Biomedical subjects

M S Jensen

Publications and source records attributed to M S Jensen.

At least 37 records · Page 2Linked to original sources

PubMed1999

Role of presynaptic L-type Ca2+ channels in GABAergic synaptic transmission in cultured hippocampal neurons.

Using dual whole cell patch-clamp recordings of monosynaptic GABAergic inhibitory postsynaptic currents (IPSCs) in cultured rat hippocampal neurons, we have previously demonstrated posttetanic potentiation (PTP) of IPSCs. Tetanic stimulation of the GABAergic neuron leads to accumulation of Ca2+ in the presynaptic terminals. This enhances the probability of GABA-vesicle release for up to 1 min, which underlies PTP. In the present study, we have examined the effect of altering the probability of release on PTP of IPSCs. Baclofen (10 microM), which depresses presynaptic Ca2+ entry through N- and P/Q-type voltage-dependent Ca2+ channels (VDCCs), caused a threefold greater enhancement of PTP than did reducing [Ca2+]o to 1.2 mM, which causes a nonspecific reduction in Ca2+ entry. This finding prompted us to investigate whether presynaptic L-type VDCCs contribute to the Ca2+ accumulation in the boutons during spike activity. The L-type VDCC antagonist, nifedipine (10 microM), had no effect on single IPSCs evoked at 0.2 Hz but reduced the PTP evoked by a train of 40 Hz for 2 s by 60%. Another L-type VDCC antagonist, isradipine (5 microM), similarly inhibited PTP by 65%. Both L-type VDCC blockers also depressed IPSCs during the stimulation (i.e., they increased tetanic depression). The L-type VDCC "agonist" (-)BayK 8644 (4 microM) had no effect on PTP evoked by a train of 40 Hz for 2 s, which probably saturated the PTP process, but enhanced PTP evoked by a train of 1 s by 91%. In conclusion, the results indicate that L-type VDCCs do not participate in low-frequency synchronous transmitter release, but contribute to presynaptic Ca2+ accumulation during high-frequency activity. This helps maintain vesicle release during tetanic stimulation and also enhances the probability of transmitter release during the posttetanic period, which is manifest as PTP. Involvement of L-type channels in these processes represents a novel presynaptic regulatory mechanism at fast CNS synapses.

Animals↗

PubMed1999

Activity-dependent depression of GABAergic IPSCs in cultured hippocampal neurons.

Short-term depression of monosynaptic GABAergic inhibitory postsynaptic currents (IPSCs) evoked between pairs of cultured rat hippocampal neurons was investigated using dual whole cell patch-clamp recordings. Paired stimuli applied to the GABAergic neuron resulted in paired-pulse depression (PPD) of the second IPSC (IPSC2) at interpulse intervals from 25 to 2,000 ms. CGP 55845A, but not CGP 35348, reduced PPD marginally. Brief paired-pulse applications of exogenous GABA indicated that postsynaptic factors made only minimal contribution to PPD of IPSCs. IPSC1 and PPD was reduced on lowering [Ca2+]o and enhanced on increasing [Ca2+]o. The potassium-channel blocker 4-aminopyridine (4-AP), which increases presynaptic Ca2+ influx, enhanced IPSC1 and PPD. Chelation of residual Ca2+ in the GABAergic boutons with EGTA-AM enhanced PPD. Stimulation of the presynaptic neuron at frequencies (f) ranging from 2.5 to 80 Hz resulted in tetanic depression of IPSCs, which declined rapidly and reached a plateau depending on f and [Ca2+]o. CGP 55845A decreased tetanic depression in the first part of the train, but this could be overcome with continued stimulation. We show that GABAergic IPSCs are robustly depressed by paired-pulse stimulation in cultured hippocampal neurons. The depression of IPSCs is mainly independent of presynaptic GABAB receptors and could be caused by depletion of releasable vesicles. Depleted synapses recover with a slow time course, depending on factors that regulate [Ca2+]i in the GABAergic boutons.

