Phase differences in electrical discharge rhythms between neuronal populations of the left and right suprachiasmatic nuclei.

The circadian pacemaker of the suprachiasmatic nuclei is a complex multioscillator system, which controls circadian and seasonal rhythmicity. A number of clock genes have been identified that play a key role in the generation of circadian rhythms. These clock genes are expressed in a circadian manner as has been shown in mice, rats and hamsters. The time at which their expression reaches peak values differs among the several genes. Expression profiles for a specific gene may also differ among subdivisions of the suprachiasmatic nuclei. It has been shown that mPer1 peaks slightly out of phase in the left and right suprachiasmatic nuclei and that the rhythm in c-fos expression is significantly different between the dorsomedial and ventrolateral regions. In the special case that the animal shows splitting of its locomotor activity pattern, mPer1 in the left and right suprachiasmatic nuclei appeared to oscillate in antiphase. Whether the molecular organization within the suprachiasmatic nuclei plays a role in seasonal rhythmicity, allowing animals to track daylength and become reproductive at the proper phase of the annual cycle, receives increasing interest (). The differences in peak expression times that exist between different genes, and the spatial differences in peak time for single genes, are suggestive of a genetic mechanism underlying the multioscillator structure. It is unknown, however, whether phase differences that are observed at the molecular level exist at the level of electrical activity rhythms in the suprachiasmatic nuclei in order to become potentially functional. In this study we investigated the presence of phase differences in neuronal discharge rhythms in the suprachiasmatic nuclei of the rat. To this purpose we combined simultaneous electrophysiological recordings of neuronal populations in the left and right suprachiasmatic nuclei with a detailed analysis of the phase relationship between them. The results demonstrate that neuronal subpopulations of the suprachiasmatic nuclei show phase differences both in their peak and half-maximum times of up to 4 h. We propose that these phase differences may play a role in the plasticity of the circadian timing system.

Membrane-controlled reagent-delivery systems--a new approach for the continuous production of reagent and standard solutions.

A new simple and robust system for the production of standard solutions, based on the mass-transfer of analytes through membranes, is described. The device consists of a cone-shaped reservoir vessel, filled with a concentrated solution of the analyte and separated from a liquid acceptor stream by a membrane. Mass-flow from donor to acceptor solution is controlled by the mass-transfer-affecting properties of the active membrane area, which is determined by the hole in a template (diameter 0.8 mm) placed between the membrane and the acceptor-channel. Using nitrate as model analyte and a track-etched membrane filter (pore size 0.1 microm) dilution factors up to 2,400,000 with long-term reproducible accuracy of < 2% have been achieved. Adjustment of a requested concentration is possible by varying either the flow rate of the acceptor stream or the concentration of the reservoir solution.

Fentanyl, a upsilon-opioid receptor agonist, phase shifts the hamster circadian pacemaker.

The phase-shifting effects of the mu-opioid receptor agonist fentanyl on the circadian timing system were investigated in the hamster. Fentanyl injections during the mid-subjective day induced phase advances of the hamsters' wheel-running activity rhythm. The shifts were not accompanied by an increase in locomotor activity but instead a decrease of activity was often observed. A dose-response curve indicated that with increasing dosage, the response probability increased, while the magnitude of the induced shift remained stable. The present data suggest that there is some role for opioid regulation of the circadian system.

Functional absence of extraocular photoreception in hamster circadian rhythm entrainment.

The mammalian circadian pacemaker is entrainable by light via the retina. The putative role of extraocular light perception was investigated in blinded hamsters. These animals were shaved and exposed to a light-emitting pad for either 30 min or 3 h. The absence of any phase-shifting effects on wheel running activity rhythms indicates that extraocular light perception plays no functional role in photic entrainment of the circadian pacemaker in the hamster.

Light responsiveness of the suprachiasmatic nucleus: long-term multiunit and single-unit recordings in freely moving rats.

