Environmental Calcium Concentrations Research Articles

Environmental calcium and variation in yolk sac size influence swimming performance in larval lake sturgeon (Acipenser fulvescens).

In many animal species, performance in the early life stages strongly affects recruitment to the adult population; however, factors that influence early life history stages are often the least understood. This is particularly relevant for lake sturgeon, Acipenser fulvescens, living in areas where environmental calcium concentrations are declining, partly due to anthropogenic activity. As calcium is important for muscle contraction and fatigue resistance, declining calcium levels could constrain swimming performance. Similarly, swimming performance could be influenced by variation in yolk sac volume, because the yolk sac is likely to affect drag forces during swimming. Testing swimming performance of larval A. fulvescens reared in four different calcium treatments spanning the range of 4-132 mg l-1 [Ca2+], this study found no treatment effects on the sprint swimming speed. A novel test of volitional swimming performance, however, revealed reduced swimming performance in the low calcium environment. Specifically, volitionally swimming larvae covered a shorter distance before swimming cessation in the low calcium environment compared with the other treatments. Moreover, sprint swimming speed in larvae with a large yolk sac was significantly slower than in larvae with a small yolk sac, regardless of body length variation. Thus, elevated maternal allocation (i.e. more yolk) was associated with reduced swimming performance. Data suggest that larvae in low calcium environments or with a large yolk sac exhibit reduced swimming performance and could be more susceptible to predation or premature downstream drift. Our study reveals how environmental factors and phenotypic variation influence locomotor performance in a larval fish.

Environmental Calcium Controls Alternate Physical States of the Caulobacter Surface Layer

Surface layers (S-layers) are paracrystalline, proteinaceous structures found in most archaea and many bacteria. Often the outermost cell envelope component, S-layers serve diverse functions including aiding pathogenicity and protecting against predators. We report that the S-layer of Caulobacter crescentus exhibits calcium-mediated structural plasticity, switching irreversibly between an amorphous aggregate state and the crystalline state. This finding invalidates the common assumption that S-layers serve only as static wall-like structures. In vitro, the Caulobacter S-layer protein, RsaA, enters the aggregate state at physiological temperatures and low divalent calcium ion concentrations. At higher concentrations, calcium ions stabilize monomeric RsaA, which can then transition to the two-dimensional crystalline state. Caulobacter requires micromolar concentrations of calcium for normal growth and development. Without an S-layer, Caulobacter is even more sensitive to changes in environmental calcium concentration. Therefore, this structurally dynamic S-layer responds to environmental conditions as an ion sensor and protects Caulobacter from calcium deficiency stress, a unique mechanism of bacterial adaptation. These findings provide a biochemical and physiological basis for RsaA’s calcium-binding behavior, which extends far beyond calcium’s commonly accepted role in aiding S-layer biogenesis or oligomerization and demonstrates a connection to cellular fitness.

The role of calcium concentration in the invasive capacity of Corbicula fluminea in crystalline basins.

The natural variation of environmental factors in freshwater basins determines their biodiversity. Among them, calcium is a key physiological compound for freshwater invertebrates. It is required for shell formation, muscle contraction, it mediates gene expression and allows counteracting acidosis during stress periods, among other functions. Although the distribution of different freshwater species has been suggested to be linked with the environmental calcium concentration, as yet, no research studies have confirmed this. Identifying whether environmental calcium concentrations might determine the invasion success of alien species would be critical in developing and implementing effective management strategies to control them. Here, a multidisciplinary approach integrating field surveys, analytical chemistry techniques, molecular biology analyses and a lab-scale experiment was taken to decipher whether the environmental calcium concentration might hamper the establishment of Corbicula fluminea in northwestern Iberian rivers. A Principal Component Analysis on water chemistry variables from 13 water bodies identified environmental calcium concentration, among others, as one key factor that best characterized the distribution area of C. fluminea. The calcium content in animals' bodies from two representative rivers was dependent on the environmental calcium concentration of freshwater basins; the lower the concentration, the lower the body's content. The expression of stress- and calcium homeostasis-related genes was higher in C. fluminea from low calcium concentration environments than in those from calcium-rich freshwater basins. Finally, under experimental conditions, lower water calcium concentrations decreased C. fluminea growth rates. The present data suggest, for the first time, that environmental calcium concentration may act as a determinant factor on the invasion success of C. fluminea in freshwater environments. Our results provide new clues for the identification of basins with increased risk of potential invasion by C. fluminea based on environmental calcium levels.

