[EFFECT OF PEPTIDE SEMAX ON SYNAPTIC ACTIVITY AND SHORT-TERM PLASTICITY OF GLUTAMATERGIC SYNAPSES OF CO-CULTURED DORSAL ROOT GANGLION AND DORSAL HORN NEURONS].

The influence of long-term culturing (12 days in vitro) of dorsal root ganglion (DRG) and dorsal horn (DH) neurons with peptide Semax on the level of synaptic activity at co-cultures, as well as short-term plasticity in sensory synapses were studied. It has been shown that neuronal culturing with peptide at concentrations of 10 and 100 µM led to increasing the frequency of spontaneous glutamatergic postsynaptic currents in DH neurons to 71.7 ± 1.8% and 93.9 ± 3.1% (n = 6; P < 0.001); Semax has a not significant effect on the amplitude and frequency of miniature glutamatergic currents, but causes an increase of the amplitudes of spontaneous postsynaptic currents, as well as elevates the quantum content. The data show the increase of multivesicular glutamate release efficiency in neural networks of co-cultures following incubation with the peptide. Also Semax (10 and 100 µM) induces changes of the basic parameters of short-term plasticity in sensory synapses: (1) increasing the paired-pulse ratio from 0.53 ± 0.028 (n = 8) to 0.91 ± 0.072 (n = 6, P < 0.01) and 0.95 ± 0.026 (n = 7; P < 0.001); (2) reducing the ratio of the coefficients of variation (CV2/ CV1) from 1.49 ± 0.11 (n = 8) to 1.02 ± 0.09 (n = 6; P < 0.05) and 1.11 ± 0.13 (n = 7; P < 0.0) respectively. The results indicate a stimulating effect of Semax on the activity of glutamatergic synapses in neural networks of co-cultures, as well as the ability of the peptide to effectively modulate the short-term plasticity in sensory synapses.

[Pre- and postsynaptic expression mechanisms of long-term depression in rats glutamatergic primary afferent synapses on dorsal horn spinal cord neurons in co-culture].

In co-culture of dorsal root ganglion (DRG) and dorsal horn (DH) neurons we studied the long-term depression (LTD) caused by 5 sec(-1) low-frequency stimulation (LFS) of DRG neurons. Dual whole-cell patch clamp recording in the pairs of DRG and DH neurons was used. The monosynaptic AMPA-receptor mediated eEPSC initiated in DH neurons by LFS of DRG neurons were analyzed. LFS with durations from 60 to 360 sec affected the LTD magnitude and involved the different LTD expression mechanisms. The amplitude and variability of miniature EPSC were not changed during LTD. LFS during 60 sec did not alter the eEPSC amplitudes, but significantly increased the coefficient of variation (CV; 56.8% +/- 11.5%, n = 5; P < 0.002) and the paired-pulse ratio (PPR; 37.8% +/- 11.4%, n = 5; P < 0.05), and decreased the release probability (p; 21.9% +/- 5.6%, n = 5; P < 0.05) calculated using binomial analysis. LFS for 120 sec led to LTD (eEPSC decreased to 14% +/- 3.3%, n = 13; P < 0.005); no changes in CV, PPR and p were found. LFS for 160, 200, 240 and 360 sec induced robust LTD: eEPSC decreased to 37.3 +/- 4.3 (n = 9); 48.1 +/- 3.5 (n = 7); 58.3 +/- 2.5 (n = 8) and 57.3% +/- 2.8% (n = 8), respectively; P < 0.001. LTD induced by such LFS durations was not accompanied by significant changing in PPR and p, but showed an increase in CV: 24.4 +/- 8.6 (n = 6); 35.1 +/- 11.2 (n = 6); 37.7 +/- 12.8 (n = 7), and 38.1% +/- 9.3% (n = 8), respectively. LTD magnitude was correlated with the enhancement in CV value at different LFS duration (r = 0.96). Our data suggest that the different mechanisms could be involved in LTD expression according to duration of LFS. Thus, 60 sec LFS induces presynaptic changes, but no change in eEPSC, whereas LTD elicited by 120 and 160 sec LFS affected the postsynaptic site. LTD initiated by longer LFS (200 - 360 sec) probably was caused by a silencing of functional synapses without changes in glutamate release probability.

[Characteristics of sensory neurotransmission in co-culture of neurons from the dorsal root ganglion and dorsal horn spinal cord in rats].

We examined properties of chemical neurotransmission at the level of primary afferent inputs into spinal cord with the new easy-to-use in vitro model of contiguously-cultivated dissociated both the dorsal root ganglion neurons (DRG) and the dorsal horn spinal cord neurons (DHSC) from newborn rats. The results of our studies showed the presence of excitatory and inhibitory DRG neurons synapses on the cells of DHSC. The excitatory afferent signaling in such synapses was mediated by presynaptic release of glutamate and a following activation of both NMDA- and non-NMDA-receptor subclasses. In these cases the activation of non-NMDA-receptors makes a main contribution to realization of excitatory postsynaptic effects. Either glycine- or GABA-ergic DRG neurons were involved in transmission of inhibitory signals to the DHSC neurons. However, in vast majority of examined neuronal pairs the inhibitory synaptic transmission was mediated by presynaptic release of glycine. As distinct from the previous similar methods, an in vitro model of co-culture of both the DRG and the DHSC neurons proposed here allows to use comprehensively modem technical approaches for examination of the transmission of somatosensory information from the periphery to the CNS. The described model could be acceptable for detailed investigation of specific properties of primary afferent synapses.