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Three-dimensional architecture of podocytes revealed by block-face scanning electron microscopy.

Ichimura K, Miyazaki N, Sadayama S, Murata K, Koike M, Nakamura K, Ohta K, Sakai T - Sci Rep (2015)

Bottom Line: Moreover, from the cell body, the foot processes were also emerged via the ridge-like prominence, as found in the primary process.The ridge-like prominence anchored the cell body and primary process to the glomerular basement membrane, and connected the foot processes to the cell body and primary process.In conclusion, serial block-face imaging is a powerful tool for clear understanding the three-dimensional architecture of podocytes through its ability to reveal novel structures which were difficult to determine by conventional transmission and scanning electron microscopes alone.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy and Life Structure, Juntendo University Graduate School of Medicine, Tokyo, Japan.

ABSTRACT
Block-face imaging is a scanning electron microscopic technique which enables easier acquisition of serial ultrastructural images directly from the surface of resin-embedded biological samples with a similar quality to transmission electron micrographs. In the present study, we analyzed the three-dimensional architecture of podocytes using serial block-face imaging. It was previously believed that podocytes are divided into three kinds of subcellular compartment: cell body, primary process, and foot process, which are simply aligned in this order. When the reconstructed podocytes were viewed from their basal side, the foot processes were branched from a ridge-like prominence, which was formed on the basal surface of the primary process and was similar to the usual foot processes in structure. Moreover, from the cell body, the foot processes were also emerged via the ridge-like prominence, as found in the primary process. The ridge-like prominence anchored the cell body and primary process to the glomerular basement membrane, and connected the foot processes to the cell body and primary process. In conclusion, serial block-face imaging is a powerful tool for clear understanding the three-dimensional architecture of podocytes through its ability to reveal novel structures which were difficult to determine by conventional transmission and scanning electron microscopes alone.

No MeSH data available.


Related in: MedlinePlus

Three-dimensional structure of podocyte foot processes.A single reconstructed podocyte based on serial FIB/SEM images is observed from luminal (a1, a2) and basal (b1, b2) sides. (a2 and b2) are the magnification of the area indicated by rectangles in (a1 and b1), respectively. In the luminal surface view, the foot processes appeared to be simply branched from the lateral aspects of primary process, as found in a plant leaf vein (a2). In the basal surface view, the most proximal portions of foot processes are connected each other via a tortuous ridge-like prominence, which was formed on the basal surface of the primary process (asterisks in b2). The ridge-like prominences was quite difficult to predict their existence only by the conventional SEM observation from the luminal side, and was structurally similar to the usual foot processes. CB, cell body; P, primary process. Bar scales, 1 μm in (b1); 200 nm in (b2). The three-dimensionally reconstructed podocyte can be observed from any direction on the reconstruction software. (See also Supplementary Movies S1, S2).
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f5: Three-dimensional structure of podocyte foot processes.A single reconstructed podocyte based on serial FIB/SEM images is observed from luminal (a1, a2) and basal (b1, b2) sides. (a2 and b2) are the magnification of the area indicated by rectangles in (a1 and b1), respectively. In the luminal surface view, the foot processes appeared to be simply branched from the lateral aspects of primary process, as found in a plant leaf vein (a2). In the basal surface view, the most proximal portions of foot processes are connected each other via a tortuous ridge-like prominence, which was formed on the basal surface of the primary process (asterisks in b2). The ridge-like prominences was quite difficult to predict their existence only by the conventional SEM observation from the luminal side, and was structurally similar to the usual foot processes. CB, cell body; P, primary process. Bar scales, 1 μm in (b1); 200 nm in (b2). The three-dimensionally reconstructed podocyte can be observed from any direction on the reconstruction software. (See also Supplementary Movies S1, S2).

Mentions: To reveal the structural relationship among the three subcellular compartments of podocytes, we next observed single reconstructed podocyte which was separated from neighboring podocytes (Fig. 5, Movies S1, S2). When viewed from the basal side of the primary processes, we found that the foot processes were branched from a ridge-like prominence, which was protruded from the basal surface of the primary process (Fig. 5b1, 5b2) and was directly attached to the GBM like the foot processes (Supplementary Fig. S2). The distal ends of foot processes made their way under the primary process, and connected to the lateral surface of the ridge-like prominence via the slit diaphragm (Figs. 3b, 3b′, 6).


