Use of Knockout Mice Reveals Involvement of M2-Muscarinic Receptors in Control of the Kinetics of Acetylcholine Release

2003 ◽  
Vol 89 (4) ◽  
pp. 1954-1967 ◽  
Author(s):  
I. Slutsky ◽  
J. Wess ◽  
J. Gomeza ◽  
J. Dudel ◽  
I. Parnas ◽  
...  
Keyword(s):  
Muscarinic Receptors ◽  
Knockout Mice ◽  
Rise Time ◽  
Time Course ◽  
Acetylcholine Release ◽  
Frog Neuromuscular Junction ◽  
Quantal Content ◽  
Experimental Treatments ◽  
Kinetics Of ◽  
M2 Muscarinic Receptors

We have previously suggested that presynaptic M2-muscarinic receptors (M2R) are involved in the control of the time course of evoked acetylcholine release in the frog neuromuscular junction. The availability of knockout mice lacking functional M2R (M2-KO) enabled us to address this issue in a more direct way. Using the phrenic diaphragm preparation, we show that in wild-type (WT) mice experimental manipulations known to affect Ca2+ entry and removal, greatly affected the amount of acetylcholine released (quantal content). However, the time course of release remained unaltered under all these experimental treatments. On the other hand, in the M2-KO mice, similar experimental treatments affected both the quantal content and the time course of release. In general, a larger quantal content was accompanied by a longer duration of release. Similarly, the rise time of the postsynaptic current produced by axon stimulation was sensitive to changes in [Ca2+]o or [Mg2+]o in M2-KO mice but not in WT mice. Measurements of Ca2+ currents revealed that the shorter rise time of the postsynaptic current seen in high [Mg2+]o in M2-KO mice was not produced by a shorter wave of the presynaptic Ca2+ current. These results support our earlier findings and provide direct evidence for the major role that presynaptic M2-muscarinic receptors play in the control of the time course of evoked acetylcholine release under physiological conditions.

Characterization of the electroantennogram in Drosophila melanogaster and its use for identifying olfactory capture and transduction mutants

1991 ◽  
Vol 65 (3) ◽  
pp. 702-714 ◽  
Author(s):  
E. Alcorta
Keyword(s):  
Time Constant ◽  
Rise Time ◽  
Time Course ◽  
Electrical Response ◽  
Fall Time ◽  
Stimulus Concentration ◽  
Olfactory Sense ◽  
Kinetics Of ◽  
Drosophila Mutant

1. Amplitude as well as time course of the electroantennogram (EAG) in Drosophila has been used for describing electrical changes produced in the antenna in response to odorous stimulation. 2. Maximal amplitude of response appears to be directly correlated to stimulus concentration but, after achieving a maximum value, is independent of stimulation duration. 3. Rise time and fall time constants have been quantified for describing kinetics of response. The rise time constant decreases, but the fall time constant increases when increasing concentrations of odorant are supplied. 4. Variation among individuals for these EAG parameters is small enough to uncover even partial defects affecting the first sensory step. This fact combined with the possibility of obtaining mutants with defects in any intermediate process producing the electrical response makes the EAG of Drosophila a very useful tool for dissecting the components of the capture and transduction processes in the olfactory sense. 5. This kind of quantitative study of the EAG has been used in a new Drosophila mutant, od A, for localizing peripheral expression of the mutation. od A has been isolated as a behavioral mutant with an abnormally enhanced olfactory response to ethyl acetate. 6. The mutant's EAG in response to this odorant displays a normal amplitude but abnormal kinetics. Rise time as well as fall time show slower kinetics than normal, suggesting some defective step in the capture and transduction process.