4-Aminopyridine↗

PubMed1998

Circadian and ultradian variation in pancreatic secretion of meal-fed pigs after weaning.

We studied the time structure of pancreatic secretion in two experiments involving seven 6- to 7-wk-old intact male pigs, surgically fitted with a jugular vein catheter for blood sampling, pancreatic catheter, and a duodenal T-cannula for chronic pancreatic juice sampling for 72 h at 30- to 60-min intervals. Pigs were kept in metabolic cages in a regimen of 12 h of light alternating with 12 h of darkness and were fed at 0800, 1500, and 2200 daily a standard diet based on barley, soybean meal, and fish meal. Beginning 4 d after surgical recovery, three 24-h collections of pancreatic juice and blood sampling were begun either at 0700 or 0800 every 2nd d for 5 d. Pancreatic secretion exhibited a pattern characterized by distinct meal-related secretions of the first phase (postprandial), containing large amounts of protein and enzymes (trypsin and chymotrypsin), and by non-food-stimulated secretions of the second phase with less protein and enzymes. During the dark span, the first phase was practically absent; the response of the pancreatic secretion to the 2200 meal was not very pronounced. Apart from the anticipated circadian rhythm demonstrable by single cosinor analysis on a group basis, a prominent 8-h component was almost invariably statistically significant. Moreover, an approximately 3.43-h component was also prominent. These data indicate that pancreatic secretions are circadian periodic and that their response to a standard meal is also circadian-stage dependent. The circadian components may have been free-running because the pigs were adjusting themselves to the changing phase and that resulted in the period being different from exactly 24 h.

Activity Cycles↗

PubMed1997

Electrophoretic separation of proteolytic enzymes in pancreatic juice collected with the pouch or catheter method.

CONCLUSION: The results of this study demonstrated that proteolytic enzymes in pancreatic juice from pigs prepared with the pouch method (PM) were nearly fully active or were fully active. When activation with enterokinase was carried out further inactivation and/or breakdown occurred for chymotrypsin C and cathodal trypsin. In addition, some inactivation and/or breakdown of proteolytic enzymes in pancreatic juice occurred during collection of pancreatic juice from PM pigs. METHODS: Samples of pancreatic juice were collected from growing pigs using either the PM or the catheter method (CM). An isolated pouch was prepared where the pancreatic duct enters the duodenum, and three pigs were fitted with a pancreatic pouch re-entrant cannula. Three different pigs had a catheter surgically inserted into the pancreatic duct. Pooled 8-h samples of pancreatic juice were analyzed before and after activation with enterokinase. Chymotrypsin, trypsin, and elastase activities were identified in pancreatic juice after separation by electrophoresis in 1% agarose gels at pH 8.6 using N-acetyl-DL-phenylalanine-beta-naphthyl ester (Ac-Phe-beta ne) as a substrate. RESULTS: This qualitative enzyme assay indicated that a considerable amount of chymotrypsin C, anodal trypsin, chymotrypsins A and B, elastase II, and cathodal trypsin were present in samples of nonactivated pancreatic juice from PM pigs. In contrast, the only active enzymes identified in pancreatic juice from CM pigs were very small amounts of chymotrypsin A and elastase II. The amounts of chymotrypsin C and cathodal trypsin were lower in activated than in nonactivated pancreatic juice from PM pigs. However, there were increases in the amounts of the other enzymes when pancreatic juice from PM pigs was activated. As expected, the activation of pancreatic juice from CM pigs resulted in the measurement of very high amounts of all the proteolytic enzymes. The amounts of anodal trypsin, chymotrypsins A and B, and elastase II were higher in activated pancreatic juice from CM pigs than from PM pigs.

Animals↗

PubMed1997

A comparison of two methods to measure amylase, lipase, trypsin, and chymotrypsin activity and the effect of freezing and thawing on enzyme activities in pancreatic juice.

