Effect of Temperature on Endplate Potential Rundown and Recovery in Rat Diaphragm

2001 ◽  
Vol 85 (5) ◽  
pp. 2070-2075 ◽  
Author(s):  
Michelle Moyer ◽  
Erik van Lunteren
Keyword(s):  
Neuromuscular Junction ◽  
Effect Of Temperature ◽  
Temperature Sensitive ◽  
Enzymatic Reactions ◽  
Rat Diaphragm ◽  
Intermittent Stimulation ◽  
Vesicle Recycling ◽  
Endplate Potential ◽  
Stimulation Period

The amplitude of neuromuscular junction end-plate potentials (EPPs) decreases quickly within a train but recovers nearly completely from train to train during intermittent stimulation. Rundown has been shown to be dependent not only on the rate of transmitter release but also on the rate of replenishment of the depleted neurotransmitter at the site of release. Two groups of processes have been proposed for synaptic vesicle recycling, both of which involve multiple energy-requiring steps and enzymatic reactions and which therefore would be expected to be very temperature-sensitive. The present study tested the hypothesis that low temperature therefore increases the rate of EPP amplitude rundown. Studies were performed in vitro on rat diaphragm and used μ-conotoxin to allow normal-sized EPPs to be recorded from intact fibers. EPP amplitude rundown during intermittent stimulation at 20 and 50 Hz (duty cycle 333 ms) was greater at 20°C than it was at 37°C. Initially, temperature affected only intra-train rundown but, over longer periods of stimulation, both intra- and inter-train rundown were significantly accelerated by cold temperature. Cumulative EPP amplitudes were calculated by successively adding the amplitudes of each EPP during the stimulation period to provide an estimate of total neurotransmitter release in the neuromuscular junction. The cumulative EPP amplitude was significantly lower at 20°C than it was at 37°C during both 20 and 50 Hz stimulation. These data indicate that the mechanism involved in EPP amplitude rundown and recovery is temperature-sensitive, with a greater decrement in EPP amplitude at cold than at warm temperatures.

Effect of Phasic Activation on Endplate Potential in Rat Diaphragm

1999 ◽  
Vol 82 (6) ◽  
pp. 3030-3040 ◽  
Author(s):  
Michelle Moyer ◽  
Erik van Lunteren
Keyword(s):  
Time Course ◽  
Complete Recovery ◽  
Stimulation Frequency ◽  
Drop Out ◽  
Rat Diaphragm ◽  
Continuous Stimulation ◽  
Intermittent Stimulation ◽  
Rate Of Decline ◽  
Phasic Activation

Neuromuscular junction endplate potentials (EPPs) decrease quickly and to a large extent during continuous stimulation. The present study examined the hypothesis that EPP rundown recovers rapidly, thereby substantially preserving neurotransmission during intermittent compared with continuous stimulation. Studies were performed in vitro on rat diaphragm, using μ-conotoxin to allow recording of normal-sized EPPs from intact fibers. During continuous 5- to 100-Hz stimulation, EPP amplitude declined with a biphasic time course. The initial fast rate of decline was modulated substantially by stimulation frequency, whereas the subsequent slow rate of decline was relatively frequency independent. During intermittent 5- to 100-Hz stimulation (duty cycle 0.33), EPP amplitude declined rapidly during each train, but recovered substantially by the onset of the following train. The intra-train declines were substantially greater than the inter-train declines in EPP amplitude. Intra-train reductions in EPP amplitude were stimulation frequency dependent, based on both the total decline and rate constant of EPP decline. In contrast, the degree of recovery from train to train was independent of stimulation frequency, indicating low frequency dependence of inter-train rundown. The substantial recovery of EPP amplitude in between trains resulted in greater cumulative EPP size during intermittent compared with continuous stimulation. During continuous stimulation, EPP drop-out was only seen during 100-Hz stimulation; this was completed mitigated during intermittent stimulation. Miniature EPP size was unaffected by either continuous or intermittent stimulation. The pattern of rapid intra-train rundown and slow inter-train rundown of EPP size during intermittent stimulation is therefore due to rapid changes in the magnitude of neurotransmitter release rather than to axonal block or postsynaptic receptor desensitization. These findings indicate considerable rundown of EPP amplitudes within a stimulus train, with near complete recovery by the onset of the next train. This substantially attenuates the decrement in EPP amplitude during intermittent compared with continuous stimulation, thereby preserving the integrity of neurotransmission during phasic activation.

scholarly journals Insulin biosynthesis in the bullhead, Ictalurus nebulosus, and the effect of temperature

