Biomedical subjects

R J Brady

Publications and source records attributed to R J Brady.

At least 19 recordsLinked to original sources

PubMed1995

Modulation of N-methyl-D-aspartic acid receptors by extracellular calcium in immature and adult hippocampal slices: whole cell recordings in CA3 pyramidal cells.

Lowering extracellular calcium concentration [Ca2+]o in rat hippocampal slices can lead to an induction of epileptiform activity. It has been shown that this effect is more pronounced in slices of neonatal rats (postnatal day, PND 8-19) than in mature slices (> PND 40) and it has been suggested that unique N-methyl-D-aspartic acid (NMDA) receptor properties of immature rat hippocampal pyramidal cells contribute to this developmental effect. In a voltage clamp experiment we tested NMDA receptor properties in hippocampal pyramidal cells by measuring NMDA receptor mediated currents evoked by iontophoretic applied NMDA in the basal dendrites of CA3 pyramidal neurons. We found that lowering extracellular calcium from 2 to 1 mM, increases NMDA evoked inward current in pyramidal cells around the resting membrane potential. However, this effect is observed in slices of neonatal as well as in slices of mature rats, suggesting that there is no difference in NMDA receptor sensitivity to extracellular Ca2+ between these two age groups. The modulation of the NMDA receptor by extracellular calcium at physiological concentrations can have important consequences in pathological conditions during which extracellular calcium reaches low levels. Because this 'hypocalcemic' condition induces a larger current influx via the NMDA receptor channel at resting membrane potentials, it can further enhance cellular excitability and contribute to sustain epileptiform activity.

Age Factors↗

PubMed1994

Developmental alterations in the sensitivity of hippocampal NMDA receptors to AP5.

Changes in the properties of the N-methyl-D-aspartate (NMDA) receptor are proposed as a factor in the decrease in synaptic plasticity during maturation of the brain. Alterations in the hippocampal NMDA receptor population were studied during development by comparing competitive antagonist efficacy during a window characterized by hyperexcitability and active synaptogenesis to that seen in a more mature period. A developmental change in the sensitivity of the N-methyl-D-aspartate (NMDA) evoked response to the competitive antagonist D(-)2-amino-5- phosphonopentanoic acid (D-AP5) was observed by whole-cell mode voltage-clamp, intracellular and extracellular recordings in hippocampal slices. These differences were observed in the portions of the hippocampus that contain the terminal axon arbors of the CA3 pyramidal neurons, the synaptic fields of the Schaffer collateral, associational and commissural pathways. The apparent antagonist efficacy was 2-3 times greater in immature slices obtained on postnatal day (PND) 10-16 than that observed in more mature tissue (> PND38). Given the possible role of the NMDA receptor in network plasticity, these observations could indicate a role for the molecular diversity of this important receptor subtype family in the maturation of hippocampal pathways.

2-Amino-5-phosphonovalerate↗

PubMed1994

NMDA receptor responses in adult hippocampal CA1 region after neonatal treatment with MK-801: comparison with NMDA receptor responses in the immature rat.

Neonatal MK-801 treatment from postnatal day 8-19 leads to long-term effects on brain function, suggesting that exposure to this drug leads to the development of a brain with immature network properties. One aspect of this hypothesis, that the NMDA receptors preserve their immature state after the treatment, has been tested by measuring the potency of the competitive antagonist D-AP5 in hippocampal slices. We have previously shown that an increased potency to D-AP5 is a characteristic property of NMDA receptors during early life. In the present study we measured field potentials in the CA1 region of rat hippocampal slices evoked by iontophoretic NMDA application in the Schaffer-commissural synaptic fields. Agonist dose-response curves were constructed, followed by bath applications of increasing concentrations of the antagonist D-AP5. The maximum NMDA evoked field response was the same in slices of mature control (PND70-90; 18.9 +/- 1.2 mV) and MK-801 treated rats (PND70-90; 19.3 +/- 0.9 mV), but significantly larger in immature slices (PND10-16; 24.0 +/- 0.6 mV). The sensitivity to NMDA in hippocampal slices from each group was estimated by quantifying the ionotophoretic ejection current (= dose) which evoked 50% of the maximum field response (EC50). A significantly higher sensitivity to NMDA was found in hippocampal slices obtained from MK-801-treated rats (EC50 = 3.6 +/- 0.2 nA) than in slices from control (EC50 = 6.1 +/- 0.7 nA) or immature (EC50 = 5.9 +/- 0.5 nA) animals.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1993

Localized excitatory synaptic interactions mediate the sustained depolarization of electrographic seizures in developing hippocampus.

