The Effects of Ghrelin on Spike Activity of the Suprachiasmatic Nucleus Neurones of the Rat

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Abstract

In one of the most important non-photic mechanisms of the circadian biological clock synchronization with environmental geophysical 24 h rhythm, information on feeding schedule, composition and calorie content of food is used. Hormone ghrelin, a product of the neuroendocrine oxyntic cells of the gastric mucosa to be a signal molecule within this mechanism. In experiments on sagittal hypothalamic slices of male Wistar rats, the effects of 25 nM ghrelin on spike activity and parameters of spike information coding were investigated. Application of ghrelin induced an increase in spike frequency and a decrease in entropy of interspike interval distribution in 32.1% of the neurones recorded. In 29.6% of the cells, opposite responses in the form of a reduction of activity and an increase in the entropy of interspike interval distribution were observed. Parameters of spike activity of the reminder 38.3% neurones of the suprachiasmatic nucleus did not change. The observed responses of the entropy of interspike interval distribution indicate the appropriate changes in a degree of irregularity of interspike interval induced by ghrelin. Application of selective high-affinity antagonist of GHS-R1a receptor, JMV 2959 (100 nM) did not induce responses of the investigated parameters of spike activity but completely prevented changes of both, spike frequency and entropy of interspike interval distribution observed in the presence of ghrelin. The obtained data show that hormone ghrelin by a direct influence on the suprachiasmatic nucleus in vitro modulates the activity level and spike code of relatively numerous population of neurones of the nucleus, wherein the effects of ghrelin are implemented via GHS-R1a receptors. The results of the current study provide additional evidence in favour of the hypothesis on the involvement of ghrelin in mechanisms of non-photic entrainment of the circadian biological clock in accordance with severity of food motivation and level of metabolism.

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A. N. Inyushkin

Samara National Research University

Email: ainyushkin@mail.ru
Russian Federation, Samara

A. Yu. Sharafutdinova

Samara National Research University

Email: ainyushkin@mail.ru
Russian Federation, Samara

E. M. Inyushkina

Samara National Research University

Email: ainyushkin@mail.ru
Russian Federation, Samara

A. A. Inyushkin

Samara National Research University

Author for correspondence.
Email: ainyushkin@mail.ru
Russian Federation, Samara

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2. Fig. 1. Examples of reactions of neurons of the suprachiasmatic nucleus to the application of 25 nM ghrelin in the form of an increase in spike activity (a), a decrease in spike activity (b) and the absence of significant changes in activity (c). For all neurons, histograms with data on changes in the level of spike activity of cells during the experiment are presented in the upper part of the figure. The period of ghrelin application is shown by a horizontal segment above the histogram. On the abscissa axis is the time in seconds (the moment of application start corresponds to 0), on the ordinate axis is the frequency of spikes (Hz). The horizontal segments under the histogram (i, ii) correspond to 100-second intervals for recording the spike activity of neurons, presented under the histogram: (i) – immediately before exposure, (ii) – at the end of ghrelin application.

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3. Fig. 2. Statistical data on the effect of 25 nM ghrelin on the average frequency of action potentials, Hz (left diagrams), the entropy of the distribution of interstitial intervals, bits (middle diagrams) and mutual information between conjugated interstitial intervals, bits (right diagrams) of neurons of the suprachiasmatic nucleus. (a) Neurons whose activity increased under the influence of ghrelin (n = 26). (b) Neurons whose activity decreased under the influence of ghrelin (n = 24). (c) The entire set of registered neurons (n = 81). The symbols under the diagrams are: Before – the initial state; Ghrelin – against the background of ghrelin action; Without – after the "laundering" of the cut of the CSF. Asterisks indicate statistically significant differences with the initial state: *p < 0.05; **p < 0.01; ***p < 0.001.

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4. 3. The selective GHS-R1a antagonist of JMV 2959 receptors had no effect on the spike activity of neurons of the suprachiasmatic nucleus, but prevented the effects of 25 nM ghrelin. (a) An example of a neuron's reaction to the application of an antagonist and subsequent application of ghrelin. The upper part of the figure shows a histogram with data on the frequency of neuron spikes during the experiment. The periods of application of the antagonist and ghrelin are shown by horizontal segments above the histogram. On the abscissa axis is the time in seconds (the moment of application start corresponds to 0), on the ordinate axis is the frequency of spikes (Hz). The horizontal segments under the histogram (i, ii, iii) correspond to 100-second intervals for recording the spike activity of neurons, presented under the histogram: (i) – immediately before exposure, (ii) – at the end of the antagonist application, (iii) – at the end of ghrelin application against the background of the antagonist action. (b) Statistical data on the effect of 100 nM JMV 2959 and 25 nM ghrelin on the background of the antagonist's action on the average

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