Search

The suprachiasmatic nucleus is regarded as the main mammalian circadian pacemaker but evidence for rhythmic firing of single units in vivo has been obtained only recently. The present study was undertaken to determine if rhythms could be seen using measures of activity in addition to the mean spike frequency. We investigated whether there were changes in the irregularity of cell activity measured by the disorder of the interspike interval distribution for neurones recorded in vivo and in vitro. By plotting the entropy of the log interval histogram that quantifies the coding capacity for each action potential against the respective zeitgeber time, we describe oscillations of spike activity in vivo. Entropy measures have the advantage over variances in that they quantify aspects of the shape of the distribution and not just the dispersion. One hundred and sixty-six cell recordings from the suprachiasmatic nucleus showed a significant rhythm in entropy with an oscillatory trend in the data (P < 0.001) showing a trough towards the end of the light period and a peak in the mid-dark period. There was a similar rhythm for the cells recorded from the peripheral zone (n = 209, P = 0.037). In separate experiments in vitro, to investigate the relationship between mean spike frequency and entropy, potassium-induced depolarization of cells recorded during the subjective night was correlated with a significant increase in mean spike frequency (r = 0.259, P = 0.011) and a decrease in entropy (r = -0.296, P = 0.004). The negative correlation between the entropy and mean spike frequency of cells recorded in vitro was significantly different from that seen in vivo (F = 15.5, P < 0.001), which may reflect differences in the balance between deterministic and stochastic influences on spike occurrence. The study shows that while there is a rhythm of mean spike frequency, parameters based on the variability of interspike interval distributions also display rhythmic changes over the day-night cycle.

Conventional extracellular recordings were made from single cells in the suprachiasmatic nucleus (SCN) region of the anaesthetized rat. Each cell was tested for its response to stimulation at three sites; the contralateral optic nerve, the ipsilateral supraoptic nucleus (SON) or the ipsilateral arcuate nucleus (ARC) to determine whether the behaviour of the synapses in the SCN was different at different times. Responses to stimulation were tested once each hour and assessed by creating peristimulus time histograms. Excitatory, inhibitory or complex (consisting of more than one component) responses were seen. The responses of some cells that were recorded for several hours changed with time. Changes were seen in the responses of SCN cells to stimulation of the ARC (31/91 cells) and the SON (26/90 cells) regions, but only rarely to stimulation of the optic nerve (2/72 cells). Such differences in proportion are unlikely to have occurred by chance (P < 0.001; chi-square test). Changes seen included the appearance of both excitatory and inhibitory responses in cells that were initially unresponsive. In some cells, one component of a complex response remained constant while another component changed with time. When the cells in the SCN were treated as a group, the proportion of excitatory, inhibitory or complex responses to ARC stimulation did not remain constant throughout the light/dark cycle (P = 0.014; chi-square test). The proportion of excitatory, inhibitory or complex responses to SON and optic nerve stimulation showed no significant variation with the light/dark cycle. If a change in response can be interpreted as a change in the behaviour of a neural connection, the results imply that some of the projections to the SCN from within the hypothalamus change at different times of the light/dark cycle, whereas no change could be seen in the input from the optic nerve. Thus, some of the connections of the SCN appear not to be hard wired, but change rapidly with time.

The electrical activity of the rat suprachiasmatic nucleus (SCN) was examined in anesthetized rats in vivo using single-unit electrophysiological techniques. The present data confirm the daily variation in the electrical activity of the SCN previously reported in vitro and in vivo using multiple-unit recording techniques. They further suggest that subpopulations of suprachiasmatic neurons with different neural connections have a different daily rhythm of activity. Neurons in the SCN region showed a significant rhythm of activity (p = 0.034; Kruskall-Wallis analysis of variance [KW-ANOVA]). The greatest activity occurred during the second part of the light period (ZT 10-12), and the lowest activity occurred in the early part of the light period (ZT 0-2). The subgroup of cells in the suprachiasmatic region with output projections to the arcuate nucleus (ARC) and/or supraoptic nucleus (SON) regions also showed a significant rhythm (p = 0.001; K-W ANOVA). Their activity appeared to show two peaks near the light-dark (ZT 10-12) and dark-light (ZT 22-24) transition periods with the lowest activity at ZT 16-18. This rhythm was significantly different (p = 0.016) from that of neurons without an output projection to the ARC and/or SON. Retinorecipient suprachiasmatic neurons appeared to have a less robust daily rhythm in their activity. The change in the firing behavior of the cells was not reflected simply by changes in mean firing rate. Examination of the coefficient of variation of the interspike interval distribution of cells at different times of day revealed changes in the firing pattern of cells in the SCN region that did not have output projections (p = 0.032; K-W ANOVA). The present results thus suggest that the SCN is composed of a heterogeneous population of neurons and that different rhythms of activity are expressed by neurons with different neural connections. There were changes in both firing pattern and firing rate.

