Biomedical subjects

F C Mohr

Publications and source records attributed to F C Mohr.

At least 19 recordsLinked to original sources

PubMed2006

Assessment of effects in mink caused by consumption of carp collected from the Saginaw River, Michigan, USA.

Polychlorinated hydrocarbons, including polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins (PCDDs), and polychlorinated dibenzofurans (PCDFs), are ubiquitous environmental contaminants that bioconcentrate in the food chain. Numerous studies have demonstrated mink (Mustela vison) to be one of the most sensitive species to this group of compounds. In recent studies, a lesion characterized by osteoinvasion of epithelial cells into the mandible and maxilla of young mink fed diets containing 3,3',4,4',5-pentachlorobiphenyl (PCB 126) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) was observed. The objective of the present study was to determine if proliferation of maxillary and mandibular squamous epithelia could be induced in ranch mink exposed to environmentally-derived polychlorinated hydrocarbons (PCBs, PCDDs, and PCDFs) in utero, during lactation, and throughout the growth period. Adult female mink were fed diets containing 0, 10, 20, or 30% carp (Cyprinus carpio) collected from the Saginaw River, Bay City, Michigan, USA, that provided 0.03, 0.83, 1.1, and 1.7 mg total PCBs (tPCBs)/kg feed and 2.5, 28, 47, and 73 ng TCDD toxic equivalents (TEQs)/kg feed, respectively, three weeks prior to breeding through weaning of the resulting offspring. Mink kits were maintained on their respective diets for up to 27 weeks of age. At 6 and 27 weeks of age, six to eight mink in each treatment group were necropsied and their jaws examined for evidence of maxillary and mandibular squamous epithelial proliferation. Results indicated that inclusion of up to 30% carp in the diet (1.7 mg tPCBs/kg feed, 73 ng TEQs/kg feed) had no effect on mink reproduction and kit survivability. However, maxillary and mandibular squamous epithelial proliferation was evident in four of the seven 27-week-old juveniles in the 20% carp group (1.1 mg tPCBs/kg feed, 47 ng TEQs/kg feed) and six of the eight juveniles in the 30% carp group (1.7 mg tPCBs/kg feed group, 73 ng TEQs/kg feed). Hepatic concentrations of tPCBs and TEQs increased in both the 6-week-old kits and the 27-week-old juveniles as the percentage of dietary carp increased. The livers of 6-week-old kits were also assessed for the presence of polybrominated diphenyl ethers, which increased as the percentage of Saginaw River carp in the diet increased.

Animal Feed↗

PubMed2005

Pathology of bartonella endocarditis in six dogs.

In a 5-year retrospective study of dogs presenting to the Veterinary Medical Teaching Hospital at the University of California, Davis, there were 31 histologic diagnoses of valvular endocarditis. By polymerase chain reaction (PCR) amplification of embedded valvular tissue, Bartonella organisms were exclusively associated with 6 out of 31 cases (19%). Confirmed Bartonella cases involved the aortic valve alone (five out of six) or in combination with the mitral valve (one of six). Microscopic features of Bartonella endocarditis were compared with valves from non-Bartonella endocarditis and with valvular change unrelated to infectious agents (endocardiosis). Features of Bartonella endocarditis included a combination of fibrosis, mineralization, endothelial proliferation, and neovascularization with variable inflammation. None of these features is specific; however, the combination is distinct both from endocarditis caused by culturable bacteria and from endocardiosis. Ultrastructural analyses revealed both extracellular and intraendothelial bacteria. Clinical history, serology, and PCR are currently necessary to establish an etiologic diagnosis of Bartonella endocarditis.

Animals↗

PubMed2004

Primary diffuse tracheobronchial amyloidosis in a dog.