An experiment was conducted to compare two methods, referred to as A and B, for measuring amylase, lipase, trypsin, and chymotrypsin activities in samples of pancreatic juice and to investigate the effect of freezing and thawing on enzyme activities. Three growing pigs were used in this study. A catheter was surgically inserted into the pancreatic duct and a simple T-cannula was placed in the duodenum to allow the return of collected pancreatic juice. Pancreatic juice was collected for a total of 24 h and 27 pooled 8-h samples were used in the analyses. Following collection and pooling, the samples were frozen at -80 degrees C, due to the large number of samples and analyses, and stored for 2 weeks. The samples were thawed and enzyme activities were determined with Method A. The samples were frozen again at -80 degrees C, stored for 2 weeks, and rethawed and enzyme activities were determined with Methods A and B. There was a direct relationship between Method A and Method B for measuring amylase, trypsin, and chymotrypsin activity (R2 = 0.95, 0.81, and 0.90, respectively). However, a poor relationship was observed between a titrimetric and a photometric method for determining lipase activity (R2 = 0.42). A second freezing, storage, and thawing did not affect (p > 0.3) the concentration of protein or amylase activity in pancreatic juice. However, the activities of chymotrypsin, trypsin, and, especially, lipase were decreased by 29.0% (p < 0.11), 40.4% (p < 0.01), and 82.9% (p < 0.001), respectively. In conclusion, either Method A or Method B can be used to measure amylase, trypsin, and chymotrypsin activities in pancreatic juice but a titrimetric method should be used to determine lipase activity. Furthermore, if the activities of lipase, trypsin, and chymotrypsin are to be determined, the sample should only be frozen and thawed once. If amylase activity will be determined, the sample can be repeatedly frozen and thawed.

Amylases↗

PubMed1997

Role of intrinsic burst firing, potassium accumulation, and electrical coupling in the elevated potassium model of hippocampal epilepsy.

Perfusing rat hippocampal slices with high-K+ (7.5 mM) saline induced brief population bursts originating in CA3 at 0.5-1 Hz and spreading synaptically into CA1. In 42% of the slices the brief bursts evoked in CA1 gave way every 0.5-2 s to sustained ictal (or seizure) episode with tonic and clonic components. Paired intra- and extracellular recordings in the CA1 pyramidal layer were used to characterize the synaptic and nonsynaptic mechanisms generating the brief and sustained epileptiform events. The interictal, tonic, or clonic primary burst response in CA1 comprised a spindle-shaped, tight cluster (170-180 Hz) of five to seven population spikes. Bursts evoked between sequential seizures (interictal bursts) were initially small and progressively increased in size. Concurrently, basal extracellular K+ concentration ([K+]o] increased from 6.5 to 7.5 mM. The tonic event emanated from a large primary burst and comprised prolonged (> 1 s), self-sustained afterdischarge, associated with a rise in [K+]o to 12 mM. Bursts generated during the subsequent [K+]o decline (clonic bursts) also were large and followed by some afterdischarge. They became small during [K+]o undershoot to 6.5 mM. Intrinsically burst firing pyramidal cells (PCs) were recruited before or at the very onset of the primary population burst and fired repetitively during its course. Nonbursters were recruited > or = 10 ms after the beginning of the primary burst and fired, on average, only one spike. The PCs depolarized during the primary burst and subsequent afterdischarge. The primary depolarizing shift was larger in bursters than in nonbursters. Both bursters and nonbursters fired repetitively, albeit intermittently, during tonic and clonic afterdischarge. Throughout the interictal-ictal cycle intracellular spikes were time-locked to population spikes, indicating that PCs fire in tight synchrony. Differential recording of transmembrane potentials unmasked rapid (4-7 ms) transmembrane depolarizing potentials of up to 10 mV, coincident with population spikes. We conclude that in the high-K+ model of hippocampal epilepsy, the local generation of population bursts in CA1 is led by intrinsic bursters, which recruit and synchronize other PCs by synaptic, electrical, and K(+)-mediated excitatory interactions. The cycling between interictal, tonic, and clonic events appears to result from feedback interactions between neuronal discharge and [K+]o.