The secretion of many hormones, including oxytocin, vasopressin and growth hormone, is not constant but shows a day-night rhythm. The suprachiasmatic nucleus (SCN) is thought to generate most mammalian biological rhythms and previous studies have reported suprachiasmatic efferents to the paraventricular nucleus (PVN) and the supraoptic nucleus (SON). We used in vivo extracellular electrophysiological techniques to show that the SCN also sends direct and indirect neural projections to the arcuate nucleus (ARC). This projection consisted of both excitatory and inhibitory components and may contribute to the entrainment of the rhythm in growth hormone secretion to the day-night cycle. Some SCN neurones appear to project to both the SON and the ARC. The SCN in turn receives excitatory and inhibitory inputs from the ARC and the peri-nuclear zone of the SON (peri-SON), which may provide feedback information, as well as allowing nonphotic entrainment of the SCN, for example, in response to feeding. Our data thus suggest extensive two-way connections between the SCN and its target nuclei which may contribute to the generation of day-night neuroendocrine rhythms. They also suggest the existence of indirect retinal projections to the ARC and PVN. We further investigated the retinal projection to the SCN. We were unable to demonstrate a significant difference in retinal input to those suprachiasmatic cells which had efferent projections to particular hypothalamic targets (SON and/or ARC), and those which did not.

1. In the rat, projections from the suprachiasmatic nucleus (SCN) to the supraoptic nucleus (SON) of the hypothalamus were characterized in vivo using extracellular recordings and in slice preparations using both extracellular and whole-cell patch clamp recording. 2. Of 117 magnocellular neurones recorded in the SON in vivo, fifteen (13%) displayed a short latency excitation, sixty-eight (58%) a short latency inhibition, six (5%) were unresponsive and twenty-eight (24%) gave long latency responses following SCN stimulation. 3. The responses of putative vasopressin cells in the SON to SCN stimulation in vivo (4 out of 61 cells, 7% excited; 49 out of 61 cells, 80% inhibited) were significantly different from those of putative oxytocin cells (10 out of 50 cells, 20% excited and 16 out of 50 cells, 32% inhibited; P < 0.02, test for differences between proportions). 4. Recordings in vitro using patch technology in whole-cell mode showed both inward and outward currents in SON cells at holding potentials near resting membrane potential following stimulation of the SCN region. The outward currents could be blocked by bicuculline (10 microM; n = 7) and the inward currents were blocked by the non-NMDA antagonist 6-nitro-7-sulphamoylbenzo(f)quinoxaline-2,3-dione (5 microM; n = 4). 5. We conclude that there is a strong projection from the SCN to the SON with both inhibitory (GABAergic) and excitatory (glutamatergic) components which may regulate the daily changes in neurohypophysial hormone secretion.

To determine whether the daily rhythms of spike activity in the supraoptic nucleus (SON) were accompanied by changes in the behaviour of its inputs, we used conventional extracellular single cell recordings from cells in the SON of anaesthetized rats while stimulating the contralateral optic nerve and the ipsilateral suprachiasmatic nucleus (SCN). Neurones in the SON region were identified by antidromic activation and classified as oxytocin or vasopressin cells, on the basis of their spontaneous firing patterns. Approximately 27% of both oxytocin (29/108) and vasopressin (39/147) neurones were excited by stimulation of the optic nerve, and the majority of responses had a long latency (>20 ms). Very few oxytocin (3/108) and vasopressin cells (2/147) were inhibited by stimulation of the optic nerve. The pattern of the responses (excitatory, inhibitory or nonresponsive) of oxytocin and vasopressin cells to stimulation of the optic nerve was significantly related to the time of day (chi-square test; P = 0.012, oxytocin cells; P = 0.006, vasopressin cells). The proportion of oxytocin cells excited by stimulation of the optic nerve was highest at ZT 4–8 and lowest at ZT 20–24. For vasopressin cells, it was highest at ZT 12–16 and lowest at ZT 20–24. The proportion of excitatory, inhibitory and complex responses seen in oxytocin and vasopressin cells following stimulation of the SCN also changed and was significantly different at different times of day (oxytocin cells: highest proportion of excitatory responses at ZT 12–16, P = 0.029; chi-square test; vasopressin cells: highest proportion of excitatory responses at ZT 0–4, P = 0.005; chi-square test). Thus, inputs to oxytocin and vasopressin neurones from the optic nerve and some outputs from the SCN changed during the light/dark cycle. Such changes may contribute to the generation of 24-h rhythms in activity of oxytocin and vasopressin neurones and release of the peptides. [ABSTRACT FROM AUTHOR]