scholarly journals ELECTRON MICROSCOPE RADIOAUTOGRAPHY AS A QUANTITATIVE TOOL IN ENZYME CYTOCHEMISTRY

1969 ◽  
Vol 42 (1) ◽  
pp. 122-134 ◽  
Author(s):  
M. M. Salpeter

The distribution of diisopropylfluorophosphate (DFP)-sensitive enzyme sites at the neuromuscular junction was determined quantitatively by electron microscope radioautography after incubation of muscle fragments in DFP-3H. Most of the sensitive sites were located in the subneural apparatus at a concentration of 90,000 sites per µ3 of cleft tissue or 12,000 sites per µ2 of postjunctional membrane surface area. A considerable concentration is also present in the teloglial cap. It has previously been demonstrated (Rogers et al., 1966) that one-third of the DFP-sensitive sites at the endplate can be reactivated by pyridine-2-aldoxime methiodide (2-PAM)—a compound which selectively reactivates phosphorylated acetylcholinesterase. In the present study, it was found that this ratio of 1:2 holds also on a fine-structural level. Muscle mast cells were found to have a heavy concentration of bound DFP.

1967 ◽  
Vol 32 (2) ◽  
pp. 379-389 ◽  
Author(s):  
Miriam M. Salpeter

Tritiated diisopropylfluorophosphate (DFP) was used to phosphorylate acetylcholinesterase (AChase) in the motor end plate of mouse sternomastoid muscle, and its distribution within the end plate was evaluated quantitatively by electron microscope radioautography. With the use of emulsion layers whose sensitivity to tritium had been calibrated, the density of AChase in different components of the end plate was calculated. The AChase was primarily localized (85%) in the junctional fold region. The concentration of AChase there was more than 20,000 active sites per cubic micron of tissue. The resolution of the technique was not sufficient to determine whether there was some AChase in the nerve end bulb; however, if there is any there, the concentration must be less than 10% of that at the junctional fold region.


1959 ◽  
Vol 6 (1) ◽  
pp. 7-10 ◽  
Author(s):  
Etienne de Harven ◽  
Christian Coërs

A preliminary electron microscope study of human neuromuscular junction is presented. The biopsy material was taken from the palmarus longus, and fixed routinely in osmium tetroxide and embedded in methacrylate. The structure of the motor endings and the relationship of the synaptic vesicles to the axolemmal membrane are described. The synaptic clefts are filled with an homogeneous material in continuity with the basement membrane covering the muscle fiber. The subneural apparatus is described, and special attention is paid to a vesicular component present in the sarcoplasm of the junctional area, which differs from synaptic vesicles and is presumed to be a derivate of the sarcoplasmic reticulum.


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.


2019 ◽  
Vol 30 (1) ◽  
pp. 108-118 ◽  
Author(s):  
Julia A. Pinette ◽  
Suli Mao ◽  
Bryan A. Millis ◽  
Evan S. Krystofiak ◽  
James J. Faust ◽  
...  

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.


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.


Perfusion ◽  
1998 ◽  
Vol 13 (2) ◽  
pp. 119-127 ◽  
Author(s):  
David W Fried ◽  
Theodore L Zombolas ◽  
Joseph J Leo ◽  
Hasratt Mohamed ◽  
Gabriel J Mattioni

The purpose of this current study was to compare the Terumo Capiox SX18 (1.8 m2) with the recently released Capiox SX25 (2.5 m2). Specifically, their oxygen transfer slopes, degree of blood shunting, extrapolated maximum oxygen transfer, blood side pressure drop and oxygen transfer consistency were compared. The lower intercept value (0.209 vs 0.236) coupled with the flatter slope (0.00171 vs 0.00225) of the oxygen transfer line for the SX25 is consistent with improved oxygen transfer performance. A lower FiO2 value would be predicted for the SX25, to achieve a specific PaO2 value, when the oxygen transfer requirement is equal for both devices. Extrapolated maximum oxygen transfer for the SX25 (462.6 ml O2/min) was 36.2% higher than that for the SX18 (339.6 ml O2/min). When indexed to membrane surface area, the SX18 (188.7 ml O2/m2/min performed 2.0% better than the SX25 (185.0 ml O2/m2/min). Both the slope (3.110 vs 3.744) and the intercept (4.595 vs 6.223) of the shunt fraction line were lower for the SX25, indicating that a lower shunt fraction would be predicted for all clinical blood flow rates. The slope (23.934 vs 22.443) and intercept values (-28.388 vs -22.650) of the pressure drop lines for the two devices indicate that the blood side pressure drop, over the range of clinical blood flows, were within 2% of each other. Oxygen transfer consistency, when expressed as the standard deviation of the oxygenator performance index and the percentage of predicted shunt, was not statistically different for the two devices.


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