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3D ultrastructure of the kidney glomerulus capillary and microfibrils imaged using SBF-SEM
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3D Ultrastructure of Kidney Glomerulus with SBF-SEM: Understanding Marfan Syndrome

byZEISS Microscopy 6 November 2023 4 min read
Introduction

3D Ultrastructure of Kidney Glomerulus with SBF-SEM: Understanding Marfan Syndrome

3D ultrastructural observations provide possible clinical insights in a mouse model of Marfan Syndrome.

Marfan syndrome (MFS) is a genetic condition that impacts connective tissue. While this can have negative consequences throughout the body and organs, it is the cardiovascular damage in particular that can cause death from this condition. Dr. Rodrigo Barbosa is a Professor of Anatomy at Faculdade Santa Marcelina as well as a Research Assistant in the group of Professor Lygia Pereira at the University of São Paulo, Brazil. Professor Pereira established the first knockout mouse model for MFS. Working with Professor Pereira, Professor Barbosa has published work characterizing the kidney in MFS, which plays an important role in the cardiovascular system. Their findings suggest that kidney phenotype may offer clinical insights for patients with Marfan Syndrome. This work incorporates in-depth imaging studies, including volume electron microscopy using SBF-SEM with a ZEISS Sigma FE-SEM with an integrated ultramicrotome.

Professor Rodrigo Barbosa

The kidney plays a critical role in the cardiovascular system through its impact on hemodynamics. We explored the possible contribution of the Fibers of Elastic Fibers System (FEFS) in the glomerulus using ultrastructural analyses, including SBF-SEM.

Professor Rodrigo Barbosa Professor of Anatomy, Faculdade Santa Marcelina & Research Assistant, University of São Paulo, Brazil

3D Ultrastructure of the Kidney Glomerulus Capillary and Microfibrils

Imaged with SBF-SEM

  • Superior Plane

  • Higher Mag

  • Lateral Plane
  • 3D ultrastructure of the Fibers of the Elastic Fiber System (FEFS) shown in red, capillary in white, and cells in blue. Superior plane. Wild type (left) presents FEFS arranged in a tubular-shaped network (white arrow) within the capillary. MFS (right) presents fractured FEFS (black arrow), and loss of capillary structure.
    3D ultrastructure of the Fibers of the Elastic Fiber System (FEFS) shown in red, capillary in white, and cells in blue. Superior plane. Wild type (left) presents FEFS arranged in a tubular-shaped network (white arrow) within the capillary. MFS (right) presents fractured FEFS (black arrow), and loss of capillary structure.

    IMAGE 1 OF 3 (Click arrows above to navigate images): The Fibers of the Elastic Fiber System (FEFS) are shown in red, capillary in white, and cells in blue. Wild type (left) presents FEFS arranged in a tubular-shaped network (white arrow) within the capillary. MFS (right) presents fractured FEFS (black arrow), and loss of capillary structure. Image 1: Figures A and B show the superior plane. Image 2: Figures C and D show the superior plane in higher magnification. Image 3: Figures E and F show the lateral plane. Data collected with ZEISS Sigma FE-SEM with integrated ultramicrotome. Data from R. Barbosa de Souza et al.

    IMAGE 1 OF 3 (Click arrows above to navigate images): The Fibers of the Elastic Fiber System (FEFS) are shown in red, capillary in white, and cells in blue. Wild type (left) presents FEFS arranged in a tubular-shaped network (white arrow) within the capillary. MFS (right) presents fractured FEFS (black arrow), and loss of capillary structure. Image 1: Figures A and B show the superior plane. Image 2: Figures C and D show the superior plane in higher magnification. Image 3: Figures E and F show the lateral plane. Data collected with ZEISS Sigma FE-SEM with integrated ultramicrotome. Data from R. Barbosa de Souza et al.

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  • 3D ultrastructure of the Fibers of the Elastic Fiber System (FEFS) shown in red, capillary in white, and cells in blue. Superior plane in higher magnification. Wild type (left) presents FEFS arranged in a tubular-shaped network (white arrow) within the capillary. MFS (right) presents fractured FEFS (black arrow), and loss of capillary structure.
    3D ultrastructure of the Fibers of the Elastic Fiber System (FEFS) shown in red, capillary in white, and cells in blue. Superior plane in higher magnification. Wild type (left) presents FEFS arranged in a tubular-shaped network (white arrow) within the capillary. MFS (right) presents fractured FEFS (black arrow), and loss of capillary structure.

    IMAGE 2 OF 3 (Click arrows above to navigate images): The Fibers of the Elastic Fiber System (FEFS) are shown in red, capillary in white, and cells in blue. Wild type (left) presents FEFS arranged in a tubular-shaped network (white arrow) within the capillary. MFS (right) presents fractured FEFS (black arrow), and loss of capillary structure. Image 1: Figures A and B show the superior plane. Image 2: Figures C and D show the superior plane in higher magnification. Image 3: Figures E and F show the lateral plane. Data collected with ZEISS Sigma FE-SEM with integrated ultramicrotome. Data from R. Barbosa de Souza et al.