Repetitive synchronized neuronal discharging that lasts for seconds and even minutes in in vitro brain slice preparations are important new models in experimental epilepsy. In hippocampal slices from 1-2-week-old rats, individual CA3 pyramidal cells undergo a sustained depolarization during such electrographic seizures, induced by GABAA receptor antagonists. In experiments reported here these events were produced in small isolated segments of the CA3 subfield, measuring only 400-500 microns along the cell body layer. In such minisclices local application of either kynurenic acid or 6-cyano-7-nitroquinoxaline-2-3-dione (CNQX) to the proximal basilar dendrites abolished the synchronized discharges of electrographic seizures. Interictal spikes appeared unaffected by this treatment. Application of these excitatory amino acid receptor antagonists to distal basilar dendrites or apical dendrites was ineffective. In "larger" minislices, measuring 700-1000 microns along the cell body layer, application of kynurenic acid, CNQX, or TTX to the proximal basilar dendrites did not abolish electrographic seizures but instead selectively suppressed the intracellularly recorded sustained depolarization and the coincident slow negative field potential recorded in proximal basilar dendrites. Results of several experiments suggest that electrographic seizures recorded under these conditions were produced by a remote network of "generator cells." Since the remote neurons were unaffected by local application of the drugs, it seemed likely that they continued to undergo a sustained depolarization. Simultaneous blockade of basilar dendritic synapses in the "generator" population abolished electrographic seizures throughout these larger minislices. These results suggest that the sustained depolarization plays a central role in seizure generation and that it does not have to be generated in every neuron, only in a critical number of "generator cells" for a seizure to occur. Taken together, results presented here suggest that the sustained depolarization of electrographic seizures is a separate physiological process from the more rapid repetitive depolarizations of the seizure discharges and is required if electrographic seizures are to occur. This slow depolarization appears to be synaptically mediated and generated exclusively in proximal basilar dendrites. Therefore, in addition to the excitatory synaptic potentials involved in paroxysmal depolarization shift generation, a second form of recurrent excitation may exist in immature hippocampus. Not only is this physiological process critical for the genesis of seizures, but it also appears to be highly partitioned within the hippocampal laminae.

6-Cyano-7-nitroquinoxaline-2,3-dione↗

PubMed1992

The ontogeny of hippocampal local circuits and focal epileptogenesis.

Our understanding of the pathophysiological mechanisms underlying the childhood epilepsies is rudimentary at this time. However, results of recent studies in animal models have lead to a number of important hypotheses concerning age-dependent alterations in seizure susceptibility. In area CA3 of hippocampus it appears that during a critical period when this brain area is particularly prone to electrographic seizures, an overabundance of recurrent excitatory synapses may exist. At the same time the synapses themselves appear to be functionally different than their mature counterpart. Age-dependent differences in the properties of postsynaptic NMDA receptors seem to contribute to enhanced seizure susceptibility. In recent years significant progress has been made in unravelling the fundamental processes that underlie the formation of connections between developing neurons. Over-production of early-formed axon collaterals appears to be common-place in the CNS. Moreover, the selection of appropriate patterns of connectivity appear to be dependent in large part on the patterning of neurophysiologic activity. In this regard the NMDA receptor seems to play a pivotal role in synapse consolidation. Based on these observations and the central role recurrent excitation appears to play in hippocampal seizures, it seems entirely plausible that excessive abnormal neuronal discharging that occurs during seizures early in life could result in the consolidation of abnormal numbers of recurrent excitatory synapses. This miswiring of hippocampal networks might be responsible for the marked seizure susceptibility into adulthood and might even contribute to complex partial epilepsy in individuals with a history of childhood seizures.

Adult↗

PubMed1991

Calcium modulation of the N-methyl-D-aspartate (NMDA) response and electrographic seizures in immature hippocampus.