Primary diffuse tracheobronchial amyloidosis was diagnosed at necropsy in an intact male Akita dog aged 11 years, a non-productive chronic cough having been the only related clinical sign. Histologically, eosinophilic hyalinized deposits were found as a band in the lamina propria underneath the epithelium of the trachea and bronchi. When stained with Congo red, apple-green birefringence was observed in the deposits viewed with polarized light. The amyloid did not lose sensitivity to Congo red staining after incubation with potassium permanganate, indicating that it was of the AL (amyloid light chain) type. Ultrastructural features of the amyloid included a typical fibrillar meshwork with individual fibrils measuring 9.5 to 10.5 nm in diameter. This is the first report of primary diffuse tracheobronchial amyloidosis in the dog.

Amyloid↗

PubMed1998

Primary hyperparathyroidism caused by a functional parathyroid adenoma in a horse.

A 14-year-old Arabian gelding had weight loss and anorexia of 3 weeks' duration. Results of repeated laboratory tests revealed persistent hypercalcemia and serum phosphorus concentration that was within or less than the reference range. Parathyroid hormone concentration was high. Histologic examination of specimens obtained at necropsy revealed parathyroid adenoma. A diagnosis of primary hyperparathyroidism attributable to a functional parathyroid adenoma was made. Abnormalities in calcium and phosphorus concentrations were similar to those seen with primary hyperparathyroidism in dogs, in which this syndrome is best described. Primary hyperparathyroidism should be considered to be a potential cause of hypercalcemia in horses in which other more common causes of hypercalcemia, such as humoral hypercalcemia of malignancy, nutritional secondary hyperparathyroidism, chronic renal failure, vitamin D toxicosis, and bony or granulomatous disease, are ruled out.

Adenoma↗

PubMed1997

Bastadins relate ryanodine-sensitive and -insensitive Ca2+ efflux pathways in skeletal SR and BC3H1 cells.

Bastadins potently interact with the FK-506-binding protein of 12 kDa (FKBP12)-ryanodine receptor (Ry1R) complex in skeletal muscle to enhance a high-affinity ryanodine binding conformation (M. M. Mack, T. F. Molinski, E. D. Buck, and I. N. Pessah. J. Biol. Chem. 269: 23236-23249, 1994). Bastadins are used to examine the relationship between ryanodine-sensitive and ryanodine-insensitive Ca2+ efflux pathways that coexist in junctional sarcoplasmic reticulum (SR) vesicles from rabbit skeletal muscle and differentiated BC3H1 cells. Complete block of caffeine-sensitive Ca2+ channels with micromolar ryanodine or ruthenium red does not alter the steady-state loading capacity of SR. Inhibition of sarco(endo)plasmic reticulum Ca2+-ATPase (SERCA) pumps with thapsigargin unmasks a ryanodine- and ruthenium red-insensitive Ca2+ efflux pathway. Bastadin 5 alone does not inhibit Ca2+ efflux unmasked by inhibition of SERCA pumps, but, in combination with blocking concentrations of ryanodine or ruthenium red, it eliminates the ryanodine-insensitive Ca2+ "leak" and enhances steady-state loading capacity of SR vesicles approximately 2.5-fold. These actions of bastadins occur in the same concentration range that enhances the number of high-affinity binding sites for [3H]ryanodine (50% effective concentration of approximately 2 microM). Similar effects on SR Ca2+ transport are found with FK-506 and ryanodine in combination. Block of Ry1R in intact BC3H1 cells with ryanodine does not eliminate the prominent Ca2+ leak unmasked by thapsigargin. A membrane-permeant mixture of bastadins in combination with ryanodine nearly eliminates the Ca2+ leak unmasked by thapsigargin, even though the Ca2+ stores are replete. The requirement of both a known Ry1R blocker and bastadins in combination provides a pharmacological link between ryanodine-sensitive Ca2+ channels and ryanodine-insensitive leak pathways in isolated junctional SR and BC3H1 cells. Together, these results strongly suggest that bastadins, through their modulatory actions on the FKBP12-Ry1R complex, convert ryanodine-insensitive leak states into ryanodine-sensitive channels that recognize [3H]ryanodine with high affinity.

