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D Lichtstein

Publications and source records attributed to D Lichtstein.

At least 19 recordsLinked to original sources
PubMed2004

Diverse effects of stress and additional adrenocorticotropic hormone on digitalis-like compounds in normal and nude mice.

Digitalis-like compounds (DLC) are steroidal hormones that are synthesized in, and released from, the adrenal gland, whose regulation may be directed by the hypothalamic-pituitary-adrenal (HPA) axis. Increasing evidence points to antitumour properties of these compounds and we hypothesized that the establishment of tumours in athymic nude mice may be facilitated by an abnormal synthesis or secretion of DLC. To explore this hypothesis, DLC concentrations were determined in the plasma, and in adrenal and hypothalamic tissues of nude compared to normal mice under basal conditions, and 30 min after a stress stimulus (i.p. injection of 100 micro l saline) with or without additional adrenocorticotropic hormone (ACTH) 1 micro g/per animal. Simultaneously, plasma corticosterone and serum adrenocorticotropic hormone (ACTH) concentrations were analysed. The basal DLC concentrations were similar in the plasma and the hypothalamus of both strains, whereas the basal adrenal DLC concentration was significantly lower in the nude mice compared to normal mice. The stress stimulus induced in normal mice a significant increase in DLC concentrations in the adrenal gland, the plasma and the hypothalamus. However, in nude mice, it caused an increase only in the adrenal gland and the hypothalamus, whereas the plasma DLC concentration was not affected. In both strains, the administration of ACTH in addition to injection stress did not provoke a further increase in DLC concentrations while inducing a significant increase in plasma corticosterone concentration. Regardless of the applied stimulus, the nude mice expressed significant lower DLC concentrations in the adrenal gland and the plasma compared to normal mice. The low basal adrenal DLC concentration in nude mice and their impaired DLC response towards stress- and ACTH stimulation both support an involvement of DLC in tumorigenesis.

Adrenal Glands↗
PubMed2001

Endogenous digitalis-like Na+, K+-ATPase inhibitors, and brain function.

Digitalis-like compounds are recently identified steroids synthesized by the adrenal gland, which resemble the structure of plant cardiac glycosides. These compounds, like the plant steroids, bind to and inhibit the activity of the Na+, K+-ATPase. The possible function of the endogenous digitalis-like compounds has to be evaluated in view of the presence of different isoforms of the Na+, K+-ATPase, which differ in their sensitivity to digitalis. This review focuses on recent published data on the Na+, K+-ATPase inhibitors, the digitalis-like compounds, regarding their structure, biosynthesis and secretion from the adrenal gland, physiological role and pathological implications in diseases such as hypertension and depression. Emphasis is given to studies describing the involvement of these compounds in brain function.

Animals↗
PubMed2001

Dietary phosphate-dependent growth is not mediated by changes in plasma phosphate concentration.

The present study was undertaken in order to assess the role of dietary phosphate in growth. A diet deficient in phosphate led to a suppression of growth in juvenile rats. The phenomenon is similar to that described for diets deficient in other essential single components such as Mg, Zn or K. However, unlike the other constituents, dietary phosphate restriction affected the growth rate much more than it altered the serum phosphate concentration; addition of phosphate to the drinking water of rats fed a low-phosphate diet restored the growth rate without a concomitant change in serum phosphate concentration. The suppression of growth rate by the deletion of phosphate was associated with a delayed decrease in food intake. This finding implies that the variation in appetite was secondary to the change in growth. The increase in body weight following phosphate supplementation was associated with a concomitant increment in food intake. The phosphate-dependent growth was, however, evident also in rats that were pair-fed with those that were not supplied with phosphate. It is concluded that dietary phosphate-dependent growth is not mediated by changes in phosphate concentrations in the extracellular fluid. It is plausible that signals arising from receptors for phosphate in the digestive system constitute part of the growth control apparatus in rats.

Animals↗
PubMed2000

Digitalis and digitalislike compounds down-regulate gene expression of the intracellular signaling protein 14-3-3 in rat lens.

