Uptake and intracellular distribution of lanthanum

The present investigation of the cellular distribution of lanthanum was undertaken in order to control the validity of the "Lanthanum method" used for the study of the cell calcium compartments. The presence of lanthanum was evaluated in the isolated guinea-pig heart and its subcellular fractions perfused with a lanthanum-containing Tyrode solution. Lanthanum was determined by instrumental neutron activation analysis. Under the adopted experimental conditions (30-min incubation in the presence of 12.5 microM lanthanum), lanthanum is carried across the cell membrane and is taken up by subcellular organelles. These results confirm the limited validity of investigations based on of the "Lanthanum method".     

Neutralized lanthanum solution: a largely noncolloidal ultrastructural tracer.

Lanthanum nitrate solution adjusted to pH 7.4 and pH 7.7 was subjected to column chromatography, ultrafiltration and conductivity measurements. Lanthanum concentration was measured by a colorimetric method employing eriochrome cyanine RC. Under these conditions, lanthanum was not excluded from the column by a packing with an exclusion limit of 1800 daltons. Ultrafiltration through a membrane with a filter limit of 500 daltons allowed approximately 75% of the lanthanum to pass. Conductivity measurements showed a decrease of charge of about 20% on adding sodium hydroxide to a solution of lanthanum nitrate up to a pH of 7.7. It is concluded that approximately 80% of the lanthanum exists as a charged particle of less than 500 daltons at pH 7.7; the other 20% consists of larger, possibly colloidal particles. Nonfiltered and ultrafiltered lanthanum have equally good staining and tracer properties in the electron microscope, suggesting that staining depends largely on the ultrafiltrable noncolloidal lanthanum ion.     

Effects of lanthanum on the heart sarcolemmal ATPase and calcium binding activities.

Effects of lanthanum on Ca2+-ATPase, Mg2+-ATPase, Na+-K+-ATPase, and calcium binding activities were studied in rat heart sarcolemma. Ten to 100 micrometers lanthanum depressed significantly the Ca2+-ATPase activity and 50--200 micrometers lanthanum inhibited the calcium binding activity. Lineweaver-Burk plots of the Ca2+-ATPase activity showed that the inhibition by lanthanum was competitive with calcium concentration. Neither Mg2+-ATPase nor Na+-K+-ATPase activities were affected by lanthanum when the assay medium contained 1 mM EDTA; however, in the absence of EDTA, these enzyme activities were significantly decreased by 10--100 micrometers lanthanum. Rat hearts perfused with HEPES buffer containing 0.5 mM lanthanum showed electron-dense deposits restricted to the outer cell surface and the sarcolemma obtained from these hearts also had the deposits, indicating that the membrane fraction isolated by the hypotonic shock--LiBr treatment method is of sarcolemmal origin. The Ca2+-ATPase activity of the sarcolemma isolated from lanthanum-perfused hearts, unlike the Mg2+-ATPase, Na+-K+-ATPase, and calcium binding activities, was significantly less than the control value. From these observations it is suggested that lanthanum may influence calcium movement across the sarcolemma by affecting sarcolemmal ATPase and calcium binding activities.     

Sites of lanthanum occlusion in the testis of the crayfish Procambarus paeninsulanus (Crustacea: Cambaridae)

The presence of stage-dependent occlusive junctions between adjacent Sertoli cells in the seminiferous epithelium of the crayfish testis was demonstrated by a lanthanum tracer study. The germinal epithelium did not appear to be compartmentalized, as evidenced by access of lanthanum to spermatogonia, spermatocytes, and spermatids. During late spermiogenesis, when encapsulated stage VI spermatids were concentrated in the center of an acinus, lanthanum was excluded apically, coincident with lumen formation. This is the first study examining occluding junctions using a barrier penetration method in the testis of a crustacean.     

Hepatitis C patients in puerto rico have an altered iron balance

The in vivo coordination structure of lanthanum ions interacting with chlorophyll-a of the fern Dicranopteris dichotoma grown in a rare earth minefield in southern China was determined by the extended x-ray absorption fine structure (EXAFS). The results show that lanthanum includes two porphyrin rings in its coordination sphere. It is postulated that the La-chlorophyll-a complex may have a bilayer structure. The analytical method may serve as a new tool to gain insight in the in vivo interactions of rare earth elements.     

Effect of lanthanum on the turnover of calcium in rat uterus.

Previous investigations suggest that lanthanum might enter uterine smooth muscle cells and work as intracellular calcium displacing agent. The present investigation had been carried out in order to confirm if lanthanum develops an intracellular effect. Experiments show that lanthanum brings about a marked increase of the intracellular calcium; the comparison of the uptake and of the wash-out curve of 45Ca shows that lanthanum induces a lowering of the rapid phase of 45Ca release from rat uterus, while the uptake of the labelled ion is not modified or is even enhanced. The present data demonstrate that the action of lanthanum in rat uterus is limited to the cell membrane, whose calcium extruding properties are inhibited.     

