Bridging the age gap in breast cancer: impact of omission of breast cancer surgery in older women with oestrogen receptor-positive early breast cancer on quality-of-life outcomes.

BACKGROUND: Primary endocrine therapy may be an alternative treatment for less fit women with oestrogen receptor (ER)-positive breast cancer. This study compared quality-of-life (QoL) outcomes in older women treated with surgery or primary endocrine therapy. METHODS: This was a multicentre, prospective, observational cohort study of surgery or primary endocrine therapy in women aged over 70 years with operable breast cancer. QoL was assessed using European Organisation for Research and Treatment of cancer QoL questionnaires QLQ-C30, -BR23, and -ELD14, and the EuroQol Five Dimensions 5L score at baseline, 6 weeks, and 6, 12, 18, and 24 months. Propensity score matching was used to adjust for baseline variation in health, fitness, and tumour stage. RESULTS: The study recruited 3416 women (median age 77 (range 69-102) years) from 56 breast units. Of these, 2979 (87.2 per cent) had ER-positive breast cancer; 2354 women had surgery and 500 received primary endocrine therapy (125 were excluded from analysis due to inadequate data or non-standard therapy). Median follow-up was 52 months. The primary endocrine therapy group was older and less fit. Baseline QoL differed between the groups; the mean(s.d.) QLQ-C30 global health status score was 66.2(21.1) in patients who received primary endocrine therapy versus 77.1(17.8) among those who had surgery plus endocrine therapy. In the unmatched analysis, changes in QoL between 6 weeks and baseline were noted in several domains, but by 24 months most scores had returned to baseline levels. In the matched analysis, major surgery (mastectomy or axillary clearance) had a more pronounced adverse impact than primary endocrine therapy in several domains. CONCLUSION: Adverse effects on QoL are seen in the first few months after surgery, but by 24 months these have largely resolved. Women considering surgery should be informed of these effects.

Neurotransmission: Chemical and electrical interneuron coupling.

Recent studies have described the coupling between pairs of neocortical interneurons involving both electrical and chemical transmission; these new results may have important implications for the mechanisms underlying neuronal synchrony and rhythmic activity in the brain.

Identification of a novel AU-Rich element in the 3' untranslated region of epidermal growth factor receptor mRNA that is the target for regulated RNA-binding proteins.

The epidermal growth factor receptor (EGF-R) plays an important role in the growth and progression of estrogen receptor-negative human breast cancers. EGF binds with high affinity to the EGF-R and activates a variety of second messenger pathways that affect cellular proliferation. However, the underlying mechanisms involved in the regulation of EGF-R expression in breast cancer cells are yet to be described. Here we show that the EGF-induced upregulation of EGF-R mRNA in two human breast cancer cell lines that overexpress EGF-R (MDA-MB-468 and BT-20) is accompanied by stabilization (>2-fold) of EGF-R mRNA. Transient transfections using a luciferase reporter identified a novel EGF-regulated approximately 260-nucleotide (nt) cis-acting element in the 3' untranslated region (3'-UTR) of EGF-R mRNA. This cis element contains two distinct AU-rich sequences (~75 nt), EGF-R1A with two AUUUA pentamers and EGF-R2A with two AUUUUUA extended pentamers. Each independently regulated the mRNA stability of the heterologous reporter. Analysis of mutants of the EGF-R2A AU-rich sequence demonstrated a role for the 3' extended pentamer in regulating basal turnover. RNA gel shift analysis identified cytoplasmic proteins (~55 to 80 kDa) from breast cancer cells that bound specifically to the EGF-R1A and EGF-R2A cis-acting elements and whose binding activity was rapidly downregulated by EGF and phorbol esters. RNA gel shift analysis of EGF-R2A mutants identified a role for the 3' extended AU pentamer, but not the 5' extended pentamer, in binding proteins. These EGF-R mRNA-binding proteins were present in multiple human breast and prostate cancer cell lines. In summary, these data demonstrate a central role for mRNA stabilization in the control of EGF-R gene expression in breast cancer cells. EGF-R mRNA contains a novel complex AU-rich 260-nt cis-acting destabilizing element in the 3'-UTR that is bound by specific and EGF-regulated trans-acting factors. Furthermore, the 3' extended AU pentamer of EGF-R2A plays a central role in regulating EGF-R mRNA stability and the binding of specific RNA-binding proteins. These findings suggest that regulated RNA-protein interactions involving this novel cis-acting element will be a major determinant of EGF-R mRNA stability.

