Skip to main content
SpringerLink
Log in

Expression and Localization of sPLA2-III in the Rat CNS

  • Original Paper
  • Published:
Neurochemical Research Aims and scope Submit manuscript

Abstract

Phospholipases A2 (PLA2) are enzymes that cleave the sn-2 bond of membrane phospholipids to yield free fatty acids and lysophospholipids. Secretory PLA2-III (sPLA2-III) has been suggested to be important for neuronal differentiation, growth and survival, and is highly expressed in the spinal cord. The aim of this study is to elucidate its expression and distribution in different regions of the adult rat CNS. Quantitative RT-PCR analyses showed high levels of sPLA2-III mRNA expression in the brainstem and spinal cord and low expression in the olfactory bulb. Western blot analyses showed high level of expression in the brainstem, spinal cord and cerebral neocortex. A dense band corresponding to the catalytically active, mature/cleaved form, and a faint band corresponding to the full length sPLA2-III were detected in post-mitochondrial supernatants, from different parts of the CNS. Subcellular fractionation of spinal cord homogenates showed that sPLA2-III protein is present in the ‘light membrane/cytosol’ fraction, but not the nucleus, synaptosomal membrane or synaptic vesicle-enriched fractions. sPLA2-III was immunolocalized to neurons in the cerebral neocortex, Purkinje neurons in the cerebellar cortex, periaqueductal gray, red nucleus, spinal trigeminal nucleus and dorsal horn of the spinal cord. Electron microscopy of the spinal cord and cerebral neocortex showed that sPLA2-III was localized in dendrites or dendritic spines, that formed asymmetrical synapses with unlabeled, putatively glutamatergic, axon terminals. The localization of mature/cleaved form of sPLA2-III in postsynaptic structures suggest a physiological role of the enzyme in neurotransmission or synaptic plasticity.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Sun GY, Shelat PB, Jensen MB, He Y, Sun AY, Simonyi A (2010) Phospholipases A2 and inflammatory responses in the central nervous system. Neuromolecular Med 12(2):133–148. doi:10.1007/s12017-009-8092-z

    Article  CAS  PubMed  Google Scholar 

  2. Dennis EA, Cao J, Hsu YH, Magrioti V, Kokotos G (2011) Phospholipase A2 enzymes: physical structure, biological function, disease implication, chemical inhibition, and therapeutic intervention. Chem Rev 111(10):6130–6185. doi:10.1021/cr200085w

    Article  CAS  PubMed  Google Scholar 

  3. Gelb MH, Valentin E, Ghomashchi F, Lazdunski M, Lambeau G (2000) Cloning and recombinant expression of a structurally novel human secreted phospholipase A2. J Biol Chem 275(51):39823–39826. doi:10.1074/jbc.C000671200

    Article  CAS  PubMed  Google Scholar 

  4. Ho IC, Arm JP, Bingham CO 3rd, Choi A, Austen KF, Glimcher LH (2001) A novel group of phospholipase A2s preferentially expressed in type 2 helper T cells. J Biol Chem 276(21):18321–18326. doi:10.1074/jbc.M008837200

    Article  CAS  PubMed  Google Scholar 

  5. Six DA, Dennis EA (2000) The expanding superfamily of phospholipase A2 enzymes: classification and characterization. Biochim Biophys Acta 1488:1–19

    Article  CAS  PubMed  Google Scholar 

  6. Murakami M, Taketomi Y, Sato H, Yamamoto K (2011) Secreted phospholipase A2 revisited. J Biochem 150(3):233–255. doi:10.1093/jb/mvr088

    Article  CAS  PubMed  Google Scholar 

  7. Goracci G, Ferrini M, Nardicchi V (2010) Low molecular weight phospholipases A2 in mammalian brain and neural cells: roles in functions and dysfunctions. Mol Neurobiol 41(2–3):274–289. doi:10.1007/s12035-010-8108-6

    Article  CAS  PubMed  Google Scholar 

  8. Valentin E, Ghomashchi F, Gelb MH, Lazdunski M, Lambeau G (2000) Novel human secreted phospholipase A2 with homology to the group III bee venom enzyme. J Biol Chem 275:7492–7496

    Article  CAS  PubMed  Google Scholar 

  9. Beck S, Beck G, Ostendorf T, Floege J, Lambeau G, Nevalainen T, Radeke HH, Gurrieri S, Haas U, Thorwart B, Pfeilschifter J, Kaszkin M (2006) Upregulation of group IB secreted phospholipase A2 and its M-type receptor in rat ANTI-THY-1 glomerulonephritis. Kidney Int 70:1251–1260