Animals↗

PubMed1996

Herpesvirus particles associated with oral and respiratory lesions in a California desert tortoise (Gopherus agassizii).

A 60-year-old captive California desert tortoise (Gopherus agassizii) which died in August 1990 at the University of California, Davis, California (USA), during treatment for colonic impaction had marked caseous necrosis of the oral cavity, choana, trachea, and lungs. Numerous intranuclear inclusion bodies and a large number of syncytial giant cells were seen in the oral cavity and respiratory tract along with bacterial granulomas. Pasteurella testudinis, Streptococcus veridans, and coagulase-negative Staphilococcus spp. were cultured from the lesions. Using electron microscopy, herpesvirus particles were observed in intranuclear inclusions and cytoplasm. Viral stomatitis, tracheitis, and bronchopneumonia complicated by bacterial infection were diagnosed. Although respiratory disease is common in desert tortoises, this is believed to be the first report of association with a viral infection.

Animals↗

PubMed1995

The delta isomer of hexachlorocyclohexane induces rapid release of the myo-inositol-1,4,5-trisphosphate-sensitive Ca2+ store and blocks capacitative Ca2+ entry in rat basophilic leukemia cells.

Antigenic stimulation of rat basophilic leukemia cells releases Ca2+ from internal stores and increases membrane permeability to Ca2+. The delta isomer of hexachlorocyclohexane (delta-HCH) is structurally similar to myo-inositol-1,4,5-trisphosphate (IP3) and is a potent releaser of stored Ca2+ from permeabilized cells. This release of Ca2+ is not mediated by a competitive interaction with the IP3 receptor on the Ca2+ release channel on the endoplasmic reticulum. In intact cells, delta-HCH and, to a lesser extent, lindane (gamma-hexachlorocyclohexane) transiently increase the intracellular Ca2+ concentration. The return to basal concentrations is mediated by the plasma membrane Ca2+ pumps and not by resequestration of Ca2+ into intracellular stores. Treatment of cells with delta-HCH (25-100 microM), but not lindane, leads to a progressive inhibition of the antigen- and thapsigargin-stimulated Ca2+ signal. Caffeine, a modulator of the ryanodine receptor Ca2+ channel, attenuates the rise in intracellular Ca2+ induced by delta-HCH, suggesting that ryanodine receptor-like Ca2+ channels may be present in RBL cells. At 25 microM delta-HCH, a concentration that does not inhibit the antigen-stimulated Ca2+ signal, the release of [3H]serotonin from antigen-stimulated cells is enhanced as is secretion of [3H]serotonin from cells pretreated with 25-100 microM lindane. The depletion of Ca2+ from intracellular stores by delta-HCH should evoke Ca2+ entry into the cells by a capacitative mechanism; however; divalent cation permeability across the plasma membrane (Mn2+ influx) is not increased but rather is decreased by delta-HCH. An understanding of the mechanism of action of delta-HCH in releasing stored Ca2+ and blocking Ca2+ influx across the plasma membrane may provide insights into the regulation of capacitative Ca2+ entry in nonexcitable cells.

Adenosine Triphosphate↗

PubMed1994

Relationships between maternal and fetal liver copper, iron, manganese, and zinc concentrations and fetal development in California Holstein dairy cows.

