Korean J Physiol Pharmacol.  2018 Sep;22(5):539-546. 10.4196/kjpp.2018.22.5.539.

Botulinum toxin type A enhances the inhibitory spontaneous postsynaptic currents on the substantia gelatinosa neurons of the subnucleus caudalis in immature mice

• Jang SH
• Park SJ
• Lee CJ
• Ahn DK
• Han SK

• KMID: 2419003
• DOI: http://doi.org/10.4196/kjpp.2018.22.5.539

Abstract

Botulinum toxin type A (BoNT/A) has been used therapeutically for various conditions including dystonia, cerebral palsy, wrinkle, hyperhidrosis and pain control. The substantia gelatinosa (SG) neurons of the trigeminal subnucleus caudalis (Vc) receive orofacial nociceptive information from primary afferents and transmit the information to higher brain center. Although many studies have shown the analgesic effects of BoNT/A, the effects of BoNT/A at the central nervous system and the action mechanism are not well understood. Therefore, the effects of BoNT/A on the spontaneous postsynaptic currents (sPSCs) in the SG neurons were investigated. In whole cell voltage clamp mode, the frequency of sPSCs was increased in 18 (37.5%) neurons, decreased in 5 (10.4%) neurons and not affected in 25 (52.1%) of 48 neurons tested by BoNT/A (3 nM). Similar proportions of frequency variation of sPSCs were observed in 1 and 10 nM BoNT/A and no significant differences were observed in the relative mean frequencies of sPSCs among 1-10 nM BoNT/A. BoNT/A-induced frequency increase of sPSCs was not affected by pretreated tetrodotoxin (0.5 µM). In addition, the frequency of sIPSCs in the presence of CNQX (10 µM) and AP5 (20 µM) was increased in 10 (53%) neurons, decreased in 1 (5%) neuron and not affected in 8 (42%) of 19 neurons tested by BoNT/A (3 nM). These results demonstrate that BoNT/A increases the frequency of sIPSCs on SG neurons of the Vc at least partly and can provide an evidence for rapid action of BoNT/A at the central nervous system.

Keyword

Botulinum toxin type A; Pain; Spontaneous postsynaptic current; Substantia gelatinosa; Whole-cell recoding

MeSH Terms

6-Cyano-7-nitroquinoxaline-2,3-dione
Animals
Botulinum Toxins*
Botulinum Toxins, Type A*
Brain
Central Nervous System
Cerebral Palsy
Dystonia
Hyperhidrosis
Mice*
Neurons*
Substantia Gelatinosa*
Synaptic Potentials*
Tetrodotoxin
6-Cyano-7-nitroquinoxaline-2,3-dione
Botulinum Toxins
Botulinum Toxins, Type A
Tetrodotoxin

Figure

• 

  Fig. 1 BoNT/A increases the frequency of sPSCs on SG neurons. (A) A representative trace showing the frequency increase of sPSCs by 3 nM BoNT/A. (B) Time course frequency histogram (bin size 20 s) of the current trace in (A). (C) Cumulative probability histogram of the inter-event interval (IEI) of sPSCs in the control (solid line) and with BoNT/A (BT, dotted line). Bar graphs showing the mean frequency (D) and mean amplitude (E) of sPSCs in control and BoNT/A, respectively. Values represent mean±SEM, *** represents p<0.001, NS (not significant), BT (BoNT/A).

• 

  Fig. 2 Concentration-response relationship of BoNT/A on the SG neurons. Each bar graphs show the relative mean frequency (A) and relative mean amplitude (B) of sPSCs by application of 1, 3, 10 nM BoNT/A compared to control, respectively. Values represent mean±SEM, (C) Stack column graph showing the proportion of frequency change by BoNT/A (1, 3, 10 nM). The numbers in parentheses mean the number of neurons recorded. D (decrease), N (no change), I (increase), * (p<0.05), NS (not significant).

• 

  Fig. 3 BoNT/A increases the frequency of mPSCs on SG neurons. Bar graphs show the mean frequency (A) and mean amplitude (B) of sPSCs by application of 3 nM BoNT/A alone (BT) and BT in the presence of 0.5 µM TTX (TTX+BT), respectively. (C) Stack column graph showing the proportion of frequency change between 3 nM BT and BT in the presence of TTX (TTX+BT). D (decrease), N (no change), I (increase), * (p<0.05), NS (not significant).

• 

  Fig. 4 BoNT/A increases the frequency of sIPSCs on SG neurons. (A) A representative trace showing the frequency increase of sIPSCs by 3 nM BoNT/A (BT) in the presence of CNQX (10 µM) and AP5 20 µM. (B) Time course frequency histogram (bin size 20 s) of the sPSCs of the current trace in (A). (C) Cumulative probability histograms of the inter-event interval (IEI, a) and amplitude (b) of the sPSCs of the current trace in (A). Solid line (CNQX+AP5), Dotted line (CNQX+AP5+BT) (D) Comparisons of the mean frequency (a) and mean amplitude (b) of sPSCs between CNQX+AP5 and 3 nM BoNT/A in the presence of CNQX+AP5, respectively. (E) Stack column graph showing the proportion of frequency change by 3 nM BoNT/A alone and BoNT/A in the presence of CNQX+AP5. D (decrease), N (no change), I (increase). ** (p<0.01), NS (not significant).

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