Effects of calcium and phosphorus limitation on the nutritional ecophysiology ofDaphnia

Declines in environmental calcium (Ca) and phosphorus (P) concentrations have occurred over the past 30 yrs in lakes across the Canadian Shield in southern Ontario, and these reductions appear to be placing strong constraints on populations of Daphnia in this region. Here, we report results from a factorial manipulation of Ca concentrations and food P content under controlled laboratory conditions where we measured resulting changes in daphnid elemental content, individual growth and survival, and life history traits related to population growth. We found significant effects of Ca- and P-limitation on all variables measured; however, dietary P explained a majority of the variation in daphnid nutrient content and growth. Dietary effects of low P [high food carbon (C): P ratios] on individual Daphnia life-history traits also translated into significant population level effects. Dietary P also explained relatively more experimental variation in population level responses than Ca concentrations. Experimental Ca concentrations most strongly altered daphnid survival partly due to the use of a lethally low Ca concentration in our experiment. Although recent work examining shifts in zooplankton communities in this region mainly focuses on the effects of Ca-limitation, we show that Ca concentrations and food nutrient content, at levels commonly found on the Canadian Shield, are both likely to strongly alter Daphnia life-history and populations dynamics. Our results underscore the need to more fully examine how multielemental limitation (e.g., Ca, N, P) affects consumer physiology and life-history given the plausible translation of these effects on the community structure of lake zooplankton.

Presence of calcium-binding motifs in PilY1 homologs correlates with Ca-mediated twitching motility and evolutionary history across diverse bacteria.

Twitching motility, involving type IV pili, is essential for host colonization and virulence of many pathogenic bacteria. Studies of PilY1, a tip-associated type IV pili protein, indicate that PilY1 functions as a switch between pilus extension and retraction, resulting in twitching motility. Recent work detected a calcium-binding motif in PilY1 of some animal bacterial pathogens and demonstrated that binding of calcium to PilY1 with this motif regulates twitching. Though studies of PilY1 in non-animal pathogens are limited, our group demonstrated that twitching motility in the plant pathogen Xylella fastidiosa, which contains three PilY1 homologs, is increased by calcium supplementation. A study was conducted to investigate the phylogenetic relationship between multiple PilY1 homologs, the presence of calcium-binding motifs therein, and calcium-mediated twitching motility across diverse bacteria. Strains analyzed contained one to three PilY1 homologs, but phylogenetic analyses indicated that PilY1 homologs containing the calcium-binding motif Dx[DN]xDGxxD are phylogenetically divergent from other PilY1 homologs. Plant-associated bacteria included in these analyses were then examined for a calcium-mediated twitching response. Results indicate that bacteria must have at least one PilY1 homolog containing the Dx[DN]xDGxxD motif to display a calcium-mediated increase in twitching motility, which likely reflects an adaption to environmental calcium concentrations.

Alpha-1 Giardin is an Annexin with Highly Unusual Calcium-Regulated Mechanisms

Alpha-giardins constitute the annexin proteome (group E annexins) in the intestinal protozoan parasite Giardia and, as such, represent the evolutionary oldest eukaryotic annexins. The dominance of alpha-giardins in the cytoskeleton of Giardia with its greatly reduced actin content emphasises the importance of the alpha-giardins for the structural integrity of the parasite, which is particularly critical in the transformation stage between cyst and trophozoite. In this study, we report the crystal structures of the apo- and calcium-bound forms of α1-giardin, a protein localised to the plasma membrane of Giardia trophozoites that has recently been identified as a vaccine target. The calcium-bound crystal structure of α1-giardin revealed the presence of a type III site in the first repeat as known from other annexin structures, as well as a novel calcium binding site situated between repeats I and IV. By means of comparison, the crystal structures of three different alpha-giardins known to date indicate that these proteins engage different calcium coordination schemes, among each other, as well as compared to annexins of groups A-D. Evaluation of the calcium-dependent binding to acidic phosphoplipid membranes revealed that this process is not only mediated but also regulated by the environmental calcium concentration. Uniquely within the large family of annexins, α1-giardin disengages from the phospholipid membrane at high calcium concentrations possibly due to formation of a dimeric species. The observed behaviour is in line with changing calcium levels experienced by the parasite during excystation and may thus provide first insights into the molecular mechanisms underpinning the transformation and survival of the parasite in the host.

