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J H Schwartz

Publications and source records attributed to J H Schwartz.

At least 19 recordsLinked to original sources
PubMed1999

Purinergic modulation of Na(+),K(+),Cl(-) cotransport and MAP kinases is limited to C11-MDCK cells resembling intercalated cells from collecting ducts.

We demonstrated recently that in renal epithelial cells from collecting ducts of Madin-Darby canine kidneys (MDCK), Na(+),K(+), Cl(-) cotransport is inhibited up to 50% by ATP via its interaction with P(2Y) purinoceptors (Biochim. Biophys. Acta 1998. 1369:233-239). In the present study we examined which type of renal epithelial cells possesses the highest sensitivity of Na(+),K(+),Cl(-) cotransport to purinergic regulation. We did not observe any effect of ATP on Na(+),K(+),Cl(-) cotransport in renal epithelial cells from proximal and distal tubules, whereas in renal epithelial cells from rabbit and rat collecting ducts ATP decreased the carrier's activity by approximately 30%. ATP did not affect Na(+),K(+),Cl(-) cotransport in C7 subtype MDCK cells possessing the properties of principal cells but led to approximately 85% inhibition of this carrier in C11-MDCK cells in which intercalated cells are highly abundant. Both C7- and C11-MDCK exhibited ATP-induced IP(3) and cAMP production and transient elevation of [Ca(2+)](i). In contrast to the above-listed signaling systems, ATP-induced phosphorylation of ERK and JNK MAP kinases was observed in C11-MDCK only. Thus, our results reveal that regulation of renal Na(+),K(+),Cl(-) cotransport by P(2Y) receptors is limited to intercalated cells from collecting ducts and indicate the involvement of the MAP kinase cascade in purinergic control of this ion carrier's activity.

Adenosine Triphosphate↗
PubMed1999

SNARE proteins regulate H(+)-ATPase redistribution to the apical membrane in rat renal inner medullary collecting duct cells.

The interaction of soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins provides the necessary steps for vesicle docking fusion. In inner medullary collecting duct (IMCD) cells, acid secretion is regulated in part by exocytotic insertion and endocytotic retrieval of an H(+)-ATPase to and from the apical membrane. We previously suggested a role for SNARE proteins in exocytotic insertion of proton pumps in IMCD cells. The purpose of the present study was to determine whether SNARE proteins are associated with the 31-kDa subunit of H(+)-ATPase in IMCD cells during exocytosis and to determine the effects of clostridial toxins on SNARE-mediated trafficking of H(+)-ATPase. Cell acidification induced a marked increment of H(+)-ATPase in the apical membrane. However, pretreating cells with clostridial toxins blocked the cellular translocation of the 31-kDa subunit. Immunoprecipitation of IMCD cell homogenate, using antibodies against either the 31-kDa subunit of H(+)-ATPase or vesicle-associated membrane protein-2, co-immunoprecipitated N-ethylmaleimide-sensitive factor, alpha-soluble NSF attachment protein (alpha-SNAP), synaptosome-associated protein-23, syntaxin, and vesicle-associated membrane protein-2. Pretreatment with clostridial toxin resulted in reduced co-immunoprecipitation of H(+)-ATPase and syntaxin. These experiments document, for the first time, a putative docking fusion complex in IMCD cells and a physical association of the H(+)-ATPase with the complex. The sensitivity to the action of clostridial toxin indicates the docking-fusion complex is a part of the exocytotic mechanism of the proton pump.

Animals↗
PubMed1999

Homeobox genes, fossils, and the origin of species.

Ever since Darwin there has been a history of debate on the tempo and mode of evolution. Is speciation a gradual process involving the accumulation of minute variations extant within a species, or is it rapid, the result of major organismal reorganization? Does one define a species on the basis of genes, morphology, or geographic or reproductive isolation? In this communication I present a model of evolutionary change that is based on the Mendelian inheritance of mutations in regulatory genes and the fact that most nonlethal mutations arise in the recessive state. Since the new recessive allele will spread through many generations without expression until there is a critical mass of heterozygotes capable of producing homozygotes for the mutation, the novel feature thus produced will appear abruptly in the population and in more than one individual. This picture of punctuation is consistent with the fossil record, which typically fails to provide evidence of smoothly transitional states of morphological change. Given that the first of their kind in the fossil record are organisms in which their novel characteristics are often more fully expressed or complex than in their descendants, it would seem that, after the mutation involving a regulatory gene is introduced, the general tendency is for its effects to become diminished. Among the implications for speciation is that this process does not depend on either reproductive isolation or genetic incompatibility. Rather, barring effects on reproductive organs or behavior, homozygotes for a novelty should be able to breed with heterozygotes and homozygotes for the wild state of the original population. This, in turn, suggests that the species barrier between individuals is probably a matter of mate recognition.