Effects of low-molecular-weight organic acids on uptake of lanthanum by wheat roots

Effects of low-molecular-weight organic acids (LMWOAs) on the uptake of lanthanum by wheat (Triticum aestivum L.) roots were studied under hydroponic conditions. Acetic and malic acids were chosen as the representatives of LMWOAs. Uptake kinetics of lanthanum indicated that when lanthanum concentrations in the uptake solutions were high or uptake time was long lanthanum uptake by roots was enhanced by LMWAOs. After wheat was cultured in the uptake solution of lanthanum containing acetic or malic acids for 48 h the uptake of lanthanum by roots increased by 57 and 44%, respectively, compared with that in the absence of acetic and malic acids. The increase in uptake of lanthanum was determined by the ratio of the concentrations of organic acids to lanthanum in the uptake solutions. The highest uptake of lanthanum was obtained at the ratio of 5:1.     

Aggregation and release reaction of washed platelets stimulated by lanthanum.

Ionized lanthanum caused clumping of washed platelets. This clumping response could be reversed by chelating agents but was not impaired by known inhibitors of platelets aggregation. Aggregation by lanthanum was not restricted to the unique clumping properties of platelets but occurred in fixed platelets and red cells and was most likely based on an electrostatic interaction.Lanthanum was able to stimulate as well as to inhibit serotonin release from platelets.At a concentration of 1 mM, lanthanum evoked a release of serotonin from washed platelets at 37°C. This release reaction was inhibited at 18°C or by prior treatment of platelets with neuraminidase or NEM.At a high concentration (10 mM), lanthanum did not stimulate the platelet release reaction but inhibited that induced by all stimuli investigated, presumably due to a fixation of membrane molecules.The release reaction promoted by thrombin or A 23187, but not that by collagen, was inhibited by a low concentration of lanthanum (0.1 mM). This inhibition is based on an interaction of lanthanum with the stimuli rather than with the platelet surface.     

Lanthanum binding to murine neuroblastoma cells

The binding of lanthanum to murine neuroblas-toma cells (clone N1E-115) was studied by means of electron microscopy and x-ray microanalysis. Lanthanum bound only to cellular membranes, in a nonuniform manner. This lanthanum binding was reduced by pharmacological agents that affect muscarinic receptors of these cells or the function of the receptors. These results suggest that this binding of lanthanum is to sites closely related to muscarinic receptors in the cells.     

On the evidence of a diffusion barrier in the outer cortex apoplast of cress-roots (Lepidium sativum), demonstrated by analytical electron microscopy

To mark the apoplastic pathway of ions in the root of the dicotyledonous plant Lepidium sativum we used the heavy element lanthanum, which can be identified by analytical electron microscopy (EELS and ESI). In the front root tip, the primary walls of all meristematic cells contained lanthanum. 10-15 mm behind the root apex, lanthanum was found in the cortex cell walls up to the endodermis, but not in the stele. 20-25 mm from the tip, lanthanum was accumulated in the radial cell walls of the hypodermis, which, however, is not a complete diffusion barrier for ions, so that traces of lanthanum also were found in the cortex cell walls up to the endodermis. This study provides evidence for the presence of two apolastic diffusion barriers in the region of highest water uptake in cress roots.     

Effects of lanthanum on calcium-dependent phenomena in human red cells.

Lanthanum (0.25 mM) does not penetrate into fresh or Mg2+-depleted cells, whereas it does into ATP-depleted or ATP + 2,3-diphosphoglycerate-depleted cells, into cells containing more than 3 mM calcium, or cells stored for more than 4 weeks in acid/citrate/dextrose solution. In fresh cells loaded with calcium, extracellular lanthanum blocks the active Ca2+-efflux completely and inhibits (Ca2+ + Mg2+)-ATPase (ATP phosphohydrolase, EC 3.6.1.3) activity to about 50%. In Mg2+-depleted cells Ca2+-Ca2+ exchange is inhibited by lanthanum. Ca2+-leak is unaffected by lanthanum up to 0.25 mM concentration; higher lanthanum concentrations reduce leak rate. In NaCl medium Ca2+-leak +/ S.D. amounts to 0.28 +/ 0.08 mumol/1 of cells per min, whereas in KC1 medium to 0.15 +/ 0.04 mumol/1 of cells per min at 2.5 mM [Ca2+]e and 0.25 mM [La3+]e pH 7.1. Lanthanum inhibits Ca2+-dependent rapid K+ transport in ATP-depleted and propranolol-treated red cells, i.e. whenever intracellular calcium is below a critical level. The inhibition of the rapid K+ transport can be attributed to protein-lanthanum interactions on the cell surface, since lanthanum is effectively detached from the membrane lipids by propranolol. Lanthanum at 0.2--0.25 mM concentration has no direct effect on the morphology of red cells. The shape regeneration of Ca2+-loaded cells, however, is blocked by lanthanum owing to Ca2+-pump inhibition. Using lanthanum the transition in cell shape can be quantitatively correlated to intracellular Ca2+ concentrations.     

