scholarly journals Altered Na+/K+-ATPase expression plays a role in rumen epithelium adaptation in sheep fed hay ad libitum or a mixed hay/concentrate diet

2011 ◽  
Vol 56 (No. 1) ◽  
pp. 36-48 ◽  
Author(s):  
J. Kuzinski ◽  
R. Zitnan ◽  
T. Viergutz ◽  
J. Legath ◽  
M. Schweigel

In this study we investigated rumen papillae morphology and the localization and expression of the<br />Na<sup>+/</sup>K<sup>+</sup>-ATPase&nbsp;in eight sheep fed hay ad libitum (h) or hay ad libitum plus additional concentrate (h/c). Four sheep were provided with the ad libitum h-diet for the complete three-week experimental period. The second group of four sheep received the h-diet for only one week and was fed the mixed hay/concentrate (h/c) diet for another two weeks. The amount of concentrate supplement was stepwise increased from 150 to 1000 g/day and given in two meals. Following slaughter rumen papillae from the atrium ruminis (AR), the rumen ventralis (RV) and the ventral blind sac (BSV) were fixed and examined for morphological changes and Na<sup>+</sup>/K<sup>+</sup>-ATPase localization by morphometric methods and immunohistochemistry. Ruminal epithelial cells (REC) originating from the strata basale to granulosum were also isolated. Cellular Na<sup>+</sup>/K<sup>+</sup>-ATPase expression (mRNA and protein) and differentiation state were determined by RT-PCR, Western blot, and flow cytometry. Compared with data from h-fed sheep, morphometric analysis revealed an increased length and width of rumen papillae in h/c-fed sheep, resulting in a marked 41% and 62% increase in rumen papillae surface in AR and RV, respectively. The rumen mucosa of h/c-fed sheep was characterized by a predominant stratum corneum (42 &plusmn; 0.7 &micro;m vs. 28 &plusmn; 0.5 &micro;m), but the thickness of the metabolically active cell layers remained unchanged. REC suspensions from sheep fed the h/c diet generally contained more cells (7.30 &plusmn; 0.83 vs. 3.49 &plusmn; 0.52 &times; 10<sup>7</sup>/ml; P &lt; 0.001) and an increased proportion of REC positive for basal cytokeratin and for the differentiation marker cytokeratin 10 (P &lt; 0.05). Cellular (cell membrane) and epithelial (stratum basale to stratum granulosum) Na<sup>+</sup>/K<sup>+</sup>-ATPase localization was similar between rumen regions and was not changed by concentrate feeding. After two weeks on the h/c-diet, a 96% increase in the absolute number of Na<sup>+</sup>/K<sup>+</sup>-ATPase-positive REC (6.56 &plusmn; 0.84 vs. 3.35 &plusmn; 0.51 &times; 10<sup>7</sup>/ml; P = 0.003) and a 61% elevation (P = 0.043) in Na<sup>+</sup>/K<sup>+</sup>-ATPase protein expression in REC from the upper third of the suprabasal cell layers were found. Moreover, a two-fold (P = 0.001) elevation in cell membrane surface area accompanied by a reduction (1.19 &times; 10<sup>&ndash;7</sup> &plusmn; 1.72 &times; 10<sup>&ndash;9</sup> arbitrary units (AU)/cm2 vs. 1.73 &times; 10<sup>&ndash;7</sup> &plusmn; 8.16 &times; 10<sup>&ndash;9</sup> AU/cm<sup>2</sup> in the h-group; P &lt; 0.001) in specific Na<sup>+</sup>/K<sup>+</sup>-ATPase fluorescence per cm<sup>2</sup> of cell membrane surface area was observed after h/c-feeding. Na<sup>+</sup>/K<sup>+</sup>-ATPase &alpha; subunit mRNA expression was also reduced (P &lt; 0.0001) from 0.154 &plusmn; 0.013 to 0.057 &plusmn; 0.004 pg per pg S18 mRNA control in the h/c-compared with the h-group. Thus, the h/c-diet led to a rapid increase in REC number and total cell membrane surface area in metabolically active and resorptive cell layers and was accompanied by a reduction in Na<sup>+</sup>/K<sup>+</sup>-ATPase mRNA expression and abundance per cell membrane surface area.

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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