Adaptation, Physiological↗
PubMed1999

Mechanisms for generating the autonomous cAMP-dependent protein kinase required for long-term facilitation in Aplysia.

The formation of a persistently active cAMP-dependent protein kinase (PKA) is critical for establishing long-term synaptic facilitation (LTF) in Aplysia. The injection of bovine catalytic (C) subunits into sensory neurons is sufficient to produce protein synthesis-dependent LTF. Early in the LTF induced by serotonin (5-HT), an autonomous PKA is generated through the ubiquitin-proteasome-mediated proteolysis of regulatory (R) subunits. The degradation of R occurs during an early time window and appears to be a key function of proteasomes in LTF. Lactacystin, a specific proteasome inhibitor, blocks the facilitation induced by 5-HT, and this block is rescued by injecting C subunits. R is degraded through an allosteric mechanism requiring an elevation of cAMP coincident with the induction of a ubiquitin carboxy-terminal hydrolase.

Animals↗
PubMed1999

Effect of acidification on the location of H+-ATPase in cultured inner medullary collecting duct cells.

In previous studies, our laboratory has utilized a cell line derived from the rat inner medullary collecting duct (IMCD) as a model system for mammalian renal epithelial cell acid secretion. We have provided evidence, from a physiological perspective, that acute cellular acidification stimulates apical exocytosis and elicits a rapid increase in proton secretion that is mediated by an H+-ATPase. The purpose of these experiments was to examine the effect of acute cellular acidification on the distribution of the vacuolar H+-ATPase in IMCD cells in vitro. We utilized the 31-kDa subunit of the H+-ATPase as a marker of the complete enzyme. The distribution of this subunit of the H+-ATPase was evaluated by immunohistochemical techniques (confocal and electron microscopy), and we found that there is a redistribution of these pumps from vesicles to the apical membrane. Immunoblot evaluation of isolated apical membrane revealed a 237 +/- 34% (P < 0.05, n = 9) increase in the 31-kDa subunit present in the membrane fraction 20 min after the induction of cellular acidification. Thus our results demonstrate the presence of this pump subunit in the IMCD cell line in vitro and that cell acidification regulates the shuttling of cytosolic vesicles containing the 31-kDa subunit into the apical membrane.

Acids↗
PubMed1999

ATP depletion increases tyrosine phosphorylation of beta-catenin and plakoglobin in renal tubular cells.

This study examines the hypothesis that the loss of integrity of the junctional complex induced by ATP depletion is related to alterations in tyrosine phosphorylation of the adherens junction proteins beta-catenin and plakoglobin. ATP depletion of cultured mouse proximal tubular (MPT) cells induces a marked increase in tyrosine phosphorylation of both beta-catenin and plakoglobin. The tyrosine phosphatase inhibitor vanadate has the same effect in ATP-replete (control) monolayers, whereas genistein, a tyrosine kinase inhibitor, reduces phosphorylation of both proteins in ATP-replete monolayers and prevents the hyperphosphorylation of these proteins with ATP depletion. This study also demonstrates that the fall in the transepithelial resistance of MPT monolayers induced by ATP depletion can be reproduced by treatment of ATP-replete monolayers with vanadate, whereas genistein substantially ameliorates the fall in transepithelial resistance induced by ATP depletion. Also, using immunofluorescence microscopy it was demonstrated that ATP depletion results in a marked diminution of E-cadherin staining in the basolateral membrane of MPT cells. Vanadate mimics this effect of ATP depletion, whereas genistein ameliorates the reduction in the intensity of E-cadherin staining induced by ATP depletion. Because it is has been well established that hyperphosphorylation of the catenins leads to dissociation of the adherens junction and to dysfunction of the junctional complex, it is proposed that the increase in tyrosine phosphorylation of catenins observed in MPT cells during ATP depletion contributes to the loss of function of the junctional complex associated with sublethal injury.

