Kidney proximal tubule cells: Epithelial cells without EGTA-extractable annexins?

2000 ◽  
Vol 78 (4) ◽  
pp. 495-502 ◽  
Author(s):  
Sandra Tribolo ◽  
Suzanne Maroux ◽  
Dominique Massey-Harroche
Keyword(s):  
Epithelial Cells ◽  
Proximal Tubule ◽  
Primary Culture ◽  
Collecting Duct ◽  
Basolateral Membrane ◽  
Isolated Cells ◽  
Vesicular Traffic ◽  
Kidney Proximal Tubule ◽  
Annexin I ◽  
Immunofluorescence Labelling

The expression and the subcellular localizations of annexins I, II, IV, VI, and XIII in renal epithelial cells were investigated, using immunological techniques with specific monoclonal antibodies. Upon performing Western blotting experiments, no annexins VI and XIII were detected in kidney, whereas annexins I, II, and IV were. Immunofluorescence labelling procedure performed on thin frozen renal sections showed the presence of these three annexins along the plasma membrane of the collecting duct cells with a restricted expression of annexin I at principal cells. Annexin I was also found present in some glomerular cells. None of these annexins, however, were detected in the proximal tubular cells upon performing immunofluorescence labelling and electrophoretic analysis on an EGTA (ethylenebis(oxyethylenenitrilo)tetraacetic acid)-extractable annexin fraction prepared from freshly isolated cells. This is the first time a mammalian epithelial cell has been found to express non-typical annexin (at least partly solubilized with EGTA). However, when these cells were grown in primary culture, they were found to express annexins I, II, IV, and V. As well as being located along the basolateral membrane, annexins I and II are also present on vesicles, which suggests that these annexins may be involved in vesicular traffic under cell culture conditions.Key words: annexin, kidney, proximal tubule, primary culture.

A network thermodynamic model of salt and water flow across the kidney proximal tubule

1978 ◽  
Vol 235 (6) ◽  
pp. F638-F648 ◽  
Author(s):  
S. R. Thomas ◽  
D. C. Mikulecky
Keyword(s):  
Proximal Tubule ◽  
Water Flow ◽  
Thermodynamic Model ◽  
Circuit Simulation ◽  
Basolateral Membrane ◽  
Kidney Proximal Tubule ◽  
Network Analyses ◽  
Pump Rate ◽  
Parameter Values ◽  
The Given

This network thermodynamic model of kidney proximal tubule epithelium treats coupled salt and water flow across each component membrane of the epithelium. We investigate the effects of various relative internal parameter values on the concentration of transepithelial flow, the concentrations in the cell and interspace, and the distribution of flows between cellular and paracellular routes. Best fit is obtaine if the apical and basolateral membrane reflection coefficients (or) are equal. The measured transepithelial filtration coefficient, Lp, is a function not only of the component Lps but also of the internal concentrations, or's, and permeabilities. For the given system topology (i.e., connectedness), parameters of component membranes must be within a narrow range to be consistent with experimental results. The dependence of the concentration of transported fluid on the balance between the solute pump rate and the transepithelial volume flow driving force is shown. This has implications for the effects of peritubular or lumen oncotic pressure on salt and water flow. With Appendix B of this paper and a user's guide for a circuit-simulation package (e.g., SPICE or PCAP) the reader can perform similar network analyses of transport models himself.

Contact behaviour during the reassociation of dissociated epithelial cells in primary culture

1987 ◽  
Vol 88 (4) ◽  
pp. 521-526
Author(s):  
R.M. Brown ◽  
C.A. Middleton
Keyword(s):  
Epithelial Cells ◽  
Chick Embryo ◽  
Primary Culture ◽  
Cell Cultures ◽  
Corneal Epithelium ◽  
Time Lapse ◽  
Cell Types ◽  
Epidermal Cells ◽  
Isolated Cells ◽  
Contact Behaviour

The behaviour in culture of dissociated epithelial cells from chick embryo pigmented retina epithelium (PRE), corneal epithelium (CE) and epidermis has been studied using time-lapse cinematography. The analysis concentrated on the contact behaviour of 60 previously isolated cells of each type during a 24 h period starting 3.5 h after the cells were plated out. During the period analysed the number of isolated cells in cultures of all three types gradually decreased as they became incorporated into islands and sheets of cells. However, there were significant differences in behaviour between the cell types during the establishment of these sheets and islands. In PRE cell cultures, islands of cells developed because, throughout the period of analysis, collisions involving previously isolated cells almost invariably resulted in the development of a stable contact. Once having established contact with another cell these cells rarely broke away again to become reisolated. In contrast the contacts formed between colliding CE and epidermal cells were, at least initially, much less stable and cells of both these types were frequently seen to break away and become reisolated after colliding with other cells. Sheets and islands of cells eventually developed in these cultures because the frequency with which isolated cells become reisolated decreased with increasing time in culture. The possible reasons underlying the different behaviour of PRE cells, when compared with that of CE and epidermal cells, are discussed. It is suggested that the decreasing tendency of isolated CE and epidermal cells to become reisolated may be related to the formation of desmosomes.