Animals↗

PubMed1997

Development of digestive enzymes in pigs with emphasis on lipolytic activity in the stomach and pancreas.

The effect of age and weaning on the activities of digestive enzymes with emphasis on the lipolytic enzymes before and after weaning was investigated. The activities of amylase, chymotrypsin, trypsin, carboxyl ester hydrolase, pancreatic lipase, and colipase in pancreatic tissue and the activity of gastric lipase in the cardiac mucosa of the stomach in 45 pigs were response variables. The activity of trypsin was not affected by weaning and the rate of increase was similar during the whole experiment. The activities of chymotrypsin and amylase decreased at weaning (P < .05). After weaning the activity of chymotrypsin increased more slowly than before weaning (P < .001), whereas the rate of increase of amylase activity remained unchanged. Lipase, colipase, and carboxyl ester hydrolase activities decreased at weaning (P < .001), whereas gastric lipase activity increased at weaning (P < .01). The development of lipase, colipase, and carboxyl ester hydrolase activity decreased postweaning (P < .01), whereas gastric lipase activity increased before weaning and remained constant after weaning. Pancreatic lipase had a considerably higher capacity for hydrolyzing tributyrin, and the total activity of pancreatic lipase was up to 600 times higher than that of gastric lipase. The lipolytic enzymes displayed a non-parallel pattern of development, and we suggest that this reflects the importance of these enzymes during the suckling and postweaning phases, respectively. However, the significance of gastric lipase for the digestion of fat in pigs remains to be elucidated.

Aging↗

PubMed1997

Collection of pancreatic juice from growing pigs. A comparative study of the pouch method and the catheter method.

CONCLUSION: The results of this study demonstrate that there are large differences in the amount of pancreatic juice secreted and in the chemical and enzymatic composition of pancreatic juice when the pouch and the catheter methods were used, and these differences must be taken into consideration in future studies with either method. METHODS: A study was performed to compare the two most commonly used methods to collect pancreatic juice from growing pigs; namely, the pouch method (PM) and the catheter method (CM). In the first part of the study, three barrows (initial weight 37 kg) were fitted with a pancreatic pouch re-entrant cannula. An isolated pouch was prepared in which the pancreatic duct enters the duodenum. In the second part of the study, also with three barrows (initial weight 32 kg), a catheter was inserted into the pancreatic duct. RESULTS: At several points during the 24-h collection, the hourly rate of pancreatic juice secretion in CM pigs was larger (p < 0.05) than for PM pigs. CM pigs also had a higher (p < 0.05) daily volume of secretion, 4.09 vs 2.63 L/24 h for PM pigs. The pH of pancreatic juice collected from CM pigs was consistently higher (p < 0.01) throughout the 24-h collection. In contrast, the concentration and daily output of bicarbonate did not differ between CM and PM pigs. The concentration of protein in pancreatic juice from PM pigs (7.21 g/L) was higher (p < 0.001) than for CM pigs (4.08 g/L). Specific amylase and lipase and total amylase activities were greater (p < 0.01) in pancreatic juice collected from PM pigs. Specific and total carboxyl ester hydrolase and colipase activities were substantially (p < 0.01) larger in pancreatic juice collected from CM pigs. A major difference between the methods was that trypsin and chymotrypsin were fully active in pancreatic juice from PM pigs, whereas virtually no trypsin or chymotrypsin activity was detected in pancreatic juice from CM pigs. Specific and total chymotrypsin activities did not differ between PM and CM pigs. Both specific and total trypsin activities were substantially higher in pancreatic juice from CM pigs: 3682 U/L and 12,752 U/24 h, respectively, vs 1031 U/L and 2639 U/24 h, respectively, in pancreatic juice from PM pigs.

Animals↗

PubMed1996

Spike after-depolarization and burst generation in adult rat hippocampal CA1 pyramidal cells.