Na+,K+-ATPase activity in the epithelial layer is fundamental to the maintenance of ionic concentration gradients and transparency of the lens. Recently we have identified endogenous digitalislike compounds (DLC), 19-norbufalin and its peptide derivatives, in human cataractous lenses (Lichtstein et al. Eur J Biochem 216: 261-268, 1993). Lenses were treated with 10 nM ouabain, bufalin or 19-norbufalin derivative for 24 h and were compared to control lenses. Differential display analysis revealed that one of the down-regulated genes was 14-3-3 theta. Down-regulation was confirmed by Northern blot and by RT-PCR analysis. RT-PCR of additional 14-3-3 isoforms revealed that the eta and gamma isoforms of 14-3-3 are also down-regulated by ouabain, bufalin and 19-norbufalin derivative, whereas the zeta isoform is down-regulated only by bufalin. These results demonstrate that one of the consequences of Na+,K+-ATPase inhibition by exogenous or endogenous inhibitors is the down-regulation of mRNA transcripts encoding several isoforms of 14-3-3. Since the 14-3-3 proteins are multifunctional regulatory proteins, the reduction in the abundance of various isoforms will have profound effects on cell function. Furthermore, These results, together with the demonstration of digitalislike compounds in the normal lens, and their increased level in human cataractous lenses, strongly suggests their involvement in the molecular mechanisms responsible for cataract formation.

14-3-3 Proteins↗
PubMed2000

The mood cycle hypothesis: possible involvement of steroid hormones in mood regulation by means of Na+, K+-ATPase inhibition.

The mood cycle hypothesis attempts to propose a model for mood regulation based on current data. The hypothesis contends that steroid hormones inhibit sodium-potassium adenosine triphosphatase (Na+, K+-ATPase; Na+ pump) in the hypothalamus, either directly or by converting into digitalis-like compounds. This inhibition stimulates beta-endorphin (beta-E) secretion, which is normally construed as elevated mood. In turn, beta-E inhibits steroid secretion, thus completing negative feedback loops. These loops are collectively termed the mood cycle.

Affect↗
PubMed2000

Use of a lipophilic cation to monitor electrical membrane potential in the intact rat lens.

PURPOSE: Tetraphenylphosphonium (TPP+) is a permeant lipophilic cation that accumulates in cultured cells and tissues as a function of the electrical membrane potential across the plasma membrane. This study was undertaken to determine whether TPP+ can be used for assessing membrane potential in intact lenses in organ culture. METHODS: Rat lenses were cultured in media containing 10 microM TPP+ and a tracer level of 3H-TPP+ for various times. 3H-TPP+ levels in whole lenses or dissected portions of lenses were determined by liquid scintillation counting. Ionophores, transport inhibitors, and neurotransmitters were also added to investigate their effects on TPP+ uptake. RESULTS. Incubation of lenses in low-K+ balanced salt solution and TC-199 medium, containing physiological concentrations of Na+ and K+, led to a biphasic accumulation of TPP+ in the lens that approached equilibrium by 12 to 16 hours of culture. The TPP+ equilibrated within 1 hour in the epithelium but penetrated more slowly into the fiber mass. The steady state level of TPP+ accumulation in the lens was depressed by 90% when the lenses were cultured in a medium containing high K+. The calculated membrane potential for the normal rat lens in TC-199 was -75 +/- 3 mV. Monensin (1 microM) and nigericin (1 microM), Na+H+ and K+H+ exchangers respectively, as well as the protonophore carbonylcyanide-m-chlorophenylhydrazone (CCCP, 10 microM) and the calcium ionophore A23187 (10 microM), abolished TPP+ accumulation and caused cloudiness of the lenses. The neurotransmitter acetylcholine at 50 microM decreased TPP+ accumulation in the lens, but this effect could be prevented by simultaneous application of 1 mM atropine. CONCLUSIONS: TPP+ accumulation can be used as an indicator of changes in membrane potential in intact lenses, but because of the long time required to reach steady state, its utility is limited. The slow accumulation of TPP+ and its slow efflux from the lens under conditions known to depolarize membranes are consistent with a diffusion barrier in the deep cortex and nucleus of the lens.

Animals↗
PubMed1999

Na+, K+-ATPase inhibitors down-regulate gene expression of the intracellular signaling protein 14-3-3 in rat lens.