The influence of environmental calcium concentrations on calcium flux, compensatory drinking and epithelial calcium channel expression in a freshwater cartilaginous fish

Calcium metabolism and mRNA levels of the epithelial calcium channel (ECaC) were examined in a freshwater cartilaginous fish, the lake sturgeon Acipenser fulvescens. Lake sturgeon were acclimated for ≥2 weeks to 0.1 (low), 0.4 (normal) or 3.3 (high) mmol l(-1) environmental calcium. Whole-body calcium flux was examined using (45)Ca as a radioactive marker. Net calcium flux was inward in all treatment groups; however, calcium influx was greatest in the low calcium environment and lowest in the high calcium environment, whereas efflux had the opposite relationship. A significant difference in the concentration of (45)Ca in the gastrointestinal tract (GIT) of fish in the low calcium environment led to the examination of drinking rate and calcium flux across the anterior-middle (mid) intestine. Drinking rate was not different between treatments; however, calcium influx across the mid-intestine in the low calcium treatment was significantly greater than that in both the normal and high calcium treatments. The lake sturgeon ECaC was 2831 bp in length, with a predicted protein sequence of 683 amino acids that shared a 66% identity with the closest sequenced ECaCs from the vertebrate phyla. ECaC mRNA levels were examined in the gills, kidney, pyloric caeca, mid-intestine and spiral intestine. Expression levels were highest in the gills, then the kidneys, and were orders of magnitude lower in the GIT. Contrary to existing models for calcium uptake in the teleost gill, ECaC expression was greatest in high calcium conditions and kidney ECaC expression was lowest in low calcium conditions, suggesting that cellular transport mechanisms for calcium may be distinctly different in these freshwater cartilaginous fishes.

Low environmental calcium blocks long-term memory formation in a freshwater pulmonate snail

The freshwater snail Lymnaea stagnalis (L.) is considered a calciphile and exhibits reduced growth and survival in environments containing less than 20mg/l environmental calcium. Although it has no apparent effect on survival at 20mg/l, reducing environmental calcium increases metabolic demand, and as such we consider that this level of calcium acts as a stressor on the snail. We exposed snails to acute periods of low environmental calcium and tested their ability to form intermediate-term memory (ITM) and long-term memory (LTM) following one trial operant conditioning (1TT) to reduce aerial respiratory activity in hypoxic conditions. We also assessed whether there were changes in the electrophysiological properties of a single neuron, right pedal dorsal 1 (RPeD1), which has been demonstrated to be necessary for LTM formation. Following training in high (80mg/l) environmental calcium, L. stagnalis formed ITM and LTM lasting 24h and demonstrated a significant reduction in all activity measured from RPeD1; however when snails were exposed to low (20mg/l) environmental calcium they were able to form ITM but not LTM. Although no behavioral LTM was formed, a partial reduction in RPeD1 activtiy measured 24h after training was observed, indicating a residual effect of training. The strong effect that environmental calcium concentration had on physiology and behavior in response to training to reduce aerial respiration in L. stagnalis suggests that it is an element of gastropod husbandry that needs to be carefully considered when studying other traits. This study also indicates that L. stagnalis found naturally in low calcium environments may be less able to adapt to novel stressors than populations found in harder waters.

Environmental calcium concentration affects long-term memory formation in a freshwater gastropod

Environmental calcium concentration affects long-term memory formation in a freshwater gastropod

Using elemental ratios of calcium and strontium to track calcium availability in the freshwater zooplankton Daphnia pulicaria

The assimilation of elements by an organism is dependent on both the environmental availability of the element and the processes of the organism. For some elements, organisms have a challenging time discriminating between nearly identical chemical analogs, for example, calcium and strontium. We tested the hypothesis that in environments where a desired element is scarce, the organism will assimilate a chemically similar analog at an increased rate. Populations of Daphnia pulicaria were manipulated using a microcosm in situ experiment and the results of that experiment tested with a field survey. Experimental results indicated a correlation between higher environmental calcium concentration and lower [Sr]/[Ca] ratios (R2 = 0.91, p < 0.05), suggesting that Daphnia in high calcium environments will assimilate more calcium relative to strontium. Field survey results across eight lakes confirmed that as lake calcium concentration increased, the value of [Sr]/[Ca] between the organism and the lake water decreased (R2 = 0.60, p < 0.05). Measurement of the elemental ratio of major and trace element analogs within organisms compared to their environments may be a useful tool for measuring the relative bioavailability of the major element, and provide insight into elemental limitation in other calcifying aquatic invertebrates.