The suprachiasmatic nuclei (SCN) of the hypothalamus contain a pacemaker that generates circadian rhythms in many functions. Light is the most important stimulus that synchronizes the circadian pacemaker to the environmental cycle. In this paper we have characterized the baseline neuronal firing patterns of the SCN as well as their response to light in freely moving rats. Multiunit and single-unit recordings showed that SCN neurons increase discharge during daytime and decrease discharge at night. Discharge levels of individual neurons that were followed throughout the circadian cycle appeared in phase with the population and were characterized by low discharge rates (often below 1 Hz), with a twofold increase during the day. The effect of light on the multiunit response was dependent on the duration of light exposure and on light intensity, with light thresholds of approximately 0.1 lux. The light response level showed a strong dependency on time of day, with large responsiveness at night and low responsiveness during day. At both phases of the circadian cycle, the response level could be raised by an increase in light intensity. Single-unit measurements revealed that the time-dependent light response of SCN neurons was present also at the level of single units. The results show that the basic light response characteristics that were observed at the multiunit level result from an integrated response of similarly behaving single units. Research at the single-unit level is therefore a useful approach for investigating the basic principles of photic entrainment.

Electrophysiologic analysis of the actions of valproate on pyramidal neurons in the rat hippocampal slice.

PURPOSE: Studies in invertebrates and cultured mammalian neurons suggested that valproate (VPA) mediates its main antiepileptic effect by slowing the recovery from inactivation of voltage-dependent sodium channels. This predicts an effect on the refractory period of the action potential and, consequently, on the bursting behavior of neurons. METHODS: We investigated this prediction using intracellular and extracellular recording techniques in hippocampal slices prepared from adult rats. The refractory period (RFP) and the ratio of the slopes (SR) of a pair of action potentials were used as indices of the recovery from inactivation of sodium channels. They were measured by injecting a series of paired depolarizing current pulses into CA1 pyramidal neurons. RESULTS: No significant changes were observed in the RFP or SR measured during a 1-h recording period when VPA was bath-applied (1 mM), or when it was present in the recording electrode (10-50 mM). Lowering the temperature from 34.5 degrees C to 26.4 degrees C resulted in an increase of the RFP by 100% and a decrease of the SR by 40%. However, VPA did not affect any of the measured action potential parameters at this lower temperature. VPA was also without effect on the presynaptic fiber volley of axons recorded extracellularly in the stratum radiatum. The antidromic population spike was unaffected by VPA (2 mM), whereas phenytoin (50 microM) clearly affected this spike in the same slices. The absence of effect of VPA on each of the measured parameters could not be attributed to poor penetration through the slice because bath-applied VPA reduced the frequency of extracellularly recorded spontaneous interictal bursts, induced by bicuculline and elevated K+, within 10 min. CONCLUSIONS: These findings suggest that at least in the hippocampal slice the drug's principal antiepileptic effect cannot be explained by its action on voltage-dependent sodium channels.

Multiunit activity recordings in the suprachiasmatic nuclei: in vivo versus in vitro models.

The suprachiasmatic nuclei (SCN) of the hypothalamus continue to oscillate when they are isolated in a brain slice preparation. We recorded multiunit activity in the SCN of the rat both in vivo and in vitro to determine the circadian discharge pattern. The variability of the discharge pattern is larger and the amplitude of the rhythm is smaller in vivo than in vitro. Moreover we found evidence for a direct effect of the animal's behavioural activity on electrical activity of the SCN in vivo. These findings may provide an electrophysiological basis for the known effects of behavioural stimuli on the circadian pacemaker. This study underscores the importance of recordings in intact preparations in addition to in vitro work when generalisations to physiological conditions are to be made.

The effects of glutamate on membrane potential and discharge rate of suprachiasmatic neurons.

