Environmental modulation of calcium and phosphorus metabolism in embryonic snapping turtles (Chelydra serpentina)

Summary Eggs of common snapping turtles (Chelydra serpentina) were incubated on wet (–150 kPa water potential) and dry (–950 kPa) substrates in a laboratory study assessing the effects of the hydric environment on patterns of mobilization of calcium and phosphorus by developing embryos. We found that embryos developing in wet environments withdrew nutrients from their yolk faster, grew more rapidly, and incubated longer than embryos exposed to dry environments. Turtles developing in both environments absorbed calcium from the yolk at similar rates and depleted the yolk of almost its entire reserve of calcium prior to hatching. Calcium withdrawn from the yolk was supplemented with calcium mobilized from the eggshell, but embryos in wet environments obtained substantially more calcium from the eggshell than did those in dry settings. Embryos obtained all of the phosphorus used in skeletogenesis from the yolk, but those incubating in wet environments mobilized phosphorus from this compartment more rapidly than did those in dry settings. Exposing embryonic snapping turtles to wet environments apparently allows them to make more efficient use of the transitory source of calcium in the eggshell than is possible in dry environments. However, the residual yolk in hatchlings from both wet and dry environments contains too little calcium to support the growth of hard and soft tissues in neonates at rates similar to those characterizing the growth phase of development in embryogenesis.     

Accumulation of calcium and phosphorus in pigeon (Columba livia) embryos

Summary Calcium and phosphorus were measured in the yolk and albumen of fertile pigeon (Columba livia) eggs incubated for 0–17 days, and in embryos and hatchlings. Shell provided most of the calcium for skeletal mineralization of the embryos, whereas phosphorus was derived from the yolk and albumen. Mobilization of calcium from the shell to the embryo commenced at approximately day 11 of incubation, accumulating both in the embryo and the yolk sac. There was 1.4 times more calcium in squab yolk sacs than that contained in newly laid egg yolks. The results suggest that whereas general patterns of calcium and phosphorus accumulation during embryogenesis in altricial birds closely resemble those of precocial birds, calcium mobilization from the shell begins later, proceeds at a slower rate and results in a less mineralized hatchling.CIDA/NSERC Visiting Research Associate Permanent address: Department of Animal Science, University of Peradeniya, Peradeniya, Sri Lanka     

Growth of yellow-headed blackbird Xanthocephalus xanthocephalus nestlings in relation to maternal body condition, egg mass, and yolk carotenoids concentrations

Condition‐dependent resource allocation to eggs can affect offspring growth and survival, with potentially different effects on male and female offspring, particularly in sexually dimorphic species. We investigated the influence of maternal body condition (i.e., mass‐tarsus residuals) and two measures of female resource allocation (i.e., egg mass, yolk carotenoid concentrations) on nestling mass and growth rates in the polygynous and highly size dimorphic yellow‐headed blackbird Xanthocephalus xanthocephalus. Egg characteristics and carotenoid concentrations were obtained from the third‐laid egg of each clutch and were correlated with the mass and growth rates of the first two asynchronously hatched nestlings. Maternal body condition was associated with the growth of first‐hatched, but not second‐hatched nestlings. Specifically, females in better body condition produced larger and faster growing first‐hatched nestlings than females in poorer body condition. As predicted for a polygynous, size‐dimorphic species, females that fledged first‐hatched sons were in better body condition than females that fledged first‐hatched daughters. Associations between egg mass, yolk carotenoid content, and nestling growth were also specific to hatching‐order. Egg mass was positively correlated with the mass and growth rates of second‐hatched nestlings, and yolk concentrations of β‐carotene were positively correlated with second‐hatched nestling mass. Surprisingly, the relationship between yolk lutein and hatchling growth differed between the sexes. Females with high concentrations of yolk lutein produced larger and faster growing first‐hatched sons, but smaller first‐hatched daughters than females with lower lutein concentrations. Mass and growth rates did not differ between first‐ and second‐hatched nestlings of the same sex, despite asynchronous hatching in the species. Results from this study suggest that maternal body condition and the allocation of resources to eggs have carotenoid‐, sex‐, and/or hatch‐order‐specific effects on yellow‐headed blackbird nestlings.     