1. Intracellular recordings in adult rat hippocampal slices were used to investigate the properties and origins of intrinsically generated bursts in the somata of CA1 pyramidal cells (PCs). The CA1 PCs were classified as either non-bursters or bursters according to the firing patterns evoked by intrasomatically applied long ( > or = 100 ms) depolarizing current pulses. Non-bursters generated stimulus-graded trains of independent action potentials, whereas bursters generated clusters of three or more closely spaced spikes riding on a distinct depolarizing envelope. 2. In all PCs fast spike repolarization was incomplete and ended at a potential approximately 10 mV more positive than resting potential. Solitary spikes were followed by a distinct after-depolarizing potential (ADP) lasting 20-40 ms. The ADP in most non-bursters declined monotonically to baseline ('passive' ADP), whereas in most bursters it remained steady or even re-depolarized before declining to baseline ('active' ADP). 3. Active, but not passive, ADPs were associated with an apparent increase in input conductance. They were maximal in amplitude when the spike was evoked from resting potential and were reduced by mild depolarization or hyperpolarization (+/- 2 mV). 4. Evoked and spontaneous burst firing was sensitive to small changes in membrane potential. In most cases maximal bursts were generated at resting potential and were curtailed by small depolarizations or hyperpolarizations (+/- 5 mV). 5. Bursts comprising clusters of spikelets ('d-spikes') were observed in 12% of the bursters. Some of the d-spikes attained threshold for triggering full somatic spikes. Gradually hyperpolarizing these neurones blocked somatic spikes before blocking d-spikes, suggesting that the latter are generated at more remote sites. 6. The data suggest that active ADPs and intrinsic bursts in the somata of adult CA1 PCs are generated by a slow, voltage-gated inward current. Bursts arise in neurones in which this current is sufficiently large to generate suprathreshold ADPs, and thereby initiate a regenerative process of spike recruitment and slow depolarization.

Action Potentials↗

PubMed1996

Ionic basis of spike after-depolarization and burst generation in adult rat hippocampal CA1 pyramidal cells.

1. Intracellular recordings in adult rat hippocampal slices were used to identify the ionic conductances underlying active spike after-depolarization (ADP) and intrinsic burst firing in the somata of CA1 pyramidal cells (PCs). To test the 'Ca2+ hypothesis', Ca2+ currents were suppressed by replacing the Ca2+ in the saline with either Mn2+ or Mg2+. Alternatively, the inorganic Ca2+ channel blockers Cd2+ (0.5 mM) or Ni2+ (2 mM) were added to the saline. To test the 'Na+ hypothesis', Na+ currents were blocked with tetrodotoxin (TTX; 0.5 microM). 2. The suppression of Ca2+ currents blocked the fast after-hyperpolarization (AHP) generated by the fast Ca(2+)-gated K+ current Ic, while enhancing the amplitude and duration of active spike ADPS. 3. Evoked and spontaneous burst firing was preserved undiminished following Ca2+ current suppression, while the propensity to fire bursts increased in many cases. The postburst medium AHP (generated primarily by the muscarine-sensitive voltage-gated K+ current, IM) was not affected by this treatment, which blocked the slow AHP (generated by the slow Ca(2+)-gated K+ current, IAHP). 4. TTX strongly suppressed active ADPs and intrinsic bursts before substantially reducing the threshold, rate of rise and amplitude of solitary spikes. 5. In Ca(2+)-free saline, caesium-filled PCs generated large, plateau ADPs following an initial burst of fast spikes. Application of TTX suppressed these ADPs before solitary fast spikes appeared to be reduced. 6. Injection of brief, just subthreshold depolarizing current pulses into bursters evoked slow depolarizing potentials lasting up to 50 ms. These persisted after suppression of Ca2+ currents and were entirely blocked by TTX. 7. We conclude that active spike ADPs and intrinsic bursts in the somata of adult CA1 PCs are generated by a low voltage-gated, persistent Na+ current. Burst termination is mediated by voltage-gated K+ currents activated during the burst (most likely IM), rather than by the Ca(2+)-gated K+ currents Ic and IAHP. The latter currents downregulate the innate tendency of CA1 PCs to burst (Ic) and limit the rate of spontaneous burst firing (IAHP).