Recordings from the CA3 region of hippocampal slices indicate a developmental change in the divalent cation sensitivity of the response elicited by N-methyl-D-aspartate (NMDA) application. In parallel experiments a developmental difference is demonstrated in the capacity of extracellular calcium to modulate electrographic seizure generation. Calcium modulation of the NMDA-elicited response may contribute to the pronounced capacity of immature hippocampus to generate electrographic seizures. Under these conditions activity dependent changes in extracellular calcium could have a greater influence on ion flow produced by activation of the NMDA receptor. The possibility that changes in the receptor isoform may occur during development would have widespread implications for normal cognitive functions and dysfunctions during brain maturation.

Age Factors↗

PubMed1991

Age-dependent alterations in the operations of hippocampal neural networks.

Results from numerous studies suggest that the functioning of rat hippocampal neural networks during the second postnatal week of life differs distinctly from that in the mature brain. During this critical period, network behavior might be considered hyperexcitable. Spontaneous network-driven bursts of synaptic potentials, which have not been reported in mature hippocampus, are commonly observed. While these events could be attributable to a late onset of GABAergic synaptic transmission, results suggest that this is not the case. In the immature hippocampus orthodromic stimulation leads to prolonged depolarizations and often repetitive synchronized discharging of the entire CA3 population. These events are in many ways reproduced by application of drugs that suppress GABAergic synaptic transmission. The synchronized discharging of the CA3 population is blocked by excitatory amino acid antagonists. This finding, coupled with our growing understanding of the role that recurrent excitation plays in CA3 network functioning, has led to the hypothesis that the differences in network behavior early in life may be largely attributable to an overabundance of local-circuit recurrent excitatory synapses. With maturation, axon collaterals and attendant synapses would regress to achieve an adult complement. Results from dual intracellular recordings as well as anatomical studies of individual CA3 pyramidal cells support this hypothesis. Unique properties of the NMDA receptor at these recurrent excitatory synapses early in life may also promote network excitability. The participation of extracellular Ca2+ in the voltage dependency of the NMDA receptor-linked iontophore could also contribute to synapse consolidation during maturation and thus in the establishment of network connectivity.

Aging↗

PubMed1990

Neural networks and synaptic transmission in immature hippocampus.

The results reviewed in this chapter indicate that local circuit synaptic interactions are surprisingly well-developed in the rat hippocampal CA3 subfield during the second postnatal week. Intracellular recordings reveal large spontaneous epsps and ipsps. Synchronized bursts of synaptic potentials are observed in most paired intracellular recordings. Antidromic and orthodromic electrical stimulation evokes synaptic responses that are reminiscent of recordings from mature hippocampus. However, following brief trains of electrical stimuli and during bath application of a GABAA receptor antagonist, large prolonged depolarizations are recorded. These are suppressed by excitatory amino acid antagonists. In comparison, slices from mature rats do not produce these events under the same conditions. Thus, a hypothesis has been presented that the degree of recurrent synaptic interaction between pyramidal cells may be enhanced during this critical period in hippocampal development. An analysis of recurrent epsps using dual intracellular recordings is consistent with this contention. The degree of excitatory synaptic interaction between CA3 neurons appears to be at least equal to and likely in excess of that reported in mature hippocampus. One possible explanation for this is that the number of recurrent excitatory synapses may increase transiently during hippocampal development, only later to regress to numbers found in the adult. Recent studies of others suggest that activation of NMDA receptors may play a key role in the maintenance of synapses during development (Cline et al., 1987; Kleinschmidt et al., 1987). In this regard it is interesting that the characteristic of the NMDA receptor-ion channel complex on immature and mature rat CA3 hippocampal pyramidal cells appear to be quite different. These differences may play a role in synapse formation and maintenance. The role of NMDA receptors in LTP and learning (for review see Cotman et al., 1989; Lynch, 1986) are widely discussed. In other circles recurrent excitatory neuronal networks are thought to be substrates for memory (Lynch, 1986; Haberly and Bower, 1989). Both NMDA receptors and recurrent excitation are well represented in the CA3 subfield of immature hippocampus. One challenging area for future study will be the clarification of the interrelations between synaptogenesis and synaptic plasticity within neural networks of the developing hippocampus.

Animals↗

PubMed1989

Postnatal development of GABA-mediated synaptic inhibition in rat hippocampus.

