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1.
X-ray microprobe analysis was used to determine concentrations (millimoles of element per kilogram dry weight) of Na, P, Cl, K, and Ca in cellular compartments of frozen, unfixed sections of rat sciatic and tibial nerves and dorsal root ganglion (DRG). Five compartments were examined in peripheral nerve (axoplasm, mitochondria, myelin, extraaxonal space, and Schwann cell cytoplasm), and four were analyzed in DRG nerve cell bodies (cytoplasm, mitochondria, nucleus, and nucleolus). Each morphological compartment exhibited characteristic concentrations of elements. The extraaxonal space contained high concentrations of Na, Cl, and Ca, whereas intraaxonal compartments exhibited lower concentrations of these elements but relatively high K contents. Nerve axoplasm and axonal mitochondria had similar elemental profiles, and both compartments displayed proximodistal gradients of decreasing levels of K, Cl, and, to some extent, Na. Myelin had a selectively high P concentration with low levels of other elements. The elemental concentrations of Schwann cell cytoplasm and DRG were similar, but both were different from that of axoplasm, in that K and Cl were markedly lower whereas P was higher. DRG cell nuclei contained substantially higher K levels than cytoplasm. The subcellular distribution of elements was clearly shown by color-coded images generated by computer-directed digital x-ray imaging. The results of this study demonstrate characteristic elemental distributions for each anatomical compartment, which doubtless reflect nerve cell structure and function.  相似文献   

2.
IONIC COMPOSITION OF THE CYTOPLASM OF NITELLA FLEXILIS   总被引:1,自引:0,他引:1  
The K, Na and Cl concentrations of the chloroplast layer andthe flowing cytoplasm of Nitella flexilis have been determinedby applying an internal perfusion technique, which enabled usto avoid contamination of ions from the cell sap. K, Na andCl concentrations of the chloroplast layer are 110, 26 and 136mM and those of the flowing cytoplasm are 125, 5 and 36 mM respectively.The cell sap contains 80 mM K, 28 mM Na and 136 mM Cl. Althoughthere are some variations in these values among samples, theflowing cytoplasm is rich in K and poor in Cl and especiallyin Na. The exchange of K and Na across the tonoplasl occursfairly easily (half-time, a few hours), while that of Cl occursextremely slowly (half-time, a few days). 1This work was supported by Research Grants from the Ministryof Education of Japan  相似文献   

3.
Hajibagheri, M. A., Gilmour, D. J., Collins, J. C. and Flowers,T. J. 1986. X-ray microanalysis and ultrastructural studiesof cell compartments of Dunaliella parva. -J. exp. Bot. 37:1725–1732. Ultrastructural studies of the unicellular green alga Dunaliellaparva showed the presence of cytoplasmic vacuoles. X-ray microanalysiswas performed on sections of cells which had been freeze substitutedin acetone. It was found that the concentrations of both Naand Cl were much higher in the vacuoles than in the cytoplasm.When cells were grown in 0·4 kmol m–3 NaCl theNa and Cl concentrations in the vacuoles were 349 and 283 molm –3 respectively, whilst cytoplasmic Na and Cl concentrationswere 37 and 26 mol m–3. Corresponding values for cellsgrown in 1·5 kmol m–3 NaCl were 392 mol m–3Na and 325 mol m–3 Cl in the vacuoles and 36 mol m–3Na and 30 mol m–3 Cl in the cytoplasm. Immediately afterexposure to an increase in external salinity Na and Q concentrationsincreased in both vacuoles and cytoplasm. The results are discussedwith reference to compartmental models for the ionic relationsof Dunaiiella. Key words: X-ray microanalysis, ultrastructural studies, Dunaliella parva  相似文献   

4.
Ion gradients across the plasma membrane, fundamentally K(+), play a pivotal role in the execution phase of apoptosis. However, little is known about other monovalent anions (Cl(-)) or cations (Na(+)) in apoptosis. In addition, the relationship between changes in total ion composition and morphological and biochemical events are poorly understood. We investigated simultaneous changes in sodium (Na), chlorine (Cl), and potassium (K) concentrations in stauroporine-induced apoptosis by quantitative electron probe X-ray microanalysis (EPXMA) in single cells. Apoptotic cells identified unequivocally from the presence of chromatin condensation in backscattered electron images were characterized by an increase in intracellular Na, a decrease in intracellular Cl and K concentrations, and a decrease in K/Na ratio. The ouabain-sensitive Rb-uptake assay demonstrated a net decrease in Na(+)/K(+)-ATPase activity, suggesting that increases in Na and decreases in K and the K/Na ratio in apoptotic cells were related with inhibition of the Na(+)/K(+)-ATPase pump. These changes in diffusible elements were associated with externalization of phosphatidyl serine and oligonucleosomal fragmentation of DNA. This alteration in ion homeostasis and morphological hallmarks of apoptosis occur in cells that have lost their inner mitochondrial transmembrane potential and before the plasma membrane becomes permeable.  相似文献   