Action Potentials↗

PubMed1996

Fast and one-step folding of closely and distantly related homologous proteins of a four-helix bundle family.

Bovine acyl-coenzyme A binding protein is a four-helix bundle protein belonging to a group of homologous eukaryote proteins that binds medium and long-chain acyl-coenzyme A esters with a very high affinity. The three-dimensional structure of both the free and the ligated protein together with the folding kinetics have been described in detail for the bovine protein and with four new sequences reported here, a total of 16 closely related sequences ranging from yeasts and plants to human are known. The kinetics of folding and unfolding in different concentrations of guanidine hydrochloride together with equilibrium unfolding have been measured for bovine, rat and yeast acyl-coenzyme A binding protein. The bovine and rat sequences are closely related whereas the yeast is more distantly related to these. In addition to the three natural variants, kinetics of a bovine mutant protein, Tyr31 --> Asn, have been studied. Both the folding and unfolding rates in water of the yeast protein are 15 times faster than those of bovine. The folding rates in water of the two mammalian forms, rat and bovine, are similar, though still significantly different. A faster unfolding rate both for rat and the bovine mutant protein results from a lower stability of the native states of these. These hydrophobic regions, mini cores, have been identified in the three-dimensional structure of the bovine protein and found to be formed primarily by residues that have been conserved throughout the entire eukaryote evolution from yeasts to both plants and mammals as seen in the sample of 16 sequences. The conserved residues are found to stabilize helix-helix interactions and serve specific functional purposes for ligand binding. The fast one-step folding mechanism of ACBP has been shown to be a feature that seems to be maintained throughout evolution despite numerous differences in sequence and even dramatic differences in folding kinetics and protein stability. The protein study raises the question to what extent does the conserved hydrophobic residues provide a scaffold for an efficient one-step folding mechanism.

Amino Acid Sequence↗

PubMed1996

Exocrine pancreatic secretions in growing pigs fed diets containing fish oil, rapeseed oil or coconut oil.

Two experiments were performed to study the effect of feeding diets containing oils with different fatty acid composition on exocrine pancreatic secretions in growing pigs using two different methods to collect pancreatic juice. In the first experiment, three barrows (initial weight 37 kg) were fitted with a pancreatic pouch re-entrant cannula. An isolated pouch was prepared where the pancreatic duct enters the duodenum. In the second experiment, also using three barrows (initial weight 32 kg), a catheter was inserted into the pancreatic duct. Three wheat starch and fish meal-based diets were formulated to contain either 15 g fish oil, rapeseed oil or coconut oil/100 g. In both experiments, the diets were fed according to a 3 times 3 Latin square design. The volume of pancreatic juice secreted, pH and secretion of bicarbonate, protein, amylase, trypsin, lipase and colipase were not significantly affected by the diets in the first experiment. In the second experiment, chymotrypsin secretion was significantly greater in pigs fed the coconut oil diet, and secretion of carboxyl ester hydrolase was significantly higher in pigs fed the fish oil diet. When compared qualitatively, pigs in Experiment 2 secreted more pancreatic juice; the pancreatic juice had a higher pH, and trypsin, carboxyl ester hydrolase and colipase secretions were substantially higher whereas amylase secretion was lower than for pigs in Experiment 1. The fatty acid composition of the different oils had minor effects on exocrine pancreatic secretion in growing pigs. However, there were considerable differences between the two surgical methods used to collect pancreatic juice, and these differences may be explained by physiological changes induced by the two methods.

Amylases↗

PubMed1996

Cooperative quizzes in the anatomy and physiology laboratory: a description and evaluation.