Developmental alterations in GABAergic synaptic transmission were examined physiologically and biochemically in hippocampus of rats from 3 days of age to adulthood. Neither antidromic nor orthodromic stimulation could elicit identifiable inhibitory postsynaptic potentials in CA1 neurons in slices from rats 5 or 6 days of age. In contrast, at this age these stimuli result in large inhibitory postsynaptic potentials in CA3 pyramidal cells. In the latter cells orthodromic stimulation produced a brief monosynaptic excitatory postsynaptic potential which was followed by a large prolonged biphasic hyperpolarization. These signals were strikingly similar to those recorded in 1-month-old rats. In addition, large recurrent inhibitory postsynaptic potentials were produced by antidromic stimulation. By postnatal day 9 similar inhibitory postsynaptic potentials could be elicited in a majority of neurons of the CA1 subfield. As in mature pyramidal cells, application of GABA antagonists, such as bicuculline, selectively eliminated the antidromic inhibitory postsynaptic potential and the first component of the biphasic inhibitory postsynaptic potential generated by stimulation of stratum radiatum. In the CA3 subfield, this blockade of GABA receptors resulted in prolonged afterdischarges in slices from immature but not month-old rats. Measurements of the equilibrium potential and the conductance of antidromic inhibitory postsynaptic potentials in CA3 neurons were very similar when made during the first postnatal week and at 1 month of age. While on days 10-11 the equilibrium potential was very similar to measurements made at these other ages, the conductance was 3-4 times greater. The activity of glutamate decarboxylase, the synthetic enzyme for GABA, was very low at 3 days in hippocampus, and increased until 30 days of age at which time adult values were obtained. By comparison, hippocampal GABA levels were high early in postnatal life. Glutamate decarboxylase activities in microdissected CA3 and CA1 subfields were similar in immature hippocampus. These results demonstrate dramatic differences in the ontogenesis of functional GABAergic inhibitory synaptic transmission in the CA1 and CA3 subfields of rat hippocampus. The late development of GABA-mediated synaptic inhibition in the CA1 subfield could play a role in the susceptibility of immature hippocampus to seizures. However, the large GABA-mediated inhibitory postsynaptic potentials present in the CA3 subfield at the same age have a critical role in dampening neuronal excitability.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗

PubMed1987

Network capitation plans: an operational perspective.

Dentistry's mission has been defined as improving the oral health of the public. In the ADA's report on the strategic plan for dentistry, the first priority addressed is to convert public unmet needs into a demand for dental services. One of the methods which the plan has outlined for action is to "expand dental prepayment and promote growth of innovative payment mechanisms." The prepaid capitation method of payment can and will provide a vehicle to service those unmet needs. Healthcare insurance companies are organizing regional and national networks of individual practices who will accept this method of prepayment. As in all systems of delivery, there are possibilities for abuse. This article addresses a prepaid capitation delivery system from an operational perspective, in order to identify the elements that should safeguard the patient, dentist, employer group, and carrier.

Capitation Fee↗

PubMed1986

Ketamine selectively suppresses synchronized afterdischarges in immature hippocampus.

The role of excitatory amino acid neurotransmission in epileptogenesis was investigated in the developing hippocampus. Bath application of ketamine blocked penicillin-induced, synchronized afterdischarges in immature rat CA3 hippocampal neurons. Ketamine also decreased the duration of the preceding intracellularly recorded depolarization shift but had no measurable effect on the resting membrane potential or input impedance of pyramidal cells. Concentrations of ketamine that blocked afterdischarge generation dramatically depressed intracellular depolarizations produced by iontophoretic application of N-methyl-D-aspartate (NMDA) but not quisqualate. The effects of the NMDA antagonist 2-amino-7-phosphonoheptanoic acid on epileptiform discharges were identical to those of ketamine. These results suggest that an endogenous excitatory amino acid acting on an NMDA receptor plays a key role in the pronounced capacity of immature hippocampus for seizures.

Aging↗

PubMed1986

Extracellular K+ accumulation during penicillin-induced epileptogenesis in the CA3 region of immature rat hippocampus.

