scholarly journals Three-dimensional ATUM-SEM reconstruction and analysis of hepatic endoplasmic reticulum-organelle interactions

2020 ◽  
Author(s):  
Yi Jiang ◽  
Linlin Li ◽  
Xi Chen ◽  
Jiazheng Liu ◽  
Jingbin Yuan ◽  
...  

AbstractThe endoplasmic reticulum (ER) is a contiguous and complicated membrane network in eukaryotic cells, and membrane contact sites (MCSs) between the ER and other organelles perform vital cellular functions, including lipid homeostasis, metabolite exchange, calcium level regulation, and organelle division. Here, we establish a whole pipeline to reconstruct all ER, mitochondria, lipid droplets, lysosomes, peroxisomes, and nuclei by automated tape-collecting ultramicrotome scanning electron microscopy (ATUM-SEM) and deep-learning techniques, which generates an unprecedented 3D model for mapping liver samples. Furthermore, the morphology of various organelles is systematically analyzed. We found that the ER presents with predominantly flat cisternae and is knitted tightly all throughout the intracellular space and around other organelles. In addition, the ER has a smaller volume-to-membrane surface area ratio than other organelles, which suggests that the ER could be more suited for functions that require a large membrane surface area. Moreover, the MCSs between the ER and other organelles are explored. Our data indicate that ER-mitochondrial contacts are particularly abundant, especially for branched mitochondria. In addition, ER contacts with lipid droplets, lysosomes, and peroxisomes are also plentiful. In summary, we design an efficient method for obtaining a 3D reconstruction of biological structures at a nanometer resolution. Our study also provides the first 3D reconstruction of various organelles in liver samples together with important information fundamental for biochemical and functional studies in the liver.

1980 ◽  
Vol 85 (3) ◽  
pp. 577-586 ◽  
Author(s):  
R P Bolender ◽  
D Paumgartner ◽  
D Muellener ◽  
G Losa ◽  
E R Weibel

The purpose of the study was to consider quantitatively the relationships between the surface area of the endoplasmic reticulum (ER) and constituent marker enzyme activities, as they occur in fractions collected from rat liver homogenates. The ER surface area was estimated in five membrane-containing fractions by use of a combined cytochemical-stereological technique (5), while, at the same time, ER marker enzymes were assayed biochemically. Fraction/homogenate recoveries for the ER enzymes averaged 100%, total membrane surface area 98%, and ER surface area 96%. Relative specific activities, which compare the relative amounts of ER marker enzyme activities to the relative ER surface area in the membrane-containing fractions, indicate variable distributions for glucose-6-phosphatase and NADPH cytochrome c reductase, but not for esterase.


2007 ◽  
Vol 3 (1) ◽  
pp. 89-113
Author(s):  
Zoltán Gillay ◽  
László Fenyvesi

There was a method developed that generates the three-dimensional model of not axisymmetric produce, based on an arbitrary number of photos. The model can serve as a basis for calculating the surface area and the volume of produce. The efficiency of the reconstruction was tested on bell peppers and artificial shapes. In case of bell peppers 3-dimensional reconstruction was created from 4 images rotated in 45° angle intervals. The surface area and the volume were estimated on the basis of the reconstructed area. Furthermore, a new and simple reference method was devised to give precise results for the surface area of bell pepper. The results show that this 3D reconstruction-based surface area and volume calculation method is suitable to determine the surface area and volume of definite bell peppers with an acceptable error.


1992 ◽  
Vol 101 (4) ◽  
pp. 907-913 ◽  
Author(s):  
G.J. Cannon ◽  
J.A. Swanson

Murine bone marrow-derived macrophages, which measure 13.8 +/− 2.3 microns diameter in suspension, can ingest IgG-opsonized latex beads greater than 20 microns diameter. A precise assay has allowed the determination of the phagocytic capacity, and of physiological parameters that limit that capacity. Ingestion of beads larger than 15 microns diameter required IgG-opsonization, and took 30 minutes to reach completion. Despite the dependence on Fc-receptors for phagocytosis of larger beads, cells reached their limit before all cell surface Fc-receptors were occupied. The maximal membrane surface area after frustrated phagocytosis of opsonized coverslips was similar to the membrane surface area required to engulf particles at the limiting diameter, indicating that the capacity was independent of particle shape. Vacuolation of the lysosomal compartment with sucrose, which expanded endocytic compartments, lowered the phagocytic capacity. This decrease was reversed when sucrose vacuoles were collapsed by incubation of cells with invertase. These experiments indicate that the phagocytic capacity is limited by the amount of available membrane, rather than by the availability of Fc-receptors. The capacity was also reduced by depolymerization of cytoplasmic microtubules with nocodazole. Nocodazole did not affect the area of maximal cell spreading during frustrated phagocytosis, but did alter the shape of the spread cells. Thus, microtubules may coordinate cytoplasm for engulfment of the largest particles.


2012 ◽  
Vol 7 (2) ◽  
pp. 9-11 ◽  
Author(s):  
NS Chowdhury ◽  
FMM Islam ◽  
F Zafreen ◽  
BA Begum ◽  
N Sultana ◽  
...  

