Korean Journal of Veterinary Research (대한수의학회지)

The Korean Society of Veterinary Science (대한수의학회)
권호 표지

Screening of active substance FS11052 as an inhibitor of neurotransmitter release from PC12 cells

PC12 세포에서 신경전달물질 방출을 저해하는 생리활성물질 FS11052의 탐색

  • Lee, Yun-Sik (Department of Infection Biology, College of Medicine, Wonkwang University) ;
  • Lee, John Hwa (Bio-Safety Research Institute, College of Veterinary Medicine, Chonbuk National University)
  • 이윤식 (원광대학교 의과대학 감염생물학교실) ;
  • 이존화 (전북대학교 수의과대학 생체안전성연구소)
  • Accepted : 2006.06.09
  • Published : 2006.06.30
Download PDF
⟨ Previous Next ⟩

Abstract

We established an in vitro experimental system using the following procedure. We first introduced tritium-labeled norepinephrine ([$^3$H]-NE) into PC12 cells, The [$^3$H]-NE incorporated into PC12 cells were then stimulated by a high concentration (60 mM) of $K^+$ buffer during 12 minutes. Then, we collected $100{\mu}l$ supernatant and counted the amount of [$^3$H]-NE release from PC12 cells with a scintillation counter. After screening fungal, Streptomyces spp. or bacterial product using this experimental sytem, we obtained FS11052 from Streptomyces spp. which inhibited [$^3$H]-NE release from PC12 cells. FS11052 also inhibits the release of ATP as a neurotransmitter of PC12 cells and rat cortical neurons, The inhibitory effect was seen even when the PC12 cells were treated with low $K^-$ buffer containing ionomycin ($1{\mu}M$) as an ionopore. This result suggests that the inhibitory action of FS11052 on neurotransmitter release appeared after the influx of $Ca^{2+}$.