Adenosine Triphosphate↗
PubMed1998

Morphology, paleoanthropology, and Neanderthals.

Morphology carries the primary signal of events in the evolutionary history of any group of organisms but has been relatively neglected by paleoanthropologists, those who study the history of the human species. Partly this is the result of historical influences, but it is also due to a rather fundamentalist adherence among paleoanthropologists to the tenets of the Neodarwinian Evolutionary Synthesis. The result has been a general paleoanthropological desire to project the species Homo sapiens back into the past as far and in as linear a manner as possible. However, it is clear that the human fossil record, like that of most other taxa, reveals a consistent pattern of systematic diversity--a diversity totally unreflected in the conventional minimalist interpretation of that record. Thus, the Neanderthals, both morphologically and behaviorally as distinctive a group of hominids as ever existed, are conventionally classified simply as a subspecies of our own species Homo sapiens--a classification that robs these extinct relatives of their evolutionary individuality. Only when we recognize the Neanderthals as a historically distinctive evolutionary entity, demanding understanding in its own terms, will we be able to do them proper justice. And we will only be able to do this by restoring morphology to its proper place of primacy in human evolutionary studies.

Animals↗
PubMed1998

Characterization of anion exchangers in an inner medullary collecting duct cell line.

Although the inner medullary collecting duct (IMCD) plays a major role in urinary acidification, the molecular identification of many of the specific components of the transport system in this nephron segment are lacking. A cultured line of rat IMCD cells was used to characterize the mediators of cellular HCO3 exit. This cell line functionally resembles alpha-intercalated cells. Physiologic experiments document that HCO3- transport is a reversible, electroneutral, Cl dependent, Na+-independent process. It can be driven by Cl-gradients and inhibited by stilbenes such as 4-acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic acid. Immunohistochemical analysis, using a rabbit polyclonal antibody against the carboxy-terminal 12 amino acids of anion exchanger 1 (AE1), revealed a distribution of immunoreactive protein that is consistent with a basolateral localization of AE in cultured cells and in alpha-intercalated cells identified in sections of rat kidney cortex. Immunoblot revealed two immunoreactive bands (approximately 100 and 180 kD in size) in membranes from cultured IMCD cells, rat renal medulla, and freshly isolated IMCD cells. The mobility of the lower molecular weight band was similar to that of AE1 in red blood cell ghosts and kidney homogenate and therefore probably represents AE1. The mobility of the 180-kD band is similar to that for rat stomach and kidney AE2 and therefore probably represents AE2. Selective biotinylation of the apical or basolateral membrane proteins in cultured IMCD cells revealed that both AE1 and AE2 are polarized to the basolateral membrane. Northern blot analysis documented the expression of mRNA for AE1 and AE2 but not AE3. Furthermore, the cDNA sequence of AE1 and AE2 expressed by these cells was found to be virtually identical to that reported for kidney AE1 and rat stomach AE2. It is concluded that this cultured line of rat IMCD cells expresses two members of the anion exchanger gene family, AE1 and AE2, and both of these exchangers probably mediate the electroneutral Cl--dependent HCO3-transport observed in this cell line.

Animals↗
PubMed1997

Identification of an 8-lipoxygenase pathway in nervous tissue of Aplysia californica.

Arachidonic acid is converted to (8R)-hydroperoxyeicosa-5,9,11, 14-tetraenoic acid (8-HPETE) during incubations with homogenates of the central nervous system of the marine mollusc, Aplysia californica. 8-HPETE can be reduced to the corresponding hydroxy acid or be enzymatically converted to a newly identified metabolite, 8-ketoeicosa-5,9,11,14-tetraenoic acid (8-KETE). These metabolites were identified by high performance liquid chromatography, UV absorbance, and gas chromatography/mass spectrometry. Stereochemical analysis of the products demonstrate that the neuronal enzyme is an (8R)-lipoxygenase. Previously we have shown that the neurotransmitters, histamine and Phe-Met-Arg-Phe-amide, activate 12-lipoxygenase metabolism in isolated identified Aplysia neurons. We now show that acetylcholine activates the (8R)-lipoxygenase pathway within intact nerve cells. Thus, both (12S)- and (8R)-lipoxygenase co-exist in intact Aplysia nervous tissue but are differentially activated by several neurotransmitters. The precise physiological role of the 8-lipoxygenase products is currently under investigation, but by analogy to the well-described 12-lipoxygenase pathway, we suggest that (8R)-HPETE and 8-KETE may serve as second messengers in Aplysia cholinoceptive neurons.