Associations between maternal trace element deficiencies and abortion have been made for many mammalian species. Objectives of this study were to estimate and correlate maternal and fetal hepatic Cu, Fe, Mn, and Zn concentrations through gestation. Additionally, aborted fetuses, stratified by cause of abortion (infectious or noninfectious), were compared to size-matched nonaborted fetuses to examine for magnitude and direction of change in hepatic trace element status. Dam and fetal liver were removed at slaughter from 103 Holstein dairy cows judged grossly normal by ante- and postmortem examination. Liver samples were collected from fetuses submitted by veterinarians for routine diagnosis of abortion (n = 80). Hepatic Cu, Fe, Mn, and Zn concentrations were determined by flame spectrophotometry. Comparisons of groups, estimations of correlations, and derived prediction equations were made by least-squares methods. Maternal liver Cu, Fe, Mn, and Zn concentrations did not vary during gestation. Compared with the dam, fetal liver Fe and Zn concentrations were higher (P < 0.05), fetal Cu concentrations were similar (P > 0.05), and fetal liver Mn concentrations were lower (P < 0.05). As fetal size increased, fetal liver Cu and Zn concentrations increased (P < 0.05), fetal liver Fe concentration decreased (P < 0.05), and fetal liver Mn did not change (P > 0.05). Aborted fetuses had lower liver Cu, Mn, and Zn concentrations than did nonaborted fetuses (P < 0.05). Liver Fe concentration was lower in aborted fetuses than in nonaborted fetuses in the second trimester only (P < 0.05). Consistently lower liver Cu, Fe, Mn, and Zn concentrations in aborted fetuses suggest a nonspecific change in trace element status, which implies an effect of abortion, not a cause of abortion.

Animals↗

PubMed1993

Regulation of inositol 1,4,5-trisphosphate receptors in rat basophilic leukemia cells. I. Multiple conformational states of the receptor in a microsomal preparation.

A detailed characterization of the inositol 1,4,5-trisphosphate (IP3) receptor in rat basophilic leukemia (RBL) cells, a neoplastic mast cell line, has been possible through the growth of solid RBL cell tumors which provide a rich source of IP3 receptor. Equilibrium binding studies show a 1.6 +/- 0.1 pmol/mg of protein maximal binding capacity for [3H]IP3 at optimal Ca2+ (10 microM). The specificity of the RBL cell IP3 receptor towards phosphoinositides, ATP and heparin parallels those previously described with excitable and nonexcitable tissues. [3H]IP3 binding is slightly enhanced from < 1 nM to 10 microM Ca2+ and inhibited by > 10 microM Ca2+. Kinetic and equilibrium studies provide evidence for at least two classes or conformational states of binding sites with pico- and nanomolar affinities. At nM concentrations of IP3, neither binding to the IP3 receptor nor IP3-induced Ca2+ efflux from permeabilized cells demonstrates cooperativity. In contrast, at pM concentrations, IP3 binding kinetics deviate from simple mass action suggesting a complex interaction among binding sites for IP3 on the receptor-channel oligomer. The mechanisms that regulate [3H]IP3 binding in RBL cells are unique when compared to what has been reported in other cells.

Animals↗

PubMed1993

Regulation of inositol 1,4,5-trisphosphate receptors in rat basophilic leukemia cells. II. Modulation of the receptor in permeabilized cells by the cytosolic compartment.

Engagement of the IP3 receptor by its ligand releases Ca2+ from intracellular stores of the rat basophilic leukemia (RBL) cell. The IP3 receptor in washed permeabilized cells has high affinity (Kd = 1.2 +/- 0.3 nM) for [3H]IP3 and is not sensitive to physiological concentrations of Ca2+. Moreover, washed permeabilized cells only release small amounts of Ca2+ when stimulated with IP3. When [3H]IP3 binding to permeabilized cells is performed in the presence of cytosolic constituents (unwashed cells), the IP3 receptor has a lower affinity for [3H]IP3 (Kd from 20 to 100 nM) and has enhanced Ca2+ release. Cytosolic supernatant, prepared by centrifugation of permeabilized cells and added back to washed permeabilized cells, decreases [3H]IP3 binding in a dose-dependent manner and increases the amount of Ca2+ released by IP3. Depletion of either MgATP or IP3 in the cytosolic supernatant does not affect the supernatant's ability to decrease [3H]IP3 binding. Though MgATP competitively inhibits [3H]IP3 binding, it cannot fully account for the shift in Kd or the modulation of IP3-stimulated Ca2+ release in the presence of cytosol. These findings suggest that components present in the cytosolic supernatant modulate the function of the IP3 receptor by maintaining it in a low affinity state capable of promoting Ca2+ release.