Glycine modulation of the NMDA receptor/channel complex.

One class of excitatory amino acid (EAA) receptor, the N-methyl-D-aspartate (NMDA) receptor, has excited enormous interest because of its unique pharmacological and physiological properties. Yet another property of this receptor/channel complex has recently emerged: its activation is greatly enhanced by and may even be dependent on, sub-micromolar concentrations of glycine.

Facilitation, augmentation and potentiation at central synapses.

Release probability (P) appears to be a major factor that influences the pattern of transmitter release. At cortical pyramidal axon inputs onto different classes of target cells, very different release patterns are observed, patterns that correlate with release probability. Simplistically, 'low P' synapses display facilitation and augmentation, whereas 'high P' synapses supplied by the same axon exhibit paired-pulse and frequency-dependent depression. Different combinations of factors probably contribute to release probability at different terminals, during development and under different experimental conditions. The recent advances made by molecular biological studies of the release machinery do, however, provide candidate proteins and protein-protein interactions whose differential distributions might be important factors in determining the patterns of transmitter release.

Temporal and spatial properties of local circuits in neocortex.

A large body of anatomical data has detailed many complexities of neocortical circuitry, and physiological studies have indicated some roles for this circuitry in the complex functions of the cortex. Until recently, however, we have little precise information about the spatio-temporal properties of synaptic connections between individual neocortical neurones. Studies of synaptic responses elicited in one neocortical neurone by action potentials in another, and parallel morphological studies that have identified these neurones and the synaptic connections between them, have now described these parameters for certain types of local circuit connection in the neocortex. Some of these studies confirmed previous observations and inferences, but others provided major surprises. Evidence indicates that the class of both the presynaptic and postsynaptic neurone together determine a wide range of synaptic properties, such as the type of postsynaptic receptors involved and the temporal pattern of transmitter release, so that each type of synapse displays unique properties. A role for retrograde diffusable messages in determining the temporal properties of these circuits is postulated.

Molecular frequency filters at central synapses.

During the 1950s to 70s most of the mechanisms that control transmitter release from presynaptic nerve terminals were described at the neuromuscular junction. It was not, however, until the 1990s that the multiplicity of protein-protein interactions that govern this process began to be identified. The sheer numbers of proteins and the complexity of their interactions at first appears excessive, even redundant. However, studies of identified central synapses indicate that this molecular diversity may underlie a important functional diversity. The task of the neuromuscular junction is to relay faithfully the rate and pattern code generated by the motoneurone. To demonstrate phenomena such as facilitation and augmentation that are apparent only when the probability of release is low, experimental manipulation is required. In the cortex, however, low probability synapses displaying facilitation can be recorded in parallel with high probability synapses displaying depression. The mechanisms are largely the same as those displayed by the neuromuscular junction, but some are differentially expressed and controlled. Central synapses demonstrate exquisitely fine tuned information transfer, each of the many types displaying its own repertoire of pattern- and frequency-dependent properties. These appear tuned to match both the discharge pattern in the presynaptic neurone and the integrative requirements of the postsynaptic cell. The molecular identification of these differentially expressed frequency filters is now just coming into sight. This review attempts to correlate these two aspects of synaptic physiology and to identify the components of the release process that are responsible for the diversity of function.

Simulating long-term and residual effects of nitrogen fertilization on corn yields, soil carbon sequestration, and soil nitrogen dynamics.