Structure and Use of Song in the Yellow-headed Blackbird (Xanthocephalus xanthocephalus)

Analysis of yellow-headed blackbird songs indicated that accenting and buzzing songs differed in note structure, energy distribution, and loudness. Buzzing songs showed a shift in energy distribution as a result of propagation through marsh habitat whereas accenting songs did not. Results suggest that accenting songs should propagate further in marsh habitat than buzzing songs. Two experiments, done to test whether song types differed in functional distance, showed that the presence of a male song of either type, or a male or female model on their territories increased use of buzzing songs by males. In the absence of auditory or visual stimulation, males used mainly accenting songs. Results supported the hypothesis that buzzing songs are used primarily for short-range interactions whereas accenting songs are used primarily for long-range general advertising.     

Chromatographic study of magnesium and calcium binding to immobilized human serum albumin

The use of immobilized human serum albumin (HSA) as a stationary phase in affinity chromatography has been shown to be useful in resolving optical antipodes or to investigate interactions between drugs and protein. However, to our knowledge, no inorganic ion binding has been studied on this immobilized protein type. To do this, the human serum albumin stationary phase was assimilated to a weak cation-exchanger by working with a mobile phase pH equal to 6.5. A study of the eluent ionic strength effect on ion retention was carried out by varying the buffer concentrations and the column temperatures. The thermodynamic parameters for magnesium and calcium transfer from the mobile to the stationary phase were determined from linear van’t Hoff plots. An enthalpy–entropy compensation study revealed that the type of interaction was independent of the mobile phase composition. A simple model based on the Gouy–Chapman theory was considered in order to describe the retention behavior of the test cations with the mobile phase ionic strength. From this theoretical approach, the relative charge densities of the human serum albumin surface implied in the binding process were estimated at different column temperatures.     

TRPM7 is involved in angiotensin II induced cardiac fibrosis development by mediating calcium and magnesium influx

Cardiac fibrosis is involved in a lot of cardiovascular pathological processes. Cardiac fibrosis can block conduction, cause hypoxia, strengthen myocardial stiffness, create electrical heterogeneity, and hamper systolic ejection, which is associated with the development of arrhythmia, heart failure and sudden cardiac death. Besides the initial stimulating factors, the cardiac fibroblasts (CFs) are the principal responsible cells in the fibrogenesis cascade of events. TRPM7, a member of the TRPM (Melastatin) subfamily, is a non-selective cation channel, which permeates both Ca²⁺ and Mg²⁺. Here we demonstrated TRPM7 expression in CFs, and 2-APB (TRPM7 inhibitor), inhibited Ang II-induced CTGF, α-SMA expression and CFs proliferation. Besides, knocking down TRPM7 by shRNA, we proved that TRPM7 mediated both calcium and magnesium changes in cardiac fibroblasts which contribute to fibrosis progress. This study suggested that TRPM7 should play a pivotal role in cardiac fibroblast functions associated to cardiac fibrosis development.     

Structural and physiological effects of calcium and magnesium in Emiliania huxleyi (Lohmann) Hay and Mohler

In organisms which perform both photosynthesis and calcification, the fact that calcification proceeds faster in the light than in the dark has led to the long-established view that photosynthesis and calcification are closely coupled. It is now clear that calcification does not promote photosynthesis, but an enhancement of calcification by photosynthesis could still explain why calcification is faster in the light. To test this, the kinetics of the two processes were monitored over a wide range of calcium concentrations (0-50 mM) in the coccolithophore Emiliania huxleyi. The addition of 50 mM calcium strongly inhibited both processes, but when incubated in lower concentrations, rates of calcification increased up to 20 mM calcium whilst those of photosynthesis remained constant over the same range of calcium concentrations. So, rates of calcification are able to rise without a concomitant increase in photosynthetic rates. In addition, calcification rate and coccolith morphology responded similarly to changes in calcium concentrations; low calcification rates were associated with poor coccolith structure (undercalcification) and high calcification rates with perfectly formed coccoliths. Calcium concentration thus strongly influences calcification affecting both crystal structure and rate of calcite deposition. A similar structural analysis of coccoliths from cells grown in different magnesium concentrations showed that this ion is also essential for calcification, since strong signs of coccolith malformation and undercalcification were apparent at both low and high magnesium concentrations. In contrast with the calcium results, coccoliths were flawless only in the normal seawater concentration of 58 mM magnesium. We conclude that photosynthesis and calcification are not closely coupled and that calcification depends on a precise balance of both calcium and magnesium concentrations.     

