Ultrastructural morphometric of gap junctions during differentiation of stratified squamous epithelium

1984 ◽  
Vol 69 (1) ◽  
pp. 67-85
Author(s):  
F.H. White ◽  
D.A. Thompson ◽  
K. Gohari

The presence of gap junctions in stratified epithelia has now been extensively documented, but there have been few attempts to quantify them. In the present report, samples of hamster cheek pouch mucosa were processed for electron microscopy and electron micrographs from defined basal, spinous and granular layers were obtained. Using a combination of direct measurement and stereological intersection counting techniques, the relative surface areas of peripheral gap junctions (i.e. those in direct contact with the epithelial plasma membrane) and annular gap junctions (i.e. those present as complete, approximately circular profiles within the epithelial cell cytoplasm) were determined. Following estimation of the plasma membrane surface area of ‘average’ epithelial cells from each of the defined strata, relative values were transformed into absolute data. Data from peripheral and annular junctions were pooled to provide an estimate of total gap junctions area. Relative surface area estimates were similar for peripheral, annular and total gap junctions, in that values were invariably highest in the spinous layer and lowest in the granular layer. Absolute data indicate that there is more than a threefold increase in the area of membrane differentiated into gap junctions in the average spinous cell when compared with the average basal cell. Values for total gap-junctional areas in the average granular cell are reduced somewhat with respect to the average spinous cell and this is effected by a decrease in the area of peripheral gap junctions. We conclude that there is synthesis of gap junctions between basal and spinous cells, which is followed by evidence of degradation between spinous and granular cells. The magnitude of the estimates of area is comparable to those obtained from other stratified and non-stratified epithelia and it would thus appear that gap junctions may play a significant role in cellular control processes in all viable epithelial strata.

1984 ◽  
Vol 66 (1) ◽  
pp. 411-429
Author(s):  
F.H. White ◽  
K. Gohari

Desmosomes in stratified squamous epithelia appear to exhibit quantitative alterations during differentiation. In this work we use stereological and other morphometric methods to quantify these structures in epithelial cells from defined basal, spinous and granular strata. Hamster cheek pouch mucosa from five animals was processed for electron microscopy using strictly standardized techniques and a stratified random sampling procedure was used to obtain micrographs of cells from basal, spinous and granular layers. Stereological intersection counting techniques were used to determine for each layer the relative surface area of plasma membrane occupied by desmosomes (Ss), the number of desmosomes per unit surface area of plasma membrane (Ns), the mean individual desmosomal diameter (delta) and the mean individual desmosomal surface area (s). In addition, estimates of nuclear volume were obtained by direct measurement of nuclear profiles and volume-to-surface ratios were obtained by a combination of point and intersection counting, which enabled estimates for the volume (Vcell) and plasma membrane surface area (SPM) of the ‘average’ cell within each stratum to be acquired. Using this information, it was then possible to calculate both the total surface area (S) and the number (N) of desmosomes on the plasma membranes of average cells. The parameters Ss and Ns showed progressive increases between basal and granular layers, whereas values for delta and s were lower in granular cells when compared with basal and spinous cells. The parameters Vcell, SPM, S and N all increased progressively and significantly during differentiation. Between basal and granular layers, the mean cell volume and surface area had each increased approximately threefold, whereas the surface area and number of desmosomes on the average cell plasma membrane had increased approximately seven- and eleven-fold, respectively. Granular cells thus possess more numerous desmosomes, which occupy a greater proportion of the plasma membrane area but which are individually smaller, when compared with basal and spinous layers.


1997 ◽  
Vol 200 (18) ◽  
pp. 2415-2423 ◽  
Author(s):  
A N Makanya ◽  
J N Maina ◽  
T M Mayhew ◽  
S A Tschanz ◽  
P H Burri