Adenosine Triphosphate↗

PubMed1992

Avian scleroderma: evidence for qualitative and quantitative T cell defects.

T cell activation is dependent upon calcium influx and protein kinase C activation, with subsequent lymphocyte proliferation dependent upon IL-2. Abnormalities in T cell proliferation, including abnormal calcium influx and defective protein kinase C activation, have been identified in aged mice and humans and many autoimmune diseases including diabetes, lupus and scleroderma. Since UCD line 200 chickens, which spontaneously develop a scleroderma-like disease, have both thymic defects and a diminished peripheral blood lymphocyte response to IL-2, we have further investigated T cell function in these birds. Interestingly, line 200 T cells respond poorly in vitro to a variety of diversely acting T cell mitogens including concanavalin A, phytohemagglutinin and anti-chicken CD3 monoclonal antibody. Moreover, they do not respond well even to phorbol myristate acetate in conjunction with ionomycin. Addition of exogenous IL-2-containing supernatant concurrently with mitogenic stimulation also had no significant effect. Analysis of intracellular free calcium demonstrated that the lymphocytes from diseased birds had a reduced influx of calcium (or release for intracellular stores) following stimulation. These data clearly reflect a unique defect in T cell activation associated with avian scleroderma. Analysis of chicken CD3, CD4 and CD8 expression revealed a 39% decrease in peripheral blood CD4+ cells in scleroderma birds, although this decrease was not sufficient to explain the 80-90% decrease observed in proliferation assays and calcium influx. Our data support the hypothesis that avian scleroderma is mediated via abnormal function of lymphocyte co-stimulatory molecules or intracellular calcium regulators.

Animals↗

PubMed1992

Stereoselective modulation of ryanodine-sensitive calcium channels by the delta isomer of hexachlorocyclohexane (delta-HCH).

delta-Hexachlorocyclohexane (delta-HCH) is shown to be 30-fold more potent as a positive inotropic agent with rat atrial strips compared with lindane (gamma-HCH). Threshold and ED50 values for enhanced contractile force at a pacing frequency of 0.5 Hz are less than 1 microM and 2.2 microM for delta-HCH and 40 microM and 63 microM for gamma-HCH, respectively. Contracture developed in atria exposed to greater than 4 microM delta-HCH (ED50 = 11 microM) but not in atria exposed to gamma-HCH. Uptake and release of Ca++ measured from actively loaded cardiac sarcoplasmic reticulum (SR) vesicles is measured with antipyrylazo III. Although delta-HCH (30 microM) decreases Ca(++)-dependent ATPase by 20%, it does not significantly alter Ca++ loading in the presence of ruthenium red. Addition of delta-HCH (5-50 microM) after loading is complete causes rapid, dose-dependent release of Ca++ from SR. Ca++ release induced by delta-HCH is markedly stereoselective. Compared with gamma-HCH (50 microM), delta-HCH (50 microM) induces a nearly 20-fold higher initial rate of Ca++ release (4.3 nmol of Ca++/mg/sec). Studies with [3H]ryanodine demonstrate that delta-HCH sharply inhibits Ca(++)- or daunorubicin-activated radioligand binding (IC50 = 37 and 25 microM, respectively, logit slope = 2). Inhibition of [3H]ryanodine-binding by delta-HCH is stereoselective inasmuch as IC50 values for alpha, beta and gamma isomers are greater than 100 microM. The delta-HCH modified Ca++ channel appears to proceed by a noncompetitive mechanism (reducing Bmax in equilibrium experiments) with respect to the conformationally sensitive binding site for [3H]ryanodine.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphatases↗

PubMed1992

Culture and initial characterization of the secretory response of neoplastic cat mast cells.