Ion-selective microelectrode techniques were used to study extracellular K+ changes associated with penicillin-induced epileptogenesis in the CA3 region of hippocampal slices. Recordings were made in slices taken from rats 9-16 days of age, which have a pronounced capacity to undergo prolonged synchronized afterdischarges. Direct comparisons were made to slices from hippocampus from more mature rats, 30-35 days old, which are much less prone to seizure-like events. The amplitude and time course of the K+ transients varied across the CA3 laminae. K+ signals were largest close to stratum pyramidale in stratum oriens (the infrapyramidal zone). Recordings from this site showed extracellular K+ accumulation to be unusually large in immature hippocampus. The ceiling [K+]o level recorded during seizure-like events ranged from 14.4 to 20.2 mM and averaged 16.9 mM. The peak amplitude of extracellular K+ transients following an epileptiform burst in slices from immature rats averaged 4.31 mM while the mean of similar recordings from mature rats was 0.97 mM. Detailed laminar distribution studies in developing hippocampus revealed that the K+ signals were large in the proximal two-thirds of the basilar dendrites and proximal half of the apical dendrites. K+ accumulation in stratum pyramidale was comparatively small even though at its very edge in stratum oriens large K+ transients were always recorded. The latter was also true in recordings from mature hippocampus. Other dendritic signals in mature tissue were comparatively small. Laminar analysis was performed of the field potentials recorded by the reference barrel of the K+ electrodes. Negative field potential for the epileptiform burst and subsequent slow potential correlated in space with sites of K+ accumulation in both immature and mature hippocampal slices. Interictal and prolonged ictal-like discharges, recorded in developing hippocampus, arose from the same baseline [K+]o. However, since [K+]o is excessively high during the course of these epileptiform events it most likely has a role in the unusual propensity of immature hippocampus for seizures.

Age Factors↗

PubMed1986

The dendritic origins of penicillin-induced epileptogenesis in CA3 hippocampal pyramidal cells.

Experiments were performed in order to identify the sites of epileptiform burst generation in rat hippocampal CA3 pyramidal cells. A subsequent slow field potential was studied, which is associated with afterdischarge generation. Laminar field potential and current source-density (CSD) methods were employed in hippocampal slices exposed to penicillin. Simultaneous intracellular and extracellular field recordings from the CA3 pyramidal cell body layer showed that whenever an epileptiform burst was recorded extracellularly, individual CA3 neurons underwent an intense depolarization shift. In extracellular records a slow negative field potential invariably followed epileptiform burst generation. In approximately 10% of slices, synchronous afterdischarges rode on the envelope of this negative field potential. Intracellularly a depolarizing afterpotential followed the depolarization shift and was coincident with the extracellular slow negative field potential. A one-dimensional CSD analysis performed perpendicular to the CA3 cell body layer showed that during epileptiform burst generation large current sinks occur simultaneously in the central portions of both the apical and basilar dendrites. The average distance of the peak amplitude for these sinks from the center of the cell body layer was 175 +/- 46.8 microns and 158 +/- 25.0 microns, respectively. A large current source was recorded in the cell body layer. Smaller current sources were observed in the distal portions of the dendritic layers. During the postburst slow field potential a current sink was recorded at the edge of the cell body layer in stratum oriens--a region referred to as the infrapyramidal zone. Simultaneous with the current sink recorded there, smaller sinks were often observed in the dendritic layers that appeared to be "tails" or prolongations of the currents underlying burst generation. Two-dimensional analyses of these field potentials were performed on planes parallel and perpendicular to the exposed surface of the slice. Isopotential contours showed that the direction of extracellular current is mainly orthogonal to the CA3 laminae. Correction of CSD estimates made perpendicular to the cell body layer for current flowing in the other direction did not alter the location of computed current sources and sinks. In order to show that the dendritic currents associated with epileptiform burst generation were active sinks, tetrodotoxin (TTX) was applied locally to the dendrites where the current sinks were recorded.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1984

Postsynaptic actions of baclofen associated with its antagonism of bicuculline-induced epileptogenesis in hippocampus.

The effect of baclofen on bicuculline-induced epileptogenesis was investigated in the CA3 region of hippocampal slices taken from rats 9 days to 8 weeks of age. Bath application of baclofen blocked all spontaneous epileptiform activity and raised the stimulus strength required for orthodromic induction of epileptiform discharges. Baclofen was equally effective in antagonizing depolarization shift generation in mature and immature rat slices. The duration of afterdischarges recorded in immature hippocampus was unaltered, yet these events were eliminated when the proceeding depolarization shift was blocked. Baclofen hyperpolarized all CA3 pyramidal cells studied with an associated decrease in membrane resistance. These effects were produced by a direct postsynaptic action.

Animals↗

PubMed1984

Penicillin-induced epileptogenesis in immature rat CA3 hippocampal pyramidal cells.