The extents of functional surfaces (villi, microvilli) have been estimated at different longitudinal sites, and in the entire small intestine, for three species of bats belonging to two feeding groups: insect- and fruit-eaters. In all species, surface areas and other structural quantities tended to be greatest at more cranial sites and to decline caudally. The entomophagous bat (Miniopterus inflatus) had a mean body mass (coefficient of variation) of 8.9 g (5%) and a mean intestinal length of 20 cm (6%). The surface area of the basic intestinal tube (primary mucosa) was 9.1 cm2 (10%) but this was amplified to 48 cm2 (13%) by villi and to 0.13 m2 (20%) by microvilli. The total number of microvilli per intestine was 4 x 10(11) (20%). The average microvillus had a diameter of 8 nm (10%), a length of 1.1 microns (22%) and a membrane surface area of 0.32 micron 2 (31%). In two species of fruit bats (Epomophorus wahlbergi and Lisonycteris angolensis), body masses were greater and intestines longer, the values being 76.0 g (18%) and 76.9 g (4%), and 73 cm (16%) and 72 cm (7%), respectively. Surface areas were also greater, amounting to 76 cm2 (26%) and 45 cm2 (8%) for the primary mucosa, 547 cm2 (29%) and 314 cm2 (16%) for villi and 2.7 m2 (23%) and 1.5 m2 (18%) for microvilli. An increase in the number of microvilli, 33 x 10(11) (19%) and 15 x 10(11) (24%) per intestine, contributed to the more extensive surface area but there were concomitant changes in the dimensions of microvilli. Mean diameters were 94 nm (8%) and 111 nm (4%), and mean lengths were 2.8 microns (12%) and 2.9 microns (10%), respectively. Thus, an increase in the surface area of the average microvillus to 0.83 micron 2 (12%) and 1.02 microns 2 (11%) also contributed to the greater total surface area of microvilli. The lifestyle-related differences in total microvillous surface areas persisted when structural quantities were normalised for the differences in body masses. The values for total microvillous surface area were 148 cm2g-1 (20%) in the entomophagous bat, 355 cm2g-1 (20%) in E. wahlbergi and 192 cm2g-1 (17%) in L. angolensis. This was true despite the fact that the insecteater possessed a greater length of intestine per unit of body mass: 22 mm g-1 (8%) versus 9-10 mm g-1 (9-10%) for the fruit-eaters.


1986 ◽  
Vol 102 (1) ◽  
pp. 289-297 ◽  
Author(s):  
M Maeda ◽  
G A Thompson

Dunaliella salina cells rapidly diluted from their normal 1.71 M NaCl-containing growth medium into medium containing 0.86 M NaCl swelled within 2--4 min to an average volume 1.76 X larger and a surface area 1.53 X larger than found in control cells. Morphometric analysis of thin section electron micrographs revealed that certain organelles, including the chloroplast, nucleus, and some types of vacuoles, also expanded in surface area as much or more than did the entire cell. It is likely that glycerol, the most important osmotically active intracellular solute, was present in high concentration within these organelles as well as in the cytoplasm itself. Thin section and freeze-fracture electron microscopy were utilized to trace the origin of membrane material whose addition permitted the large increase in plasma membrane surface area and the equally large growth of the chloroplast outer envelope. The findings indicated that the plasma membrane's expansion resulted from its selective fusion with numerous small (less than or equal to 0.25 micron diam) vesicles prevalent throughout the cytoplasm. In contrast, new membrane added to the chloroplast outer envelope was drawn from an entirely different source, namely, elements of the endoplasmic reticulum.


Membranes ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 538
Author(s):  
María Soto-Herranz ◽  
Mercedes Sánchez-Báscones ◽  
Juan Manuel Antolín-Rodríguez ◽  
Matías B. Vanotti ◽  
Pablo Martín-Ramos