Quantal release and facilitation at frog neuromuscular junctions at about 0 degrees C

1991 ◽  
Vol 65 (4) ◽  
pp. 834-840 ◽  
Author(s):  
J. Molgo ◽  
W. Van der Kloot
Keyword(s):  
Time Course ◽  
Neuromuscular Junctions ◽  
Frog Neuromuscular Junction ◽  
Deconvolution Method ◽  
Release Process ◽  
Quantal Release ◽  
Probabilistic Nature ◽  
Kinetics Of ◽  
Insight Into ◽  
Plate Current

1. It has been reported that at the frog neuromuscular junction at temperatures around 0 degrees C the release of transmitter quanta following nerve stimulation becomes disrupted, and the facilitation obtained after a second stimulus is no longer detectable. We thought that further investigation might give insight into the mechanism of quantal release, so we undertook experiments on Rana pipiens and Rana berlanieri. 2. In these species neuromuscular transmission occurs at temperatures as low as -0.8 degrees C. As the temperature is decreased further, transmission fails, apparently by a block in nerve conduction. The number of quanta released per stimulus decreases as temperature is lowered, with a Q10 of approximately 2.4. Owing to the decrease in the quantal output and the probabilistic nature of the release process, in occasional single records of an end-plate current (EPC), the pattern of release appeared disrupted. The kinetics of quantal release was studied by the use of a deconvolution method, which requires recording of EPCs and miniature EPCs (MEPCs) in preparations in high Mg(2+)-low Ca2+ solution. At approximately 0 degrees C the pattern of quantal release was similar to that at higher temperatures, although with a slower time course. At 0 degrees C the peak of release occurred approximately 3.5 ms after onset. 3. In our experiments there was almost no difference in the frequency of MEPCs at 22 degrees C and at 0 degree C. 4. We observed as much facilitation to a second stimulus at 0 degree C as at 10 degrees C. The Q10 for the decay of facilitation with time was between 1.9 and 2.3.(ABSTRACT TRUNCATED AT 250 WORDS)

Differences in presynaptic action of 4-aminopyridine and tetraethylammonium at frog neuromuscular junction

1987 ◽  
Vol 65 (5) ◽  
pp. 747-752 ◽  
Author(s):  
M. I. Glavinović
Keyword(s):  
Neuromuscular Junction ◽  
Transmitter Release ◽  
Action Potentials ◽  
Time Course ◽  
Potassium Conductance ◽  
Frog Neuromuscular Junction ◽  
Quantal Content ◽  
Low Concentrations ◽  
Voltage Dependent ◽  
Intracellular Ca

4-Aminopyridine markedly potentiates transmitter release at the frog cutaneous pectoris neuromuscular junction by increasing the quantal content even when applied at low concentrations (5–20 μM). This enhancement of transmitter release is associated with greater minimum synaptic latency, but the dispersion of the synaptic latencies does not appear much affected. This is in contrast with the action of tetraethylammonium (0.2–0.5 mM) in which case similar enhancement of transmitter release results not only in larger minimum synaptic latency but also in greater dispersion of the synaptic latencies. The time course of transmitter release associated with enhanced transmitter output is hence much more prolonged in the presence of tetraethylammonium than 4-aminopyridine, at least for low concentrations of 4-aminopyridine (5–20 μM). This indicates that their presynaptic actions differ significantly. This conclusion is further strengthened by the finding that unlike tetraethylammonium, 4-aminopyridine induces bursts of release, presumably by producing multiple action potentials in the nerve terminal. Tetraethylammonium probably acts by blocking the delayed potassium conductance, but the blockade of Ca2+-activated K+ conductance cannot be excluded. 4-Aminopyridine, however, probably blocks the fast inactivating (IA) K+ current, but it also may be acting directly on the voltage-dependent Ca2+ conductance or on the intracellular Ca2+ buffering.