Soil carbon sequestration (SCS) has the potential to attenuate increasing atmospheric CO2 and mitigate greenhouse warming. Understanding of this potential can be assisted by the use of simulation models. We evaluated the ability of the EPIC model to simulate corn (Zea mays L.) yields and soil organic carbon (SOC) at Arlington, WI, during 1958-1991. Corn was grown continuously on a Typic Argiudoll with three N levels: LTN1 (control), LTN2 (medium), and LTN3 (high). The LTN2 N rate started at 56 kg ha(-1) (1958), increased to 92 kg ha(-1) (1963), and reached 140 kg ha(-1) (1973). The LTN3 N rate was maintained at twice the LTN2 level. In 1984, each plot was divided into four subplots receiving N at 0, 84, 168, and 252 kg ha(-1). Five treatments were used for model evaluation. Percent errors of mean yield predictions during 1958-1983 decreased as N rate increased (LTN1 = -5.0%, LTN2 = 3.5%, and LTN3 = 1.0%). Percent errors of mean yield predictions during 1985-1991 were larger than during the first period. Simulated and observed mean yields during 1958-1991 were highly correlated (R2 = 0.961, p < 0.01). Simulated SOC agreed well with observed values with percent errors from -5.8 to 0.5% in 1984 and from -5.1 to 0.7% in 1990. EPIC captured the dynamics of SOC, SCS, and microbial biomass. Simulated net N mineralization rates were lower than those from laboratory incubations. Improvements in EPIC's ability to predict annual variability of crop yields may lead to improved estimates of SCS.

Factors affecting slow regular firing in the suprachiasmatic nucleus in vitro.

In isolated slices of hypothalamus, suprachiasmatic nucleus (SCN) neurons were recorded intracellularly. Blockade of Ca++ channels increased spike duration, eliminating an early component of the afterhyperpolarization (AHP) that followed evoked spikes. The duration and reversal potential of AHPs were, however, unaffected, suggesting that only an early, fast component of the AHP was Ca(++)-dependent. Unlike other central neurons that exhibit pacemaker activity, therefore, SCN neurons do not display a pronounced, long-lasting Ca(++)-dependent AHP. Extracellular Ba++ and intracellular Cs+ both revealed slow depolarizing potentials evoked either by depolarizing current injection, or by repolarization following large hyperpolarizations. They had different effects on the shape of spikes and the AHPs that followed them, however. Cs+, which blocks almost all K+ channels, dramatically reduced resting potential, greatly increased spike duration (to tens of milliseconds), and blocked AHPs completely. In contrast, Ba++ had little effect on resting potential and produced only a small increase in spike duration, depressing an early Ca(++)-dependent component and a later Ca(++)-independent component of the AHP. The relatively weak pacemaker activity of SCN neurons appears to involve voltage-dependent activation of at least one slowly inactivating inward current, which brings the cells to firing threshold and maintains tonic firing; both Ca(++)-dependent and Ca(++)-independent K+ channels, which repolarize cells after spikes and maintain interspike intervals; and Ca++ channels, which contribute to activation of Ca(++)-activated K+ currents and may also contribute to slow depolarizing potentials. In the absence of powerful synaptic inputs, SCN neurons express a pacemaker activity that is sufficient to maintain an impressively regular firing pattern. Slow, repetitive activation of optic input, however, increases local circuit activity to such an extent that the normal pacemaker potentials are overridden and firing patterns are altered. Since SCN neurons are very small and have large input resistances, they are particularly susceptible to synaptic input.

Iron-regulatory proteins, iron-responsive elements and ferritin mRNA translation.

Iron plays a central role in the metabolism of all cells. This is evident by its major contribution to many diverse functions, such as DNA replication, bacterial pathogenicity, photosynthesis, oxidative stress control and cell proliferation. In mammalian systems, control of intracellular iron homeostasis is largely due to posttranscriptional regulation of binding by iron-regulatory RNA-binding proteins (IRPs) to iron-responsive elements (IREs) within ferritin and transferrin receptor (TfR) mRNAs. the TfR transports iron into cells and the iron is subsequently stored within ferritin. IRP binding is under tight control so that it responds to changes in intracellular iron requirements in a coordinate manner by differentially regulating ferritin mRNA translational efficiency and TfR mRNA stability. Several different stimuli, as well as intracellular iron levels and oxidative stress, are capable of regulating these RNA-protein interactions. In this mini-review, we shall concentrate on the mechanisms underlying modulation of the interaction of IRPs and the ferritin IRE and its role in regulating ferritin gene expression.

Hormonal regulation of mRNA stability and RNA-protein interactions in the pituitary.

