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D Cremaschi

Publications and source records attributed to D Cremaschi.

At least 37 records · Page 2Linked to original sources

Beta-agonist activation of an amiloride-insensitive transport mechanism in rabbit pleura.

The beta-agonist terbutaline increases the net rate of liquid absorption from hydrothoraces with albumin-Ringer solution: since beta-agonists decrease lymphatic drainage, the effect of terbutaline seems due to an increase in solute-coupled liquid absorption, (Zocchi et al. 1994 Respir. Physiol. 97:347-356). In this research we determined in anesthetized rabbits the rate of volume change in albumin-Ringer hydrothoraces of different size with amiloride plus terbutaline, and compared it with that previously obtained in hydrothoraces with amiloride alone. The net rate of liquid absorption was 0.09 ml/h greater (P < 0.01) with amiloride plus terbutaline than with amiloride alone. This indicates that terbutaline activates an amiloride-insensitive mechanism of Na+ transport. The increase in net rate of liquid absorption produced by terbutaline persisted with bumetanide 10(-6) M and SITS 10(-4) M, disappeared almost completely with bumetanide 10(-5) M, and completely with furosemide 10(-3) M. These findings suggest that the mechanism activated by terbutaline, when the amiloride-sensitive mechanisms of the pleura have been blocked, is a Na(+)-K(+)-2 Cl- or Na(+)-Cl- symport little sensitive to bumetanide.

Amiloride↗

Selective transport of microparticles across Peyer's patch follicle-associated M cells from mice and rats.

M cells are specialized structures in the Peyer's patch follicle-associated epithelium capable of taking up bacteria, viruses and other pathogens for later presentation to the gut-associated lymphoid tissue. The present work studies how coating microspheres with different proteins affects their ability to be taken up by M cells under near physiological conditions in vivo. The later appearance of microspheres in intestinal lymph has also been measured by flow cytometry. The protein preparations used in these experiments included bovine serum albumin (bSA), human immunoglobulin G (hIgG), secretory immunoglobulin A (hIgA), bovine growth hormone (bGH) and bGH complexed with an IgG antibody raised against bGH (bGH-Ab). Selectivity in binding of these microspheres to M cells, determined by confocal microscopy, was bGH < bSA < hIgG (mice) and bGH < bGH-Ab (rats and mice). A similar selectivity was seen for microsphere entry into M cells (bGH < bSA < hIgG; bGH < bGH-Ab). The appearance of protein-coated microspheres in rat mesenteric lymph showed a similar selectivity to that found for binding and entry into M cells (bGH < bGH-Ab). This latter selectivity was also found for hIgA-coated microspheres (bSA < hIgA). Preservation of transport selectivity throughout transcytosis highlights the unique importance of the M cell surface as being the primary site determining which type of antigen can be presented subsequently to the gut immune system. The possibility that this is a transient or phasic property of the M cell surface and that this could have physiological relevance is also discussed.

Animals↗

Hydrochlorothiazide enhances the apical Cl- backflux in rabbit gallbladder epithelium: radiochemical analysis.