Effects of Lanthanum Chloride Administration on Detouring Learning in Chicks

Lanthanum cations are well known for their inhibitory actions on calcium channels, and calcium cations are indispensable for the development of brain. Lanthanum may interfere with the developing central nervous system. Detour learning task in chicks is an excellent model to study the development of central nervous system. In the present study, we examined the effects of lanthanum chloride exposure on the development of spatial cognition using the detour learning task. The data suggest that the chicks injected with lanthanum chloride (10 or 5 mM) had significantly delayed the response latency of detour learning but not the chicks injected with lanthanum chloride (1 mM). The effect of lanthanum exposure on the development of spatial cognition is dose relative.     

The chemoreceptor surface of the taste disc in the frog,Rana esculenta. An ultrastructural study with lanthanum nitrate

Summary The distribution of lanthanum on the taste disc of the frog,Rana esculenta, afteren bloc staining of the tongue with lanthanum nitrate was studied at the ultrastructural level by means of scanning electron microscopy (in the secondary electron mode or in the back scattered electron mode), energy-dispersive X-ray microanalysis, transmission electron microscopy and electron spectroscopic imaging. It was consistently found that lanthanum distribution on the surface of the taste disc is not homogeneous and that the surface of putative receptor cells is in contact with strongly lanthanum-positive material. Calcium co-localizes with lanthanum at that level. These results suggest that different microenvironments exist at the surface of the taste disc and that this could be relevant to the receptor function.     

Opening of blood-brain barrier in Triturus cristatus carnifex by hyperosmolar mannitol solutions

The permeability of the newt cerebral capillaries to lanthanum ion has been studied after perfusion with mannitol solutions of increasing molarity. In the control specimens lanthanum deposits were limited to the luminal side of the capillaries and tracer did not spread to the pericapillary spaces due to the tight junctions. Treatment with hypertonic solutions of mannitol (0.25M, 0.5M, 1M) caused opening of the blood brain barrier with a progressive increase in lanthanum between the endothelial cell edges, in the basal lamina and in the extracellular spaces of the nervous parenchyma in relation to the molarity of the mannitol solution. The spread of lanthanum is probably due to opening of the tight junctions between the endothelial cells, since pinocytotic vesicles labelled with tracer were not evident.     

Stomatal Response to Humidity and Lanthanum

Lanthanum fed to the base of excised leaves of Sesamum indicum L. and Helianthus annuus L. was used as a tracer to investigate by electron microscopy the path of water in the apoplast of leaves. The generally random distribution of lanthanum in cell walls provided no support for the hypothesis that cuticular transpiration may be greater for guard cells than for adjacent epidermal cells. Occasionally, accumulations of lanthanum were observed in anticlinal walls of epidermal cells and at the outer surface of the plasma membrane but lanthanum was not observed in the symplast. The influx of ⁸⁶Rb to excised roots of sesame and sunflower was inhibited during incubation with 0.5 mM lanthanum or calcium for 15 or for 180 min. Stomata of sunflower partially closed when 2.5 mM lanthanum was supplied to the base of excised shoots in a potometer, whereas this treatment had little effect on stomatal conductance of sesame shoots maintained in a constant environment. Supplying 2.5 mM lanthanum to the base of sesame shoots strongly inhibited stomatal opening response to increase in ambient humidity but had little effect on stomatal opening response to light. It was concluded that stomatal opening response to increased humidity may be dependent upon some process, such as ion influx, that is inhibited by lanthanum, and that opening response to humidity may differ in mechanism from stomatal opening response to increased irradiance.     

A rapid radiochemical assay for hypoxanthine-guanine phosphoribosyltransferase

A simple radiochemical method is described for assay of hypoxanthine-guanine phosphoribosyltransferase. ¹⁴C-Hypoxanthine is incubated with enzyme PRPP. The labelled product is precipitated on strips of Whatman No. 1 paper by the addition of lanthanum nitrate. Unreacted substrate is eluted with distilled water. The major advantages of this method are speed, reproducibility, ability to process many samples and low blank values.     