1973 ◽  
Vol 134 (3) ◽  
pp. 753-761 ◽  
Author(s):  
Margaret L. Moule ◽  
Cecil C. Yip
Keyword(s):  
Qualitative Difference ◽  
Gel Filtration ◽  
Effect Of Temperature ◽  
Insulin Biosynthesis ◽  
Temperature Sensitive ◽  
Bicarbonate Buffer ◽  
Control Tissue ◽  
Ictalurus Nebulosus ◽  
Lower Temperature

Insulin biosynthesis in the brown bullhead, Ictalurus nebulosus (Le Sueur), was studied by measuring the incorporation in vitro of [3H]leucine into proteins of the principal islet. The tissue was incubated for 6–15h in Krebs–Ringer bicarbonate buffer with [3H]leucine, supplemented with amino acids and glucose. Proteins, precipitated with trichloroacetic acid and extracted with acid ethanol, were separated by gel-filtration on Biogel P-30 in 3m-acetic acid. Three major components were found after incubation of the islets at 22°C. On the basis of the results of sulphitolysis, biological activity and the demonstrated precursor–product relationship, components I and II were identified as proinsulin and insulin respectively. The third component was not identified. At 12°C, [3H]leucine was incorporated only into proinsulin. No radioactivity was found in insulin or the unidentified component III at 12°C as was found after incubation at 22°C. When the temperature was lowered from 22° to 12°C after 3h of a 15h incubation, decreased conversion of proinsulin into insulin resulted at the lower temperature compared with the control tissue maintained at 22°C. When the temperature was raised from 12° to 22°C at 3h of a 15h incubation, conversion of proinsulin into insulin occurred. No conversion occurred in the control tissue with the temperature maintained at 12°C. No qualitative difference in the incorporation of [3H]leucine into proinsulin and its conversion into insulin at 12° and 22°C could be demonstrated between islet tissue from fish acclimated to less than 12°C or to 22°C. The results suggest that the enzyme(s) responsible for converting proinsulin into insulin in the bullhead may be temperature sensitive with low activity at 12°C.

scholarly journals An internal GAP domain negatively regulates presynaptic dynamin in vivo

2005 ◽  
Vol 169 (1) ◽  
pp. 117-126 ◽  
Author(s):  
Radhakrishnan Narayanan ◽  
Marilyn Leonard ◽  
Byeong Doo Song ◽  
Sandra L. Schmid ◽  
Mani Ramaswami
Keyword(s):  
Point Mutations ◽  
Temperature Sensitive ◽  
Vesicle Formation ◽  
Gtpase Domain ◽  
Vesicle Recycling ◽  
Self Assembling ◽  
Step Model ◽  
Internal Gap

The mechanism by which the self-assembling GTPase dynamin functions in vesicle formation remains controversial. Point mutations in shibire, the Drosophila dynamin, cause temperature-sensitive (ts) defects in endocytosis. We show that the ts2 mutation, which occurs in the switch 2 region of dynamin's GTPase domain, compromises GTP binding affinity. Three second-site suppressor mutations, one in the switch 1 region of the GTPase domain and two in the GTPase effector domain (GED), dynamin's putative GAP, fully rescue the shits2 defects in synaptic vesicle recycling. The functional rescue in vivo correlates with a reduction in both the basal and assembly-stimulated GTPase activity in vitro. These findings demonstrate that GED is indeed an internal dynamin GAP and establish that, as for other GTPase superfamily members, dynamin's function in vivo is negatively regulated by its GAP activity. Based on these and other observations, we propose a two-step model for dynamin during vesicle formation in which an early regulatory GTPase-like function precedes late, assembly-dependent steps during which GTP hydrolysis is required for vesicle release.

scholarly journals Temperature sensitivity differs between heart and red muscle mitochondria in mahi-mahi (Coryphaena hippurus)

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Gigi Y. Lau ◽  
Georgina K. Cox ◽  
John D. Stieglitz ◽  
Daniel D. Benetti ◽  
Martin Grosell
Keyword(s):  
Heart Mitochondria ◽  
Effect Of Temperature ◽  
Temperature Sensitive ◽  
Temperature Changes ◽  
Coryphaena Hippurus ◽  
Wide Range ◽  
Irreversible Effects ◽  
Effects Of Temperature ◽  
Lower Temperature