Changes in sodium,calcium and magnesium ion concentrations in sturgeon (Huso huso) urine and in kidney morphology

During adaptation to brackish water the young great sturgeon Huso huso is able to regulate its serum osmolarity and ion concentrations. After transfer from fresh water to brackish water the ion concentrations in the urine increase and the urine becomes isoosmotic to the blood serum after 24h. The Na⁺ and K⁺ concentrations in the urine increase during the first 12 h by 4.4 and 7.7 times, respectively, later decreasing again. The Mg²⁺ and Ca²⁺ concentrations in the urine increase by 3.4 and 14 times during the first 72h in brackish water and remain high thereafter. These results suggest that the kidneys play an important part in the regulation of serum osmolarity and in the removal of Ca²⁺; however, in contrast to teleosts, Mg²⁺ must be removed extrarenally. During adaptation to a hyperosmotic medium the diameters of the Malpighian bodies, the glomeruli and the diameter of the tubules initially all decrease, but the distal tubules become morphologically differentiated into two regions and the diameter of the distal section later increases again. It is suggested that this is the site of Ca²⁺ secretion into the urine.     

Influence of maternal dietary calcium levels on milk zinc, calcium and phosphorus contents and milk production in rats

The aim of this study was to analyze zinc (Zn), calcium (Ca) and phosphorus (P) contents in milk and the lactational performance in rats fed different Ca levels. Female Wistar rats were fed during pregnancy and lactation with experimental diets containing 20% protein and high (0.90%, HCa), normal (0.60%, NCa) or low (0.20%, LCa) Ca levels. Milk samples were collected after 15 days to determine the milk mineral composition. Pup weight was recorded from birth to weaning (litter size: 6-8 pups) to determine weight gain and calculate milk production. At delivery there were no significant differences in the body weight of the pups between the groups, but at day 15, the LCa group showed lower values than both NCa and HCa groups (p<0.05). The weight gain of the LCa group was significantly lower than of the HCa and NCa groups, between delivery and day 5 (p<0.05). This reduced rate of weight gain led to the LCa group reaching weaning weight later than the other groups. Milk production (g/pup/day) was significantly lower when dams were fed the LCa than the NCa and HCa diets (p<0.05). There were no significant differences among the groups in milk Ca, P and Zn levels and Ca/P ratio. The body mineral composition of the pups at birth did not differ between the groups; at weaning, however, both LCa and HCa groups had lower element contents than the NCa group (p<0.05). In conclusion, dams fed with a diet containing low Ca levels produced smaller volumes of milk and their pups reached weaning weights later than the other groups. As the milk mineral composition was not affected, it can be hypothesized that in dams fed low dietary Ca, the smaller milk yield might have been a way of maintaining milk quality. High Ca levels affected neither pregnancy outcome nor lactational performance.     

Supply and uptake of potassium,calcium and magnesium of spray carnations (Dianthus caryophyllus) grown in rockwool

Summary In two experiments with carnations grown in rockwool the effects of different cation ratios in the nutrient solution were studied. The results showed that carnations need a high calcium supply. The crop did not appear to be sensitive to different potassium-magnesium ratios in the nutrient solution. In the nutrient solution added mole ratios K∶Ca∶Mg=55∶35∶10 seemed to be optimal. Such ratios in addition led to ratios of 55∶30∶15 in the root environment. Tissue analysis showed that in younger leaves of peduncles harvested a potassium content of 900 mmol per kg dry matter was optimal. For calcium a content of 350 and for magnesium 100–150 mmol per kg was needed. Analytical data of plant-sap analyses were closely correlated with data gained by digestion of dried material. For potassium and magnesium the relationships were linear. However, for calcium a curvilinear relationship was found. In the experiments indications were obtained that a sufficient calcium supply suppressed foot rot in carnations.     

Localization of calcium and phosphorus in early predentin-matrix components by electron spectroscopic imaging (ESI)-analysis in rat molars

Summary The subcellular distribution of the inorganic elements calcium (Ca) and phosphorus (P) was studied in the first-formed dentin matrix during initial mineralization in neonatal rat molars. This most peripheral matrix region is comprised of a proteoglycan-rich ground substance, interwoven by a collagenous network, matrix vesicles, aperiodic fibrils derived from the dental basal lamina, and apical odontoblastic cell processes. All matrix components may possibly serve as templets for mineral deposition during initial calcification of first-formed mantle dentin and predentin. By means of the very sensitive ESI-analysis we studied the subcellular localization of Ca and P and their possible association with distinct organic extracellular matrix components and odontoblasts. Ca-signals were found in the ground substance, at striated collagen fibrils and plasma membranes of odontoblasts in the cuspal early matrix region, but occurred only sparsely in the ground substance of the more distal matrix region where odontoblast processes attach to aperiodic fibrils of the dental basal lamina. Ca was generally absent in matrix vesicles. In contrast, P-signals were found in matrix vesicles, at aperiodic fibrils and at the plasma membranes of odontoblasts. Ca and P co-localized at striated collagen fibrils (type I or II). These results suggest that striated collagen fibrils might serve as primary deposition sites for calcium phosphate during early biological calcification of organic extracellular macromolecules.     