Regulating gene expression from DNA to protein is a complex multistage process with multiple control mechanisms. Transcriptional regulation has been considered the major control point of protein production in eukaryotic cells; however, there is growing evidence of pivotal posttranscriptional regulation for many genes. This has prompted extensive investigations to elucidate the mechanisms controlling RNA processing, mRNA nuclear export and localization, mRNA stability and turnover, in addition to translational rates and posttranslational events. The regulation of mRNA stability has emerged as a critical control step in determining the cellular mRNA level, with individual mRNAs displaying a wide range of stability that has been linked to discrete sequence elements and specific RNA-protein interactions. This review will focus on current knowledge of the determinants of mRNA stability and RNA-protein interactions in the pituitary. This field is rapidly expanding with the identification of regulated cis-acting stability-modifying elements within many mRNAs, and the cloning and characterization of trans-acting proteins that specifically bind to their cognate cis elements. We will present evidence for regulation of multiple pituitary genes at the level of mRNA stability and some examples of the emerging data characterizing RNA-protein interactions.

Inhibitory postsynaptic potentials evoked in thalamic neurons by stimulation of the reticularis nucleus evoke slow spikes in isolated rat brain slices--I.

In isolated slices of rat thalamus, inhibitory postsynaptic potentials evoked by electrical stimulation of the nucleus reticularis, were recorded intracellularly in relay neurons in the anterior part of the thalamus. These inhibitory postsynaptic potentials were found to have reversal potentials close to the resting potential of the recorded cell, to reduce neuronal excitability and to be sensitive to electrophoretic application of the GABA antagonists bicuculline and picrotoxin, indicating that they were GABA-activated, chloride mediated events. Voltage sensitive responses of relay neurons evoked by current injection and by inhibitory postsynaptic potentials were then compared. Hyperpolarizing current pulses and hyperpolarizing inhibitory postsynaptic potential trains elicited from membrane potentials positive to -70 mV resulted in rebound slow spike activation on repolarization. Depolarizing current pulses and depolarizing inhibitory postsynaptic potential trains evoked slow spikes when elicited from membrane potentials negative to -60 mV. There was, however, one major difference, the slow spikes evoked by inhibitory postsynaptic potentials were always delayed to the end of the train. Reversal potentials of evoked inhibitory postsynaptic potentials were found to depend on the potential at which the membrane was held immediately before the inhibitory postsynaptic potential was evoked, indicating that passive distribution of chloride ions contributes to their equilibrium potential. Evoked inhibitory postsynaptic potentials consisted of at least two components with different reversal potentials although current voltage relations indicated that similar decreases in membrane resistance were associated with both components and that they shifted approximately in parallel when inhibitory postsynaptic potentials were evoked from different holding potentials. Trains of GABA-mediated inhibitory postsynaptic potentials, similar to those recorded during spindling, will evoke slow spikes in almost all thalamic relay neurons irrespective of other synaptic inputs. This response will effectively synchronize burst firing in all cells receiving the same inhibitory input.

Biphasic responses of thalamic neurons to GABA in isolated rat brain slices--II.

In isolated thalamic slices, responses of relay neurons to electrophoretically applied GABA were recorded intracellularly and compared with inhibitory postsynaptic potentials evoked by electrical stimulation of the reticularis nucleus of the thalamus. Both reduced the excitability of thalamic neurons and were biphasic in the majority of neurons studied, consisting of an early, negative-going and a later, positive-going component, when recorded close to reversal potential (mean reversal potentials -66.6 and -57.7 mV). Bicuculline and picrotoxin applied electrophoretically reduced conductance increases evoked by GABA in all neurons. The later, positive-going component was more sensitive to these antagonists (applied with submaximal doses) than the early component. Current-voltage relations for responses to GABA, like those for inhibitory postsynaptic potentials, were non-linear in the majority of neurons. In particular, there was a region of reduced slope resistance close to the reversal potential. Holding the membrane at a conditioning potential was found to change the subsequent response and its reversal potential. Positive holding potentials shifted reversal potentials in the positive direction only when GABA was applied during the conditioning period. Negative holding potentials were effective whether GABA was applied during the conditioning period or not. Recovery from these effects followed a similar time course at all membrane potentials tested. Injection of Cl- produced a positive shift in the reversal potential for both components of the response to GABA and of the evoked inhibitory postsynaptic potential. Inhibitory postsynaptic potentials evoked in thalamic relay neurons by stimulation of the nucleus reticularis resembled responses to GABA in their biphasic nature, reversal potentials and sensitivity to antagonists and to changes in intracellular chloride.

