Innervation of Epi- and Endoneurial Compartments of Rat Facial, Vagus and Sciatic Nerves as Studied by Double-Labeling Immunofluorescence

1994 ◽  
Vol 149 (4) ◽  
pp. 264-271 ◽  
Author(s):  
W. Kummer ◽  
E. Seifert ◽  
A. Schadel
Author(s):  
Frank A. Rawlins

Several speculations exist as to the site of incorporation of preformed molecules into myelin. The possibility that an autoradiographic analysis of cholesterol-1,2-H3 incorporation at very short times after injection might shed some light in the solution of that problem led to the present experiment.Cholesterol-1,2-H3 was injected intraperitoneally into 24 tenday old mice. The animals were then sacrificed at 10,20,30,40,60,90,120 and 180 min after the injection and the sciatic nerves were processed for electron microscope autoradiography. To analyze the grain distribution in the autoradiograms of cross and longitudinal sections from each sciatic nerve myelin sheaths were subdivided into three compartments named: outer 1/3, middle 1/3 and inner 1/3 compartments.It was found that twenty min. after the injection of cholesterol -1.2-H3 (Figs. 1 and 2), 55% of the total number of grains (t.n.g) found in myelin were within the outer 1/3 compartment, 9% were within the middle 1/3 and 36% within the inner 1/3 compartment


Author(s):  
Margaret Hukee

Gold labeling of two antigens (double labeling) is often done on two section surfaces separated by section thickness. Whether labeling is done on both sides of the same section or on two parallel surfaces separated by section thickness (PSSST), comparable results are dependent on an equal number of epitopes being exposed at each surface. We propose a method to study protein labeling within the same field of proteins, by examining two directly adjacent surfaces that were split during sectioning. The number of labeling sites on adjacent surfaces (AS) were compared to sites on PSSST surfaces in individual bacteria.Since each bacteria needed to be recognizable in all three section surfaces, one-hole grids were used for labeling. One-hole grids require a supporting membrane and excessive handling during labeling often ruptures the membrane. To minimize handling, a labeling chamber was designed that is inexpensive, disposable, minimizes contamination, and uses a minimal amount of solution.


1999 ◽  
Vol 277 (2) ◽  
pp. G487-G494 ◽  
Author(s):  
Christopher R. Marino ◽  
Virginia Jeanes ◽  
Walter F. Boron ◽  
Bernhard M. Schmitt

The cellular mechanisms of [Formula: see text] secretion in the human pancreas are unclear. Expression of a Na+-[Formula: see text]cotransporter (NBC) mRNA has been observed recently, but the distribution and physiological role of the NBC protein are not known. Here we examined the expression and localization of NBC in human pancreas by Northern blot, immunoblot, and immunofluorescence microscopy. Rat kidney NBC probes detected a single 9.5-kb band by Northern blot. On immunoblots, two polyclonal antisera directed against different epitopes of rat kidney NBC identified a single ∼130-kDa protein. In cryosections of normal human pancreas, both antisera labeled basolateral membranes of large, morphologically identifiable ducts and produced a distinct labeling pattern in the remainder of the parenchyma. In double-labeling experiments, NBC immunoreactivity in the parenchyma colocalized with the Na+-K+pump, a basolateral marker. In contrast, NBC and cystic fibrosis transmembrane conductance regulator, an apical membrane marker, were detected within the same histological structures but at different subcellular localizations. The NBC antisera did not label acinar or islet cells. Our observations suggest that secretion of[Formula: see text] by human pancreatic duct cells involves the basolateral uptake of Na+and[Formula: see text] via NBC, an electrogenic Na+-[Formula: see text]cotransporter.


Cells ◽  
2017 ◽  
Vol 6 (3) ◽  
pp. 30 ◽  
Author(s):  
Susana Brun ◽  
Nicolas Schall ◽  
Hélène Jeltsch-David ◽  
Jérôme de Sèze ◽  
Sylviane Muller

2014 ◽  
Vol 32 (5) ◽  
pp. 164-170 ◽  
Author(s):  
Vanessa Cristina Pereira Barreiros ◽  
Fernando José Dias ◽  
Mamie Mizusaki Iyomasa ◽  
Joaquim Coutinho-Netto ◽  
Luiz Gustavo de Sousa ◽  
...  

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