The effects of thermally-induced activity in vivo upon the ultrastructure and Na, K and Cl composition of the epithelial cells of sweat glands from patients with cystic fibrosis

The secretory cells of the fundus of sweat glands from cystic fibrosis (CF) patients had higher Na and Cl contents and showed more granule depletion, cellular disruption and dilated intercellular canaliculi than normal. The cells of the coiled duct also had higher cytoplasmic levels of Na and Cl but were structurally normal. Thermal stimulation produced ultrastructural changes in the CF fundus comparable to normal, including further dilatation of the basolateral clefts, but did not induce the marked changes in the coiled duct which normally occur. The elevated Na and fall in K in the fundus and raised Na and Cl in the coiled duct upon activation, were not observed in the CF glands in which no significant changes were detected.     

Some Effects of Root Anatomy on K, Na and Cl Loading of Citrus Roots and Leaves

Loading of K, Na and Cl into fibrous roots of salt-treated citrusgenotypes, Rangpur lime (Citrus reticulata var. austera hybrid?)and Etrog citron (C. medica L.) was investigated in relationto root anatomy, in particular, the differentiation of the epidermal-hypodermallayers with distance from the root tip. The influence of durationof salinity treatment on the characteristics of K, Na and Claccumulation in leaves of the two genotypes was explored intwo experiments respectively, covering the short term (14 d)and long term (12 weeks). This study focused on two regions of the fibrous root, a segment2–12 mm from the root tip, immediately basipetal to thezone of elongation and a differentiated region of the maturesuberized, fibrous root, 40–50 mm from the root tip. Inthe distal root segment (2–12 mm) the epidermis and hypodermisof both genotypes was observed as two closely packed, uniseriatelayers of living cells. In the proximal root segment (40–50mm) the differentiated hypodermis was evident as a uniseriatelayer of thick-walled lumina interspersed with ‘passagecells’ which were frequently associated with clustersof viable epidermal cells. The characteristics of Na and Cl loading in the two root zonesdiffered profoundly during the short term loading (acclimation)phase. Attainment of quasi-steady-states for Na and Cl in thedistal region (with the exception of Na in Etrog citron) wasrapid as was Na equilibration of the proximal root segmentsin both genotypes. In contrast, Cl loading in the proximal regiontook c. 14 d to reach a quasi-steady-state by which time Cllevels were 2 to 3 times higher in the proximal than in thedistal root segments. The superior tolerance of Rangpur lime to long term salinitywas highly correlated to Cl exclusion from the leaves. However,during the first 14 d of acclimation to 50 mol m^–3 NaClthere was no segregation of the two genotypes based upon leafCl levels. Expression of differential accumulation of Cl inleaves appeared to be a time dependent process and was manifestonly after Cl saturation of the proximal root which representsthe bulk of the fibrous root system. The salt tolerance of Rangpurwas also associated with high selectivity of fibrous roots forK. over Na. A pronounced loss of K from cortical cells in theproximal root segment of salt-stressed Etrog citron was alsoevident by X-ray microanalysis. Key words: Citrus, anatomy, salinity, roots, X-ray microanalysis     

Effects of insulin on Na and K transporters in the rat CCD

We tested the effects of insulin (2 nM, 30-60 min) on principal cells of isolated split-open rat cortical collecting ducts (CCD) using whole-cell current measurements. Insulin addition to the superfusate of the tubules enhanced Na pump (ouabain-sensitive) current from 18 ± 3 to 31 ± 3 pA/cell in control and from 74 ± 9 to 126 ± 11 pA/cell in high K-fed animals. It also more than doubled ROMK (tertiapin-Q-sensitive) K(+) currents in control CCD from 320 ± 40 to 700 ± 80 pA/cell, although it did not affect this current in tubules from K-loaded rats. Insulin did not induce the appearance of amiloride-sensitive Na(+) current in control animals, while in high K-fed animals the currents were similar in the presence (140 ± 30) and the absence (180 ± 70 pA/cell) of insulin. Intraperitoneal injection of insulin plus hypertonic dextrose decreased Na excretion, as previously reported. However, injection of dextrose alone, or the nonmetabolized sugar mannose, had similar effects, suggesting that they were largely the result of vascular volume depletion rather than specific actions of the hormone. In summary, we find no evidence for acute upregulation of the epithelial Na channel (ENaC) by physiological concentrations of insulin in the mammalian CCD. However, the hormone does activate both the Na/K pump and apical K(+) channels and could, under some conditions, enhance renal K(+) secretion.     