Hydrochlorothiazide (HCTZ) was shown to inhibit the transepithelial NaCl transport and the apical Na(+)-Cl- symport and to depolarize the apical membrane potential in the rabbit gallbladder epithelium. The depolarization was likely related to the opening of a Cl- conductance. To better understand whether an apical Cl- leak is involved in the mechanism of action of HCTZ, the transapical Cl- backflux was measured radiochemically by the washout technique. The gallbladder wall, pretreated with pronase on the serosal side to homogenize the subepithelium, was loaded with 36Cl- on the luminal side; mucosal and serosal 36Cl- effluxes (Jm, Js) were then measured every 2 min. The pretreatment with pronase did not alter the membrane potentials and the selectivity of the epithelium. Under control conditions and the tissue in steady-state, Jm and Js time courses were each described by two exponential decays (A, B); the rate constants, kA and kB, were 0.71 +/- 0.03 and 0.16 +/- 0.01 min-1, respectively, and correspondingly the half-times (tA1/2, tB1/2) were 1.01 +/- 0.05 and 5.00 +/- 0.44 min (n = 10); these parameters were not significantly different for Jm and Js time courses. Js was always greater than Jm (Js/Jm = 2.02 +/- 0.22 and 1.43 +/- 0.17 for A and B decays). Under SCN- treatment in steady-state conditions, both Jm and Js time courses were described by only one exponential decay, the component B being abolished. Moreover tA1/2 was similar to that predictable for the subepithelium. It follows that it is the component B which exits the epithelial compartment. Based on the intracellular specific activity and 36Cl- JBm at 0 min time of the washout experiment, the cell-lumen Cl- backflux in steady-state was calculated to be equal to about 2 mumol cm-2hr-1, in agreement with the value indirectly computable by other techniques. The experimental model was well responsive to different external challenges (increases in media osmolalities; luminal treatment with nystatin). HCTZ (2.5 x 10(-4) M) largely increased 36Cl- JBm. The increase was abolished by luminal treatment with 10(-4) M SITS, which not only brought back the efflux time courses to the ones observed under control conditions but even increased Js/Jm of the cellular component, an indication of a reduced JBm. It is concluded that HCTZ opens an apical, SITS-sensitive Cl- leak, which contributes to dissipate the intracellular Cl- accumulation and to inhibit the NaCl transepithelial transport.(ABSTRACT TRUNCATED AT 400 WORDS)

4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfo↗

Stimulation of Cl- secretion by the mucoactive drug S-carboxymethylcysteine-lysine salt in the isolated rabbit trachea.

Ion transport by the airway epithelium contributes to the regulation of the quantity and composition of respiratory tract fluid, thereby affecting mucociliary clearance. We have investigated the effect of the mucoactive drug S-carboxymethylcysteine-lysine salt (S-CMC-Lys) on the transepithelial bioelectric properties of isolated rabbit trachea. Transepithelial potential difference (Vms), short-circuit current (Isc) and resistance (R) were measured in the isolated rabbit trachea mounted between flux half-chambers, in the presence and in the absence of S-CMC-Lys (100 microM), added to the mucosal or submucosal chamber. In some experiments, tissues were also exposed to ion channel-inhibitors, in order to evaluate the contribution of Na+ and Cl- active transport to Isc. The excised rabbit trachea expressed transepithelial bioelectric properties based on an active ion transport supported by the Na(+)-K(+)-adenosine triphosphatase (ATPase) activity, since ouabain (500 microM) completely abolished the transepithelial potential difference. In control preparations, Vms and Isc declined significantly during 300 min recording, whereas R remained constant. The Isc decline was essentially attributable to a decrease in Cl- transport. Bumetanide (100 microM) almost completely abolished the Isc fraction related to Cl- transport. Treatment of the tissues with S-CMC-Lys reduced the progressive fall in Isc, with the most clear-cut and significant effect observed for the mucosal treatment. In parallel, S-CMC-Lys significantly lowered R, without affecting Vms. Either mucosal or submucosal exposure to S-CMC-Lys significantly increased Cl- secretion to normal values, whilst Na+ absorption was not modified.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Nature of the neutral Na(+)-Cl- coupled entry at the apical membrane of rabbit gallbladder epithelium: IV. Na+/H+, Cl-/HCO3- double exchange, hydrochlorothiazide-sensitive Na(+)-Cl- symport and Na(+)-K(+)-2Cl- cotransport are all involved.