Physiology educators read journals, such as this one, to gather ideas about curriculum and instruction. Most articles focus on curriculum (i.e., what is taught), but this paper will focus on instruction (i.e., how curriculum is implemented). Just as there are different types of curricula, there are different types of instruction. The most common strategy is lecture. Lectures are extremely efficient for delivering large amounts of information in a short period of time. A common laboratory strategy is discovery or inquiry-based learning (i.e., giving students tools, cognitive and physical, to deduce new information via investigations). A third instructional strategy is the use of cooperative learning. Proper conditions are required for each instructional strategy, and problems arise when the wrong combinations are put together. This paper will describe how a cooperative learning environment can be created in the anatomy and physiology laboratory through the use of cooperative quizzes. It will include a brief introduction to the pedagogical theory behind cooperative learning and an evaluation of the effectiveness of quizzes compared with more traditional methods.

Anatomy↗

PubMed1995

The pattern of the circadian rhythm of pancreatic secretion in fed pigs.

The pattern of the circadian rhythm of pancreatic secretion was studied in four 6- to 7-wk-old intact male pigs that were kept in metabolic cages under 12 h light:12 h dark cycles and fed three times a day at 0800, 1500, and 2200. Three 24-h collections of pancreatic juice and blood were begun at 0800 every 2nd day during 5 d. A biphasic pattern of the exocrine pancreas was detected. Secretions during the first phase (postprandial peak) contained large amounts of protein and enzymes, and secretions appeared immediately after feed ingestion. Secretions during the second phase (between meals) had lesser amounts (P < .001) of protein and enzymes, but were of longer duration (P < .001). The output of protein and trypsin activity were generally correlated (P < .001) with each other, but the correlation of either with the volume outflow were small. However, these variables were correlated (P < .01) with the volume outflow at night during the second phase. Plasma concentrations of insulin and glucose increased after each meal, but the increase was significant (P < .05) only for insulin. Insulin and glucose concentrations were correlated (P < .01) with each other after the meals at 0800 and 1500. The results indicated a biphasic pattern of the exocrine pancreas and a monophasic pattern of the endocrine pancreas in pigs. The regulation of these patterns seems to vary during the 24-h period. Furthermore, feed enhanced secretion, but it did not seem to be the only factor controlling the circadian pattern.

Analysis of Variance↗

PubMed1994

Muscarinic modulation of intrinsic burst firing in rat hippocampal neurons.

Intracellular recordings in rat hippocampal slices were used to examine how exogenous and endogenous cholinergic agonists modulate the firing pattern of intrinsically burst-firing pyramidal cells. About 24% of CA1 pyramidal cells generated all-or-none, high-frequency bursts of fast action potentials in response to intracellular injection of long positive current pulses. Application of carbachol (5 microM) converted burst firing in these neurons into regular trains of independent spikes. Acetylcholine (5 microM) exerted a similar effect, provided acetylcholine esterase activity was blocked with neostigmine (2 microM). Atropine (1 microM) reversed this cholinergic effect, indicating its mediation by muscarinic receptors. Cholinergic agonists also caused mild neuronal depolarization but the block of intrinsic burst firing was independent of this effect. Repetitive stimulation of cholinergic fibres in the presence of neostigmine (2 microM) evoked a slow cholinergic excitatory postsynaptic potential (EPSP) lasting about a minute. During the slow EPSP, burst firing could not be evoked by depolarizing pulses and the neurons fired in regular mode. These effects were prevented by pretreatment with atropine (1 microM). Exogenously applied cholinergic agonists and endogenously released acetylcholine also reduced spike frequency accommodation and suppressed the long-duration afterhyperpolarization in burst-firing pyramidal cells in an atropine-sensitive manner. A membrane-permeable cAMP analogue (8-bromo-cAMP; 1 microM) also reduced frequency accommodation and blocked the long-duration afterhyperpolarization, but did not affect intrinsic burst firing at all. Taken together, the data show that muscarinic receptor stimulation transforms the stereotyped, phasic response of burst-firing neurons into stimulus-graded, tonic discharge.

8-Bromo Cyclic Adenosine Monophosphate↗

PubMed1994

Variant firing patterns in rat hippocampal pyramidal cells modulated by extracellular potassium.