To identify genes that are differentially expressed by Na+, K+-ATPase inhibitors, we used the differential display technique to compare mRNA expression patterns in rat lens. Lenses were treated with 10 microM ouabain, bufalin, or 19-norbufalin derivative for 24 h and were compared with control lenses. Differential display analysis revealed that one of the down-regulated genes was 14-3-3 theta. Down-regulation was confirmed by Northern blot and by reverse transcription-polymerase chain reaction analysis. Reverse transcription-polymerase chain reaction of additional 14-3-3 isoforms revealed that the eta and gamma isoforms of 14-3-3 are also down-regulated by ouabain, bufalin, and 19-norbufalin derivative, whereas the zeta isoform is down-regulated only by bufalin. Down-regulation of the 14-3-3 isoforms occurred without a significant change in gamma-crystallin gene expression. These results demonstrate that one of the consequences of Na+, K+-ATPase inhibition by exogenous or endogenous inhibitors is the down-regulation of mRNA transcripts encoding several isoforms of 14-3-3. Because the 14-3-3 proteins are multifunctional regulatory proteins, the reduction in the abundance of various isoforms will have profound effects on cell function.

14-3-3 Proteins↗
PubMed1999

The effects of digitalis-like compounds on rat lenses.

PURPOSE: Fundamental to the maintenance of ionic concentration gradients and transparency of the lens is the activity of Na+,K+-adenosine triphosphatase (ATPase) in the epithelial layer. Recent studies have identified endogenous digitalis-like compounds (DLCs) and 19-norbufalin and its peptide derivatives in human cataractous lenses. These compounds inhibit the activity of Na+,K+-ATPase and have been suggested to be involved in cataract formation. The present experiments were designed to test this hypothesis by determining the ability of digitalis and DLCs to induce changes in protein composition and leakage from rat lenses in organ culture. METHODS: DLCs were determined in rat lenses using three independent assays: interaction with ouabain antibodies, interaction with bufalin antibodies, and inhibition of [3H]-ouabain binding to red blood cells. Rat lenses were incubated in modified TC-199 medium in 5% CO2 atmosphere at 37 degrees C for the time of the experiment. The onset of cataractogenesis was assessed by measuring protein leakage from lenses and by crystallin composition in the lens and media. RESULTS: DLCs were present in rat lens with concentrations 7 to 30 times higher in the capsular-epithelial layer than in the lens fibers regions. Ouabain, bufalin, digoxin, and DLC induced dose- and time-dependent leakage of protein from rat lenses. Lenses incubated with these compounds showed alterations in crystallin content consistent with changes that initiate opacity. All the compounds caused a multilayering of epithelial cells in the region surrounding the mitotic area and, at the same time, cell death in the central anterior region. CONCLUSIONS: Digitalis and endogenous DLCs are cataractogenic factors. These results, together with the demonstration of DLCs in the normal lens and their increased levels in human cataractous lenses, strongly suggest their involvement in the molecular mechanisms responsible for cataract formation.

Animals↗
PubMed1998

Biosynthesis of digitalis-like compounds in rat adrenal cells: hydroxycholesterol as possible precursor.

The biosynthesis of digitalis-like compounds (DLC) was determined in bovine and rat adrenal homogenates, as well as in primary rat adrenal cells, by following changes in the concentration of DLC using three independent sensitive bioassays: inhibition of [3H]-ouabain binding to red blood cells and competitive ouabain and bufalin ELISA. The amounts of DLC in bovine and rat adrenal homogenates, as measured by the two first bioassays, increased with time when the mixtures were incubated under tissue culture conditions. Rat primary adrenal cells were incubated in the presence of [1,2-(3)H]-25-hydroxycholesterol, [26,27-(3)H]-25-hydroxycholesterol or [7-(3)H]-pregnenolone. The radioactive products, as well as the digitalis-like activity, were fractionated by three sequential chromatography systems. When [1,2-(3)H]-25-hydroxycholesterol or [7-(3)H]-pregnenolone was added to the culture medium, the radioactivity was co-eluted with digitalis-like activity, suggesting that at least one of the DLC might originate in hydroxycholesterol. In contrast, when the culture medium was supplemented with [26,27-(3)H]-25-hydroxycholesterol, the radioactivity was not co-eluted with the digitalis-like activity, indicating that side chain cleavage is the first step in the synthesis of digitalis-like compounds by rat adrenal.

Adrenal Cortex↗
PubMed1997

Sodium-dependent transport of phosphate in neuronal and related cells.

Sodium-dependent phosphate entry into neuronal cells was demonstrated in synaptic plasma membrane vesicles and synaptosomes prepared from rat brains, in PC12 cells and in primary culture of pituitary cells. The extent of the sodium-dependent phosphate transport in the synaptic plasma membrane preparation, at [Na]out = 110 mM and [P(i)]out = 0.1 mM, varied between 0.28 to 1.02 nmol phosphate/mg membrane protein/min. In pituitary cells the value was only about 0.05 nmol P(i)/mg protein/min. In PC12 cells the activity increased from 0.0085 to 0.26 nmol P(i)/mg protein/min in the transit from undifferentiated to differentiated cells. The dependence of phosphate on sodium concentrations fits a model in which two sodium ions are required to transfer the phosphate into the cells with a K[Na]0.5 of 43 mM. The K(m) for the phosphate transport in the synaptic plasma membrane preparations was between 0.1 and 0.45 mM. It is concluded that sodium-driven active transport of phosphate is a ubiquitous activity in various types of neuronal cells.