Distribution of Na+, K+ and Cl− between nucleus and cytoplasm inChironomus salivary gland cells

Summary Previous studies of the electrochemical activity coefficients of intracellular Na⁺ and K⁺ have suggested that the free form of these ions may be unevenly distributed within the intracellular fluids. One possible site of such subcellular compartmentalization is the cell nucleus. In order to examine this possibility, the cells ofChironomus salivary glands were studied with conventional liquid ion-exchange microelectrodes sensitive to K⁺ and Cl^–, with a new liquid ion-exchange microelectrode sensitive to Na⁺, and with open-tipped micropipets. Both the electrochemical activities for Na⁺, K⁺ and Cl^–, and the electrical potential were the same on both sides of the nuclear membrane. The possibility was considered that a difference in the junction potentials within the nucleoplasm and cytoplasm might have masked a real difference in electrical potential between these two phases. To study that possibility, changes were induced in the junction potentials by altering the composition of the fluid filling the exploring micropipets. The results have suggested that the magnitudes of the junction potentials are the same on both sides of the nuclear envelope. The simplest interpretation of the data is that the chemical activities of Na⁺, K⁺ and Cl^– are the same within the nucleus and cytoplasm. This suggests that other subcellular structures, possibly the endoplasmic reticulum and mitochondria, are responsible for subcellular compartmentalization.     

Effects of saliva from chronically reserpinized rat on Na and K transport in perfused main excretory duct of submandibular gland of normal rat

Reserpine (RES) (0.5 mg/kg body wt, ip) was administered to rats for 7 days. On Day 8 saliva was evoked from these animals by intraperitoneal injection of pilocarpine nitrate (10 mg/kg body wt) and saliva from submandibular and parotid glands was collected separately. These collected salivas were used to perfuse through the main ducts of the submandibular glands of normal rats. After a control period of perfusion of the main duct with bicarbonate saline solution, parotid saliva from RES rats was perfused through the duct followed by regular perfusion. There was inhibition of Na absorption (22%) and K secretion (23%). Moreover, when submandibular saliva from treated rat was perfused through the main duct prior to regular perfusion, there was a decrease in Na absorption (31%) and K secretion (28%). In contrast, perfusion of the main duct with either parotid or submandibular saliva from normal rats caused no significant changes in Na and K transport. The present experiments confirm previous studies that there is some Na-inhibitory factor(s) present in saliva of the chronically RES-treated rat.     

Effects of short-term exercise in the heat on thermoregulation, blood parameters, sweat secretion and sweat composition of tropic-dwelling subjects

This study investigates the effects of a short-term aerobic training program in a hot environment on thermoregulation, blood parameters, sweat secretion and composition in tropic-dwellers who have been exposed to passive heat. Sixteen healthy Malaysian-Malay male volunteers underwent heat acclimation (HA) by exercising on a bicycle ergometer at 60% of VO2max for 60 min each day in a hot environment (Ta: 31.1+/-0.1 degrees C, rh: 70.0+/-4.4%) for 14 days. All parameters mentioned above were recorded on Day 1 and at the end of HA (Day 16). On these two days, subjects rested for 10 min, then cycled at 60% of VO2max for 60 min and rested again for 20 min (recovery) in an improvised heat chamber. Rectal temperature (Tre), mean skin temperature (Tsk) heart rate (HR), ratings of perceived exertion (RPE), thermal sensation (TS), local sweat rate and percent dehydration were recorded during the test. Sweat concentration was analysed for sodium [Na+]sweat and potassium. Blood samples were analysed for biochemical changes, electrolytes and hematologic indices. Urine samples were collected before and after each test and analysed for electrolytes.After the period of acclimation the percent dehydration during exercise significantly increased from 1.77+/-0.09% (Day 1) to 2.14+/-0.07% (Day 16). Resting levels of hemoglobin, hematocrit and red blood cells decreased significantly while [Na+]sweat increased significantly. For Tre and Tsk there were no differences at rest. Tre, HR, RPE, TS, plasma lactate concentration, hemoglobin and hematocrit at the 40th min of exercise were significantly lower after the period of acclimation but mean corpuscular hemoglobin and serum osmolality were significantly higher while no difference was seen in [Na+]sweat and Tsk. It can be concluded that tropic-dwelling subjects, although exposed to prolonged passive heat exposure, were not fully heat acclimatized. To achieve further HA, they should gradually expose themselves to exercise-heat stress in a hot environment.     