Introduction: Patients with end stage renal disease require 12 hours of haemodialysis per week in three equal sessions (4 hours/day for 3 days/week). But the duration and frequency of treatment can be reduced by increasing the surface area of the dialyzer membrane. Methods: In this prospective study 40 patients of end stage renal disease receiving haemodialysis for more than six months were included to observe the effects of increment in the surface area of the dialyzer membrane on the adequacy of haemodialysis. Result: It was observed that 20 patients receiving haemodialysis on a dialyzer with membrane surface area of 1.2 m² did not have satisfactory solute clearance index. Urea reduction ratio was 45.9 ± 3.03 and fractional urea clearance (Kt/V) was 0.76 ± 0.09. On the other hand patients (20 cases) receiving haemodialysis on a dialyzer with membrane surface area of 1.3 m² had a urea reduction ratio 50.76± 5.16 and fractional urea clearance (Kt/V) 0.91 ± 0.16. All the patients of both groups received dialysis for 8 hours/week in two equal sessions (4 hours/day for 2 days/week). Statistically the increment was significant (p<0.001). Conclusion: This study reveals, adequacy of dialysis can be increased by increasing the surface area of the dialyzer membrane. So, considering the poor socioeconomic condition of Bangladesh and patients' convenience, a short duration, low cost dialysis regime can be tried by increasing the surface area of dialyzer membrane. DOI: http://dx.doi.org/10.3329/jafmc.v7i2.10387 JAFMC 2011; 7(2): 9-11


1983 ◽  
Vol 217 (1207) ◽  
pp. 191-213 ◽  

Purkinje strands from both ventricles of adult mongrel dogs were excised, and electrical properties were studied by the voltage-clamp technique. The strands were then examined with light and electron microscopy and structural properties were analysed by morphometric techniques. The canine Purkinje strand contains (by volume) about 28% myocyte and 55% dense outer connective tissue. The remainder of the volume is taken up by the inner shell of loosely packed connective tissue within 10 μm of a myocyte membrane. These volume fractions vary considerably from one strand to another. Clefts less than 10 μm wide occupy 18% of the myocyte volume and clefts less than 1 μm wide occupy 1%. The membrane surface area of the myocytes can be divided into three categories by reference to the size of the adjacent cleft. About 47.8% of the membrane surface area faces clefts wider than 1 μm, another 22.2% faces clefts between 0.1 and 1 μm wide, and the final 30% faces clefts less than 0.1 μm wide. The surface area facing the narrowest clefts (less than 0.1 μm wide) is divided between nexuses 3%, desmosomes 10%, and unspecialized membrane 17% (each figure is expressed as a percentage of the total surface area of myocyte membrane). The canine Purkinje strand has a more favourable anatomy than the sheep Purkinje strand for most physiological experiments. We expect that the complicating effects of series resistance and change in the concentration of extracellular ions will be much smaller than in sheep strands, but still not negligible.


1994 ◽  
Vol 72 (2) ◽  
pp. 861-871 ◽  
Author(s):  
D. Ulrich ◽  
R. Quadroni ◽  
H. R. Luscher

1. Voltage-clamp, current-clamp, and morphological data were obtained from visually identified motoneurons in organotypic cocultures of rat embryonic spinal cord, dorsal root ganglia, and skeletal muscle. The cells were injected with Biocytin during whole-cell patch-clamp recordings and stained with horseradish peroxidase. 2. The somata and dendritic trees of the cells were reconstructed with a semiautomatic reconstruction system. The motoneurons had a common multipolar shape. An elliptic soma gave rise to 3-9 stem dendrites with a mean diameter of 2.5 +/- 0.9 (SD) micron terminating in 24 +/- 7 dendritic endings. The mean total dendritic path length was 3,306 +/- 1,075 microns. The mean total membrane surface area was 15,594 +/- 10,404 microns 2 with a dendritic to somatic membrane surface area ratio of 3.4 +/- 1.4 (n = 7 cells). 3. The ratio between the sum of the diameters of the two daughter branches and the diameter of the parental branch each raised to the 3/2 power at all branch points was 1.3 +/- 0.28 (n = 8 cells). The dendritic trees of the cells tapered continuously from the soma to the distal ends. The mean normalized dendritic trunk parameter of all cells was 0.62 +/- 0.22. 4. The motoneurons had a mean input resistance RN of 498 +/- 374 M delta, a mean membrane time constant (tau m) of 22 +/- 4.6 ms, and a mean dendritic dominance (rho) of 2.7 +/- 0.86 (n = 5 cells). The mean electronic length (L) calculated from tau m and the slowest voltage-clamp time constant (tau VC1) was 0.7 +/- 0.04 (n = 7 cells). 5. The specific membrane capacitance (Cm) estimated from the charge of the capacitive current during a voltage step and the total membrane surface area was 1.08 +/- 0.3 microF/cm2 (n = 6 cells). 6. Compartmental computer models were constructed of individual cells. Experimental and simulated voltage transients were matched with Cm = 1 microF/cm2, a uniform membrane resistivity (Rm) = tau m/Cm and a cytosolic resistivity (Ri) of 308 +/- 39 omega.cm (n = 3 cells). 7. The mean electrotonic length of the dendritic paths was 0.83 +/- 0.2 (n = 5 cells). The mean input resistance at the dendritic terminals (RT) was 1,413 +/- 260 M omega. Synaptic conductances were applied at all distal dendritic compartments of the model cells. The resulting synaptic currents were calculated at the input site and at the soma. The mean transient current attenuation ratio was 4.7 +/- 1.7 under idealized voltage-clamp conditions.(ABSTRACT TRUNCATED AT 400 WORDS)


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