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J A DeSimone

Publications and source records attributed to J A DeSimone.

At least 37 records · Page 2Linked to original sources
PubMed1992

Dialysis disequilibrium syndrome (DDS) in the rat: role of the "reverse urea effect".

DDS is characterized by neurologic deterioration and cerebral edema which occurs after hemodialysis. To investigate the pathogenesis of DDDS, we studied the effects of rapid hemodialysis on plasma and brain electrolytes, urea, and osmolality in the rat. Forty-two hours after bilateral nephrectomy, nine uremic rats were hemodialyzed for 90 minutes against dialysate without urea (model of DDS), yielding a decrease in plasma urea from 72 +/- 2 mM to 34 +/- 2 mM (P less than 0.01) and an 8% (29 mOsm/kg) decrease in plasma osmolality. This group was compared to three control groups: 11 uremic animals dialyzed against a bath with urea added so that no fall in plasma urea occurred, and 15 uremic and 12 nonuremic animals that were not dialyzed. In animals dialyzed without urea, compared to uremic non-dialyzed animals, there was a 6% increase in brain water (3.89 +/- 0.04 liter/kg dry wt vs. 3.67 +/- 0.03, P less than 0.01) and an increase in the brain to plasma (urea) ratio (1.30 +/- 0.06 vs. 0.79 +/- 0.05, P less than 0.01). Comparison of these parameters in animals dialyzed without urea versus other control groups yielded similar results. In animals dialyzed without urea, the 53% decrease in plasma urea was associated with only a 13% decrease in brain urea content. Brain content of sodium and potassium was not significantly different among groups. Retention of brain urea despite the large decrease in plasma urea was able to account for the increased brain water observed in animals dialyzed without urea.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

The anion paradox in sodium taste reception: resolution by voltage-clamp studies.

Sodium salts are potent taste stimuli, but their effectiveness is markedly dependent on the anion, with chloride yielding the greatest response. The cellular mechanisms that mediate this phenomenon are not known. This "anion paradox" has been resolved by considering the field potential that is generated by restricted electrodiffusion of the anion through paracellular shunts between taste-bud cells. Neural responses to sodium chloride, sodium acetate, and sodium gluconate were studied while the field potential was voltage-clamped. Clamping at electronegative values eliminated the anion effect, whereas clamping at electropositive potentials exaggerated it. Thus, field potentials across the lingual epithelium modulate taste reception, indicating that the functional unit of taste reception includes the taste cell and its paracellular microenvironment.

Animals↗
PubMed1990

Membrane currents in taste cells of the rat fungiform papilla. Evidence for two types of Ca currents and inhibition of K currents by saccharin.

Taste buds were isolated from the fungiform papilla of the rat tongue and the receptor cells (TRCs) were patch clamped. Seals were obtained on the basolateral membrane of 281 TRCs, protruding from the intact taste buds or isolated by micro-dissection. In whole-cell configuration 72% of the cells had a TTX blockable transient Na inward current (mean peak amplitude 0.74 nA). All cells had outward K currents. Their activation was slower than for the Na current and a slow inactivation was also noticeable. The K currents were blocked by tetraethylammonium, Ba, and 4-aminopyridine, and were absent when the pipette contained Cs instead of K. With 100 mM Ba or 100 mM Ca in the bath, two types of inward current were observed. An L-type Ca current (ICaL) activated at -20 mV had a mean peak amplitude of 440 pA and inactivated very slowly. At 3 mM Ca the activation threshold of ICaL was near -40 mV. A transient T-type current (ICaT) activated at -50 mV had an average peak amplitude of 53 pA and inactivated with a time constant of 36 ms at -30 mV. ICaL was blocked more efficiently by Cd and D600 than ICaT. ICaT was blocked by 0.2 mM Ni and half blocked by 200 microM amiloride. In whole-cell voltage clamp, Na-saccharin caused (in 34% of 55 cells tested) a decrease in outward K currents by 21%, which may be expected to depolarize the TRCs. Also, Na-saccharin caused some taste cells to fire action potentials (on-cell, 7 out of 24 cells; whole-cell, 2 out of 38 cells responding to saccharin) of amplitudes sufficient to activate ICaL. Thus the action potentials will cause Ca inflow, which may trigger release of transmitter.