Imitational modeling of process of evolution: From organic macromolecules to protocell and animal cell

A dynamic imitational model of initial stages of cell evolution has been developed based on role of environmental calcium concentration. The model is designed from our hypothesis about the medium of the appearance of protocells, which could be potassium water reservoirs rather than sea salt water with its predominance of sodium salts. The necessary elements of the appearance of the protocells served organic molecules, code of their synthesis, and formation of macromolecules under favorable ion concentration in environment: a high K+ and Mg2+ and a low Na+ concentration. The model is based on an assumption that one of the first stages in evolution of life was the appearance in the potassium-magnesium water reservoirs of organic molecules capable for self-replication on the basis of genetic code and formation of protocell with the potassium cytoplasm. The model has demonstrated necessity of formation of cell envelope for development of the protocell. Replacement of the dominant cation in water reservoirs—potassium by sodium—required the appearance of ion-transporting devices in plasma membrane and their participation in adaptation of cells to environment. This stage of evolution was accompanied by the most important morphofunctional event—formation of the plasma membrane instead of cell envelope. The membrane provided the ion asymmetry in the cell (preservation of K+ in it) relatively to the sodium external medium for maintaining optimal intracellular medium. In the model system, predecessors of animal cells elaborated mechanism of maintenance of the potassium cytoplasm with the sodium counterion dominating in the environment.

Transcriptional changes in response to growth of Arabidopsis in high external calcium

Transcriptional responses to growth in high environmental calcium concentrations were characterized and compared between wild-type and mutant Arabidopsis plants containing a knockout mutation in the gene encoding a cyclic nucleotide-gated channel (CNGC2). We show that the transcriptional profile of cngc2 plants grown in normal media resembled that from wild-type plants grown under elevated exogenous calcium conditions. The mutant grown in high-calcium media exhibited transcriptional changes not seen in the wild-type. The pattern of transcription suggests that adaptation to high external calcium overlaps with responses towards various biotic and abiotic stresses.

Environmental salinity regulates the in vitro production of [ 3H]-1,25-dihydroxyvitamin D 3 and [ 3H]-24,25 dihydroxyvitamin D 3 in rainbow trout ( Oncorhynchus mykiss)

Previous studies have shown that specific binding of 1,25-dihydroxyvitamin D 3 (1,25(OH) 2D 3) and 24,25-dihydroxyvitamin D 3 (24,25(OH) 2D 3) to enterocyte basolateral membranes (BLM), as well as circulating concentrations, is affected in response to changes in environmental salinity. It is not known if the production of 1,25(OH) 2D 3 and 24,25(OH) 2D 3 is affected by environmental salinity. The aim of the present study was to measure the in vitro production of [ 3H]-1,25(OH) 2D 3 and [ 3H]-24,25(OH) 2D 3 in fresh water (FW) and after 1, 2, 3, and 7 days after transfer to seawater (SW). Pooled sub-cellular fractions (mitochondria and microsomes) from liver or kidney was incubated with [ 3H]-25(OH)D 3 and the produced metabolites were separated using HPLC. Hepatic production of [ 3H]-1,25(OH) 2D 3 was decreased after 24 h in SW. This was followed by an up-regulation after 48 h and a second, slower decrease in production rate which leveled out after 7 days in SW. The production rate in SW was lower than the original rate in FW-adapted fish. For hepatic [ 3H]-24,25(OH) 2D 3 production the pattern was reversed. Renal production of [ 3H]-24,25(OH) 2D 3 increased significantly during the period of SW acclimation. These results suggest that environmental salinity regulates the production rate of the two antagonizing calcium regulatory hormones; 1,25(OH) 2D 3 and 24,25(OH) 2D 3. This gives further evidence to the hypothesis that there is a physiological regulation and a differentiated importance of 1,25(OH) 2D 3 and 24,25(OH) 2D 3 in relation to environmental calcium concentrations.

Survival and water balance of black sea bass held in a range of salinities and calcium-enhanced environments after abrupt salinity change

Black sea bass Centropristis striata juveniles survived abrupt transfer from 15 g/L salinity to 8 g/L, but not to 6 g/L salinity. Seven days after abrupt transfer, the plasma osmolality was proportional to environmental osmolality. Increasing environmental calcium concentration in receiving environments did not have an effect on either survival or plasma osmolality after abrupt transfer. Abrupt transfer from 15 g/L salinity to 4.5 g/L artificial sea salt + 4.5 g/L calcium chloride resulted in 100% mortality. Abrupt transfer of black sea bass to salinities below ∼ 15 g/L will have osmotic and perhaps survival consequences.

Regulated secretion of YopN by the type III machinery of Yersinia enterocolitica.