Augmentation by glycine and blockade by 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) of responses to excitatory amino acids in slices of rat neocortex.

Responses of neocortical pyramidal cells to excitatory amino acids were recorded intracellularly. Agonists and antagonists were applied electrophoretically from a separate multibarrel pipette and care taken to ensure that the pipette was positioned to evoke optimal responses to N-methyl-D-aspartate (NMDA), or homocysteic acid, before control responses were recorded. Responses to NMDA, but not those to alpha-amino-3-hydroxy-5-methyl-4-isoxazdepropionic acid (AMPA) or quisqualate, were enhanced when glycine was co-applied. Responses to AMPA, quisqualate and NMDA were reduced by 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) applied either electrophoretically, or in the bathing medium, with responses to quisqualate being the least and those to AMPA being the most sensitive to CNQX. The blockade of NMDA responses by CNQX was selectively reversed by additional glycine confirming that CNQX blocks NMDA receptor-channel complexes at the glycine, rather than at the NMDA site. Under control conditions, responses to glutamate resembled responses to quisqualate, and were relatively insensitive to CNQX, 3-((+/-)-2-carboxypiperazin-4-yl)-propyl-l-phosphonic acid and 2-amino-5-phosphonovalerate, while responses to homocysteic acid resembled responses to NMDA and were blocked by these antagonists. This suggested that homocysteic acid acted at NMDA receptors, while glutamate acted primarily at non-NMDA receptors. However, responses to both glutamate and homocysteic acid were augmented by additional glycine when these transmitter candidates were applied close to a "hot spot" for NMDA receptor activation. The glycine enhancement of responses to glutamate was sensitive to NMDA antagonists, indicating that glutamate can activate NMDA receptors in an intact preparation if glycine levels are high enough.

Supraoptic neurons sustain high frequency firing when extracellular Ca2+ is replaced with other divalent cations in rat brain slices.

In slices of hypothalamus, maintained in vitro, the discharge of supraoptic neurons in standard artificial cerebrospinal fluid was compared with that produced when extracellular Ca2+ was replaced with Mg2+, Co2+ or Mn2+. Interspike interval histograms were constructed for periods before, during and after replacement of extracellular Ca2+. Of the 31 cells recorded in normal medium, 16 fired slowly and irregularly, 9 were phasic, 3 fired continuously at more than 3 spikes/s and 3 produced short, high frequency bursts of activity that were separated by slow irregular discharge. Interspike interval distributions were broad showing little preference for any one interval and intervals shorter than 30-50 ms were rare. The cell firing rate could be increased by the electrophoretic application of glutamate and under these conditions, the interval distributions became narrower as shorter intervals predominated. However, when cells discharged above 20 spikes/s, the spike amplitude declined rapidly and became indistinguishable from the noise. Replacement of extracellular Ca2+ with Mg2+, Co2+ or Mn2+ produced reversible changes in interspike interval distribution, although no consistent change in mean firing frequency was observed. Supraoptic neurons were now able to maintain relatively high frequency discharge (15-25 spikes/s) for longer periods; firing either continuously or periodically, and interspike intervals became grouped more closely at the shorter end of the normal distribution. However, no very short interspike were recorded. Less than 2% of all recorded intervals were shorter than 30 ms, even in cells exposed to test medium for 1-3 h and excited by application of glutamate.(ABSTRACT TRUNCATED AT 250 WORDS)

Slow, regular discharge in suprachiasmatic neurones is calcium dependent, in slices of rat brain.