Action Potentials↗
PubMed1989

Direct measurement of translingual epithelial NaCl and KCl currents during the chorda tympani taste response.

We have measured the NaCl or KCl currents under voltage clamp across the dorsal lingual epithelium of the rat and simultaneously the response of the taste nerves. Under short-circuit conditions a NaCl stimulus evoked an inward current (first current) that coincided with excitation of the chorda tympani. This was followed by a slower inward current (second current) that matched the kinetics of taste nerve adaptation. The peak first current and the coincident neural response satisfied the same saturating NaCl concentration dependence. Both first and second currents were partially blocked by amiloride as were the phasic and tonic components of the neural response. The NaCl-evoked second current was completely blocked by ouabain. Investigation of the NaCl-evoked current and the neural response over a range of clamped voltages showed that inward negative potentials enhanced the inward current and the neural response to 0.3 M NaCl. Sufficiently high inward positive potentials reversed the current, and made the neural response independent of further changes in voltage. Therefore, one of the NaCl taste transduction mechanisms is voltage dependent while the other is voltage independent. A KCl stimulus also evoked an inward short-circuit current, but this and the neural response were not amiloride-sensitive. The data indicate that neural adaptation to a NaCl stimulus, but not a KCl stimulus, is mediated by cell Na/K pumps. A model is proposed in which the connection between the NaCl-evoked second current and cell repolarization is demonstrated.

Amiloride↗
PubMed1988

Ion transport across the frog olfactory mucosa: the action of cyclic nucleotides on the basal and odorant-stimulated states.

The action of cyclic nucleotides on the short-circuit current across the isolated bullfrog olfactory mucosa was studied both in the absence and presence of odorants. 8-Bromo-cAMP applied to the ciliated side of the mucosa caused a concentration-dependent, reversible increase in the basal short-circuit current, but not when it was applied to the submucosal side. The current had a sigmoidal concentration dependence described by the Hill equation. The magnitude of the odorant-evoked current was enhanced after bathing the ciliated side with cAMP analogs or modulators of intracellular cAMP. GTP gamma S added to the ciliated side increased the odorant-evoked current, while GDP beta S caused a decrease. Current transients induced by stimulating the ciliated side with either pulses of odorant or 8-bromo-cAMP were partially suppressed by amiloride, but only when amiloride and stimulant were presented simultaneously. Pulses of 8-bromo-cAMP and odorant presented simultaneously resulted in currents that added nonlinearly. In the absence of odorant, 8-bromo-cGMP caused a concentration-dependent decrease in net inward current that was reversed by 8-bromo-cAMP. Odorant-evoked currents were also reduced by 8-bromo-cGMP, and these could not be reversed by 8-bromo-cAMP. The results indicate that one type of olfactory transduction process involves the activation by cAMP of an inward current through an amiloride-sensitive apical ion channel and that this mechanism is mediated by a stimulatory G-protein.

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1988

Sugar-activated ion transport in canine lingual epithelium. Implications for sugar taste transduction.