Keywords

References

  1. Alder J, Xie ZP, Valtorta F, Greenfard P, Poo MM. Antibodies to synaptophysin enter with transmitter secretion at neiuromuscular synapses. Neuron 1992, 9, 759-768 https://doi.org/10.1016/0896-6273(92)90038-F
  2. Boomert K, Charlton MP, DeBello WM, Chin GJ, Betz H. Augustine 0.1. Inhibition of neurotransmitter release by C2-domain peptides implicates synaptotamin in exocytosis. Nature 1993, 363, 163-165 https://doi.org/10.1038/363163a0
  3. Brennwald P, Kearns B, Champion K, Keranen S, Bankaitis V, Novick P. Sec9 is sSNAP-25-like component of a yeast SNARE complex that may be the effector of Sec4 function in exocytosis. Cell 1994, 79, 245-258 https://doi.org/10.1016/0092-8674(94)90194-5
  4. Brose N, Petrenko AG, Sudhof TC, Jahn R. Synaptotagmina: a calcium sensor on the synaptic vesicle surface. Science 1992, 256, 1021-1025 https://doi.org/10.1126/science.1589771
  5. Cutler DF, Cramer LP. Sorting during transport to the surface of PC 12 cells: Divergence of synaptic vesicle and secretory granule proteins. J Cell Biol 1990, 110, 721-730 https://doi.org/10.1083/jcb.110.3.721
  6. Daikoku S, Hisano S, Kagotani Y. Neuronal associations in the rat suprachiasmatic nucleus demonstrated by immunoelectron microscopy. J Comp Neurol 1992, 325, 559-571 https://doi.org/10.1002/cne.903250408
  7. Delay-Goyet P, Lundberg JM. Actinomycino is a competitive neurokinin receptor anragotist. Biochem Biophs Res Commun 1991, 180, 1342-1349 https://doi.org/10.1016/S0006-291X(05)81343-9
  8. Hirling H, Steiner P, Chaperon C, Marsault R, Regazzi R, Catsicas S. Syntaxin 13 is a developmentally regulated SNARE involved in neurite outgrowth and endosomal trafficking. Eur J Neurosci 2000, 12,1913-1923 https://doi.org/10.1046/j.1460-9568.2000.00076.x
  9. Hunter T. Protein kinase and phosphatases The Yin and Yang of protein phosphorylation and signaling. Cell 1995, 80, 225-236 https://doi.org/10.1016/0092-8674(95)90405-0
  10. Kikuchi A, Yamashita T, Kawata M, Yamamoto K, Ikeda K, Tanimoto T Takai Y. Purification and characterization of a novel GTP-binding protein with a molecular weight of 24,000 from bovine brain membranes. J Biol Chem 1988, 263, 2897-2904
  11. Kishida S, Shirataki H, Sasaki T, Kato M, Kaibuchi K, Takai Y. Rab3A GTPaseactivating protein-inhibiting activity of rabphilin3A, a putative rat3A target protein. J Biol Chem 2000, 268, 22259-22261
  12. Li MF, Shi YL. Toosendanin interferes with pore formation of botulinum toxin type A in PC 12 cell membrane. Acta Pharmacol Sin 2006, 27, 66-70 https://doi.org/10.1111/j.1745-7254.2006.00236.x
  13. Liu J, Misler S. Alpha-Latrotoxin alters spontaneous and depolarization-evoked quantal release from rat adrenal chromaffin cells: evidence for multiple modes. J of Neurosci 1998, 18, 6113-6125 https://doi.org/10.1523/JNEUROSCI.18-16-06113.1998
  14. Moriyama Y, Futai M. $H^--ATPase$, a Primary pump for accumulation of neurotransmitter, is a major constituent of brain synaptic vesicles. Biochem Biophys Commun 1990, 173, 443-448 https://doi.org/10.1016/S0006-291X(05)81078-2
  15. O'Connor VM, Shamotienko O, Grishin E, Betz H. On the structure of the 'synaptosecretosome'-Evidence for a $neuexin/synaptntagmin/syntaxin/Ca^{2+}$ channel complex. FEBS 1993, 326, 255-260 https://doi.org/10.1016/0014-5793(93)81802-7
  16. Petrenko AG, Perin MS, Davletov BA, Ushkaryov YA, Geppert M, Sudhof TC. Binding of synaptotagmin to the alpha-latrotoxin receptor implicates both in synaptic vesicle exocytosis. Nature 1991, 353, 65-68 https://doi.org/10.1038/353065a0
  17. Popov SV, Poo M. Synaptotagmin: a calcium-sensitive inhibitor of exocytosis? Cell 1993, 73, 1247-1249 https://doi.org/10.1016/0092-8674(93)90352-Q
  18. Sakaba T, Stein A, Jahn R, Neher E. Distinct kinetic changes in neurotransmitter release after SNARE protein cleavage. Science 2005, 309, 491-494 https://doi.org/10.1126/science.1112645
  19. Schiavo G, Benfenati F, Poulain B, Rossetto O, Polverino de L, DasGupta BR, Montecucco C. Tetanus and botulinum-B neurotoxins block neurotransmitter release by proteolytic cleavage of synaptobrevin. Nature 1992, 359, 832-835 https://doi.org/10.1038/359832a0
  20. Shin OH, Rhee JS, Tang J, Sugita S, Rosenmund C, Sudhof TC. $Sr^{2+}$ binding to the $Ca^{2+}$ binding site of the synaptotagmin 1 C2B domain triggers fast exocytosis without stimulating SNARE interactions. Neuron 37, 99-108 https://doi.org/10.1016/S0896-6273(02)01145-5
  21. Shoji-Kasai Y, Yoshida A, Sato K, Hoshino T, Ogura A, Kondo S, Fujimoro Y, Kuwahara R, Kato R, Takahasahi M. Neurotransmitter release form synaptotagmin-deficient clonal variants of PC12 cells. Science 1992, 256, 1821-1823
  22. Tuz K, Pasantes-Morales H. Hyposmolarity evokes norepinephrine efflux from synaptosomes by a depolarization-and $Ca^{2+}$ -dependent exocytotic mechanism. Eur J Neurosci 2005, 22, 1636-1642 https://doi.org/10.1111/j.1460-9568.2005.04344.x
  23. Vitale ML, Seward EP, Trifao JM. Chromaffin cell cortical actin network dynamics control the size of the release-ready vesicle pool and the initial rate of exocytosis. Neuron 1995, 14, 353-363 https://doi.org/10.1016/0896-6273(95)90291-0
  24. Wang CT, Bai J, Chang PY, Chapman ER, Jackson MB. $Synaptotagmin-Ca^{2+}$ triggers two sequential steps in regulated exocytosis in rat PC12 cells: fusion pore opening and fusion pore dilation. J Physiol 2006, 570, 295-307 https://doi.org/10.1113/jphysiol.2005.097378
  25. Wang P, Chicka MC, Bhalla A, Richards DA, Chapman ER. Synaptotagmin VII is targeted to secretory organelles in PC12 cells, where it functions as a high-affinity calcium sensor. Mol Cell Biol 2005, 25, 8693-8702 https://doi.org/10.1128/MCB.25.19.8693-8702.2005
  26. Wang Y, Okamoto M, Schmitz F, Hofmann K, Sudhof TC. Rim is a putative Rab3 effector in regulating synaptic-vesicle fusion. Nature 1997, 388, 593-398 https://doi.org/10.1038/41580
  27. Yamaguchi T, Shirataki H, Kishida S, Miyazaki M, Nishikawa J, Wada K, Numata S, Kaibuchi K, Takai Y. Two functionally different domains of rabphilin-3A, rab 3Ap25/smg p25A-binding and phospholipids and $Ca^{2+}-binding$ domains. J Biol Chem 1993, 268, 27164-27170
  28. Yelamanchili SV, Reisinger C, Becher A, Sikorra S, Bigalke H, Binz T, Ahnert-Hilger G. The C-terminal transmembrane region of synaptobrevin binds synaptophysin from adult synaptic vesicles. Eur J Cell Biol 2005, 84, 467-475 https://doi.org/10.1016/j.ejcb.2004.11.007
  29. Zhang YQ, Rodesch CK, Broadie K. Living synaptic vesicle marker: synaptotagmin-GFP. Genesis 2002, 34, 142-145 https://doi.org/10.1002/gene.10144
  30. Zhou Q, Xiao J, Liu Y. Participation of syntaxin 1A in membrane trafficking involving neurite elongation and membrane expansion. J Neurosci Res 2000, 61, 321-328 https://doi.org/10.1002/1097-4547(20000801)61:3<321::AID-JNR10>3.0.CO;2-L