Mast cells isolated from feline splenic mastocytomas were cultured to study their structural and functional properties. Isolated cells from various cats were grown as monolayer cultures for a mean of 56 days (range, 30 to 76 days). Cat mast cells released allergic mediators in response to compound 48/80, anti-cat serum antibodies, and concanavalin A. On the basis of the finding that secretion from cat mast cells was stimulated by anti-cat serum antibodies and concanavalin A, these cells contain surface-bound immunoglobulins. The presence of mast cell-sensitizing antibodies has been suspected in cats, but never before directly demonstrated. Cultured cat mast cells have cytochemical and functional characteristics common to connective tissue-type mast cells and provide one of the few non-rodent models of cultured cells for the study of this type of mast cell.

Animals↗

PubMed1990

The effect of mitochondrial inhibitors on calcium homeostasis in tumor mast cells.

The depletion of intracellular ATP by mitochondrial inhibitors in a glucose-free saline solution inhibited antigen-stimulated 45Ca uptake, the rise in cytoplasmic calcium, measured by fura-2, and secretion in rat basophilic leukemia cells. Lowering the intracellular ATP concentration also released calcium from an intracellular store and made further 45Ca efflux from the cells unresponsive to subsequent antigen stimulation. Antigen-stimulated 45Ca efflux could be restored by the addition of glucose. The ATP-sensitive calcium store appeared to be the same store that releases calcium in response to antigen. In contrast, intracellular ATP was not lowered, and antigen-stimulated secretion was unaffected by mitochondrial inhibitors, provided that glucose was present in the bathing solution. Similarly, antigen-stimulated 45Ca uptake, 45Ca efflux, and the rise in free ionized calcium were unaffected by individual mitochondrial inhibitors in the presence of glucose. However, when the respiratory chain inhibitor antimycin A was used in combination with the ATP synthetase inhibitor oligomycin in the presence of glucose, antigen-stimulated 45Ca uptake was inhibited, whereas the rise in free ionized calcium and secretion were unaffected. Also, antigen-induced depolarization (an indirect measurement of Ca2+ influx across the plasma membrane) was not affected. The inhibition of antigen-stimulated 45Ca uptake could, however, be overcome if a high concentration of the Ca2+ buffer quin2 was present in the cells to buffer the incoming 45Ca. These results suggest that in fully functional rat basophilic leukemia cells the majority of the calcium entering in response to antigen stimulation is initially buffered by a calcium store sensitive to antimycin A and oligomycin, presumably the mitochondria.

Adenosine Triphosphate↗

PubMed1989

Calcium: an important second messenger in mast cells.

Recently there has been considerable controversy over the mechanism(s) by which intracellular Ca2+ is elevated when receptors for IgE on the surface of mast cells are aggregated by antigen. The central role played by calcium in the initiation of secretion from these cells has also been called into question. In a mast cell line which has been widely used to study stimulus-secretion coupling in non-excitable cells it is now clear that calcium is indeed important in the physiological response of the cells but that other intracellular messengers are also involved. In addition it has been shown that while the increase in intracellular Ca2+ probably originates from intracellular stores it can only be sustained by the influx of calcium across the plasma membrane. The nature of the Ca2+ permeability pathway has yet to be elucidated although a number of candidates for the calcium channel in mast cells have been proposed. Significant oscillations and spatial gradients of Ca2+ are often seen when the responses of individual antigen-stimulated cells are measured using digital imaging microscopy. The complexity of these responses highlights the importance of single-cell measurements in elucidating the relationship between IgE receptor activation, Ca2+ movements and exocytosis.

Calcium↗

PubMed1987

The relative contributions of extracellular and intracellular calcium to secretion from tumor mast cells. Multiple effects of the proton ionophore carbonyl cyanide m-chlorophenylhydrazone.