There is good evidence indicating that ion-transport pathways in the apical regions of lingual epithelial cells, including taste bud cells, may play a role in salt taste reception. In this article, we present evidence that, in the case of the dog, there also exists a sugar-activated ion-transport pathway that is linked to sugar taste transduction. Evidence was drawn from two parallel lines of experiments: (a) ion-transport studies on the isolated canine lingual epithelium, and (b) recordings from the canine chorda tympani. The results in vitro showed that both mono- and disaccharides in the mucosal bath stimulate a dose-dependent increase in the short-circuit current over the concentration range coincident with mammalian sugar taste responses. Transepithelial current evoked by glucose, fructose, or sucrose in either 30 mM NaCl or in Krebs-Henseleit buffer (K-H) was partially blocked by amiloride. Among current carriers activated by saccharides, the current response was greater with Na than with K. Ion flux measurements in K-H during stimulation with 3-O-methylglucose showed that the sugar-evoked current was due to an increase in the Na influx. Ouabain or amiloride reduced the sugar-evoked Na influx without effect on sugar transport as measured with tritiated 3-O-methylglucose. Amiloride inhibited the canine chorda tympani response to 0.5 M NaCl by 70-80% and the response to 0.5 M KCl by approximately 40%. This agreed with the percent inhibition by amiloride of the short-circuit current supported in vitro by NaCl and KCl. Amiloride also partially inhibited the chorda tympani responses to sucrose and to fructose. The results indicate that in the dog: (a) the ion transporter subserving Na taste also subserves part of the response to K, and (b) a sugar-activated, Na-preferring ion-transport system is one mechanism mediating sugar taste transduction. Results in the literature indicate a similar sweet taste mechanism for humans.

Amiloride↗
PubMed1988

Lingual epithelium of spontaneously hypertensive rats has decreased short-circuit current in response to NaCl.

Alterations in ion transport associated with hypertension have been found in a variety of organs. We used a modified Ussing chamber to compare the NaCl dependence of the short-circuit current across the dorsal lingual epithelium in vitro from spontaneously hypertensive rats (SHR) with that from Wistar-Kyoto rats (WKY). The short-circuit current in response to mucosal NaCl was less in SHR than in WKY at hyperosmotic concentrations (above 0.15 M and up to 2.0 M). Since ion transport in the lingual epithelium has been found to play a role in early events of salt taste transduction, the attenuation in the short-circuit current in hypertensive animals may be a factor in the enhanced salt preference of SHR compared with WKY.

Analysis of Variance↗
PubMed1987

Ion transport across the frog olfactory mucosa: the basal and odorant-stimulated states.

The Ussing method was adapted to study the basal electrolyte transfer as well as the events that occur upon odorant stimulation in frog olfactory mucosa. The unstimulated short-circuit current was due mainly to a furosemide-sensitive ion transport system on the apical side of the olfactory mucosa. This current was not amiloride sensitive. The current-voltage relationship of the unstimulated state was linear. That of the odorant-evoked current was non-linear and amiloride-sensitive. Ouabain caused collapse of both the unstimulated and odorant-stimulated short-circuit current. In this case, voltage-clamping the tissue to non-zero values restored the odorant-evoked current with polarity depending on that of the clamping voltage. This suggested that the direction of the current is determined by that of the sodium electrochemical potential difference. Our results indicate that the unstimulated short-circuit current occurs through an apical sodium cotransport system, while the odorant-evoked current is due to odorant-activated, passive sodium channels that are amiloride sensitive.

Amiloride↗
PubMed1987

Odorant stimulation of secretory and neural processes in the salamander olfactory mucosa.