During infection, Yersinia enterocolitica exports Yop proteins via a type III secretion pathway. Secretion is activated when the environmental concentration of calcium ions is below 100 microM (low-calcium response). Yersiniae lacking yopN (lcrE), yscB, sycN, or tyeA do not inactivate the type III pathway even when the concentration of calcium is above 100 microM (calcium-blind phenotype). Purified YscB and SycN proteins form cytoplasmic complexes that bind a region including amino acids 16 to 100 of YopN, whereas TyeA binds YopN residues 101 to 294. Translational fusion of yopN gene sequences to the 5' end of the npt reporter generates hybrid proteins that are transported by the type III pathway. The signal necessary and sufficient for the type III secretion of hybrid proteins is located within the first 15 codons of yopN. Expression of plasmid-borne yopN, but not of yopN(1-294)-npt, complements the calcium-blind phenotype of yopN mutants. Surprisingly, yopN mutants respond to environmental changes in calcium concentration and secrete YopN(1-294)-Npt in the absence but not in the presence of calcium. tyeA is required for the low-calcium regulation of YopN(1-294)-Npt secretion, whereas sycN and yscB mutants fail to secrete YopN(1-294)-Npt in the presence of calcium. Experiments with yopN-npt fusions identified two other signals that regulate the secretion of YopN. yopN codons 16 to 100 prevent the entry of YopN into the type III pathway, a negative regulatory effect that is overcome by expression of yscB and sycN. The portion of YopN encoded by codons 101 to 294 prevents transport of the polypeptide across the bacterial double membrane envelope in the presence of functional tyeA. These data support a model whereby YopN transport may serve as a regulatory mechanism for the activity of the type III pathway. YscB/SycN binding facilitates the initiation of YopN into the type III pathway, whereas TyeA binding prevents transport of the polypeptide across the bacterial envelope. Changes in the environmental calcium concentration relieve the TyeA-mediated regulation, triggering YopN transport and activating the type III pathway.

Effect of environmental calcium levels on calcium uptake in tilapia larvae Oreochromis mossambicus

Effects of environmental calcium concentrations on the survival, growth, body calcium content and calcium uptake kinetics in developing tilapia (Oreochromis mossambicus) larvae were studied. Fertilized eggs were incubated in high- and low-calcium artificial freshwater (0.88-0.96 mmol l(-1) vs. 0.02-0.03 mmol l(-1) CaCl2 or CaSO4) until 3 days after hatching. Tilapia larvae showed similar hatching rates and wet weights in either high- or low-calcium medium, indicating neither the development nor the growth in tilapia larvae was affected by the environmental calcium levels. The body calcium content in low-calcium groups was about 90-95% that of high-calcium groups, No matter what calcium source was used (CaCl2 or CaSO4), acclimation to low calcium medium caused a stimulation of calcium uptake (measured in 0.2 mmol l(-1) calcium),i.e., 1.2-1.3 fold higher than that of high calcium groups. This enhanced calcium uptake capacity was characterized by a 50% decrease in Km and a 25% increase in Jmax. Effect of different calcium salts on calcium influx was significant only in low calcium level,i.e., calcium influx in low-CaCl2 group higher than that in low-CaSO4 group. These results suggest that tilapia larvae are able to modulate their calcium uptake mechanism to maintain normal body calcium content and growth in environments with different levels of calcium.

Characterization of Proteoglycans Synthesized by a Rat Parathyroid Cell Line

The structure, biosynthesis, and distribution of cell-associated proteoglycans in a clonal line of parathyroid cells, which exhibit differentiated characteristics such as calcium-regulated hormone secretion and cell growth, were studied by metabolic labeling with [3H] glucosamine and [35S]sulfate as precursors. Proteoglycans were isolated by two consecutive ion exchange chromatography steps and then analyzed by gel filtration, polyacrylamide gel electrophoresis, and specific enzyme and chemical reactions. The cells synthesize almost exclusively (greater than 95%) heparan sulfate (HS) proteoglycans with a glycosaminoglycan synthesis rate of approximately 0.5 micrograms/10(6) cells/24 h. Two major HS proteoglycan species were identified. HS proteoglycan-I has a mass of approximately kDa with a single HS chain (approximately 12 kDa) and a core protein of approximately 150 kDa including oligosaccharides. HS proteoglycan-II has a mass of approximately 170 kDa with 3-4 HS chains (approximately 30 kDa) and a core protein of 70-80 kDa including oligosaccharides. In the medium with low ionized calcium (0.05 mM), HS proteoglycan-I is synthesized at approximately 1.6 times the rate and HS proteoglycan-II at a similar rate as for cells cultured in the medium with high ionized calcium (2.1 mM). The distribution of proteoglycans, examined by the accessibility of the molecules to trypsin, was dramatically influenced by environmental calcium concentration; at low calcium levels 70-80% of the HS proteoglycans are trypsin-accessible while only 20-30% are accessible at high calcium levels. This suggests that the proteoglycans are primarily on the cell surface in low calcium and in trypsin-inaccessible compartments in high calcium conditions.