Acetylcholine↗
PubMed1997

Ubiquitin C-terminal hydrolase is an immediate-early gene essential for long-term facilitation in Aplysia.

The switch from short-term to long-term facilitation of the synapses between sensory and motor neurons mediating gill and tail withdrawal reflexes in Aplysia requires CREB-mediated transcription and new protein synthesis. We isolated several downstream genes, one of which encodes a neuron-specific ubiquitin C-terminal hydrolase. This rapidly induced gene encodes an enzyme that associates with the proteasome and increases its proteolytic activity. This regulated proteolysis is essential for long-term facilitation. Inhibiting the expression or function of the hydrolase blocks induction of long-term but not short-term facilitation. We suggest that the enhanced proteasome activity increases degradation of substrates that normally inhibit long-term facilitation. Thus, through induction of the hydrolase and the resulting up-regulation of the ubiquitin pathway, learning recruits a regulated form of proteolysis that removes inhibitory constraints on long-term memory storage.

Amino Acid Sequence↗
PubMed1997

Fatty acid-induced cytotoxicity: differences in susceptibility between MDCK cells and primary cultures of proximal tubular cells.

We have compared the cytotoxicity of exogenously added fatty acid (oleic acid) and that of endogenous free fatty acids released from cell membranes by phospholipase A2 in primary cultures of mouse proximal tubular (MPT) cells and in Madine-Darby canine kidney (MDCK) cells. Exposure of MPT cell monolayers to oleic acid (125 mmol/L) for 2 hours resulted in severe irreversible injury to 70% +/- 4% of MPT cells. In striking contrast, only 8% +/- 3% of MDCK cells were killed by the same insult. This striking difference in the response to exogenous oleate by MPT and MDCK cells was associated with modest and comparable reductions in cell adenosine triphosphate (ATP) content in both cell types. Chemical anoxia induced by cyanide plus deoxyglucose (CN-DOG) in the absence of glucose was associated with greater injury in MPT cells (45% +/- 6% killed) than in MDCK cells (16% +/- 5% cells killed) despite severe and comparable depletion of cell ATP content in both MPT cells (96.0% +/- 0.6% reduction) and MDCK cells (96.0% +/- 0.5% reduction). The release of endogenous fatty acids by the exposure of cells to exogenous phospholipase A2 caused mild injury in both cell types that was more severe in MPT cells than in MDCK cells. The combined insult of phospholipase A2 and chemical anoxia for 2 hours caused substantially greater cell injury in both MPT and MDCK cells than either intervention alone, but the combined insult was still more damaging to MPT cells (73% +/- 4% killed) than to MDCK cells (30% +/- 4% killed). We conclude that the cell membrane in MDCK cells is intrinsically more resistant to fatty acid-induced injury than the lipid membrane in MPT cells.

Adenosine Triphosphate↗
PubMed1997

Mechanisms of host wasting induced by administration of cytokines in rats.

This study determined the effects of chronic administration of the two principal proximate cytokines, interleukin-1 (IL-1) and tumor necrosis factor (TNF), on host wasting in rats. The effects were compared with those observed in a matched pair-fed group to distinguish the contribution from anorexia. Both TNF and IL-1 produced weight loss, net nitrogen loss, and skeletal muscle catabolism and increased liver weight. Such effects were independent from and additive to those resulting from semistarvation. However, under equivalent nutritional conditions, TNF infusion led to a greater effect on muscle protein catabolism and liver weight and caused liver protein anabolism, whereas only the group receiving IL-1 had altered glucose metabolism in the postabsorptive state. Tachyphylaxis was seen in the response of food intake over time after administration of IL-1. These actions define the two principal mechanisms for the development of protein calorie malnutrition that occur because of cytokine action, anorexia producing semistarvation and an increased net protein catabolic rate reducing anabolic efficiency.

Animals↗
PubMed1997

Heat stress ameliorates ATP depletion-induced sublethal injury in mouse proximal tubule cells.