Three-dimensional architecture of podocytes revealed by block-face scanning electron microscopy.

Ichimura K, Miyazaki N, Sadayama S, Murata K, Koike M, Nakamura K, Ohta K, Sakai T - Sci Rep (2015)

Three-dimensional structure of podocyte foot processes.A single reconstructed podocyte based on serial FIB/SEM images is observed from luminal (a1, a2) and basal (b1, b2) sides. (a2 and b2) are the magnification of the area indicated by rectangles in (a1 and b1), respectively. In the luminal surface view, the foot processes appeared to be simply branched from the lateral aspects of primary process, as found in a plant leaf vein (a2). In the basal surface view, the most proximal portions of foot processes are connected each other via a tortuous ridge-like prominence, which was formed on the basal surface of the primary process (asterisks in b2). The ridge-like prominences was quite difficult to predict their existence only by the conventional SEM observation from the luminal side, and was structurally similar to the usual foot processes. CB, cell body; P, primary process. Bar scales, 1 μm in (b1); 200 nm in (b2). The three-dimensionally reconstructed podocyte can be observed from any direction on the reconstruction software. (See also Supplementary Movies S1, S2).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4355681&req=5

f5: Three-dimensional structure of podocyte foot processes.A single reconstructed podocyte based on serial FIB/SEM images is observed from luminal (a1, a2) and basal (b1, b2) sides. (a2 and b2) are the magnification of the area indicated by rectangles in (a1 and b1), respectively. In the luminal surface view, the foot processes appeared to be simply branched from the lateral aspects of primary process, as found in a plant leaf vein (a2). In the basal surface view, the most proximal portions of foot processes are connected each other via a tortuous ridge-like prominence, which was formed on the basal surface of the primary process (asterisks in b2). The ridge-like prominences was quite difficult to predict their existence only by the conventional SEM observation from the luminal side, and was structurally similar to the usual foot processes. CB, cell body; P, primary process. Bar scales, 1 μm in (b1); 200 nm in (b2). The three-dimensionally reconstructed podocyte can be observed from any direction on the reconstruction software. (See also Supplementary Movies S1, S2).
Mentions: To reveal the structural relationship among the three subcellular compartments of podocytes, we next observed single reconstructed podocyte which was separated from neighboring podocytes (Fig. 5, Movies S1, S2). When viewed from the basal side of the primary processes, we found that the foot processes were branched from a ridge-like prominence, which was protruded from the basal surface of the primary process (Fig. 5b1, 5b2) and was directly attached to the GBM like the foot processes (Supplementary Fig. S2). The distal ends of foot processes made their way under the primary process, and connected to the lateral surface of the ridge-like prominence via the slit diaphragm (Figs. 3b, 3b′, 6).

Bottom Line: Moreover, from the cell body, the foot processes were also emerged via the ridge-like prominence, as found in the primary process.The ridge-like prominence anchored the cell body and primary process to the glomerular basement membrane, and connected the foot processes to the cell body and primary process.In conclusion, serial block-face imaging is a powerful tool for clear understanding the three-dimensional architecture of podocytes through its ability to reveal novel structures which were difficult to determine by conventional transmission and scanning electron microscopes alone.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomy and Life Structure, Juntendo University Graduate School of Medicine, Tokyo, Japan.

ABSTRACT
Block-face imaging is a scanning electron microscopic technique which enables easier acquisition of serial ultrastructural images directly from the surface of resin-embedded biological samples with a similar quality to transmission electron micrographs. In the present study, we analyzed the three-dimensional architecture of podocytes using serial block-face imaging. It was previously believed that podocytes are divided into three kinds of subcellular compartment: cell body, primary process, and foot process, which are simply aligned in this order. When the reconstructed podocytes were viewed from their basal side, the foot processes were branched from a ridge-like prominence, which was formed on the basal surface of the primary process and was similar to the usual foot processes in structure. Moreover, from the cell body, the foot processes were also emerged via the ridge-like prominence, as found in the primary process. The ridge-like prominence anchored the cell body and primary process to the glomerular basement membrane, and connected the foot processes to the cell body and primary process. In conclusion, serial block-face imaging is a powerful tool for clear understanding the three-dimensional architecture of podocytes through its ability to reveal novel structures which were difficult to determine by conventional transmission and scanning electron microscopes alone.

No MeSH data available.


Related in: MedlinePlus