Ammonia losses from manure pose serious problems for ecosystems and human and animal health. Gas-permeable membranes (GPMs) constitute a promising approach to address the challenge of reducing farm ammonia emissions and to attain the EU’s Clean Air Package goals. In this study, the effect of NH3-N concentration, membrane surface area, acid flux, and type of capture solution on ammonia recovery was investigated for a suspended GPM system through three experiments, in which ammonia was released from a synthetic solution (NH4Cl + NaHCO3 + allylthiourea). The effect of two surface areas (81.7 and 163.4 cm2) was first evaluated using three different synthetic N emitting concentrations (3000, 6000, and 12,000 mg NH3-N∙L−1) and keeping the flow of acidic solution (1N H2SO4) constant (0.8 L·h−1). A direct relationship was found between the amount of NH3 captured and the NH3-N concentration in the N-emitting solution, and between the amount of NH3 captured and the membrane surface area at the two lowest concentrations. Nonetheless, the use of a larger membrane surface barely improved ammonia capture at the highest concentration, pointing to the existence of other limiting factors. Hence, ammonia capture was then studied using different acid flow rates (0.8, 1.3, 1.6, and 2.1 L∙h−1) at a fixed N emitting concentration of 6000 mg NH3-N∙L−1 and a surface area of 122.5 cm2. A higher acid flow rate (0.8–2.1 L∙h−1) resulted in a substantial increase in ammonia absorption, from 165 to 262 mg of NH3∙d−1 over a 14-day period. Taking the parameters that led to the best results in experiments 1 and 2, different types of ammonia capture solutions (H2SO4, water and carbonated water) were finally compared under refrigeration conditions (at 2 °C). A high NH3 recovery (81% in 7 days), comparable to that obtained with the H2SO4 solution (88%), was attained when chilled water was used as the capture solution. The presented results point to the need to carefully optimize the emitter concentration, flow rate, and type of capture solution to maximize the effectiveness of suspended GPM systems, and suggest that chilled water may be used as an alternative to conventional acidic solutions, with associated savings.


1987 ◽  
Author(s):  
Mony M Frojmovic ◽  
Truman Wong ◽  
Jane Wylie ◽  
J G White

Osmotic swelling can double the external plasma membrane surface area of human platelets independently of size, proposed to recruit the open surface-connected canalicular system ((SCCS) (Thrombos. Res. Suppl. VI: 119, 1986). As bovine (B) platelets have been reported to lack SCCS, we compared osmotic swelling for B and human (H) cells. Addition of water to piatelet-rich-plasma (10-90% v/v) caused sequential shape change and osmotic spherocyte (OS) formation, analyzed for size and surface area changes from time-dependent phase-contrast videomicroscopic images. Selected samples were fixed and stained with tannic acid prior to osmic acid fixation for visualization of open SCCS by transmission electron microscopy. B platelets required 3-4x less water dilution of PRP than H platelets, with significant OS forming at 20% water addition. Continued water dilution converted 50% of platelets to OS, with maximally stable swelling and no significant lysis for bovine OS up to 60% dilution. Electron micrographs of unactivated discocytes (D) and of optimally-swollen OS showed open SCCS in human D not detectable in any of the swollen platelets, though granules, mitochondria and a small number of vesicles and vacuoles persisted; no evidence for any open SCCS was found for bovine D or OS, though the OS otherwise appeared similar to H-0S. Geometric measurements of D and nonlysed OS showed a stable, maximal 2.1±0.1 fold increase in external plasma membrane surface area with osmotic swelling, identical for different-sized H platelets (mean volume = 2.8-6.8 f1) or for B platelets (3.6 f1 ). B platelets show equal or greater sensitivity for ADP-induced activation as H platelets, with 2-fold slower maximal rates of recruitment in early aggregation. As osmotic swelling appears to primarily externalize SCCS in H platelets, the identical relative amounts of internal membrane externalized for B platelets is hypothesized to arise from an osmotically more labile, “closed”, and structurally simpler SCCS or from a distinct membrane source tnan in H platelets.


1985 ◽  
Vol 78 (1) ◽  
pp. 233-246 ◽  
Author(s):  
J.L. Merchant ◽  
D.S. Papermaster ◽  
R.J. Barrnett

During salt-water adaptation, an increase occurs in Na+,K+-ATPase content and surface area of the basolateral plasma membrane of the principal cell of the duck salt gland. To determine the degree to which these changes are correlated, accepted morphometric methods were used to determine numerical cell densities and plasma membrane surface densities of peripheral and principal cells. After adaptation, the plasma membrane surface area per principal cell was five times greater than in controls. Following de-adaptation, the plasma membrane content in principal cells returned to 1.9 times control levels. Two other cell constituents, mitochondria and lipid droplets, displayed similar quantitative changes. Na+,K+-ATPase content increased about fourfold with adaptation and decreased to near control levels with de-adaptation. Thus, changes in Na+,K+-ATPase content and basolateral plasma membrane surface area in adapting and de-adapting secretory epithelia of the salt gland occur nearly in parallel. These quantitative data enable Na+,K+-ATPase synthesis and degradation to be investigated in relation to membrane biogenesis.


Sign in / Sign up

Export Citation Format

Share Document