Antigen retrieval reveals widespread basolateral expression of syntaxin 3 in renal epithelia

2002 ◽  
Vol 282 (3) ◽  
pp. F523-F529 ◽  
Author(s):  
Sylvie Breton ◽  
Takeaki Inoue ◽  
Mark A. Knepper ◽  
Dennis Brown
Keyword(s):  
Epithelial Cells ◽  
Intracellular Transport ◽  
Collecting Duct ◽  
Basolateral Membrane ◽  
Antigen Retrieval ◽  
Surface Epithelium ◽  
Transitional Epithelium ◽  
Intercalated Cells ◽  
Snare Protein ◽  
Syntaxin 3

Soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins play a key role in docking and fusion of intracellular transport vesicles and may regulate apical and basolateral membrane protein delivery in epithelial cells. In a previous study, syntaxin 3 (a target SNARE) protein was detectable in the kidney only in intercalated cells. We now report a more widespread distribution of syntaxin 3 in a variety of renal epithelial cells after antigen retrieval. Sections of rat kidney were treated with SDS and incubated with antisyntaxin 3 antibodies. Strong basolateral membrane staining was seen in descending and ascending thin limbs of Henle, thick ascending limbs of Henle, the macula densa, distal and connecting tubules, and all cells of the collecting duct including A- and B-intercalated cells. The papillary surface epithelium and the transitional epithelium of the ureter were also stained, but proximal tubules were negative. Western blotting revealed a strong signal at 37 kDa in all regions, and the antigen was restricted to membrane fractions. SDS treatment was not necessary to reveal syntaxin 3 in intercalated cells. These data show that syntaxin 3 might be involved in basolateral trafficking pathways in most renal epithelial cell types. The exclusive basolateral location of syntaxin 3 in situ, however, contrasts with the apical location of this SNARE protein in some kidney epithelial cells in culture.

scholarly journals Basolateral membrane K+ channels in renal epithelial cells

2012 ◽  
Vol 302 (9) ◽  
pp. F1069-F1081 ◽  
Author(s):  
Kirk L. Hamilton ◽  
Daniel C. Devor
Keyword(s):  
Epithelial Cells ◽  
Collecting Duct ◽  
Basolateral Membrane ◽  
Gene Families ◽  
Proper Function ◽  
Major Function ◽  
Basolateral Membranes ◽  
Family Approach ◽  
Membrane Transport Proteins ◽  
Significant Disease

The major function of epithelial tissues is to maintain proper ion, solute, and water homeostasis. The tubule of the renal nephron has an amazingly simple structure, lined by epithelial cells, yet the segments (i.e., proximal tubule vs. collecting duct) of the nephron have unique transport functions. The functional differences are because epithelial cells are polarized and thus possess different patterns (distributions) of membrane transport proteins in the apical and basolateral membranes of the cell. K+ channels play critical roles in normal physiology. Over 90 different genes for K+ channels have been identified in the human genome. Epithelial K+ channels can be located within either or both the apical and basolateral membranes of the cell. One of the primary functions of basolateral K+ channels is to recycle K+ across the basolateral membrane for proper function of the Na+-K+-ATPase, among other functions. Mutations of these channels can cause significant disease. The focus of this review is to provide an overview of the basolateral K+ channels of the nephron, providing potential physiological functions and pathophysiology of these channels, where appropriate. We have taken a “K+ channel gene family” approach in presenting the representative basolateral K+ channels of the nephron. The basolateral K+ channels of the renal epithelia are represented by members of the KCNK, KCNJ, KCNQ, KCNE, and SLO gene families.

Differential localization of two glucose transporter isoforms in rat kidney

1990 ◽  
Vol 259 (2) ◽  
pp. C286-C294 ◽  
Author(s):  
B. Thorens ◽  
H. F. Lodish ◽  
D. Brown
Keyword(s):  
Proximal Tubule ◽  
Glucose Transporter ◽  
Collecting Duct ◽  
Rat Kidney ◽  
Basolateral Membrane ◽  
Staining Intensity ◽  
Intercalated Cells ◽  
Brain Glucose ◽  
Nephron Segments ◽  
Glucose Transporter Isoforms

The localization of two glucose transporter isoforms was mapped in the rat kidney: the high-Michaelis constant (Km; 15-20 mM) low-affinity "liver" transporter and the low-Km (1-2 mM) high-affinity "erythroid/brain" transporter. Both are basolateral membrane proteins, but the liver transporter was present exclusively in the S1 part of the proximal tubule, whereas the erythroid/brain transporter was expressed at variable levels in different nephron segments. Staining intensity was low in the straight proximal tubule (S3), intermediate in the medullary thin and thick ascending limbs, and highest in connecting segments and collecting ducts. In the collecting duct, the erythroid/brain glucose transporter was expressed at the highest level in intercalated cells; less was present in principal cells. In the papilla, only intercalated cells expressed this transporter isoform. These results suggest specific involvements of each transporter isoform in transepithelial glucose reabsorption by different segments of the proximal tubule. They also indicate that while the liver glucose transporter is present in gluconeogenic cells, there is a good correlation between the level of expression of the erythroid/brain glucose transporter and the glycolytic activity of the different nephron segments.

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