5.
X-ray microprobe analysis was used to determine the effects of axotomy on distribution and concentration (millimoles of element per kilogram dry weight) of Na, P, Cl, K, and Ca in frozen, unfixed sections of rat sciatic nerve. Elemental concentrations were measured in axoplasm, mitochondria, and myelin at 8, 16, and 48 h after transection in small-, medium-, and large-diameter fibers. In addition, elemental composition was determined in extraaxonal space (EAS) and Schwann cell cytoplasm. During the initial 16 h following transection, axoplasm of small fibers exhibited a decrease in dry weight concentrations of K and Cl, whereas Na and P increased compared to control values. Similar changes were observed in mitochondria of small axons, except for an early, large increase in Ca content. In contrast, intraaxonal compartments of larger fibers showed increased dry weight levels of K and P, with no changes in Na or Ca concentrations. Both Schwann cell cytoplasm and EAS at 8 and 16 h after injury had significant increases in Na, K, and Cl dry weight concentrations, whereas no changes, other than an increase in Ca, were observed in myelin. Regardless of fiber size, 48 h after transection, axoplasm and mitochondria displayed marked increases in Na, Cl, and Ca concentrations associated with decreased K. Also at 48 h, both Schwann cell cytoplasm and EAS had increased dry weight concentrations of Na, Cl, and K. The results of this study indicate that, in response to nerve transection, elemental content and distribution are altered according to a specific temporal pattern. This sequence of change, which occurs first in small axons, precedes the onset of Wallerian degeneration in transected nerves.  相似文献   

6.
Summary The extracellular and intracellular fluid volumes of pondwater acclimatedLigumia subrostrata are equal (3.9 ml/g dry tissue). Total blood solute is 47 mOsm and is composed primarily of Na (19.1 mM), Cl (10.6 mM), HCO3 (12.7 mM), Ca (4.3 mM), and K (0.5 mM). Major intracellular solutes are K (14.0 mM), Na (7.0 mM) and Cl (2.4 mM).L. subrostrata continuously exposed to deionized water at 20°C exhibit a maximum decrease of 23% in extracellular fluid total solute within 30 days. The maximum [Na] and [Cl] losses are 40% and 76% respectively, while [Ca] and [HCO3] increase by 44% and 37% respectively. No apparent change in extracellular [K] occurs. Intracellular [Na] decreases 53% and [Cl] decreases 79%, but [K] declines only 15%. Intracellular fluid volume, extracellular fluid volume, and total body water decrease 17%, 31%, and 22% respectively. Inulin clearance is 0.41 ml/g dry tissue·h for pondwater acclimated mussels and declines to 0.24 ml/g dry tissue·h during salt depletion. When salt depleted mussels are returned to solutions containing Na or Cl, they experience a net uptake of salt. The accumulated ions are about equally distributed in the extra- and intracellular compartments.  相似文献   

7.
Summary Intracellular concentrations of Na, K, Cl ([Na], [K] and [Cl], respectively) and other elements were determined in isolated monkey eccrine sweat secretory coil cells using quantitative electron probe X-ray microanalysis of freeze dried cryosections. The validity of the methodology was partially supported by qualitative agreement of the X-ray microanalysis data with those obtained by micro-titration with a helium glow spectrophotometer. [Na], [K] and [Cl] of the cytoplasm were the same as those in the nucleus in both clear and dark cells. [Na], [K], and [Cl] of the clear cells were also the same as those of the dark cells at rest and after stimulation with methacholine (MCh), suggesting that these two cell types behave like a functional syncytium. MCh stimulation induced a pharmacologically specific, dose-dependent decrease in [K] and [Cl] (as much as 65%), and a 3.7-fold increase in [Na]. In myoepithelial cells, a similar change in [Na] and [K] was noted after MCh stimulation although the decrease in [Cl] was only 20%. The MCh-induced change in [Na], [K] and [Cl] was almost completely inhibited by removal of Ca2+ from the medium. 10–4 m bumetanide inhibited the MCh-induced increase in [Na], reduced the decrease in [K] by about 50%, but slightly augmented the MCh-induced decrease in [Cl]. 10–4 m ouabain increased [Na] and decreased [K] as did MCh; however, unlike MCh, ouabain increased [Cl] by 56% after 30 min of incubation. Thus the data may be best interpreted to indicate that Ca-dependent K efflux and (perhaps also Ca-dependent) Cl efflux are the predominat initial ionic movement in muscarinic cholinergic stimulation of the eccrine sweat secretory coils and that the ouabain-sensitive Na pump plays an important role in maintenance of intracellular ions and sweat secretion.  相似文献   