The effect of zinc and magnesium on calcium uptake into the rat duodenum slices

Summary The experiments were carried out on 80 male Wistar rats, divided into four groups as follows: group 1, treated orally with ZnCl₂ at a dose of 10 mg Zn²⁺/kg for 14 days; group 2, control; group 3, MgCl₂-treated at a dose of 5 mg Mg²⁺/kg; group 4, treated with ZnCl₂ plus MgCl₂ in the same manner as groups 1 and 3. The influx of calcium into the rat duodenum slices was investigated in vitro by the method of Papworth and Patrick. Over a range of calcium concentrations (0–10 mM) the influx of this element was defined as a sum of a saturable term (active transport) and a linear term dependent on concentration (passive transport). In the zinc-treated rats only the saturable term was affected. The study of this term by Lineweaver-Burk plots showed a decrease of the half-saturation constant,K _t, while the maximal value,J _m, remained unchanged. Moreover, magnesium was shown to interact with zinc at gut level because simultaneous oral administration of Mg²⁺ and Zn²⁺ to rats protected them against the inhibition of calcium uptake observed when Zn²⁺ was given alone.     

Synthesis and solid-state structure of magnesium and calcium imidazole complexes

Three octahedral complexes built from N-alkylsubstituted imidazoles and magnesium as well as calcium chloride are reported. The obtained solid-state structures differ significantly from each other, depending on the size of the metal ion and the substituent on the imidazole. A chloro-bridged structure is found in the case of tert-butylimidazole, while for the iso-propylimidazole the calcium ion is coordinated by six imidazoles. For the smaller magnesium cation, we also found a sixfold coordination, but here only three of the iso-propylimidazoles coordinate to the magnesium, the three other positions are taken by water molecules.     

Influence of copper on proton efflux from Saccharomyces cerevisiae and the protective effect of calcium and magnesium

Abstract The inhibitory effect of Cu on glucose-dependent H⁺ efflux from Saccharomyces cerevisiae was manifest at low (micromolar) concentrations, with the time period between the addition of glucose and commencement of H⁺ efflux, H⁺ efflux rate and duration all being affected with increasing Cu concentration (5–100 μM). Ca, at a concentration of 0.5 mM, completely removed the inhibitory effect of Cu at concentrations up to 50 μM and considerably reduced it at higher concentrations (up to 150 μM). Mg exhibited a similar but weaker protective effect against the influence of Cu. The protective effect of Ca against 50 μM Cu was evident at low Ca concentrations (2.5–5 μM), whereas Mg was effective at ≥50 μ M. In order to prevent the inhibitory effect of Cu, it was necessary to add Ca or Mg to the cell suspension before Cu addition. It is concluded that the protective effect of Ca and Mg is mediated by competitive and stabilizing interactions at the cell surface as well as physiological functions of Ca and Mg.     

Age-dependent Increases of Calcium, Phosphorus, and Sodium in the Uterine Arteries of Thai

To elucidate compositional changes of the uterine artery with aging, the authors investigated age-related changes of elements in the uterine arteries of Thai. After ordinary dissection by medical students at Chiang Mai University was finished, the uterine arteries were resected from Thai subjects. Thai subjects ranged in age from 27 to 86 years (average age = 63.3 +/- 17.7 years). The element content of the uterine arteries was analyzed by inductively coupled plasma-atomic emission spectrometry. It was found that the Ca, P, and Na contents increased progressively in the uterine arteries of Thai with aging. A significant accumulation of Ca and P in the uterine arteries of Thai was found in the sixties patients, and the accumulation increased markedly in the seventies. Regarding the uterine arteries in subjects more than 60 years, the extent of accumulation of Ca and P in the uterine arteries of Thai was one half of that in the uterine arteries of Japanese. Regarding the relationships among elements, extremely significant direct correlations were found among the contents of Ca, P, Mg, Zn, and Na in the uterine arteries of Thai. As Ca increased in the uterine arteries of Thai, P, Mg, Zn, and Na increased simultaneously in the arteries.     