Animals↗
PubMed1997

Digitalis-like compounds and Na+, K+-ATPase activity in bovine lens.

Digitalis-like compounds in bovine lens capsule, cortex and nucleus were determined quantitatively, following extraction, by their ability to inhibit [3H]ouabain binding to red blood cells. These compounds were found to be highly concentrated in the epithelium capsule and were significantly diminished in the cortex and nucleus. Na+, K+-ATPase density in the different regions was determined by [3H]ouabain binding to membranes and by autoradiography of lens slices. The highest concentration of [3H]ouabain-binding sites was observed to occur in membranes prepared from the epithelial cells of the capsule, and was almost 100- and 200-fold higher than the concentrations observed in membranes prepared from fiber cells of the cortex and nucleus, respectively. In the autoradiography studies, strong labeling of [3H]ouabain appeared in the epithelial cell zone, and only weak specific labeling appeared in the lens cortex and nucleus. Almost all (99%) of the Na+,K+-ATPase specific activity was found to be in the capsule epithelium and only 0.5% was measured in the cortex and no activity was detected in the nucleus. These results indicate that the digitalis-like compounds and Na+, K+-ATPase are concentrated in the lens capsule epithelium and are present only at low levels in the cortex and nucleus, thus implying that the lens capsular epithelial layer is the major region of the lens responsible for the homeostasis of ions and water in this tissue.

Animals↗
PubMed1997

Ouabain-sensitive Na+,K(+)-ATPase activity in toad brain.

Toads of the genus Bufo are highly resistant to the toxic effects of digitalis glycosides, and the Na+,K(+)-ATPase of all toad tissues studied to date has been relatively insensitive to inhibition by digitalis and related compounds. In studies of brain microsomal preparations from two toad species, Bufo marinus and Bufo viridis, inhibition of ATPase activity and displacement of [3H]ouabain from Na+,K(+)-ATPase occurred over broad ranges of ouabain or bufalin concentrations, consistent with the possibility that more than one Na+,K(+)-ATPase isoform may be present in toad brain. The data could be fitted to one- or two-site models, both of which were consistent with the presence of Na+,K(+)-ATPase activity with high sensitivity to ouabain and bufalin. Ki (concentration capable of producing 50% inhibition of activity) values for ouabain in the one-site model were in the 0.2 to 3.7 microM range, whereas Ki1 values in the two-site model ranged from 0.085 to 0.85 microM, indicating that brain ATPase was at least three orders of magnitude more sensitive to ouabain than B. marinus bladder ATPase (Ki = 5940 microM). Ouabain was also an effective inhibitor of 86Rb+ uptake in B. marinus brain tissue slices (Ki = 3.1 microM in the one-site model; Ki1 = 0.03 microM in the two-site model). However, the relative contribution of the high ouabain-sensitivity site to the total activity was 17% in the transport assay as compared with 63% in the Na+,K(+)-ATPase enzymatic assay. We conclude that a highly ouabain-sensitive Na+,K(+)-ATPase activity is present and functional in toad brain but that its function may be partially inhibited in vivo.

Animals↗
PubMed1995

Possible involvement of atrial natriuretic peptides in olfaction.

Atrial natriuretic peptides (ANP) are a family of humoral compounds released from the heart atria and involved in water and salt homeostasis. ANP immunoreactivity and ANP-binding sites were also found in several areas of the central nervous system including the olfactory bulb. In the present study, the possible involvement of ANP in olfaction was tested by measuring the content and distribution of IR-ANP and ANP-binding sites in rat olfactory bulb in control and rats. The results implicate ANP in the processes leading to olfactory perception.

Animals↗
PubMed1995

Multiple types of binding sites for atrial natriuretic peptide in rat olfactory bulb membranes and synaptosomes.