Abstract Maintaining energy balance over a wide range of temperatures is critical for an active pelagic fish species such as the mahi-mahi (Coryphaena hippurus), which can experience rapid changes in temperature during vertical migrations. Due to the profound effect of temperature on mitochondrial function, this study was designed to investigate the effects of temperature on mitochondrial respiration in permeabilized heart and red skeletal muscle (RM) fibres isolated from mahi-mahi. As RM is thought to be more anatomically isolated from rapid ambient temperature changes compared to the myocardium, it was hypothesized that heart mitochondria would be more tolerant of temperature changes through a greater ability to match respiratory capacity to an increase in temperature and to maintain coupling, when compared to RM mitochondria. Results show that heart fibres were more temperature sensitive and increased respiration rate with temperature increases to a greater degree than RM. Respiratory coupling ratios at the three assay temperatures (20, 26, and 30 °C), revealed that heart mitochondria were less coupled at a lower temperature (26 °C) compared to RM mitochondria (30 °C). In response to an in vitro acute temperature challenge, both tissues showed irreversible effects, where both heart and RM increased uncoupling whether the assay temperature was acutely changed from 20 to 30 °C or 30 to 20 °C. The findings from this study indicate that mahi-mahi heart mitochondria were more temperature sensitive compared to those from RM.

scholarly journals Fever and immunoregulation. III. Hyperthermia augments the primary in vitro humoral immune response.

1983 ◽  
Vol 157 (4) ◽  
pp. 1229-1238 ◽  
Author(s):  
H D Jampel ◽  
G W Duff ◽  
R K Gershon ◽  
E Atkins ◽  
S K Durum
Keyword(s):  
Immune Response ◽  
Cell Response ◽  
Direct Action ◽  
Interleukin 1 ◽  
Effect Of Temperature ◽  
Temperature Sensitive ◽  
Humoral Immune ◽  
Fever Response ◽  
And Function

We have examined the possibility that hyperthermia, such as that occurring during fever, may benefit the immune response. The effect of temperature on the in vitro immune response of unprimed murine spleen cells against the antigen sheep erythrocytes was tested. Hyperthermia potently augmented the plaque-forming cell response. Temperature-sensitive events occurred early in the culture period. Subsets of lymphocytes were independently assessed for effects of temperature on their activation and function. We showed that the beneficial effect of elevated temperature on the plaque-forming cell response probably occurs during the priming stage of T helper cells, and neither improves the delivery of help or the activation of B cells, nor impairs suppressor T cell generation or function. We propose that this powerful immunopotentiating effect of hyperthermia may account for the selective value of the fever response. This suggests taht the monokine interleukin 1, which is the endogenous mediator of fever, may promote immune responses both through a direct action on lymphocytes, and indirectly by an action on the central nervous system resulting in fever.

Effects of DAP on diaphragm force and fatigue, including fatigue due to neurotransmission failure

1996 ◽  
Vol 81 (5) ◽  
pp. 2214-2220 ◽  
Author(s):  
Erik Van Lunteren ◽  
Michelle Moyer
Keyword(s):  
Muscle Force ◽  
Channel Blocker ◽  
Force Frequency ◽  
Rat Diaphragm ◽  
Muscle Contractility ◽  
Tetanic Force ◽  
Intermittent Stimulation ◽  
Relative Contribution ◽  
Frequency Relationship

Van Lunteren, Erik, and Michelle Moyer. Effects of DAP on diaphragm force and fatigue, including fatigue due to neurotransmission failure. J. Appl. Physiol. 81(5): 2214–2220, 1996.—Among the aminopyridines, 3,4-diaminopyridine (DAP) is a more effective K+ channel blocker than is 4-aminopyridine (4-AP), and, furthermore, DAP enhances neuromuscular transmission. Because 4-AP improves muscle contractility, we hypothesized that DAP would also increase force and, in addition, ameliorate fatigue and improve the neurotransmission failure component of fatigue. Rat diaphragm strips were studied in vitro (37°C). In field-stimulated muscle, 0.3 mM DAP significantly increased diaphragm twitch force, prolonged contraction time, and shifted the force-frequency relationship to the left without altering peak tetanic force, resulting in increased force at stimulation frequencies ≤50 Hz. During 20-Hz intermittent stimulation, DAP increased diaphragm peak force compared with control during a 150-s fatigue run and, furthermore, significantly improved maintenance of intratrain force. The relative contribution of neurotransmission failure to fatigue was estimated by comparing the force generated by phrenic nerve-stimulated muscles with that generated by curare-treated field-stimulated muscles. DAP significantly increased force in nerve-stimulated muscles and, in addition, reduced the neurotransmission failure contribution to diaphragm fatigue. Thus DAP increases muscle force at low-to-intermediate stimulation frequencies, improves overall force and intratrain fatigue during 20-Hz intermittent stimulation, and reduces neurotransmission failure.