Depolarization Initiates Phasic Acetylcholine Release by Relief of a Tonic Block Imposed by Presynaptic M2 Muscarinic Receptors

2005 ◽  
Vol 93 (6) ◽  
pp. 3257-3269 ◽  
Author(s):  
H. Parnas ◽  
I. Slutsky ◽  
G. Rashkovan ◽  
I. Silman ◽  
J. Wess ◽  
...  
Keyword(s):  
Muscarinic Receptors ◽  
Knockout Mice ◽  
Neuromuscular Junctions ◽  
Pulse Amplitude ◽  
Dependent Manner ◽  
Wild Type ◽  
Ach Release ◽  
Voltage Dependent ◽  
M2 Muscarinic Receptors

The role of presynaptic muscarinic autoreceptors in the initiation of phasic acetylcholine (ACh) release at frog and mouse neuromuscular junctions was studied by measuring the dependency of the amount (m) of ACh release on the level of presynaptic depolarization. Addition of methoctramine (a blocker of M2 muscarinic receptors), or of acetylcholinesterase (AChE), increased release in a voltage-dependent manner; enhancement of release declined as the depolarizing pulse amplitude increased. In frogs and wild-type mice the slope of log m/log pulse amplitude (PA) was reduced from about 7 in the control to about 4 in the presence of methoctramine or AChE. In M2 muscarinic receptor knockout mice, the slope of log m/log PA was much smaller (about 4) and was not further reduced by addition of either methoctramine or AChE. The effect of a brief (0.1 ms), but strong (−1.2 μA) depolarizing prepulse on the dependency of m on PA was also studied. The depolarizing prepulse had effects similar to those of methoctramine and AChE. In particular, it enhanced release of test pulses in a voltage-dependent manner and reduced the slope of log m/log PA from about 7 to about 4. Methoctramine + AChE occluded the prepulse effects. In knockout mice, the depolarizing prepulse had no effects. The cumulative results suggest that initiation of phasic ACh release is achieved by depolarization-mediated relief of a tonic block imposed by presynaptic M2 muscarinic receptors.

Influence of Radioiodine Therapy on Urinary Iodine Excretion

1998 ◽  
Vol 37 (03) ◽  
pp. 107-112 ◽  
Author(s):  
I. Lauer ◽  
M. Bähre ◽  
E. Richter ◽  
B. Melier
Keyword(s):  
Time Course ◽  
Radioiodine Therapy ◽  
Urinary Iodine ◽  
Target Volume ◽  
Urinary Iodine Excretion ◽  
Urinary Creatinine ◽  
Iodine Excretion ◽  
Kinetics Of ◽  
Persistent Elevation ◽  
Benign Thyroid

Summary Aim: In 214 patients with benign thyroid diseases the time-course of urinary iodine excretion (UIE) was investigated in order to identify changes after radioiodine therapy (RITh). Method: UIE was measured photometrically (cerium-arsenite method) and related to urinary creatinine on the first and last day of the radioiodine test and then three days, seven days, four weeks, and six months after 1311 administration. Results: As compared with the level found immediately before radioiodine therapy, median UIE had almost doubled four weeks after therapy and was still significantly elevated six months after therapy. This increase correlated significantly with the target volume as measured by scintigraphy and sonography. Conclusions: The persistent elevation of UIE for months after RITh is a measure of treatment-induced damage to thyrocytes. Therefore, in view of the unfavourable kinetics of iodine that follow it, RITh should if possible be given via a single-dose regime.

Paired-pulse facilitation in the dentate gyrus: a patch-clamp study in rat hippocampus in vitro

1994 ◽  
Vol 72 (1) ◽  
pp. 326-336 ◽  
Author(s):  
M. Andreasen ◽  
J. J. Hablitz
Keyword(s):  
Patch Clamp ◽  
Dentate Gyrus ◽  
Time Constant ◽  
Rise Time ◽  
Time Course ◽  
Nmda Antagonist ◽  
Paired Pulse ◽  
Stimulation Intensity ◽  
Paired Pulse Facilitation ◽  
Epsc Amplitude