The suprachiasmatic nucleus (SCN) is a major pacemaker for circadian rhythms in mammals. Photic entrainment of the circadian pacemaker is mediated by the retinohypothalamic tract (RHT). Most likely, excitatory amino acids function as neurotransmitters in this pathway. We have now investigated the effect of glutamate on the membrane potential of cultured SCN cells of the rat with the aid of the patch clamp technique. It was found that 1 mM glutamate depolarizes the cells by about +44 mV. In spontaneously active neurons, the glutamate induced depolarization caused either an increase in discharge or a depolarization block. We then investigated the effect of 1 mM glutamate on SCN discharge in the acutely prepared hypothalamic slice of the hamster. In most cells glutamate induced an increase in discharge whilst in a few cells discharge was suppressed. Both series of experiments indicate that glutamate in the used dosage was effective and its effect reversible. The data are discussed with respect to the failure of 1 mM glutamate injections to mimic the effect of light on the circadian activity rhythm of the hamster.

[Parvovirus B 19 infection and associated arthritis in children and adolescents]. / Parvovirus B 19-Infektion und assoziierte Arthritis bei Kindern und Jugendlichen.

The commonest clinical manifestation of an acute parvovirus B 19 infection is "infectious erythema (E.i.)". Both in connection with "E.i." and without typical skin symptoms, transient arthralgias can be observed relatively frequently. Moreover, patients suffering from juvenile chronic arthritis (jcA) have now been described, in whom the initial symptoms were closely related to an acute parvo B 19 infection. Fife of our own patients with arthropathies of different kinds and a well documented acute parvovirus B 19 infection are presented here and all the relevant contemporary scientific problems are discussed.

Enhancement of recurrent inhibition by angular bundle kindling is retained in hippocampal slices.

Rats were kindled in the right angular bundle. EEG and monosynaptically evoked responses were monitored in the ipsilateral fascia dentata. Although every animal was kindled, Long Term Potentiation (LTP) of monosynaptic responses was observed only in part of the kindling sessions, suggesting that LTP is not required for kindling. Paired pulse inhibition of granule cell discharge was progressively enhanced by kindling. Transverse hippocampal slices (400-700 micron) of fully kindled rats were prepared 1 hour after the last seizure. Field potentials evoked by stimulation appeared completely normal. Spontaneous epileptiform discharges were not observed in control solution, in low [Ca++]0, in high [K+]0 or after high frequency stimulation. Paired pulse inhibition was enhanced in fascia dentata, but not in area CA1. Enhancement of inhibition may be caused by increased activity of inhibitory synapses or by a nonsynaptic hyperpolarizing current. The relation between the absence of spontaneous activity and enhanced inhibition in the fascia dentata is unclear.

[Ventral intercorporal spondylodesis of the lumbar spine in segmental instability]. / Die ventrale interkorporelle Spondylodese der Lendenwirbelsäule bei Segmentinstabilität.

Ventral intercorporal spondylodesis or anterior interbody fusion was applied via the extraperitoneal route to the lumbar spine of 25 adult patients. The approach had in all cases been indicated for segmental instability in spondylolisthesis or degeneration of vertebral discs with low-back pain in the wake of surgery. Described are the surgical technique used as well as intra-operative and postoperative complications. Solid union was recorded from 21 patients, while non-union occurred to four. Follow-up checks within six months to four and a half years from surgery (2.2 years on average) revealed good clinical success in 19 patients. Four of them were absolutely free of low-back pain, while the condition of 15 had changed to the better. Pain symptoms were not affected in six patients.

Spontaneous epileptiform discharges in hippocampal slices induced by 4-aminopyridine.

4-Aminopyridine (4-AP) induced 2 types of spontaneous field potentials (SFPs) in the hippocampal slice. Type I resembled spontaneous activity induced by other convulsants. They occurred at a rate of approximately 1 Hz, started in the CA2/CA3 region and spread at a velocity of 0.3 m/s to area CA1. Transsection experiments and laminar profiles indicated that they spread synaptically along the Schaffer collateral pathway. Synaptic blockade by low Ca2+/high Mg2+ or kynurenic acid reversibly abolished type I SFPs. Increasing [Ca2+]o lowered the rate and slightly increased the amplitude. Possibly, increased spontaneous transmitter release, and not disinhibition, is responsible for the generation of type I SFPs. Type II occurred at a rate of about 0.15 Hz and travelled in the same direction, but a factor 10 slower. They could not be blocked by separation of the CA1 and CA3 region; coupling remained until stratum moleculare was severed. Type II could not be suppressed by blockade of synaptic transmission. The laminar profile is similar in shape to that of type I but not identical. Increasing [Ca2+]o had the same but stronger effect as on type I. Type II SFPs depressed evoked population spikes up to a second and delayed the next type I SFP. The mechanisms involved remain largely speculative; further analysis is needed to help understand the epileptogenic action of 4-AP.