The proton ionophore carbonyl cyanide m-chlorophenylhydrazone (CCCP) inhibited antigen-stimulated secretion and calcium influx in rat basophilic leukemia cells. In a glucose-free solution the inhibitory effects of CCCP were due to a decrease in the intracellular ATP concentration; however, when glucose was present there was no decrease in ATP. Instead, we found that in a glucose-containing saline solution, CCCP inhibited antigen-stimulated calcium uptake because it depolarized the plasma membrane, which in rat basophilic leukemia cells inhibits antigen-stimulated calcium uptake. In the presence of glucose, relatively low concentrations of CCCP inhibited calcium uptake while higher concentrations were required to inhibit secretion. In contrast, the initial antigen-stimulated rise in cytoplasmic calcium, measured with the fluorescent calcium indicator quin2, was not inhibited by CCCP. This suggests that the release of calcium from intracellular stores might, in some cases, be sufficient to support antigen-stimulated secretion. In the presence of CCCP the pH gradient becomes important for regulating the membrane potential across the plasma membrane. When cells were depolarized with CCCP and the external pH was increased, the membrane potential returned to resting levels and antigen-stimulated calcium uptake was restored. Inhibition of antigen-stimulated secretion by higher concentrations of CCCP could also be reversed by increasing the external pH.

Adenosine Triphosphate↗

PubMed1987

IgE receptor-mediated depolarization of rat basophilic leukemia cells measured with the fluorescent probe bis-oxonol.

Receptor-mediated changes in plasma membrane potential were recorded in rat basophilic leukemia (RBL) cells with the potential-sensitive fluorescent indicator bis-oxonol. Depolarization of the mitochondria with metabolic inhibitors was not detected by bis-oxonol, suggesting that only potential changes across the plasma membrane were being measured. The resting membrane potential of RBL cells was largely generated by the equilibrium distribution of K+ and not through electrogenic activity of the sodium pump. Depolarization was maintained as long as IgE receptors remained aggregated. We believe that at physiologic calcium concentrations a large portion of the measured potential change may be due to calcium influx across the plasma membrane. Prevention of calcium influx by lanthanum, disruption of aggregated receptors, or prior depolarization in a high K+ saline solution completely inhibited the antigen-induced depolarization. The time course of the antigen-stimulated increase in bis-oxonol fluorescence was similar, but not identical, to the antigen-stimulated rise in cytoplasmic free ionized calcium measured with fura-2. Antigen-stimulated depolarization was inhibited by removing both calcium and sodium and could be restored by the addition of either ion. Reduction of total cellular adenosine triphosphate inhibited depolarization in response to antigen stimulation.

Adenosine Triphosphate↗

PubMed1987

Depolarization of rat basophilic leukemia cells inhibits calcium uptake and exocytosis.

We have investigated the unusual observation that depolarization of rat basophilic leukemia cells in high potassium not only fails to induce secretion, but also inhibits the secretion induced when receptors for IgE are aggregated by antigen. Antigen-stimulated 45Ca uptake and the rise in cytoplasmic free ionized calcium measured with the fluorescent indicator quin2 were both inhibited in depolarized cells. 45Ca efflux, on the other hand, was unaffected, which confirms that IgE receptor activation was not impaired in high potassium. Unlike the large increase in total cell calcium seen when cells in normal saline solution were stimulated with antigen, there was a decrease in total cell calcium when depolarized cells were stimulated. This is consistent with our finding that 45Ca uptake was inhibited while 45Ca efflux was unaffected. Inhibition of 45Ca uptake and secretion closely paralleled the decrease in membrane potential, and could be overcome by increasing the extracellular calcium concentration. We conclude that changes in the electrochemical gradient for calcium are important in determining calcium influx and the magnitude of antigen-stimulated secretion from rat basophilic leukemia cells, while the release of calcium from intracellular stores is unaffected.

Animals↗