Topical application of the odorants guaiacol (10(-3) mol/l, 1-30 min) and 2-isobutyl-3-methoxypyrazine (IBMP, 10(-5)-10(-3) mol/l, 15 min) caused time- and concentration-dependent reductions in the secretory granule content of acinar cells of the superficial Bowman's glands (sBG) and moderate to extensive vacuolation in acinar cells of sBG and deep olfactory glands (dG). Topical application of 9.8 mg/ml scopolamine 10 min before 10(-4) mol/l IBMP significantly reduced the amount of secretory granule depletion from sBG compared to that seen with IBMP alone and resulted in less extensive vacuolation in sBG and dG acinar cells. The i.p. injection of 42 mg/kg propranolol 10 min before topical application of 10(-4) mol/l IBMP had no effect on the action of IBMP. Guaiacol and IBMP also had time- and concentration-dependent effects on the secretory activity of sustentacular cells in the olfactory epithelium. The protrusion of secretory material into the mucociliary matrix that covers the epithelial surface and vacuolation within the secretory material resulted from odorant application. Scopolamine and propranolol had no effects on the action of IBMP on sustentacular cell secretory activity. When applied in the vapor phase, guaiacol elicited action potentials recorded from individual olfactory receptor neurons; the impulse frequency was concentration-dependent and showed tonic and phasic components when the duration of stimulation was varied. Low to moderate concentrations of IBMP delivered in the vapor phase evoked monophasic negative slow voltage transients recorded from the surface of the olfactory mucosa. The amplitudes of these transients increased with increasing stimulus concentrations. Higher concentrations or longer stimulus durations evoked longer-latency positive-voltage generating processes and negative afterpotentials. The properties of the electrophysiological responses to both odorants were characteristic of responses evoked by a wide variety of 'typical' odorants.

Animals↗
PubMed1986

Potential dependence of unidirectional chloride fluxes across isolated frog skin.

Isolated frog skins were voltage clamped at transepithelial potentials (Vt) ranging from -60 mV to 60 mV to measure transepithelial 36Cl- fluxes from the apical to the basolateral bathing solution (J13) and in the opposite direction (J31). The potential dependence of fluxes obtained in Na+-free choline Ringer's indicates the presence of conductive and nonconductive components that probably correspond to fluxes through paracellular and cellular pathways, respectively. Rectification of fluxes with reversal of the potential reflects a structural asymmetry, presumably in surface charge density. The data are consistent with a charge density of one negative charge per 280 A2 on the apical side. A new model for passive Cl- transport was developed that includes surface charge asymmetry and specifically accounts for the observed variation of conductance with potential. In normal frog Ringer's, J13 was larger than J31 at zero potential (active Cl- transport), J13 rose exponentially with increasing positive potential to reach a maximum at 40 mV (approximately open-circuit), and the predicted partial Cl- conductance exceeded the measured conductance leading to the conclusion that when J13 is largely driven by Na+ transport, much of the coupling occurs via nonconductive pathways. Theophylline stimulates Cl- transport that also occurs via nonconductive pathways as Vt becomes more positive.

Amiloride↗
PubMed1985

The identity of the current carriers in canine lingual epithelium in vitro.

Ion transport across the lingual epithelium has been implicated as an early event in gustatory transduction. The fluxes of isotopically labelled Na+ and Cl- were measured across isolated canine dorsal lingual epithelium under short-circuit conditions. The epithelium actively absorbs Na+ and to a lesser extent actively secretes Cl-. Under symmetrical conditions with Krebs-Henseleit buffer on both sides, (1) Na+ absorption accounts for 46% of the short-circuit current (Isc); (2) there are two transcellular Na+ pathways, one amiloride-sensitive and one amiloride-insensitive; (3) ouabain, added to the serosal solution, inhibits both Isc and active Na+ absorption. When hyperosmotic (0.25 M) NaCl is placed in the mucosal bath, both Isc and Na+ absorption increase; net Na+ absorption is at least as much as Isc. Ion substitution studies indicate that the tissue may transport a variety of larger ions, though not as effectively as Na+ and Cl-. Thus we have shown that the lingual epithelium, like other epithelia of the gastrointestinal tract, actively transports ions. However, it is unusual both in its response to hyperosmotic solutions and in the variety of ions that support a transepithelial short-circuit current. Since sodium ion transport under hyperosmotic conditions has been shown to correlate well with the gustatory neural response, the variety of ions transported may likewise indicate a wider role for transport in taste transduction.

Amiloride↗
PubMed1985

Analysis of amiloride inhibition of chorda tympani taste response of rat to NaCl.