1. The distribution of distinctive firing modes within the population of CA1 pyramidal cells and their modulation by the extracellular concentration of potassium ([K+]o) were investigated with intracellular recordings in rat hippocampal slices. 2. Pyramidal cells were injected with long (> 250 ms) and brief (3-5 ms) positive current pulses of increasing intensity. In normal [K+]o (3.5 mM), most cells (38 of 46 cells; 83%) were regular spiking neurons (generating accommodating trains of independent action potentials during long depolarizations and a single spike in response to brief stimuli). The remaining pyramidal cells (8 of 46; 17%) displayed differential tendencies to generate stereotyped clusters of action potentials, or bursts, according to which they were grouped into three subsets of endogenous bursters: grade I, bursting only when stimulated with long depolarizing current pulses (6 of 46; 13%); grade II, bursting also in response to brief stimulation (1 of 46; 2%); grade III, bursting also spontaneously even in absence of synaptic transmission (1 of 46; 2%). 3. Raising [K+]o from 3.5 to 7.5 mM (high [K+]o) significantly reduced resting membrane potential and input impedance but did not change the threshold potential for eliciting an action potential. 4. Raising [K+]o to 7.5 mM reversibly converted many regular spiking cells to bursters. Likewise, the burst tendency of normally bursting pyramidal cells increased to a higher grade in high [K+]o. Consequently, the fraction of bursters in high [K+]o (17 of 41 cells; 42%) was approximately 2.5-fold higher than in normal [K+]o and their differential distribution was shifted toward higher grades of bursting.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1993

Opponent effects of potassium on GABAA-mediated postsynaptic inhibition in the rat hippocampus.

1. The effects of raising the concentration of extracellular potassium ([K+]o) on gamma aminobutyric acid (GABA)-mediated inhibitory postsynaptic potentials (IPSPs) were investigated in adult rat hippocampal slices using intracellular recording techniques. IPSPs were evoked in CA1 pyramidal neurons by direct activation of inhibitory interneurons in slices treated with glutamatergic antagonists to block excitatory synaptic transmission. The fast (Cl(-)-dependent, GABAA receptor-mediated) IPSPs (fIPSPs) were isolated from the slow (K(+)-dependent; GABAB receptor-mediated) IPSPs (sIPSPs) by intracellular injection of QX-314, which also suppressed fast (Na(+)-dependent) action potentials. 2. In normal (3.5 mM) and in high (7.5 mM) [K+]o, the peak fIPSP amplitude changed nonlinearly with membrane potential (VM) in a way consistent with outward rectification of the underlying conductance. The input conductance of the fIPSP (GfIPSP) measured around resting VM (about -67 mV) increased 1.7-fold on changing from normal to high-K+ saline, whereas resting VM depolarized 6.8 mV. Repolarizing VM reversed the increase in GfIPSP, suggesting that it was due to outward rectification. The resting input conductance of the neurons increased 1.4-fold in high K+. 3. The time course of fIPSPs was prolonged by high K+. The half time of fIPSP decay increased 1.4-fold, and in half of the neurons the decay became conspicuously multipeaked, suggesting that neurally evoked GABA release from inhibitory interneurons was prolonged. 4. In normal K+, the reversal potentials of fIPSPs (EfIPSP; -76.5 mV) was 9.7 mV more negative than resting VM. Polarizing VM 10-20 mV for 15 min with current injection did not change EfIPSP appreciably.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1992

Purification and characterization of variants of acyl-CoA-binding protein in the bovine liver.

Four differently modified forms of acyl-CoA-binding protein (ACBP) were identified in ACBP purified from bovine liver. The majority of the purified ACBP was focused at pH 5.9 in isoelectric focusing and could be shown to be N-acetylated ACBP without any further modifications. Two minor peaks were focused at pH 5.25 and 4.85 respectively. Mass spectrometry and sequence determination showed that the pI 5.25 form was acetylated at Lys18 and that the pI 4.85 form was malonylated in the same position. Furthermore, it could be shown that non-enzymic glycosylation occurred during purification. The acetylated and malonylated variants of ACBP were only found in adult cattle.

Amino Acid Sequence↗