Transepithelial fluid transport was measured gravimetrically in rabbit gallbladder (and net Na+ transport was calculated from it), at 27 degrees C, in HCO(3-)-free bathing media containing 10(-4) M acetazolamide. Whereas luminal 10(-4) M bumetanide or 10(-4) M 4-acetamido-4'-iso-thiocyanostilbene-2,2'-disulfonate (SITS) did not affect fluid absorption, 25 mM SCN- abolished it; hydrochlorothiazide (HCTZ) in the luminal medium reduced fluid absorption from 28.3 +/- 1.6 (n = 21) to 8.6 +/- 1.6 microliters cm-2 hr-1 (n = 10), i.e., to about 30%. This maximum effect was already obtained at 10(-3) M concentration; the apparent IC50 was about 2 x 10(-4) M. The residual fluid absorption, again insensitive to SITS, was completely inhibited by SCN- or bumetanide. Cl- influx at the luminal border of the epithelium, measured under the same conditions and corrected for the extracellular space and paracellular influx, proved insensitive to 10(-4) M bumetanide, but was slowly inhibited by 10(-3) M HCTZ, with maximum inhibition (about 54%) reached after a 10-min treatment; it subsequently rose again, in spite of the presence of HCTZ. However, if the epithelium, treated with HCTZ, was exposed to 10(-4) M bumetanide during the measuring time (45 sec), inhibition was completed and the subsequent rise of Cl- influx eliminated. Intracellular Cl- accumulation with respect to the predicted activity value at equilibrium decreased significantly upon exposure to 10(-3) M HCTZ, reached a minimum within 15-30 min of treatment, then rose again significantly at 60 min. Simultaneous exposure to HCTZ and bumetanide decreased the accumulation to a significantly larger extent as compared to HCTZ alone, already in 15 min, and impeded the subsequent rise. Intracellular K+ activity rose significantly within 30 min treatment with HCTZ; the increase proved bumetanide dependent. The results obtained show that Na(+)-Cl- symport, previously detected under control conditions, is the HCTZ-sensitive type; its inhibition elicits bumetanide-sensitive Na(+)-K(+)-2Cl- cotransport. Thus, the three forms of neutral Na(+)-Cl(-)-coupled transport so far evidenced in epithelia, Na+/H+, Cl-/HCO3- double exchange (in the presence of exogenous bicarbonate), HCTZ-sensitive Na(+)-Cl- symport and bumetanide-sensitive Na(+)-K(+)-2Cl- cotransport, are all present in the apical membrane of rabbit gallbladder.

Animals↗

Confocal microscopical analysis of epithelial cell heterogeneity in mouse Peyer's patches.

Cyanine dye fluorescence and alkaline phosphatase activities have been compared directly by confocal microscopy in a wide variety of cells present in the follicle-associated epithelium of the mouse Peyer's patch to test the hypothesis that antigen-transporting M cells have a low membrane potential. In order to make these comparisons it was first necessary to equilibrate living tissue with the membrane potential sensitive dye DIOC5(3), fix with glutaraldehyde and then incubate the fixed tissue with naphthol AS-BI phosphate, a substrate which is hydrolysed by alkaline phosphatase present in the luminal membrane of these epithelial cells. Naphthol AS-BI produced by this reaction is then coupled to Fast Red TR diazonium salt at the site of hydrolysis. Selecting the 488 nm wavelength of the argon laser source then allows one to measure alkaline phosphatase activities as Fast Red absorbance and membrane potentials by DIOC5(3) fluorescence. Results obtained show a linear correlation between membrane potential and alkaline phosphatase activity. Relative lack of alkaline phosphatase activity, determined in fixed tissue, has been used previously to identify antigen-transporting M cells (Smith et al., 1987). The present work shows that it is now possible to recognize these cells in living tissue by measurement of DIOC5(3) fluorescence. The possible importance of this finding in providing a way to study cell surface-antigen interactions taking place in living tissue is discussed.

Alkaline Phosphatase↗

Liquid volume, Na+ and mannitol concentration in a hypertonic mannitol-Ringer hydrothorax.