The Ca2+ dependence of suprachiasmatic firing patterns was studied in isolated hypothalamic slices. Single neurones were first recorded in a standard artificial cerebrospinal fluid and then followed through a change to a medium in which the Ca2+ was replaced with Mg2+, Co2+ or Mn2+. These test media caused disruption of the normal, slow, regular firing patterns of suprachiasmatic neurones. Some interspike intervals were shorter and some longer than any recorded under normal conditions, for a given firing frequency. In the absence of Ca2+, these cells could not be driven to fire more regularly as they fired more rapidly during glutamate applications. In contrast, in the presence of Ca2+ more slowly and irregularly firing suprachiasmatic cells can be driven to fire regularly if their firing rate is increased. The effects of these test media were reversible. When Ca2+ was replaced with Ba2+, a bursting pattern of discharge resulted. Periods of high frequency discharge, possibly superimposed on Ba2+ spikes, alternated with periods of slow, regular firing or silence. It is concluded that Ca2+ is necessary for the maintenance of regular firing in suprachiasmatic neurones. The possibility that Ca2+ channels similar to those present in other slow, regularly firing central neurones, play an important role in suprachiasmatic activity, is discussed.

Comparison of responses to transmitter candidates at an N-methylaspartate receptor mediated synapse, in slices of rat cerebral cortex.

Intracellular recordings from pyramidal neurones in isolated slices of rat cerebral cortex allowed a comparison of postsynaptic potentials and responses to electrophoretic application of excitatory amino acids. The responses of all neurones to N-methylaspartate displayed an unusual voltage relation and were associated with an apparent increase in membrane resistance, properties that were dependent on the presence of extracellular Mg2+. Responses to N-methylaspartate could be elicited only when the electrophoretic pipette was positioned close to the cell soma and were associated with generation of slow spikes which triggered bursts of fast spikes. In contrast, in the majority of neurones, responses to glutamate, aspartate, cysteate and cysteine sulphinate demonstrated a conventional voltage relation, were associated with a decrease in membrane resistance, evoked no slow spikes, were insensitive to extracellular Mg2+ concentrations between 1 and near 0 mM and could be evoked with small currents of amino acids when the electrophoretic pipette was greater than 100 micron from the cell soma. Responses to the five amino acids were tested with the N-methylaspartate antagonist 2-amino-5-phosphonovaleric acid. In the majority of cells, this antagonist blocked responses to N-methylaspartate at doses that had only a small effect on responses to the other amino acids. In these experiments it was also possible to confirm previous reports that ketamine and cyclazocine act as selective N-methylaspartate antagonists. In 4/63 neurones, responses to glutamate and aspartate and in 1/5 neurones one component of the response to cysteate, displayed properties similar to those of responses to N-methylaspartate. In one other neurone, large applications of cysteine sulphinate or glutamate could evoke slow spikes and fast spike bursts, a firing pattern that was sensitive to 2-amino-5-phosphonovalerate. The present experiments therefore demonstrate that all four naturally occurring amino acids tested can activate N-methylaspartate receptors and indicate that any one of these could be the transmitter at the N-methylaspartate receptor-mediated synapse on cortical pyramidal neurones. However, in the majority of neurones, these putative transmitters activated other receptor types preferentially. The possibility that this may result from the greater accessibility of non-N-methylaspartate receptors to electrophoretically applied agonists, is discussed.

Postsynaptic pyramidal target selection by descending layer III pyramidal axons: dual intracellular recordings and biocytin filling in slices of rat neocortex.

Paired intracellular recordings in slices of adult rat neocortex with biocytin filling of synaptically connected neurons were used to investigate the pyramidal targets, in layer V, of layer III pyramidal axons. The time-course and sensitivity of excitatory postsynaptic potentials to current injected at the soma, and locations of close appositions between presynaptic axons and postsynaptic dendrites, indicated that the majority of contributory synapses were located in layer V. Within a "column" of tissue, radius < or = 250 microm, the probability that a randomly selected layer III pyramid innervated a layer V pyramid was 1 in 4 if the target cell was a burst firing pyramid with an apical dendritic tuft in layers II/I. If, however, the potential target was a regular spiking pyramid, the probability of connectivity was only 1 in 40, and none of the 13 anatomically identified postsynaptic layer V targets had a slender apical dendrite terminating in layers IV/III. Morphological reconstructions indicated that layer III pyramids select target layer V cells whose apical dendrites pass within 50-100 microm of the soma of the presynaptic pyramid in layer III and which have overlapping apical dendritic tufts in the superficial layers. The probability that a layer V cell would innervate a layer III pyramid lying within 250 microm of its apical dendrite was much lower (one in 58). Both presynaptic layer III pyramids and their large postsynaptic layer V targets could therefore access similar inputs in layers I/II, while small layer V pyramids could not. One prediction from the present data would be that neither descending layer V inputs to the striatum or thalamus, nor transcallosal connections would be readily activated by longer distance cortico-cortical "feedback" connections that terminated in layers I/II. These could, however, activate corticofugal pathways to the superior colliculus or pons, both directly and via layer III.