Nutrient mobilization by plasmodium of myxomycete Physarum rigidum in deadwood

To assess the nutrient mobilization ability of myxomycete plasmodia in deadwood, a microcosm experiment was conducted. Alive or dead plasmodia of a common lignicolous myxomycete, Physarum rigidum, were inoculated on field-collected crushed wood powder of white-rot or brown-rot pine wood and incubated for 24 d. The activity of living plasmodia led to increased concentrations of Ca²⁺, K⁺, Mg²⁺, Na⁺, and Cl⁻, but lower PO₄^3– concentrations in the wood powder. For NO₃⁻ concentration, the effect of living plasmodia was negative in white-rotted wood but positive in brown-rotted wood. These results suggest that the plasmodium of P. rigidum has the ability to mobilize nutrients in deadwood.     

钙离子、镁离子对戊型肝炎病毒感染PLC/PRF/5细胞的影响

为检测钙离子、镁离子对戊型肝炎病毒(hepatitis E virus,HEV)感染PLC/PRF/5细胞(人肝癌亚历山大细胞)的影响,本研究在各实验组PLC/PRF/5细胞的培养体系中加入等量HEV毒种进行孵育,利用实时荧光定量反转录聚合酶链反应以及酶联免疫法,监测HEV核酸和抗原含量;在HEV感染细胞实验组的维持培养液中分别加入钙离子、镁离子、乙二胺四乙酸(ethylene diamine tetraacetic acid, EDTA)和乙二醇二乙醚二胺四乙酸(ethylene glycol tetraacetic acid, EGTA),观察钙离子、镁离子、EDTA和EGTA分别存在以及钙离子、镁离子同时存在的情况下,HEV感染后不同时间PLC/PRF/5细胞内及培养上清液中HEV含量。结果显示,HEV接种细胞后1～24 h,钙离子、镁离子的加入能够促进病毒与细胞的结合,而金属离子鏊合剂的加入抑制了病毒与细胞的结合。HEV感染后2～5周,钙离子、镁离子均能增加PLC/PRF/5细胞培养上清液中产生的病毒,其中钙离子的促进作用更加显著。本研究结果表明钙离子、镁离子能够促进HEV感染细胞,在HEV接种细胞后的培养过程中,添加钙离子、镁离子有助于病毒产生。     

Seasonal variation of calcium,magnesium, potassium,and manganese contents in xylem sap of beech(Fagus sylvatica L.) in a 35-year-old limestone beech forest stand

Summary In a 35-year-old calcareous beech forest stand five beech trees (Fagus sylvatica L.) were felled every 2 weeks, and xylem sap was obtained by means of water displacement from the lowest trunk sections, each 100 cm in length. From mid-October 1988 to mid-October 1989 a total of 130 trees were investigated. The seasonal variations of the Ca, Mg, K and Mn contents, as well as those of pH, show four characteristic phases. Additionally, distribution of the mineral contents along the trunk was studied in four trees. The seasonal increase and decrease of xylem sap mineral contents along the trunk is shown for the characteristic phenophases. The Ca, Mg, K, and Mn contents of xylem saps were determined by means of atomicabsorptionspectrophotometry.     

Insights into modulation of calcium signaling by magnesium in calmodulin, troponin C and related EF-hand proteins

The Ca²⁺-binding helix-loop-helix structural motif called “EF-hand” is a common building block of a large family of proteins that function as intracellular Ca²⁺-receptors. These proteins respond specifically to micromolar concentrations of Ca²⁺ in the presence of ~1000-fold excess of the chemically similar divalent cation Mg²⁺. The intracellular free Mg²⁺ concentration is tightly controlled in a narrow range of 0.5-1.0 mM, which at the resting Ca²⁺ levels is sufficient to fully or partially saturate the Ca²⁺-binding sites of many EF-hand proteins. Thus, to convey Ca²⁺ signals, EF-hand proteins must respond differently to Ca²⁺ than to Mg²⁺. In this review the structural aspects of Mg²⁺ binding to EF-hand proteins are considered and interpreted in light of the recently proposed two-step Ca²⁺-binding mechanism (Grabarek, Z., J. Mol. Biol., 2005, 346, 1351). It is proposed that, due to stereochemical constraints imposed by the two-EF-hand domain structure, the smaller Mg²⁺ ion cannot engage the ligands of an EF-hand in the same way as Ca²⁺ and defaults to stabilizing the apo-like conformation of the EF-hand. It is proposed that Mg²⁺ plays an active role in the Ca²⁺-dependent regulation of cellular processes by stabilizing the “off state” of some EF-hand proteins, thereby facilitating switching off their respective target enzymes at the resting Ca²⁺ levels. Therefore, some pathological conditions attributed to Mg²⁺ deficiency might be related to excessive activation of underlying Ca²⁺-regulated cellular processes. This article is part of a Special Issue entitled: 11th European Symposium on Calcium.