In anesthetised rabbits with a 2 ml hypertonic mannitol-Ringer hydrothorax in the right space 30 mM/L mannitol were required for an unchanged volume of the hydrothorax after 60 min. [Na+] in the pleural liquid 10, 30 and 60 min after this hydrothorax was 8, 7 and 5 mEq/L, respectively, lower (P less than 0.01) than the initial one and that in a Ringer-hydrothorax. This seems due to the active transport of Na+ out of the pleural space followed by little water because of the osmotic pressure exerted by mannitol. This finding provides further evidence for an active transport without using inhibitors, and implies that the mesothelium offers an appreciable resistance to small solute diffusion. Mannitol concentration, measured at corresponding times from the activity of labeled mannitol, was 76, 68 and 56%, respectively, of the initial one (24.5 mM/L). From 30 to 60 min 6.5 microM of mannitol left the right space mainly by diffusion. The diffusional permeability of the mesothelium was indirectly assessed from the diffusional outflux of mannitol, the surface of the pleural space, and an estimate of mannitol concentration in the interstitium next to the mesothelium: it is smaller than that found in vitro.

Animals↗

Confocal analysis of fluorescent bead uptake by mouse Peyer's patch follicle-associated M cells.

Latex beads coated with secretory immunoglobulin (IgA) instilled into mouse intestinal loops are taken up by M cells present in Peyer's patch follicle-associated epithelial tissue. Bead adsorption and uptake is greater at the edge compared with the apex of follicle domes. Coating beads with bovine serum albumin (BSA) causes a fourfold reduction in adsorption and a twentyfold reduction in uptake. Results demonstrate selectivity between adsorption and uptake and between the ability of different proteins to facilitate uptake.

Adsorption↗

Active transport of polypeptides in rabbit nasal mucosa: possible role in the sampling of potential antigens.

Transepithelial pathways of macromolecule transport have been studied in vitro in rabbit nasal respiratory mucosa, maintained at 27 degrees C. Transepithelial electrical potential difference, short-circuit current and resistance were 3.4 +/- 0.5 mV (submucosa positive), 65.0 +/- 6.7 microA cm-2 and 52.1 +/- 5.6 omega cm-2 respectively (n = 15). These electrical characteristics are those of a leaky epithelium allowing macromolecules to permeate paracellularly. A detailed permeation study of a polypeptide (elcatonin, Mw = 3362) was also undertaken. Elcatonin mucosa-submucosa (Jms) and submucosa-mucosa (Jsm) fluxes were measured by radioimmunoassay. With 10 micrograms/ml elcatonin, Jms was significantly larger than Jsm for the whole 120-min period of observation; net flux showed a maximum in the first 30 min (Jms = 13.6 +/- 1.0 ng cm-2 h-1, Jsm = 1.4 +/- 0.1 ng cm-2 h-1, n = 10). Jms fell towards the value of Jsm if the temperature was reduced to 4 degrees C or if the mucosa was simultaneously treated with 0.1 mM dinitrophenol and 3 mM monoiodoacetate. Jms and Jnet followed saturation kinetics with increasing elcatonin concentrations. Adrenocorticotropic hormone (Mr = 4500) produced a similar pattern to elcatonin. However, Jms and Jsm were not significantly different from each other at any time either for [3H]sucrose (Mw = 342) or for [14C]polyethyleneglycol-4000 (Mw = 4000) when present in the bathing medium at 500 microM concentration. The results show active transport of polypeptides in parallel with passive permeation (possibly through leaky intercellular junctions). Active transport does not appear to be related to nonspecific pinocytosis but to receptor-mediated endocytosis.(ABSTRACT TRUNCATED AT 250 WORDS)

Adrenocorticotropic Hormone↗

Electrolyte transport across the pleura of rabbits.