Sodium-dependent sugar and amino acid transport in isolated goldfish intestinal epithelium: electrophysiological evidence against direct interactions at the carrier level.

The effects of mucosal application of monosaccharides and amino acids on transepithelial and membrane potentials in isolated goldfish intestinal epithelium were investigated. Isosmotic replacement of mucosal mannitol by sugars or L-amino acids resulted in a rapid depolarization of the mucosal membrane potential psi mc followed by a slow repolarization. Phlorizin inhibited the responses to sugar but not those to amino acids. D-Amino acids did not induce any electrical response in the epithelium. Dose-response curves for L-amino acids showed simple saturation. Simultaneous application of L-amino acid and glucose induced transepithelial responses of about 80% of the sum of the separate responses to the application of amino acid or glucose alone. Simultaneous application of different amino acids in saturating concentrations did not increase the magnitude of the electrical responses. From the measured changes in potentials we calculated the change in electromotive force across the mucosal (delta Em) and serosal (delta Es) membrane. The change in Em induced by combined application of alanine and glucose was 90% of the sum of the calculated values induced by glucose and alanine alone. The response of Es to both substrates was accelerated with respect to that of separate substrates alone. We conclude that by application of glucose in addition to alanine the influx of sodium is increased, thereby stimulating the basolaterally located electrogenic Na+/K+-pump. There are no indications for direct interaction of sugars and amino acids at the mucosal membrane of the intestinal epithelial cell.

Analysis of the ouabain-induced increase in transepithelial electrical resistance in the goldfish intestinal mucosa.

1. The ouabain-induced increase in transmural resistance of goldfish intestinal mucosa stripped free from underlying muscular layers is analysed by comparing the resistance increase in normal and in low chloride saline, the resistance increased induced by anaerobic conditions and the resistance increase provoked by hypotonicity. 2. It is concluded that the collapse of the lateral intercellular space is the prime reason for the resistance increase and that the lateral intercellular space is maintained dilated by a ouabain-sensitive solute transport mechanism. 3. This mechanism can be either a rheogenic or a neutral Na/K-pump. In the latter case additional conditions have to be specified concerning values for ion concentrations in the lateral intercellular space and in the unstirred layer adjacent to the luminal membrane. 4. There are no indications for a chloride dependent mechanism involved in the maintenance of the width of the lateral intercellular spaces in the goldfish intestinal mucosa.

Microscopical determination of the filtration permeability of the mucosal surface of the goldfish intestinal epithelium.

The rate of shrinkage of the mucosal folds of goldfish intestine in response to mucosal hypertonicity was measured by microscopic means. Because of the geometry of the intestinal folds the rate of shrinkage could be directly related to the loss of volume from the fold through the brush border membranes and tight junctions. Experimentally a wide range of velocities was observed, reflecting the difficulty of rapidly establishing a uniform osmotic gradient at the preparation's mucosal surface. The initial velocity of volume loss provided a measure of the filtration permeability (Pf) of the mucosal surface. From the highest velocities observed the filtration permeability was estimated to be approximately 14 X 10(-3) cm/sec related to the folded mucosal surface and 65 X 10(-3) cm/sec related to the straight serosal surface. Consideration of the experimental errors and unstirred layer effects make it probable that the latter value is still an underestimate of the true Pf. The series barriers of the epithelium cause the total tissue Pf to be less than the Pf of the mucosal surface alone. In addition the Pf measured in the presence of an osmotic gradient may differ substantially from the tissue filtration permeability which exists in the absence of a change in osmolarity.