):  
Functional Capacity ◽  
Brush Border ◽  
Intestinal Tract ◽  
Membrane Surface ◽  
Careful Examination ◽  
Tissue Cell ◽  
Apical Surface ◽  
Membrane Surface Area ◽  
Solute Uptake

Transporting epithelial cells optimize their morphology for solute uptake by building an apical specialization: a dense array of microvilli that serves to increase membrane surface area. In the intestinal tract, individual cells build thousands of microvilli, which pack tightly to form the brush border. Recent studies implicate adhesion molecule CDHR2 in the regulation of microvillar packing via the formation of adhesion complexes between the tips of adjacent protrusions. To gain insight on how CDHR2 contributes to brush border morphogenesis and enterocyte function under native in vivo conditions, we generated mice lacking CDHR2 expression in the intestinal tract. Although CDHR2 knockout (KO) mice are viable, body weight trends lower and careful examination of tissue, cell, and brush border morphology revealed several perturbations that likely contribute to reduced functional capacity of KO intestine. In the absence of CDHR2, microvilli are significantly shorter, and exhibit disordered packing and a 30% decrease in packing density. These structural perturbations are linked to decreased levels of key solute processing and transporting factors in the brush border. Thus, CDHR2 functions to elongate microvilli and maximize their numbers on the apical surface, which together serve to increase the functional capacity of enterocyte.

scholarly journals Genetic engineering for in vivo optical interrogation of neuronal responses to cell type-specific silencing

2021 ◽  
Author(s):  
Firat Terzi ◽  
Johannes Knabbe ◽  
Sidney B. Cambridge
Keyword(s):  
High Resolution ◽  
Genetic Engineering ◽  
Transgene Expression ◽  
Neuronal Morphology ◽  
Dependent Manner ◽  
Cell Type ◽  
Fluorescent Marker ◽  
Genetic Perturbations ◽  
Cell Type Specific

SummaryGenetic engineering of quintuple transgenic brain tissue was used to establish a low background, Cre-dependent version of the inducible Tet-On system for fast, cell type-specific transgene expression in vivo. Co-expression of a constitutive, Cre-dependent fluorescent marker selectively allowed single cell analyses before and after inducible, tet-dependent transgene expression. Here, we used this method for acute, high-resolution manipulation of neuronal activity in the living brain. Single induction of the potassium channel Kir2.1 produced cell type-specific silencing within hours that lasted for at least three days. Longitudinal in vivo imaging of spontaneous calcium transients and neuronal morphology demonstrated that prolonged silencing did not alter spine densities or synaptic input strength. Furthermore, selective induction of Kir2.1 in parvalbumin interneurons increased the activity of surrounding neurons in a distance-dependent manner. This high-resolution, inducible interference and interval imaging of individual cells (high I5, ‘HighFive’) method thus allows visualizing temporally precise, genetic perturbations of defined cells.

scholarly journals Correction: Long-term C. elegans immobilization enables high resolution developmental studies in vivo

Lab on a Chip ◽  
2018 ◽  
Vol 18 (12) ◽  
pp. 1802-1802
Author(s):  
Simon Berger ◽  
Evelyn Lattmann ◽  
Tinri Aegerter-Wilmsen ◽  
Michael Hengartner ◽  
Alex Hajnal ◽  
...  
Keyword(s):  
High Resolution ◽  
Developmental Studies ◽  
C Elegans

Correction for ‘Long-term C. elegans immobilization enables high resolution developmental studies in vivo’ by Simon Berger et al., Lab Chip, 2018, 18, 1359–1368.

scholarly journals Long-term C. elegans immobilization enables high resolution developmental studies in vivo

Lab on a Chip ◽  
2018 ◽  
Vol 18 (9) ◽  
pp. 1359-1368 ◽  
Author(s):  
Simon Berger ◽  
Evelyn Lattmann ◽  
Tinri Aegerter-Wilmsen ◽  
Michael Hengartner ◽  
Alex Hajnal ◽  
...  
Keyword(s):  
High Resolution ◽  
Developmental Studies ◽  
Developmental Processes ◽  
C Elegans

Microfluidics enables the interference free observation of sensitive developmental processes in C. elegans.