The amounts of Na+ and Cl- in the right pleural space of anesthetized rabbits were determined 10 and 60 min after a 2 ml hydrothorax with the following solutions: Ringer, Ringer with an inhibitor of the Na(+)-Cl- coupled transport or of the Na+/K+ pump, Ringer with gluconate instead of Cl- or with methylglucamine instead of Na+. During the 10-60 min period: (a) with Ringer Na+ and Cl- decreased (P less than 0.01) along with an iso-osmotic liquid absorption, (b) with disulfonic-stilbene (0.1 mM), amiloride (0.7 mM), acetazolamide (0.1 mM), or ouabain (0.5 mM) Na+ did not change and Cl- decreased less (P less than 0.01) than with Ringer. With gluconate-Ringer or methylglucamine-Ringer the liquid flow reversed: in the former case Cl- and, to a smaller extent, Na+ increased (P less than 0.01); in the latter only Na+ increased (P less than 0.01). These findings suggest: (1) the occurrence of a Na+/H+ and Cl-/HCO3- double exchange on the serosal side and of a Na+/K+ pump on the interstitial side of the pleural mesothelium; (2) a slow efflux from the pleural space of gluconate or methylglucamine relative to the corresponding influx of Cl- or Na+, respectively; this drags liquid into the space by osmotic gradient.

Acetazolamide↗

Transepithelial electrophysiological parameters in rabbit respiratory nasal mucosa isolated in vitro.

1. Transepithelial electrical p.d. (Vms), short circuit current (Isc) and transepithelial resistance (Rep) were determined in rabbit nasal mucosa and were compared with the equivalent parameters of human nasal mucosa. 2. Vms was about 1 mV (submucosa positive) in the absence of glucose, but increased continuously in the presence of glucose. Since Isc also increased in parallel and Rep remained constant (about 40 omega cm2), glucose effect was to power pumping. 3. Diffusion potentials raised by reducing luminal Cl- or Na+ or Cl- and Na+ concentrations were also measured and compared with values obtained in trachea and gallbladder. 4. Evidence is produced, with sounder basis than in trachea, that junctional pathways in airway epithelia are lined with fixed positive charges (which make anions more permeable than cations), unlike junctional pathways of the gastrointestinal tract which are lined with fixed negative charges.

Animals↗

The nature of the neutral Na(+)-Cl- coupled entry at the apical membrane of rabbit gallbladder epithelium: III. Analysis of transports on membrane vesicles.

In rabbit gallbladder epithelium, a Na+/H+, Cl-/HCO3- double exchange and a Na(+)-Cl- symport are both present, but experiments on intact tissue cannot resolve whether the two transport systems operate simultaneously. Thus, isolated apical plasma membrane vesicles were prepared. After preloading with Na+, injection into a sodium-free medium caused a stable intravesicular acidification (monitored with the acridine orange fluorescence quenching method) that was reversed by Na+ addition to the external solution. Although to a lesser extent, acidification took place also in experiments with an electric potential difference (PD) equal to 0. If a preset pH difference (delta pH) was imposed [( H+]in greater than [H+]out, PD = 0), the addition of Na-gluconate to the external solution caused delta pH dissipation at a rate that followed saturation kinetics. Amiloride (10(-4) M) reduced the delta pH dissipation rate. Taken together, these data indicate the presence of Na+ and H+ conductances in addition to an amiloride-sensitive, electroneutral Na+/H+ exchange. An inwardly directed [Cl-] gradient (PD = 0) did not induce intravesicular acidification. Therefore, in this preparation, there was no evidence for the presence of a Cl-/OH- exchange. When both [Na+] and [Cl-] gradients (outwardly directed, PD = 0) were present, fluorescence quenching reached a maximum 20-30 sec after vesicle injection and then quickly decreased. The decrease was not observed in the presence of a [Na+] gradient alone or the same [Na+] gradient with Cl- at equal concentrations at both sides. Similarly, the decrease was abolished in the presence of both Na+ and Cl- concentration gradients and hydrochlorothiazide (5 x 10(-4) M). The decrease was not influenced by an inhibitor of Cl-/OH- exchange (10(-4) M furosemide) or of Na(+)-K(+)-2Cl- symport (10(-5) M bumetanide). We conclude that a Na+/H+ exchange and a Na(+)-Cl- symport are present and act simultaneously. This suggests that in intact tissue the Na(+)-Cl- symport is also likely to work in parallel with the Na+/H+ exchange and does not represent an induced homeostatic reaction of the epithelium when Na+/H+ exchange is inhibited.

