Ultrastructural localization of glucose-6-phosphatase activity in proximal convoluted tubule cells of rat kidney

Histochemie ◽  
1971 ◽  
Vol 28 (4) ◽  
pp. 288-295 ◽  
Author(s):  
Shinsuke Kanamura
Keyword(s):  
Phosphatase Activity ◽  
Rat Kidney ◽  
Ultrastructural Localization ◽  
Proximal Convoluted Tubule ◽  
Tubule Cells

scholarly journals A new method based on cobalt for histochemical and cytochemical demonstration of glucose-6-phosphatase activity.

1990 ◽  
Vol 38 (10) ◽  
pp. 1503-1509 ◽  
Author(s):  
G Thiéry ◽  
J Bernier ◽  
M Bergeron
Keyword(s):  
Phosphatase Activity ◽  
Rat Kidney ◽  
Strong Reaction ◽  
Proximal Tubule Cells ◽  
Ammonium Sulfide ◽  
Cytochemical Demonstration ◽  
Impregnation Technique ◽  
Osmium Impregnation ◽  
Tubule Cells ◽  
Collecting Tubules

We describe a novel technique for the histochemical and cytochemical demonstration of glucose-6-phosphatase activity. In this method, lead is replaced by cobalt. After activity of glucose-6-phosphatase, cobalt phosphate Co3(PO4)2 is formed, and in the presence of ammonium sulfide (NH4)2S, the precipitate is transformed into a sulfide that fixes osmium and provides good electron density. Glucose-6-phosphatase activity was determined mostly in rat kidney cells, but controls were also performed in liver cells. A strong reaction was seen in proximal tubule cells, but the reaction was weak in distal convoluted tubule cells. This technique showed the same endoplasmic reticulum (ER) organization in proximal and distal nephron as that seen with the osmium impregnation technique. In collecting tubules, intercalated cells had irregular reactivity, while principal cells had none. Our results indicate that the cobalt technique is valid, reliable, and sensitive enough to detect low glucose-6-phosphatase activity. Moreover, the technique can be used with 1-mm-thick specimens and obviates the need for use of frozen tissue sections.

scholarly journals THE BASE OF THE PROXIMAL CONVOLUTED TUBULE CELLS OF RAT KIDNEY

1957 ◽  
Vol 3 (2) ◽  
pp. 249-254 ◽  
Author(s):  
Helmut Ruska ◽  
Dan H. Moore ◽  
John Weinstock
Keyword(s):  
Hydrostatic Pressure ◽  
Capillary Pressure ◽  
Functional Unit ◽  
Rat Kidney ◽  
Three Dimensional ◽  
Blood Stream ◽  
Proximal Convoluted Tubule ◽  
Basal Labyrinth ◽  
Tubule Cells

The three dimensional arrangement of the compartments on the base of proximal convoluted tubule cells of rat kidney is described. An extracellular basal labyrinth is found to be enclosed by these compartments. The compartments with their mitochondria and the basal labyrinth are regarded as forming a functional unit. It is supposed that this basal unit serves for excretion of reabsorbed fluid from the cell into the labyrinth and for the development of hydrostatic pressure in the labyrinth to overcome the capillary pressure and to pass the reabsorbed fluid into the blood stream.

Dependence of ion fluxes on fluid transport by rat proximal tubule

1986 ◽  
Vol 250 (4) ◽  
pp. F680-F689 ◽  
Author(s):  
K. Bomsztyk ◽  
F. S. Wright
Keyword(s):  
Proximal Tubule ◽  
Fluid Transport ◽  
Rat Kidney ◽  
Water Flux ◽  
Fluid Secretion ◽  
Proximal Convoluted Tubule ◽  
Ion Fluxes ◽  
Fluid Absorption ◽  
Fluid Flux ◽  
K Transport