    Article  CAS  PubMed  Google Scholar 

  10. Murakami M, Masuda S, Shimbara S, Bezzine S, Lazdunski M, Lambeau G, Gelb MH, Matsukura S, Kokubu F, Adachi M, Kudo I (2003) Cellular arachidonate-releasing function of novel classes of secretory phospholipase A2s (groups III and XII). J Biol Chem 278(12):10657–10667. doi:10.1074/jbc.M211325200

    Article  CAS  PubMed  Google Scholar 

  11. Masuda S, Yamamoto K, Hirabayashi T, Ishikawa Y, Ishii T, Kudo I, Murakami M (2008) Human group III secreted phospholipase A2 promotes neuronal outgrowth and survival. Biochem J 409(2):429–438. doi:10.1042/BJ20070844

    Article  CAS  PubMed  Google Scholar 

  12. Titsworth WL, Onifer SM, Liu NK, Xu XM (2007) Focal phospholipases A2 group III injections induce cervical white matter injury and functional deficits with delayed recovery concomitant with Schwann cell remyelination. Exp Neurol 207(1):150–162. doi:10.1016/j.expneurol.2007.06.010

    Article  CAS  PubMed  Google Scholar 

  13. Sato H, Kato R, Isogai Y, Saka G, Ohtsuki M, Taketomi Y, Yamamoto K, Tsutsumi K, Yamada J, Masuda S, Ishikawa Y, Ishii T, Kobayashi T, Ikeda K, Taguchi R, Hatakeyama S, Hara S, Kudo I, Itabe H, Murakami M (2008) Analyses of group III secreted phospholipase A2 transgenic mice reveal potential participation of this enzyme in plasma lipoprotein modification, macrophage foam cell formation, and atherosclerosis. J Biol Chem 283:33483–33497

    Article  CAS  PubMed  Google Scholar 

  14. Sato H, Taketomi Y, Isogai Y, Masuda S, Kobayashi T, Yamamoto K, Murakami M (2009) Group III secreted phospholipase A2 transgenic mice spontaneously develop inflammation. Biochem J 421(1):17–27. doi:10.1042/BJ20082429

    Article  CAS  PubMed  Google Scholar 

  15. Nicolas JP, Lin Y, Lambeau G, Ghomashchi F, Lazdunski M, Gelb MH (1997) Localization of structural elements of bee venom phospholipase A2 involved in N-type receptor binding and neurotoxicity. J Biol Chem 272(11):7173–7181

    Article  CAS  PubMed  Google Scholar 

  16. Thwin MM, Ong WY, Fong CW, Sato K, Kodama K, Farooqui AA, Gopalakrishnakone P (2003) Secretory phospholipase A2 activity in the normal and kainate injected rat brain, and inhibition by a peptide derived from python serum. Exp Brain Res 150(4):427–433. doi:10.1007/s00221-003-1476-7

    CAS  PubMed  Google Scholar 

  17. Ma MT, Yeo JF, Shui G, Wenk MR, Ong WY (2012) Systems wide analyses of lipids in the brainstem during inflammatory orofacial pain—evidence of increased phospholipase A2 activity. Eur J Pain 16(1):38–48

    Article  CAS  PubMed  Google Scholar 

  18. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25:402–408

    Article  CAS  PubMed  Google Scholar 

  19. Murakami M, Masuda S, Shimbara S, Ishikawa Y, Ishii T, Kudo I (2005) Cellular distribution, post-translational modification, and tumorigenic potential of human group III secreted phospholipase A2. J Biol Chem 280:24987–24998

    Article  CAS  PubMed  Google Scholar 

  20. Hallett PJ, Collins TL, Standaert DG, Dunah AW (2008) Biochemical fractionation of brain tissue for studies of receptor distribution and trafficking. Current protocols in neuroscience/editorial board, Jacqueline N Crawley [et al] Chapter 1:Unit 1, vol 16. doi:10.1002/0471142301.ns0116s42

  21. Edwards FA (1995) Anatomy and electrophysiology of fast central synapses lead to a structural model for long-term potentiation. Physiol Rev 75:759–787

    CAS  PubMed  Google Scholar 

  22. Macchioni L, Corazzi L, Nardicchi V et al (2004) Rat brain cortex mitochondria release group II secretory phospholipase A2 under reduced membrane potential. J Biol Chem 279:37860–37869

    Article  CAS  PubMed  Google Scholar 

  23. Nardicchi V, Macchioni L, Ferrini M et al (2007) The presence of a secretory phospholipase A2 in the nuclei of neuronal and glial cells of rat brain cortex. Biochim Biophys Acta 1771:1345–1352