Turnover of intestinal brush-border proteins during postnatal development in rat

1980 ◽  
Vol 239 (6) ◽  
pp. G524-G531
Author(s):  
B. Seetharam ◽  
K. Y. Yeh ◽  
D. H. Alpers
Keyword(s):  
Postnatal Development ◽  
Brush Border ◽  
Cathepsin B ◽  
Leucine Incorporation ◽  
Turnover Rates ◽  
Distal Intestine ◽  
Rapid Turnover ◽  
Intestinal Brush Border ◽  
Proximal Intestine

Protein turnover in brush-border membranes of rats during postnatal development has been studied by the double isotope technique. Unlike adult animals where only large proteins (mol wt > 140,000) show relatively rapid turnover, most brush-border proteins in 12-day-old rats show high 3H-to-14C ratios of leucine incorporation, consistent with rapid turnover. Lysosomal proteases, including cathepsin B, are partly responsible for this rapid turnover. This conclusion is based on the following findings: 1) In vivo treatment of 12-day-old animals with leupeptin, an inhibitor of cathepsin B, alters relative turnover rates, enzyme activity, and content of many brush-border proteins. Activities of maltase and trehalase rise while lactase falls. 2) Cathepsin B activity falls rapidly in intestine after the animals are 16 days of age, at a time when luminal pancreatic proteases are rising. Moreover, cathepsin B activity shows less latency in distal intestine at 12 and 16 days than at later ages or in proximal intestine. It is suggested that during postnatal development lysosomal enzymes, e.g., cathepsin B, play an important role in the turnover of intestinal brush-border proteins.

scholarly journals Plastin 1 Binds to Keratin and Is Required for Terminal Web Assembly in the Intestinal Epithelium

2009 ◽  
Vol 20 (10) ◽  
pp. 2549-2562 ◽  
Author(s):  
Eva-Maria S. Grimm-Günter ◽  
Céline Revenu ◽  
Sonia Ramos ◽  
Ilse Hurbain ◽  
Neil Smyth ◽  
...  
Keyword(s):  
Brush Border ◽  
Actin Filaments ◽  
Keratin 19 ◽  
Terminal Web ◽  
Intestinal Brush Border ◽  
Important Regulator ◽  
Increased Sensitivity ◽  
Filament Network ◽  
Deficient Mice

Plastin 1 (I-plastin, fimbrin) along with villin and espin is a prominent actin-bundling protein of the intestinal brush border microvilli. We demonstrate here that plastin 1 accumulates in the terminal web and interacts with keratin 19, possibly contributing to anchoring the rootlets to the keratin network. This prompted us to investigate the importance of plastin 1 in brush border assembly. Although in vivo neither villin nor espin is required for brush border structure, plastin 1-deficient mice have conspicuous ultrastructural alterations: microvilli are shorter and constricted at their base, and, strikingly, their core actin bundles lack true rootlets. The composition of the microvilli themselves is apparently normal, whereas that of the terminal web is profoundly altered. Although the plastin 1 knockout mice do not show any overt gross phenotype and present a normal intestinal microanatomy, the alterations result in increased fragility of the epithelium. This is seen as an increased sensitivity of the brush border to biochemical manipulations, decreased transepithelial resistance, and increased sensitivity to dextran sodium sulfate-induced colitis. Plastin 1 thus emerges as an important regulator of brush border morphology and stability through a novel role in the organization of the terminal web, possibly by connecting actin filaments to the underlying intermediate filament network.

scholarly journals Live-cell vibrational imaging of choline metabolites by stimulated Raman scattering coupled with isotope-based metabolic labeling

The Analyst ◽  
2014 ◽  
Vol 139 (10) ◽  
pp. 2312-2317 ◽  
Author(s):  
Fanghao Hu ◽  
Lu Wei ◽  
Chaogu Zheng ◽  
Yihui Shen ◽  
Wei Min
Keyword(s):  
High Resolution ◽  
Raman Scattering ◽  
Mammalian Cells ◽  
Stimulated Raman Scattering ◽  
Primary Neurons ◽  
High Resolution Imaging ◽  
Metabolic Labeling ◽  
C Elegans ◽  
Resolution Imaging

High-resolution imaging of choline metabolites in living mammalian cells, primary neurons andC. eleganshas been demonstrated with the potential forin vivodisease detection and developmental monitoring.

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