Cortisol stimulates whole body calcium uptake and the branchial calcium pump in freshwater rainbow trout

Exposure of rainbow trout to a reduced ambient calcium level (from 490 to 25 mumol Ca2+/l) caused hypocalcaemia and induced a rapid increase (within 1 h) in systemic cortisol levels. Under conditions of low environmental calcium concentrations, cortisol levels remained increased for at least 8 days. After this time the in-vitro Ca2+-transport capacity of branchial basolateral membrane vesicles was increased due to stimulation of Ca2+-ATPase activity, presumably as a result of chloride cell proliferation. Pituitary prolactin cells were unaffected by low ambient calcium levels. Fish kept in water containing 490 mumol Ca2+/l and treated with cortisol for 7 days displayed an increase in whole body calcium uptake and an enhancement of the branchial calcium transport capacity; concomitantly, hypercalcaemia was observed. We conclude that, in the rainbow trout, cortisol exerts hypercalcaemic effects by stimulating Ca2+ uptake from the water and that this effect forms an intrinsic part of the established mineralocorticoid action of cortisol in fish.

The Effect of Calcium on Mycelial Growth and Calcium Oxalate Crystal Formation inGilbertella Persicaria(Mucorales)

Patterns of calcium oxalate crystal development on sporangiophores and sporangia of Gilbertella persicaria are a consistent feature of this fungus when grown on a variety of standard mycological media. When grown on a liquid synthetic medium consisting of dextrose and salts, initial growth of the fungus and production of calcium oxalate crystals mimics growth on standard solid media. Soon, however, crystal production becomes aberrant, then ceases. Concomitant with the change in crystal morphology, growth of aerial portions of the colony increases dramatically. Normal colony morphology and crystal production are maintained when calcium salts are added to the synthetic medium. Significantly more fungal dry weight is produced on the synthetic medium when no calcium is added. Data gathered suggest that: 1) mineralized deposits may provide structural support; 2) early production of crystal-bearing sporangia lowers calcium concentrations in the medium sufficiently to alter and eventually stop calcium oxalate crystal production; 3) calcium may inhibit growth; 4) calcium oxalate crystal production in G. persicaria may regulate environmental calcium concentrations.

Abnormal calcium distribution in human parathyroid adenomas as possible cause of primary hyperparathyroidism.

Current knowledge suggests that normal parathyroid glands and parathyroid adenomas have different sensitivities to environmental calcium. In search for morphological equivalents, 5 normal human and 10 porcine parathyroid glands, as well as 10 human parathyroid adenomas were investigated with regard to intracellular and extracellular calcium distribution. The glands were incubated for 2, 4, 6 and 20 h in tissue cultures using HAM's F10 medium with various calcium concentrations. For visualization of the calcium distribution in the tissue the method of pyroantimonate precipitation was applied. Specificity of the reaction was controlled by X-ray microanalysis. Shifts of the calcium pyroantimonate precipitates were quantitated by morphometry using an area-counting system. The results demonstrate that in normal parathyroid glands calcium precipitates are distributed randomly. Incubation of normal glands in medium with low calcium concentration (0.6 mM) provoked reduced amounts of intracellular and extracellular calcium complexes. When the incubations were performed in medium with high calcium content (2.6 mM), calcium accumulated inside parathyroid and stroma cells. In contrast to normal parathyroid glands, parathyroid adenomas fixed immediately after surgery showed an atypical calcium distribution with low amounts of intracellular and high amounts of extracellular calcium grains. The data suggest that in normal parathyroid glands the intracellular calcium concentration follows the extracellular environmental calcium concentration. Thus, calcium modulates parathyroid hormone (PTH) secretion via intracellular regulatory mechanisms. In parathyroid adenomas the calcium transport via the tumor cell membrane appears to be disturbed resulting in lowered intracellular calcium levels. This is remarkable since the environmental calcium concentration is elevated due to the hypercalcemia of primary hyperparathyroidism.(ABSTRACT TRUNCATED AT 250 WORDS)