N-methylaspartate receptors mediate epileptiform activity evoked in some, but not all, conditions in rat neocortical slices.

Using intracellular recordings from pyramidal neurons in isolated slices of rat cerebral cortex epileptiform discharges evoked (1) in the presence of gamma-aminobutyric acid antagonists, and (2) in the absence of Mg2+ were compared. Depolarization shift responses recorded in the presence of bath applied picrotoxin, or electrophoretically applied picrotoxin or bicuculline, were similar in many respects to depolarization shifts reported previously, except that they could be evoked by stimuli subthreshold for evoking discernible postsynaptic potentials in these experiments. Large depolarizations evoked by repetitive activation of an N-methylaspartate receptor mediated synapse in the absence of Mg2+, displayed several properties similar to those of depolarization shifts evoked in the presence of gamma-aminobutyric acid antagonists, i.e. similar shape, latency, inability to follow high repetition rates and a similar voltage relation, suggesting activation of the same cellular mechanism. "Slow spikes" evoked as part of the response to electrophoretically applied N-methylaspartate were augmented, i.e. they were replaced by larger, longer, more complex events, when gamma-aminobutyric acid antagonists were applied. The potentiated response, evoked in the absence of Mg2+, was dependent on the activation of an N-methylaspartate receptor mediated synapse and was blocked by N-methylaspartate antagonists. In contrast, depolarization shifts could be evoked in the presence of large doses of N-methylaspartate antagonists, when gamma-aminobutyric acid antagonists were applied. Spontaneous depolarizations similar to depolarization shifts were recorded when cells were exposed to low, tonic, electrophoretic applications of excitatory amino acids under control conditions. In addition, some potentiation of the N-methylaspartate receptor mediated excitatory postsynaptic potential was achieved in the presence of Mg2+ when cells were depolarized by 10-20 mV. Depolarization shifts evoked when bicuculline was applied electrophoretically to different parts of the dendritic field, some hundreds of microns from the soma, differed in shape, latency and time course and the depolarization shift evoked when bicuculline was applied at one site summed with the depolarization shift evoked when it was applied elsewhere. We conclude that different inputs are required to activate the responses evoked in the presence of gamma-aminobutyric acid antagonists and in the absence of Mg2+. The possibility that both involve activation of dendritic Ca2+ currents and that the magnitude of the response depends on the proportion of the dendritic field activated, is discussed.

The electrical properties of neurones of the rat suprachiasmatic nucleus recorded intracellularly in vitro.

Stable intracellular recordings were obtained from 22 suprachiasmatic neurones in isolated brain slices. These cells were characterized by resting potentials of about -60 mV, high input resistances, relatively short time constants and action potentials of short duration. The action potentials were preceded by a slow depolarization and followed by a relatively brief afterhyperpolarization and long-lasting increase in membrane conductance. Current-voltage relations were usually linear between 0 and 80 mV negative to the resting potential. Postsynaptic potentials were evoked in these cells by electrical stimulation of the optic chiasm or contralateral suprachiasmatic nucleus. Both excitatory postsynaptic potentials, which evoked action potentials, and inhibitory postsynaptic potentials were recorded. Synaptic potentials were associated with an increase in membrane conductance. Action potentials evoked by synaptic activation were sometimes followed by up to three small, fast potentials. Small fast potentials were not seen to occur spontaneously, or to follow spontaneous, or current-evoked spikes, nor were they evoked by synaptic potentials that failed to evoke action potentials. The suprachiasmatic nucleus is essential for the generation of normal biological rhythms in mammals. The input it receives from the optic nerve is thought to be important in this role. It is hoped that these preliminary intracellular studies will form a basis for further work on the inherent properties of suprachiasmatic neurones and their responses to visual input.