Effect of temperature and divalent cations on the substratum attachment of rat hepatocytes in vitro

1978 ◽  
Vol 34 (1) ◽  
pp. 117-131
Author(s):  
P.O. Seglen ◽  
R. Gjessing
Keyword(s):  
Cytochalasin B ◽  
Arrhenius Plot ◽  
Divalent Cations ◽  
Rat Hepatocytes ◽  
Effect Of Temperature ◽  
Temperature Sensitive ◽  
Dextran Sulphate ◽  
Cell Binding ◽  
Better Than

The attachment of rat hepatocytes to polystyrene-adsorbed serum protein is relatively insensitive to inhibitors such as dextran sulphate, cycloheximide, colchicine and cytochalasin B, and enzymes like trypsin and neuraminidase, but it is strongly dependent on divalent cations. Mg2+ supports attachment better than Ca2+, but a combination of both is required for maximal attachment. The attachment is very temperature-sensitive, with a biphasic Arrhenius plot indicating an activation energy of 123 kJ/mol above 34 degrees C and 374 kJ/mol below 34 degrees C. The adsorbed attachment-promoting serum factor is inactivated by trypsin, or by Ca2+-dependent proteases which contaminate commercial preparations of collagenase. The adsorbed factor is resistant to treatment with glutaraldehyde, neuraminidase and heating to 90 degrees C, whereas the same factor in the unadsorbed state (in serum) is destroyed by heating to 70 degrees C. The factor in serum is unable to compete with the adsorbed factor for cell binding, hence it would appear that adsorption to polystyrene induces the active, heat-resistant conformation of the factor.

Effect of temperature on metabolism of glucose in vitro

1964 ◽  
Vol 207 (4) ◽  
pp. 849-852 ◽  
Author(s):  
Geraldine J. Fuhrman ◽  
Frederick A. Fuhrman
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
Low Temperatures ◽  
Arrhenius Equation ◽  
Effect Of Temperature ◽  
Rat Diaphragm ◽  
Abnormal Metabolism ◽  
Effects Of Temperature ◽  
Straight Lines

We recently showed that hypothermic rats fail to remove glucose from the extracellular phase. This information led to these studies on the uptake of glucose at low temperatures by rat diaphragm and red blood cells and its phosphorylation by hexokinase. It is shown here that rat diaphragm and red blood cells utilize glucose at all temperatures from 1 to 38 C. Both processes follow the Arrhenius equation and give µ equal to 18,000 and 21,900, respectively. The velocities of the phosphorylation of glucose by hexokinase from yeast and from rat muscle both yield straight lines on an Arrhenius plot with µ equal to 13,300 and 14,900. The temperature coefficient of the velocity of action of hexokinase is consistent with the effects of temperature on other enzymes. Penetration of glucose into cells at low temperatures apparently equals or exceeds the rate of phosphorylation. Thus the abnormal metabolism of glucose in hypothermia is not caused by failure of glucose to penetrate cells, and there is no unusual sensitivity of hexokinase to changes in temperature.

The development of dormancy in bulblets of Lilium speciosum generated in vitro. II. The effect of temperature

1990 ◽  
Vol 80 (3) ◽  
pp. 431-436 ◽  
Author(s):  
Isabelle Delvallee ◽  
Annie Paffen ◽  
Geert-Jan De Klerk
Keyword(s):  
Effect Of Temperature ◽  
Lilium Speciosum

Effect of Temperature on ADP-Induced Platelet Aggregation

1973 ◽  
Vol 29 (01) ◽  
pp. 183-189
Author(s):  
C. A Praga ◽  
E. M Pogliani
Keyword(s):  
Platelet Aggregation ◽  
Incubation Period ◽  
Room Temperature ◽  
Important Variable ◽  
Practical Significance ◽  
Effect Of Temperature ◽  
Induce Platelet Aggregation ◽  
Cuvette Holder ◽  
Exact Temperature

SummaryTemperature represents a very important variable in ADP-induced platelet aggregation.When low doses of ADP ( < 1 (μM) are used to induce platelet aggregation, the length of the incubation period of PRP in the cuvette holder of the aggregometer, thermostatted at 37° C, is very critical. Samples of the same PRP previously kept at room temperature, were incubated for increasing periods of time in the cuvette of the aggregometer before adding ADP, and a significant decrease of aggregation, proportional to the length of incubation, was observed. Stirring of the PRP during the incubation period made these changes more evident.To measure the exact temperature of the PRP during incubation in the aggre- gometer, a thermocouple device was used. While the temperature of the cuvette holder was stable at 37° C, the PRP temperature itself increased exponentially, taking about ten minutes from the beginning of the incubation to reach the value of 37° C. The above results have a practical significance in the reproducibility of the platelet aggregation test in vitro and acquire particular value when the effect of inhibitors of ADP induced platelet aggregation is studied.Experiments carried out with three anti-aggregating agents (acetyl salicyclic acid, dipyridamole and metergoline) have shown that the incubation conditions which influence both the effect of the drugs on platelets and the ADP breakdown in plasma must be strictly controlled.

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