The role of prior heat stress (HS) in ameliorating changes in the actin cytoskeleton and the loss of tight junction integrity that accompany ATP depletion was examined. Mouse proximal tubule cells in primary culture were exposed to sodium cyanide (CN) in the absence of dextrose for 1 h, a maneuver that produced equivalent degrees of ATP depletion in control and in HS cells. After ATP depletion, actin stress fibers were completely disrupted in control cells. In contrast, HS cells with elevated HSP-72 content showed preservation of stress fibers after CN exposure. ATP depletion in control and HS cells produced similar and reversible depletion of the G-actin pool without altering total actin content. Integrity of the tight junction was assessed by transepithelial electrical resistance (TER) and unidirectional flux of lucifer yellow (LY, mol wt 482). After CN alone, the nadir in TER was lower than that of HS + CN cells (51.6 +/- 2.5 vs. 96.2 +/- 3.2 omega x cm2, respectively; P < 0.05). After 30-min recovery, TER of HS + CN recovered to control values (277 +/- 7.2 vs. 227 +/- 6.6 omega x cm2; P > 0.05), whereas CN did not (165 +/- 7.3 vs. 227 +/- 6.6 omega x cm2; P < 0.05). Changes in LY flux paralleled those in TER. HS is associated with preservation of the actin cytoskeleton and improved integrity of the tight junction after sublethal ATP depletion injury. These protective effects may contribute to the preservation of epithelial cell polarity and function following an ischemic insult.

Actins↗
PubMed1997

H+ secretion is inhibited by clostridial toxins in an inner medullary collecting duct cell line.

Renal epithelial cell H+ secretion is an exocytic-endocytic phenomenon. In the inner medullary collecting duct (IMCD) cell line, which we have utilized as a model of renal epithelial cell acid secretion, we found previously that acidification increased exocytosis and alkalinization increased endocytosis. It is likely, therefore, that the rate of proton secretion is regulated by the membrane insertion and retrieval of proton pumps. There is abundant evidence from studies in the nerve terminal and the chromaffin cell that vesicle docking, membrane fusion, and discharge of vesicular contents (exocytosis) involve a series of interactions among so-called trafficking proteins. The clostridial toxins, botulinum and tetanus are proteases that specifically inactivate some of these proteins. In these experiments we demonstrated, by immunoblot and immunoprecipitation, the presence in this IMCD cell line of the specific protein targets of these toxins, synaptobrevin/vesicle-associated membrane proteins (VAMP), syntaxin, and synaptosomal-associated protein-25 (SNAP-25). Furthermore, we showed that these toxins markedly inhibit the capacity of these cells to realkalinize after an acid load. Thus these data provide new insight into the mechanism for H+ secretion in the IMCD.

Ammonium Chloride↗
PubMed1996

Significance of some previously unrecognized apomorphies in the nasal region of Homo neanderthalensis.

For many years, the Neanderthals have been recognized as a distinctive extinct hominid group that occupied Europe and western Asia between about 200,000 and 30,000 years ago. It is still debated, however, whether these hominids belong in their own species, Homo neanderthalensis, or represent an extinct variant of Homo sapiens. Our ongoing studies indicate that the Neanderthals differ from modern humans in their skeletal anatomy in more ways than have been recognized up to now. The purpose of this contribution is to describe specializations of the Neanderthal internal nasal region that make them unique not only among hominids but possibly among terrestrial mammals in general as well. These features lend additional weight to the suggestion that Neanderthals are specifically distinct from Homo sapiens.

Animals↗
PubMed1996

Whose teeth?

Explore the source record for details and available documents.

Animals↗
PubMed1996

Insulin receptor in Aplysia neurons: characterization, molecular cloning, and modulation of ion currents.

We have isolated the cDNA for a tyrosine kinase receptor that is expressed in the nervous system of Aplysia californica and that is similar to the vertebrate insulin receptor. Binding studies and immunocytochemical staining show that the receptor is abundant in the bag cell neurons. Application of vertebrate insulin to clusters of bag cell neurons stimulates the phosphorylation of the receptor on tyrosine residues, and exposure of isolated bag cell neurons to insulin produces an increase in height and a decrease in duration of the action potentials that can be detected within 15-30 min. These effects were not seen with insulin-like growth factor-1. In voltage-clamped neurons, insulin produces an increase in the amplitude of the voltage-dependent Ca2+ current that can be blocked by preincubation with herbimycin A, an inhibitor of tyrosine kinases. Insulin also enhances a delayed K+ current. We suggest that insulin-like peptides regulate the excitability of the bag cell neurons.

Action Potentials↗