In Utero and Lactational Lanthanum Exposure Induces Olfactory Dysfunction Associated with Downregulation of βIII-tubulin and Olfactory Marker Protein in Young Rats

We evaluated the role of βIII-tubulin in the morphology of olfactory receptor neuron (ORN) and olfactory dysfunction in offspring caused by prenatal and postnatal lanthanum exposure. Pregnant rats were exposed to 0.25% lanthanum chloride in drinking water from gestational day (GD) 7 until postnatal day 21. From postnatal day 23 until postnatal day 28, pups were examined with buried food pellet and olfactory maze test. The ultrastructural features of ORNs in the olfactory epithelium (OE) were observed by transmission electron microscope. The expression of βIII-tubulin and olfactory marker protein (OMP) in the tissue sections and homogenates of OE were, respectively, measured by immunodetection and western blot. Behavioral analysis of olfaction showed that lanthanum chloride exposure induced olfactory dysfunction. Offsprings exposed to lanthanum chloride showed enlarged ORN knobs and a decreased number of cilia. In addition, the levels of OMP and βIII-tubulin expression in lanthanum chloride exposure offsprings significantly decreased. Developmental lanthanum exposure could impair olfaction, and this deficit may be attributed to the downregulation of βIII-tubulin and OMP in the OE.     

The action of lanthanum ions and formaldehyde on the proton-pumping function of bacteriorhodopsin

The photochemical cycle and the proton-pumping function of bacteriorhodopsin modified with lanthanum and formaldehyde has been studied. In both preparations, the M412 leads to BR570 transition time has been found to increase considerably. The deceleration of the photochemical cycle has been shown to be accompanied by inhibition of the millisecond phase of the photoelectrical response of bacteriorhodopsin membranes associated with phospholipid-impregnated collodion film. Photoelectrogenic activity measured with permeable ion probe in proteoliposomes was also inhibited. Effects of lanthanum were reversed by EDTA. Formation of M412 was slightly accelerated and the microsecond electrogenic phase was not affected by lanthanum and by formaldehyde. It is concluded that lanthanum, but not formaldehyde, can be used as a specific reversible inhibitor of the second half of the bacteriorhodopsin photocycle and of the associated H+ uptake on the cytoplasmic side of the halobacterial membrane. Possible mechanisms of these effects are discussed.     

PERMEABILITY OF SERTOLI CELL TIGHT JUNCTIONS TO LANTHANUM AFTER LIGATION OF DUCTUS DEFERENS AND DUCTULI EFFERENTES

The permeability of Sertoli cell tight junctions to lanthanum administered during fixation has been compared in rats after ligation of the ductus deferens and after ligation of the ductuli efferentes. In both control and vasoligated testes, lanthanum penetrated only short distances into the Sertoli cell tight junctions before stopping abruptly. The tight junction, consisting of numerous pentalaminar fusions of contiguous Sertoli cell membranes, prevented diffusion of lanthanum into the adluminal compartment of the seminiferous epithelium. In rats with ligated ductuli efferentes, lanthanum completely permeated many Sertoli cell tight junctions and occupied intercellular spaces of the adluminal compartment. In spite of their newly acquired permeability to lanthanum, tight junctions retained characteristic ultrastructural features, including numerous membrane fusions. When lanthanum-filled tight junctions were sectioned en face, membrane fusions appeared as pale lines in lakes of electron-opaque tracer. These linearly extensive fasciae occludentes occasionally ended blindly, suggesting that lanthanum may have traversed the junction by diffusing around such incomplete barriers. The increased permeability of Sertoli cell tight junctions after efferent ductule ligation, which caused rapid testicular weight gain followed by atrophy, indicates that tight junctions are sensitive to enforced retention of testicular secretions inside the seminiferous tubules. The apparent normalcy of Sertoli cell tight junctions after vasoligation, which had no effect on testis weight, supports the view that blockage of testicular secretions distal to the epididymis is relatively innocuous.     

A calcium-sensitive lanthanum inhibition of amoeboid movement

The effects of lanthanum ions upon amoeboid movement and upon the redistribution of dense cytoplasmic inclusions following centrifugation were investigated in Amoeba discoides. Treatment with low lanthanum concentrations produced a characteristic inhibited condition in which locomotion ceased while saltatory movements continued. The resorting of stratified cytoplasm by local internal cytoplasmic flow was unaffected. This contrasted with p-hydroxymercuribenzoate treatment which inhibited both locomotion and saltatory movements. The inhibitory effects of lanthanum ions were antagonised by calcium ions but not by magnesium ions. It is reasoned that lanthanum competes for a peripheral binding site thereby disrupting a calcium-mediated regulatory mechanism of amoeboid movement.