Amiloride↗

Chloride conductance and intracellular chloride accumulation in mouse Peyer's patch enterocytes.

1. Measurements of membrane potential, Cl- conductance and intracellular Cl-, K+, Na+ and H+ activities have been carried out in the follicle-associated epithelium of the mouse Peyer's patch to characterize further the physiological properties of antigen-transporting M cells, enterocytes and intraepithelial lymphocytes. 2. Intraepithelial lymphocytes, identified in random impalements as a second negative jump in membrane potential (Vm), were found to have higher K+ and H+ activities, lower Na+ and Cl- activities and higher negative values for Vm than either of the other two epithelial cell types. Cl- activity in these cells was higher than that predicted from the Nernst equation. 3. M cells identified as having the first negative jump in Vm in impalements involving intraepithelial lymphocytes were unable to accumulate Cl-. They also had a lower Vm than that found in apparently fully differentiated enterocytes. This Vm was partly depolarized at a low Cl- concentration. 4. Apparently fully differentiated enterocytes could be further divided into two populations depending on whether the Vm could or could not be partly depolarized by a low Cl- concentration. The mean Vm and intracellular Cl- activity of Cl(-)-sensitive cells, measured in high Cl- medium, were less than values found for Cl(-)-insensitive enterocytes. Cl- was only accumulated by enterocytes showing no depolarization at low external Cl- concentration. 5. The Vm of mature villus enterocytes was the same as that determined for Cl(-)-insensitive follicle-associated enterocytes. Villus enterocyte Vm was not depolarized at a low Cl- concentration. 6. The present ability to distinguish two apparently large populations of follicle-associated enterocytes having M cell-like or villus enterocyte-like properties is discussed in relation to current theories describing how M cells might be formed. The presence of a Cl- conductance in M cell enterocytes, the inability to accumulate Cl- and the maintenance of a low Vm could aid endocytosis of macromolecules across the apical membrane.

Animals↗

Brush-border membrane alkaline phosphatase activity in mouse Peyer's patch follicle-associated enterocytes.

1. Using a new method of quantitative cytochemistry to determine alkaline phosphatase activity, it was possible to identify M cells and distinguish two further types of enterocytes present in mouse Peyer's patch follicle-associated epithelial tissue. 2. A quantitative description of these three cell types was provided by making direct absorbance measurements of enzyme reaction product on the luminal surface of individual follicle-associated enterocytes. M cells having a negligible alkaline phosphatase activity had a mean absorbance of 6 +/- 2 arbitrary units. The other two populations of enterocytes had overlapping but significantly different mean absorbances of 20 +/- 2 and 30 +/- 3 arbitrary units. 3. M cells accounted for 3.2 +/- 1% of the total number of cells present in the apical part of the follicle-associated epithelium. This percentage is a minimum value describing an extreme example of cells having M cell-like characteristics. The remaining epithelial cells were divided equally between those having high and low alkaline phosphatase activities in their brush-border membranes. A small percentage of cells having low alkaline phosphatase activities could perform M cell functions. 4. Results obtained suggest that reduced alkaline phosphatase activity and brush-border membrane Cl- conductance can both be used as differentiation markers to identify a large population of poorly differentiated enterocytes present in mouse follicle-associated epithelial tissue.

Alkaline Phosphatase↗

Measurement with microelectrodes of intracellular Na+, K+, H+ and Cl- activities and of membrane potential in normal rat liver slices and during the 4-dimethylaminoazobenzene-induced rat hepatocarcinogenesis.