The binding of atrial natriuretic peptide (ANP) to rat olfactory bulb membranes and synaptosomes was examined. [125I]ANP (rat, 99-126) bound specifically to a single class of binding site on olfactory bulb membrane preparation with dissociation constant (Kd) of 106 pM and maximum binding capacity (Bmax) of 13.6 fmol/mg protein. Comparable results were obtained when the binding was characterized using displacement and kinetic experiments. The ring deleted analog of ANP, C-ANP (rat, 4-23) displaced [125I]ANP only minimally from its binding site in the membrane preparation. Saturation, displacement and blocking experiments on [125I]ANP binding to rat olfactory bulb synaptosomes revealed the presence of two distinct binding sites. Simultaneous analysis of homogeneous and heterogeneous displacement curves and blocking experiments revealed the quantitative characteristics of these receptors to be: Kd1 = 44 pM, Bmax1 = 42 fmol/mg protein and Kd2 = 1050 pM, Bmax2 = 173 fmol/mg protein, for the high and low affinity binding sites, respectively. Kinetic experiments further confirmed the differences between the receptors present in the membranes and the synaptosomes preparations. The demonstration of multiple ANP binding sites in olfactory bulb synaptosomes but not membrane preparations raises the possibility of a particular function of ANP in nerve terminals.

Animals↗
PubMed1995

Na+, K(+)-ATPase and heart excitability.

The Na+, K(+)-activated adenosine triphosphatase (ATPase) is present in the membrane of eukaryotic cells and represent a major pathway for Na+ and K+ transport across the plasma membrane. Cardiac glycosides, such as digoxin or ouabain, inhibit this enzyme activity by binding to a specific receptor on the membrane. Studies conducted in this and other laboratories have proven the existence of digitalis-like compounds in animal and human tissues which may serve as regulators, in vivo, of the Na+, K(+)-pump activity. The levels of digitalis-like compounds in the plasma are increased in hypertension and other illnesses. A possible link at the cellular and molecular level between these compounds and etiology of arrhythmias, an important cause of morbidity and mortality in patients with various diseases of the heart, can be postulated: Na+, K(+)-ATPase activity contributes directly and indirectly to the electrical membrane potential of cardiac cells. The inhibition of this pump by the endogenous digitalis-like compounds, in discrete areas of the heart, can induce changes of the membrane potential of these cells. These changes may cause an increase in excitability of the particular cells and contribute to the generation of arrhythmias.

Animals↗
PubMed1993

Identification of digitalis-like compounds in human cataractous lenses.

Human cataractous lens nuclei extract inhibited, in a dose-dependent fashion, [3H]ouabain binding to rat brain synaptosomes and microsomal Na(+)- and K(+)-dependent adenosine triphosphate (Na+, K(+)-ATPase) activity and interacted with anti-digoxin antibodies. The compounds responsible for these activities, termed digitalis-like compounds (DLC), were also detected in bovine, rat, cat and rabbit, normal, transparent lenses, but the levels were only 0.7-5.4% of the average levels in the cataractous human lenses. DLC from the human cataractous lenses were purified by a procedure consisting of organic extractions and batch chromatography followed by filtration through a 3000 Da cut-off filter and subsequent separations using reverse-phase high-performance liquid chromatography. The presence of DLC in the different fractions obtained in the chromatograms was monitored by their ability to inhibit [3H]ouabain binding and Na+, K(+)-ATPase activity. Based on chemical ionization mass spectrometry together with ultraviolet spectrometry and biological characterization, it is suggested that new bufodienolides, 19-norbufalin and 19-norbufalin peptide derivatives are responsible for the endogenous DLC activity. It is proposed that these compounds may regulate Na+, K(+)-ATPase activity in the lens under some physiological and pathological conditions.

Animals↗
PubMed1993

Digitalis-like compounds in the toad Bufo viridis: interactions with plasma proteins.

Digitalis-like compounds (DLC), normal constituents of animal tissues, are possible regulators of the Na+,K(+)-ATPase implicated in water and salt homeostasis. DLC are present in toad (Bufo viridis) tissues. Although DLC highest levels were found in toad skin, it was also detected in plasma and many internal organs. The abundant distribution and the different levels of DLC in various tissues exclude the possibility that toxicity is the only function of these compounds in the toad. The concentration of DLC in toad plasma is 30 microM, out of which 25-30% is bound to plasma proteins. Fractionation of toad plasma proteins on a G-100 Sephadex column followed by the extraction of DLC from the plasma proteins revealed that DLC are bound primarily to proteins of 48,000-53,000 Da. These results establish the existence of bufodienolide-binding protein(s) in animal plasma.

Animals↗