Penicillin's ability to produce epileptiform discharges in the CA3 region of hippocampus was examined both extracellularly and intracellularly in slices taken from immature rats 3-25 days of age. Comparisons were made to similar recordings from slices taken from mature rats. Between postnatal days 9 and 19 penicillin treatment resulted in spontaneous extracellular epileptiform bursts and coincident intracellular depolarization shifts. These events were more prolonged and less frequent than in slices from mature rats, and the bursts were followed by prolonged afterdischarges, often 20-30 s in duration. Intracellularly these afterdischarges consisted of large, rhythmic slow depolarizing potentials, which resulted in one or more action potentials in individual CA3 pyramidal cells. Extracellular field recordings showed these events to be simultaneous with synchronous discharges of a large population of CA3 pyramidal cells. In pups 1-2 weeks of age the ability of hippocampus to produce prolonged afterdischarges was associated with a slow depolarizing afterpotential, which followed the downstroke of the depolarization shift. Coincident with this afterpotential was a prolonged negative field in the CA3 pyramidal cell body layer. By postnatal days 24 and 25 the tendency to generate afterdischarges was greatly reduced. In addition, afterdischarges were observed infrequently in slices taken during the first postnatal week. Spike trains produced by prolonged intracellular current injection in slices taken on postnatal days 9-19 were followed by large afterhyperpolarizations and were unable to produce afterdischarges in individual CA3 pyramidal cells. Intracellular recordings from presumed glial cells suggest that extracellular K+ accumulation may play a role in the pronounced capacity of hippocampus from 1- and 2-week-old rat pups to generate prolonged afterdischarges.

Age Factors↗

PubMed1983

The design and implementation of a regional economic-demographic simulation model.

The management of urban growth patterns in the United States in order to maximize the benefits of public and private investments in urban areas is examined. The authors present "the results of a modelling design effort to link a hybrid regional input-output model to demographic, labor force and energy models for use in planning in the San Francisco Bay Region. Extensive use of system feedback is a major component of this research effort. Summary results of the model outputs are presented...."

Americas↗

PubMed1983

Pressure effects on the ADH-induced initiation of water flow in toad bladder.

Earlier studies employed colchicine to demonstrate the need for microtubules in the ADH-induced initiation of increased water permeability in toad bladder. We have used colchicine and hydrostatic pressure together to determine whether formed or growing microtubules are required for initiation of the ADH response in Bufo marinus. When ADH and 8,000 psi were administered simultaneously, the ADH-induced increase in water flow was inhibited while under pressure by 107 +/- 7% (n = 6). Application of 8,000 psi for 10 min before ADH administration resulted in an increased initiation of the ADH osmotic response over the non-pressure-treated control (average acceleration, rate of water flow increase during first 3 min after ADH stimulation, 1.25 +/- 0.27 vs. 0.43 +/- 0.12 mg X cm-2 X min-2, n = 6). In addition, the inhibition of the ADH response brought about by colchicine incubation was overcome with pretreatment of the colchicine-incubated bladders with 8,000 psi for 10 min (average 3-min acceleration, 0.18 +/- 0.04 vs. 1.13 +/- 0.06 mg X cm-2 X min-2, respectively). Repeating the experiments with dibutyryl cAMP gave similar results. We interpret these data as suggesting that growing microtubules are required for initiation. The proposed model is as follows. Pressure removes colchicine inhibition by introducing, through disassembly of formed microtubules, more colchicine-free tubulin subunits. These subunits are then available following decompression to reassemble when the tissue is challenged with hormone.

Animals↗

PubMed1981

Nocodazole inhibition of the vasopressin-induced water permeability increase in toad urinary bladder.

Nocodazole is a synthetic antitumor drug that binds rapidly to tubulin. When this drug is applied to toad bladder prior to vasopressin stimulation it inhibits the vasopressin response. A maximum inhibition (68%) is reached with a dose level of 10 micrograms/ml applied one-half hour prior to vasopressin stimulation (20 mU/ml). This compares with an inhibition of 50% seen with a 3-h exposure of the tissue to colchicine (0.1 mM) prior to stimulation with vasopressin. Application of nocodazole (1 microgram/ml) 3 min after hormonal stimulation shows no inhibition of the response at one-half hour past stimulation. These data support the view that microtubules are involved in the vasopressin-induced increase in water permeability in toad bladder and also indicate that this involvement is limited to the period prior to or directly after stimulation.

Animals↗