Steady-state membrane potentials (Vm) and intracellular Na+ (aiNa), K+ (aiK), H+ (aiH) and Cl- (aiCl) activities were measured with double-barrelled ion-selective microelectrodes in liver slices from normal rats and during the 4-dimethylaminoazobenzene-induced (DAB) hepatocarcinogenesis. Rats fed with the experimental regimen without the carcinogen were used as control animals. In Krebs-Henseleit bicarbonate saline containing 5.5 mM glucose as bathing solution at 37 degrees C, Vm was found to be significantly lower in neoplastic hepatocytes, compared to normal liver cells. Vm decreased also in control rat liver cells. Increased Na+/K+ ratios and Na+ + K+ activities were found in cancerous hepatocytes whereas H+ and Cl- activities decreased. Therefore, the intracellular pH increased significantly in neoplastic cells, compared to normal and control cells. This could reflect activation of the Na+/H+ exchange system during the DAB-induced hepatocarcinogenesis, leading to a stimulation of cell metabolism with increased rate of protein and DNA synthesis and loss of growth control, under these conditions.

Animals↗

Effect of [D-Ala2,MePhe4,Gly-ol5]enkephalin and [D-Ala2,D-Leu5]enkephalin on ion and amino acid transport in rabbit ileum.

The selective mu-opioid agonist [D-Ala2,MePhe4,Gly-ol5]enkephalin (DAGO) (10 microM) reduced the short circuit (Isc) and the L-valine induced increase of the transepithelial potential difference and Isc(delta Vms and delta Isc) measured in-vitro in rabbit ileum, with a mechanism antagonized by naloxone (1 microM). [D-Ala2,D-Leu5]enkephalin (DADLE) (10 microM) had no significant effect on the transepithelial potential difference (Vms), Isc, delta Vms and delta Isc. In the ileum deprived of the serosa and muscolaris, DAGO reduced the delta Vms and delta Isc, but not the Vms and Isc, suggesting localization of the receptors responsible for this latter effect in the myenteric plexus and/or the muscularis mucosae. These preliminary results suggest that in the rabbit ileum opioids influence electrolyte and amino acid transport and these effects may be at least partly mediated by mu-receptors.

Amino Acids↗

Enterocyte-lymphocyte interactions in the follicle-associated epithelium of the mouse Peyer's patch.

1. Measurements of membrane potential (Vm) have been carried out in the follicle-associated epithelium of the mouse Peyer's patch to investigate possible site-dependent anomalies in enterocyte development. Initial heterogeneity in Vm values obtained from carrying out random impalements in the upper third of the follicle-associated epithelium could be described as arising from the presence of three different cell populations. 2. The predominant cell type in this epithelium (T1) had a mean Vm of -35.3 mV which was partly depolarized by increasing concentrations of K+. These cells were not stained with the vital dye Neutral Red. Two other types of cell (T2L and T2H) were recorded together as consecutive negative jumps in Vm, but only in areas of the epithelium previously stained with Neutral Red. T2L behaved like T1 cells to K+ but with a low Vm of -23.3 mV. T2H cells had a high mean Vm of -47.5 mV showing transient hyperpolarization to increasing K+ concentration. 3. The electrophysiological and non-staining properties of T1 cells are similar to those expected from mature enterocytes. The anatomical size, site, staining properties and Vm of T2H cells correspond to those expected for intraepithelial lymphocytes. The low Vm, K+ depolarization and close association of T2L with T2H cells are typical of properties predicted for an immature-type of antigen-transporting enterocyte known to be present in this type of epithelium. 4. The possibility that intraepithelial lymphocytes either slow or reverse a more normal process of development of enterocytes in their immediate vicinity is discussed along with wider aspects of enterocyte-lymphocyte interactions leading to the transfer to enteric antigens across these modified cells.

Animals↗