The kinetics of inhibition by amiloride of the integrated chorda tympani response were investigated in rats subjected to lingual stimulation with NaCl. In one series of experiments the time of exposure to amiloride was varied at fixed amiloride concentration. Exposure to 10(-4) M amiloride for 2 s reduced the response to 0.5 M NaCl by approximately 50%. The time course of recovery from amiloride inhibition was first order (relaxation time approximately equal to 4 min) for all exposure times. For exposure to 10(-4) M amiloride for less than or equal to 30 s recovery was better than 90% in 20 min. Not all of the chorda tympani response was inhibited by amiloride. With 0.5 M NaCl there was a 70% reduction in response, whereas at 0.05 M NaCl the reduction was only 30%. Parallel effects of amiloride were seen in the short-circuit current of an in vitro preparation of canine lingual epithelium. Amiloride reduced the short-circuit current by the same percentage as it inhibited the chorda tympani response. These results suggest that gustatory transduction is mediated in part by an apical membrane transport system that can be inhibited by amiloride. There exists, however, a second transducing element that is amiloride insensitive. A model is developed, assuming, in part, that the neural response reflects the flows of Na through amiloride-sensitive apical pathways.

Amiloride↗
PubMed1984

Salt taste transduction occurs through an amiloride-sensitive sodium transport pathway.

An important early event in mammalian gustatory transduction with respect to sodium chloride has been found to be the passage of sodium ions through specific transport pathways in the apical region of the taste bud. The inward current caused by sodium chloride placed on the mucosal surface of an in vitro preparation of rat dorsal lingual epithelium can be substantially reduced by the blocker of sodium ion transport, amiloride. The data show (i) that amiloride is a specific blocker of the chorda tympani response to sodium chloride, but not to potassium chloride, (ii) that the sodium and potassium gustatory systems are largely independent at the peripheral level, and (iii) that the classical ion taste "receptor" is actually a specific transport pathway permitting the cation to enter the taste-bud cell and thereby to spread depolarizing current.

Amiloride↗
PubMed1981

Active ion transport in dog tongue: a possible role in taste.

An in vitro preparation of the dorsal epithelium of the dog tongue actively transports ions, producing a transepithelial potential difference characteristic of the ions and their concentration. Hypertonic sodium chloride solutions generally cause increased potentials and short-circuit currents and reduced resistances when placed on the mucosal surface. This hypertonic flux is eliminated by ouabain and is not found in ventral lingual epithelia. When either sodium acetate or tetramethylammonium chloride is substituted for sodium chloride in the mucosal medium, the currents are diminished but their sum at a given concentration approximates that for sodium chloride at the same concentration. This result suggests a current composed of inward sodium ion movement and outward chloride ion movement. Actively regulated potentials and currents, whether generated in the taste buds or in supporting cells, may be important in both normal chemotransduction and in taste responses evoked by currents passing through the tongue.

Animals↗
PubMed1980

The location of olfactory receptor sites. Inferences from latency measurements.

Excitatory responses recorded from vertebrate olfactory sensory neurons are characterized by long latencies compared with those from other sensory receptors. Explanations which assume free access of the stimuli to receptor molecules presumably located on the olfactory cilia necessarily imply an intrinsic delay in the transduction mechanism. In contrast, the possibility of restricted or delayed access due to diffusion of the stimulus to molecular receptors located on the dendritic know or proximal portions of the cilia suggests transduction processes having time courses similar to those in other sensory systems. We show that the threshold stimulus concentrations and the latency of the excitatory response of the salamander can be predicted primarily on the basis of a diffusional delay and that the receptor molecules are well below the surface of the mucus. Examination of response latencies for other species reported in the literature support the generality of diffusional delay. The predicted location of molecular receptor sites is largely insensitive to assumptions based on the mode of clearance of the stimuli. Additional access restrictions are discussed but are shown to generate qualitatively different latency functions than does diffusion, suggesting that they exert only minor influences on latency and threshold characteristics.

Action Potentials↗