    IMAGE 2 OF 3 (Click arrows above to navigate images): The Fibers of the Elastic Fiber System (FEFS) are shown in red, capillary in white, and cells in blue. Wild type (left) presents FEFS arranged in a tubular-shaped network (white arrow) within the capillary. MFS (right) presents fractured FEFS (black arrow), and loss of capillary structure. Image 1: Figures A and B show the superior plane. Image 2: Figures C and D show the superior plane in higher magnification. Image 3: Figures E and F show the lateral plane. Data collected with ZEISS Sigma FE-SEM with integrated ultramicrotome. Data from R. Barbosa de Souza et al.

     Download image (211 KB)
  • 3D ultrastructure of the Fibers of the Elastic Fiber System (FEFS) shown in red, capillary in white, and cells in blue. Lateral plane. Wild type (left) presents FEFS arranged in a tubular-shaped network (white arrow) within the capillary. MFS (right) presents fractured FEFS (black arrow), and loss of capillary structure.

    IMAGE 3 OF 3 (Click arrows above to navigate images): The Fibers of the Elastic Fiber System (FEFS) are shown in red, capillary in white, and cells in blue. Wild type (left) presents FEFS arranged in a tubular-shaped network (white arrow) within the capillary. MFS (right) presents fractured FEFS (black arrow), and loss of capillary structure. Image 1: Figures A and B show the superior plane. Image 2: Figures C and D show the superior plane in higher magnification. Image 3: Figures E and F show the lateral plane. Data collected with ZEISS Sigma FE-SEM with integrated ultramicrotome. Data from R. Barbosa de Souza et al.

     Download image (189 KB)

SBF-SEM Reveals Differences in Kidney FEFS Networks

For ultrastructural analyses, Professor Barbosa was supported by Professor Keith Meek and Dr. Philip Lewis from Cardiff University, United Kingdom. Serial Block-Face Scanning Electron Microscopy (SBF-SEM) enabled visualization of the 3D structural organization of the FEFS, capillary walls, and cells. In particular, the ultrastructure analysis facilitated the localization of the FEFS, the fiber shape, and the relationship of the FEFS with other components. FEFS from wild type mice were arranged in tubular-shaped networks within the capillary. In comparison, the MFS mice FEFS were fractured with a loss of capillary structure. The MFS mice capillary wall also appeared substantially closer to the intracapillary cells compared to the wild type mice. 

Videos Illustrates the Complex 3D Arrangement of the Kidney Glomerulus and Microfiber

Comparison of Wild Type and MFS Mice

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Wild Type Mice: The Fibers of the Elastic Fiber System (FEFS) are shown in red, capillary in white, and cells in blue. Imaged with ZEISS Sigma FE-SEM with integrated ultramicrotome. Data from R. Barbosa de Souza et al.

Wild Type Mice

In wild type mice, the FEFS (red) is arranged in a tubular-shaped network within the capillary (white).

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Mouse Model for Marfan Syndrome (MFS): The Fibers of the Elastic Fiber System (FEFS) are shown in red, capillary in white, and cells in blue. Imaged with ZEISS Sigma FE-SEM with integrated ultramicrotome. Data from R. Barbosa de Souza et al.

Mouse Model for Marfan Syndrome (MFS)

In a mouse model for Marfan Syndrome (MFS) the FEFS (red) were fractured with a loss of capillary structure (white). The MFS mice capillary wall also appeared substantially closer to the intracapillary cells (blue) compared to the wild type mice.

Professor Rodrigo Barbosa

The ultrastructural observations lead us to suspect a loss of capillary elasticity and reduced hemodynamic flow in MFS mice. We went on to confirm decreased blood flow velocity with live imaging.

Professor Rodrigo Barbosa Professor of Anatomy, Faculdade Santa Marcelina & Research Assistant, University of São Paulo, Brazil

Kidney Phenotype in Marfan Syndrome

PLOS One article: Extracellular matrix and vascular dynamics in the kidney of a murine model for Marfan syndrome
PLOS One article: Extracellular matrix and vascular dynamics in the kidney of a murine model for Marfan syndrome

Potential New Clinical Aspects

Professor Barbosa and colleagues propose that alterations in kidney vessels and tissues resulting from Marfan Syndrome could be important to understanding patients with this genetic condition and may be something to consider in the clinical management of the disease.

Read the full article in PLOS One and see their additional work using histology, transmission electron microscopy (TEM), fluorescence microscopy, and macro and micro blood flow measurements in the kidney.

Read the full article.
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