8.
Cell shrinkageis an early prerequisite in programmed cell death, and cytoplasmicK+ is a dominant cation that controls intracellular ionhomeostasis and cell volume. Blockade of K+ channelsinhibits apoptotic cell shrinkage and attenuates apoptosis. We examined whether apoptotic repressor with caspase recruitment domain (ARC), an antiapoptotic protein, inhibits cardiomyocyte apoptosis by reducing K+ efflux throughvoltage-gated K+ (Kv) channels. In heart-derived H9c2cells, whole cell Kv currents (IK(V)) wereisolated by using Ca2+-free extracellular (bath) solutionand including 5 mM ATP and 10 mM EGTA in the intracellular (pipette)solution. Extracellular application of 5 mM 4-aminopyridine (4-AP), ablocker of Kv channels, reversibly reduced IK(V)by 50-60% in H9c2 cells. The remaining currents during 4-APtreatment may be generated by K+ efflux through4-AP-insensitive K+ channels. Overexpression of ARC inheart-derived H9c2 cells significantly decreasedIK(V), whereas treatment with staurosporine, apotent apoptosis inducer, enhanced IK(V)in wild-type cells. The staurosporine-induced increase inIK(V) was significantly suppressed and thestaurosporine-mediated apoptosis was markedly inhibited incells overexpressing ARC compared with cells transfected with thecontrol neomycin vector. These results suggest that theantiapoptotic effect of ARC is, in part, due to inhibition of Kvchannels in cardiomyocytes.

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9.
Summary Methacholine (MCh)-induced changes in intracellular concentrations of Na, K, and Cl ([Na]i, [K]i, and [Cl]i, respectively) and in cellular dry mass (a measure of cell shrinkage) were examined in isolated monkey eccrine sweat secretory coils by electron probe X-ray microanalysis using the peripheral standard method. To further confirm the occurrence of cell shrinkage during MCh stimulation, the change in cell volume of dissociated clear and dark cells were directly determined under a light microscope equipped with differential interference contrast (DIC) optics. X-ray microanalysis revealed a biphasic increase in cellular dry mass in clear cells during continuous MCh stimulation; an initial increase of dry mass to 158% (of control) followed by a plateau at 140%, which correspond to the decrease in cell volume of 37 and 29%, respectively. The latter agrees with the MCh-induced cell shrinkage of 29% in dissociated clear cells. The MCh-induced increase in dry mass in myoepithelial cells was less than half that of clear cells. During the steady state of MCh stimulation, both [K]i and [Cl]i of clear cells decreased by about 45%, whereas [Na]i increased in such a way as to maintain the sum of [Na]i+[K]i constant. There was a small (12–15mm) increse in [Na]i and a decrease in [K]i in myoepithelial cells during stimulation with MCh. Dissociated dark cells failed to significantly shrink during MCh stimulation. The decrease in [Cl]i in the face of constant [Na]i+[K]i suggests the accumulation of unknown anion(s) inside the clear cell during MCh stimulation. While the decrease in [K]i and [Cl]i may be instrumental in facilitating influx of ions via Na–K–2Cl cotransporters, the functional significance of MCh-induced cell shrinkage remains unknown.  相似文献   

10.
Salt Tolerance in the Triticeae: K/Na Discrimination in Barley   总被引:5,自引:4,他引:1  
Concentrations of ions were measured in the youngest fully-expandedleaves of Triticum aestivum, T. durum, Hordeum vulgare, H. spontaneum,Secale cereale, and Aegilops squarrosa accessions grown in hydroponicculture in the presence of salt (NaCl+CaCl2). Triticum aestivum,Secale cereale, and Ae. squarrosa had the low leaf Na and highleaf K concentrations typical of plants which contain the enhancedK/Na discrimination character originally found in Ae. squarrosa.T. durum and the Hordeum species did not have this character.The better growth of H. vulgare than of T. durum with similarsalt concentrations in the youngest fully-expanded leaves maybe a result of better compartmentation of Na, Cl, and K betweendifferent tisssues or between different compartments withincells. The enhanced K/Na discrimination character was expressedin disomic addition lines of H. vulgare chromosomes in Triticumaestivum. The H. vulgare variety Herta and its slender mutantboth had similar leaf cation concentrations, although they differedin growth rate when grown at 60 mol m–3 NaCl. H. vulgareand T. durum seedlings grown in the absence of monovalent cationsaccumulated more 22Na in their shoots than seedlings of otherspecies when incubated in 1.0 mol m–3 NaCl labelled with22Na. Key words: Salt, ion transport, I genome, barley, wheat  相似文献   

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