1. Whole-cell patch-clamp recordings were used to study paired-pulse facilitation (PPF) of the lateral perforant path input to the dentate gyrus in thin hippocampal slices. 2. Orthodromic stimulation of the lateral perforant pathway evoked a excitatory postsynaptic current (EPSC) with a latency of 3.3 +/- 0.1 ms (mean +/- SE) that fluctuated in amplitude. The EPSC had a rise time (10-90%) of 2.79 +/- 0.06 ms (n = 35) and decayed with a single exponential time course with a time-constant of 9.14 +/- 0.24 ms (n = 35). No correlation was found between the amplitude of the EPSC and the rise time or decay time-constant. The non-N-methyl-D-aspartate (NMDA) antagonist 6-cyano-7-nitroquinoxaline-2,3-dione completely blocked the EPSC whereas the NMDA antagonist D-aminophosphonovaleric acid (APV) had modest effects. 3. When a test (T-)EPSC was preceded at an interval of 100 ms by a conditioning (C-)EPSC, a significant increase in the amplitude of the T-EPSC was seen in 38 out of 44 trials analyzed from a total of 27 granule cells. The average amount of PPF was 35.7 +/- 2.1%. There was no apparent correlation between the amount of PPF and the stimulation intensity or mean amplitude of the C-EPSC. The time course of the facilitated T-EPSC was not significantly different from that of the C-EPSC. 4. No correlation was found between the amplitude of the C-EPSC and that of the T-EPSC. Estimates of quantal content (mcv) were determined by calculating the ratio of the squared averaged EPSC amplitude (from 48 responses) to the variance of these responses (M2/sigma 2) whereas quantal amplitudes (qcv) were estimated by calculating the ratio of the response variance to average EPSC amplitude (sigma 2/M). PPF was found to be associated with an average increase in mcv of 64.8 +/- 7.2% (n = 38) whereas qcv was decreased by 12.1 +/- 3.8%. 5. The time course of PPF was studied by varying the interval between the C- and T-pulse from 10 to 400 ms while keeping the stimulation intensity constant. Maximal facilitation of the T-EPSC was obtained with interpulse intervals < or = 25 ms where the average facilitation amounted to approximately 70% (n = 6). The decline of facilitation was nearly exponential and was no longer evident with intervals > 350 ms.(ABSTRACT TRUNCATED AT 400 WORDS

scholarly journals Changes in J chain and mu chain RNA expression as a function of B cell differentiation.

1984 ◽  
Vol 160 (3) ◽  
pp. 877-892 ◽  
Author(s):  
G Lamson ◽  
M E Koshland
Keyword(s):  
B Cell ◽  
Time Course ◽  
Rna Synthesis ◽  
Rna Expression ◽  
B Cell Differentiation ◽  
Activated Lymphocytes ◽  
J Chain ◽  
Pattern Characteristic ◽  
Kinetics Of ◽  
Lymphoid Cell Lines

The time course of differentiative events in the pentamer IgM response was examined by following the expression of J chain and mu chain RNA and their protein products in mitogen-stimulated lymphocytes. The analyses showed that the shift to mus RNA synthesis begins shortly after stimulation and precedes proliferation of the cells and any increase in mu RNA levels. In contrast, expression of J chain RNA and the amplification of J chain and mus message are late events that coincide with a phase of rapid proliferation and with the secretion of pentamer IgM antibody. The kinetics of J and mu chain RNA expression observed in normal lymphocytes were supported by analyses of lymphoid cell lines. B lymphomas were found to display the RNA pattern characteristic of early-activated lymphocytes, i.e., a partial shift to mus RNA production and no J chain RNA, whereas IgM-secreting lines resembled late-activated lymphocytes in their expression of high levels of both mus and J chain mRNA. Moreover, the kinetics of J and mus chain RNA expression correlates with the sequential action of B cell lymphokines in the induction of the pentamer IgM response. This correlation suggests that the successive differentiative changes are triggered by successive membrane stimuli.

Sign in / Sign up

Export Citation Format

Share Document