The effect of an endogenous Na+, K+-ATPase inhibitor on rat lens transparency and ultrastructure

1. The purpose of the present study was to analyze the possible effect of ouabain and an endogenous ouabain-like substance (endobain E), on lenses of 100- and 400-g body weight rats.2. Lenses were incubated with ouabain or endobain E for 120 min, either at room temperature or in the cold; opalescence was checked by gross examination and ultrastructure by electron microscopy.3. Lenses from 400-g rats invariably remained translucent whereas those from 100-g rats presented variable opalescence.4. As disclosed with the electron microscope, lenses of 100-g rats incubated at room temperature, with or without ouabain or endobain E, presented variable degrees of ultrastructural changes: with ouabain, there was fiber separation and vacuole formation but with endobain E, no vacuoles were found and fibers, though disorganized, appeared attached. After incubation in an ice bath, lenses were markedly altered in all conditions assayed.5. It is concluded that ouabain and endobain E effect on lens transparency depends on the rat age and that in young animals, it is crucial incubation temperature during experimental procedure.     

Effect of angiotensin on the ultrastructure of the rat thyroid gland

Four hours after administering angiotensin to rats accumulation of radioactive iodine by the thyroid glands is inhibited. Angiotensin produces the contraction of thyroid arterioles. The cytoplasm of endotheliocytes of the perifollicular exchange microvessels becomes dense and luminar surfaces very involuted. Administration of angiotensin also leads to the changes in the ultrastructure of thyrocytes. The cisternae of the granular endoplasmic network get enlarged, the number of membrane-associated ribosome and mitochondria diminishes, that of lysosomes increases.     

Effects of vasoactive intestinal polypeptide on acetylcholine stimulation of rat submandibular gland

Studies were carried out on the role of vasoactive intestinal polypeptide (VIP) in the regulation of secretion and blood flow in the rat salivary gland. The first experiments to investigate the spontaneous secretory pattern revealed a clear diurnal fluctuation with a significant increase at night, so that the subsequent experiments were performed during the daytime where the secretion was consistently low. Intravenous administration of VIP at a dose smaller than 40 pmole caused a dose-dependent vasodilatory response, but at a high dose such a local effect was hampered by a decrease in systemic blood pressure. VIP potentiated the acetylcholine chloride (AcCho)--evoked salivary secretion, but VIP (0-100 pmole/kg) alone did not cause salivary secretion. Atropine reduced the salivary secretion evoked by AcCho and VIP, and the blood flow change evoked by AcCho. However, the blood flow change evoked by VIP was not affected by atropine. Hexamethonium exerted no significant effect on the response to administration of AcCho or VIP. The results indicate that VIP has a significant vasodilatory action and cooperates with AcCho in the regulation of salivary secretion in the rat, and VIP effects are atropine resistant, as in other species of animals.     

Effects of bromocriptine on sweat gland function during heat acclimatization

This study examined the involvement of the hormones aldosterone and prolactin in sweat gland function during heat acclimatization. Two groups of male subjects (n = 8) were tested - one receiving a placebo (control), the other receiving bromocriptine. Both groups performed cycle ergometer exercise at 50% of maximal oxygen uptake over 10 consecutive days in an environmental chamber maintained at 39 degrees C and 30% relative humidity. Duration of exercise was 90 min on days 2-4 and 6-9, and 45 min on test days 1, 5 and 10. Electrolyte concentrations were determined by total body washdown. Prolactin increased (p less than 0.001) during exercise on day 1 in the control group but not on days 5 and 10. In contrast, prolactin was suppressed by bromocriptine and did not rise in response to exercise or heat exposure. Plasma aldosterone increased during exercise in both groups, showing no differences between groups. The sodium concentration in sweat decreased significantly (p less than 0.05) in the control group from day 1 to 10 but was unchanged in the treatment group. These data suggest that acclimatization-related changes in sweat gland function may be attenuated by increases in central dopaminergic activity and implicate prolactin in control of sweat gland function.     