The effects of changes in transepithelial water flux (Jv) on sodium, chloride, calcium, and potassium transport by the proximal convoluted tubule were examined by applying a microperfusion technique to surface segments in kidneys of anesthetized rats. Perfusion solutions were prepared with ion concentrations similar to those in fluid normally present in the later parts of the proximal tubule. Osmolality of the perfusate was adjusted with mannitol. With no mannitol in the perfusates, net fluid absorption was observed. Addition of increasing amounts of mannitol first reduced Jv to zero and then reversed net fluid flux. At the maximal rates of fluid absorption, net absorption of Na, Cl, Ca, and K was observed. When Jv was reduced to zero, Na, Cl, and Ca absorption were reduced and K entered the lumen. Na, Cl, and Ca secretion occurred in association with the highest rates of net fluid secretion. The lumen-positive transepithelial potential progressively increased as the net fluid flux was reduced to zero and then reversed. The results demonstrate that changes in net water flux can affect Na, Cl, Ca, and K transport by the proximal convoluted tubule of the rat kidney. These changes in net ion fluxes are not entirely accounted for by changes in bulk-phase transepithelial electrochemical gradients.

Immunolocalization of NHE8 in rat kidney

2005 ◽  
Vol 288 (3) ◽  
pp. F530-F538 ◽  
Author(s):  
Sunita Goyal ◽  
SueAnn Mentone ◽  
Peter S. Aronson
Keyword(s):  
Proximal Tubule ◽  
Brush Border ◽  
Brush Border Membrane ◽  
Rat Kidney ◽  
Surface Membrane ◽  
Proximal Tubules ◽  
Intense Staining ◽  
Proximal Tubule Cells ◽  
Nephron Segment ◽  
Tubule Cells

In situ hybridization studies demonstrated that Na+/H+ exchanger NHE8 is expressed in kidney proximal tubules. Although membrane fractionation studies suggested apical brush-border localization, precise membrane localization could not be definitively established. The goal of the present study was to develop isoform-specific NHE8 antibodies as a tool to directly establish the localization of NHE8 protein in the kidney by immunocytochemistry. Toward this goal, two sets of antibodies that label different NHE8 epitopes were developed. Monoclonal antibody 7A11 and polyclonal antibody Rab65 both specifically labeled NHE8 by Western blotting as well as by immunofluorescence microscopy. The immunolocalization pattern in the kidney seen with both antibodies was the same, thereby validating NHE8 specificity. In particular, NHE8 expression was observed on the apical brush-border membrane of all proximal tubules from S1 to S3. The most intense staining was evident in proximal tubules in the deeper cortex and medulla with a significant but somewhat weaker staining in superficial proximal tubules. Colocalization studies with γ-glutamyltranspeptidase and megalin indicated expression of NHE8 on both the microvillar surface membrane and the coated-pit region of proximal tubule cells, suggesting that NHE8 may be subject to endocytic retrieval and recycling. Although colocalizing in the proximal tubule with NHE3, no significant alteration in NHE8 protein expression was evident in NHE3-null mice. We conclude that NHE8 is expressed on the apical brush-border membrane of proximal tubule cells, where it may play a role in mediating or regulating ion transport in this nephron segment.

scholarly journals Ultrastructural localization of type V collagen in rat kidney.

1982 ◽  
Vol 92 (2) ◽  
pp. 343-349 ◽  
Author(s):  
A Martinez-Hernandez ◽  
S Gay ◽  
E J Miller
Keyword(s):  
Type I Collagen ◽  
Rat Kidney ◽  
Ultrastructural Localization ◽  
Particulate Material ◽  
Basement Membranes ◽  
Type I ◽  
Type V Collagen ◽  
Collagen Molecules ◽  
Immunohistochemical Studies ◽  
Type V

Antibodies specific for the alpha 1 (V) chain and native collagen molecules containing the alpha 1 (V) chain have been used in electron immunohistochemical studies of rat kidney to determine the ultrastructural distribution of this class of collagen molecules. In addition, antibodies against type I collagen and whole basement membrane were used as markers for interstitial collagen and authentic basement membranes. Our results indicate that type V collagen is present in the renal interstitium in different forms: in close apposition to interstitial collagen fibers; in the stromal aspect of vascular basement membranes; and as particulate material not bound to other structures. On the basis of these findings, we postulate a binding or connecting function for this collagen type.

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