    Article  CAS  PubMed  Google Scholar 

  24. Ferrini M, Nardicchi V, Mannucci R et al (2010) Effects of NGF on the subcellular localization of group IIA secretory phospholipase A2 (GIIA) in PC12 cells: role in neuritogenesis. Neurochem Res 35:2168–2174

    Article  CAS  PubMed  Google Scholar 

  25. Ma MT, Nevalainen TJ, Yeo JF, Ong WY (2010) Expression profile of multiple secretory phospholipase A2 isoforms in the rat CNS: enriched expression of sPLA2-IIA in brainstem and spinal cord. J Chem Neuroanat 39:242–247

    Article  CAS  PubMed  Google Scholar 

  26. Than A, Tan Y, Ong WY, Farooqui AA, Chen P (2012) Kainate receptors mediate regulated exocytosis of secretory phospholipase A2 in SH-SY5Y neuroblastoma cells. Neurosignals 20(2):72–85

    Article  CAS  PubMed  Google Scholar 

  27. Wei S, Ong WY, Thwin MM, Fong CW, Farooqui AA, Gopalakrishnakone P, Hong W (2003) Group IIA secretory phospholipase A2 stimulates exocytosis and neurotransmitter release in pheochromocytoma-12 cells and cultured rat hippocampal neurons. Neuroscience 121(4):891–898

    Article  CAS  PubMed  Google Scholar 

  28. Dorandeu F, Pernot-Marino I, Veyret J, Perrichon C, Lallement G (1998) Secreted phospholipase A2-induced neurotoxicity and epileptic seizures after intracerebral administration: an unexplained heterogeneity as emphasized with paradoxin and crotoxin. J Neurosci Res 54:848–862

    Article  CAS  PubMed  Google Scholar 

  29. Florea A, Puica C, Vintan M, Benga I, Craciun C (2011) Electrophysiological and structural aspects in the frontal cortex after the bee (Apis mellifera) venom experimental treatment. Cell Mol Neurobiol 31(5):701–714. doi:10.1007/s10571-011-9667-4

    Article  CAS  PubMed  Google Scholar 

  30. DeCoster MA, Lambeau G, Lazdunski M, Bazan NG (2002) Secreted phospholipase A2 potentiates glutamate-induced calcium increase and cell death in primary neuronal cultures. J Neurosci Res 67(5):634–645

    Article  CAS  PubMed  Google Scholar 

  31. Martínez-García A, Sastre I, Recuero M, Aldudo J, Vilella E, Mateo I, Sánchez-Juan P, Vargas T, Carro E, Bermejo-Pareja F, Rodríguez-Rodriguez E, Combarros O, Rosich-Estrago M, Frank A, Valdivieso F, Bullido MJ (2010) PLA2G3, a gene involved in oxidative stress induced death, is associated with Alzheimer’s disease. J Alzheimers Dis 22(4):1181–1187

    PubMed  Google Scholar 

  32. Yang X, Sheng W, He Y, Cui J, Haidekker M, Sun GY, Lee JC (2010) Secretory phospholipase A2 type III enhances alpha-secretase-dependent amyloid precursor protein processing through alterations in membrane fluidity. J Lipid Res 51(5):957–966

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by the Neurobiology and Ageing Research Programme and the National University Health System of the National University of Singapore. The authors extend their appreciation to the Deanship of Scientific Research, King Saud University, for funding this work through the Visiting Professorship Program to WYO. We thank Professor Makoto Murakami, from the Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan, for his kind gift of sPLA2-III antibody.

Author information

Authors and Affiliations

  1. Department of Anatomy, National University of Singapore, Singapore, 119260, Singapore

    Hui Yang & Wei-Yi Ong

  2. Neurobiology and Ageing Research Programme, National University of Singapore, Singapore, 119260, Singapore

    Wei-Yi Ong

  3. Department of Biochemistry, College of Science, King Saud University, Riyadh, Saudi Arabia

    Nikhat J. Siddiqi & A. S. Alhomida

Authors
  1. Hui Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Nikhat J. Siddiqi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. A. S. Alhomida
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Wei-Yi Ong
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Wei-Yi Ong.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Yang, H., Siddiqi, N.J., Alhomida, A.S. et al. Expression and Localization of sPLA2-III in the Rat CNS. Neurochem Res 38, 753–760 (2013). https://doi.org/10.1007/s11064-013-0974-7

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11064-013-0974-7

Keywords

Search

Navigation