Cyclic AMP-dependent stimulation of Na,K-ATPase in shark rectal gland

Summary Scatchard analysis of³H ouabain bound to isolated rectal gland cells as a function of increasing ouabain concentrations produced a concave curvilinear plot that was resolved into two specific sites with either a high (I) or low (II) affinity for ouabain. Cyclic cAMP/theophylline (±furosemide, 10^–4 m) increased the amount of³H ouabain bound to the high-affinity site I. Vanadate, a phosphate congener which promotes formation of the ouabain-binding state of the enzyme, mimicked the effects of cAMP/theophylline at low concentrations of ouabain, suggesting that cAMP/theophylline increases binding to site I by enhancing the rate of turnover of resident enzyme. Enhanced⁸⁶Rb uptake seen following cAMP/theophylline administration was primarily associated with increased flux through the high-affinity ouabain site, and this stimulation was not obliterated by the co-administration of furosemide. A model was presented which suggested the presence of two noninteracting pools of enzyme or isozymes which exhibit either a high or low affinity for ouabain. Cyclic AMP both stimulated turnover via site I, and modified the kinetics of binding of³H ouabain to site II. The (ave)K _d of³H ouabain for site II was increased from 3.6 m (controls) to 0.5 m (cAMP/theophylline) and the Hill coefficient was modified from 0.45 (controls) to 1.12 (caMP/theophylline), suggesting a transition from a negative- to a noncooperative binding state. While furosemide reversed the effects of cAMP/theophylline on site II kinetics, it did not obliterate cAMP/theophylline effects on site I. This suggests that cAMP may alter the intrinsic turnover rate of this particular pool of Na,K-ATPase in shark rectal gland.     

Effects of cadmium on the ultrastructure of the mouse parathyroid gland

Electron-microscopic studies were made on the parathyroid gland of cadmium-treated mice. Most chief cells in treated mice were rich in free ribosomes and secretory granules compared to the control mice. In the parathyroid gland after 3 and 4 weeks of administration of cadmium, interdigitations between adjacent chief cells became more complex than in the control mice. In most chief cells of the parathyroid gland after 2, 3 and 4 weeks of administration of cadmium, the Golgi complexes associated with numerous prosecretory granules were better developed than in the control mice. These ultrastructural appearances suggest that parathyroid gland cellular activity was stimulated in response to cadmium treatment.     

Effects of ionizing radiations on the ultrastructure of the thymus gland

Different doses of gamma rays induce ultrastructural alterations in limphoid and epithelial cells of the chick thymus which exhibit a different temporal evolution after the treatment. The main changes which occur in lymphoid cells treated with 50 rads disappear 48 hours after the irradiation, while with 100 rads the radiation damage is still evident in epithelial cells.     

Effect of postganglionic sympathectomy on the ultrastructure of the rat parotid gland

Summary Following two weeks of superior cervical ganglionectomy, the parotid glands of adult rats were removed and studied by electron microscopy. Sympathectomy induced striking alterations of acini, resulting in a heterogeneous population of acinar cells, but it had no obvious effect on the duct system. Most of the altered cells could be classified on a cytological basis as dark cells or light cells. Dark cells predominated and contained more secretory granules, less granular endoplasmic reticulum, fewer Golgi membranes, and smaller lumina and intercellular canaliculi than normal acinar cells. The synthesis and extrusion of secretory products appeared to be minimal in these cells. Light cells possessed ultrastructural features, such as dilated cisternae of granular endoplasmic reticulum and prominent Golgi membranes, which were opposite to those of dark cells and indicative of a high degree of secretory activity.The heterogeneous population of cells following sympathectomy indicates that the sympathetic nervous system may play an important role in regulating the secretory synchrony of acinar cells.Supported by U.S.P.H.S. Grant DE 02110.