Advertisement

Neurochemical Research

, Volume 41, Issue 4, pp 795–803 | Cite as

Protein Interacting C-Kinase 1 Modulates Surface Expression of P2Y6 Purinoreceptor, Actin Polymerization and Phagocytosis in Microglia

  • Jia Zhu
  • Zhen Wang
  • Nan Zhang
  • Jiao Ma
  • Shui-Lin Xu
  • Yin Wang
  • Ying Shen
  • Yun-Hong Li
Original Paper

Abstract

Microglia clean up dead cells and debris through phagocytosis in the central nervous system. UDP-activated P2Y6 receptors (P2Y6Rs) induce the formation of phagocytic cup-like structure and P2Y6R expression is increased during the phagocytosis. However, it remains unclear how surface expression of P2Y6R is increased. PICK1 (protein interacting with C-kinase-1) interacts with various neurotransmitter receptors, transporters, and enzymes. We here report that PICK1 might interact with P2Y6R. Surface P2Y6R was reduced in microglia from PICK1-knockout mice and PICK1-knockdown BV2 cells, which was also confirmed by electrophysiological recordings, showing that P2Y6R-mediated current was increased by PICK1 overexpression but was reduced by PICK1-knockdown in BV2 microglia. Finally, PICK1 was sufficient to affect cytoskeletal aggregation and phagocytosis both in primary microglia and BV2 cells. These results indicate that PICK1 is an important regulator of P2Y6R expression and microglial phagocytosis.

Keywords

PICK1 P2Y6 receptor BV2 Microglia 

Abbreviations

AMPAR

Α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid receptor

BAR

Bin-Amphiphysin-Rvs

Iba1

Ca2+-binding adapter molecule 1

PICK1

Protein interacting with C-kinase 1

PDZ

PSD-95/DlgA/ZO-1

PKC

Protein kinase C

PBS

Phosphate-buffered saline

Notes

Acknowledgments

This work was supported by grants from National Natural Science Foundation of China (81501043, 31460257, 31471024, and 81571098).

Compliance with Ethical Standards

Conflict of interest

All authors declare that there are no conflicts of interest on the paper.

Supplementary material

11064_2015_1754_MOESM1_ESM.tif (175 kb)
Supplementary material 1 (TIFF 174 kb)
11064_2015_1754_MOESM2_ESM.tif (1.5 mb)
Supplementary material 2 (TIFF 1583 kb)

References

  1. 1.
    Aloisi F (2001) Immune function of microglia. Glia 36:165–179CrossRefPubMedGoogle Scholar
  2. 2.
    Hanisch UK, Kettenmann H (2007) Microglia: active sensor and versatile effector cells in the normal and pathologic brain. Nat Neurosci 10:1387–1394CrossRefPubMedGoogle Scholar
  3. 3.
    Ji K, Akgul G, Wollmuth LP, Tsirka SE (2013) Microglia actively regulate the number of functional synapses. PLoS One 8:e56293CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Sierra A, Abiega O, Shahraz A, Neumann H (2013) Janus-faced microglia: beneficial and detrimental consequences of microglial phagocytosis. Front Cell Neurosci 7:6CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Chan A, Hummel V, Weilbach FX, Kieseier BC, Gold R (2006) Phagocytosis of apoptotic inflammatory cells downregulates microglial chemoattractive function and migration of encephalitogenic T cells. J Neurosci Res 84:1217–1224CrossRefPubMedGoogle Scholar
  6. 6.
    Koizumi S, Ohsawa K, Inoue K, Kohsaka S (2013) Purinergic receptors in microglia: functional modal shifts of microglia mediated by P2 and P1 receptors. Glia 61:47–54CrossRefPubMedGoogle Scholar
  7. 7.
    Mika T, Prochnow N (2012) Functions of connexins and large pore channels on microglial cells: the gates to environment. Brain Res 1487:16–24CrossRefPubMedGoogle Scholar
  8. 8.
    Koizumi S, Shigemoto-Mogami Y, Nasu-Tada K, Shinozaki Y, Ohsawa K, Tsuda M, Joshi BV, Jacobson KA, Kohsaka S, Inoue K (2007) UDP acting at P2Y6 receptors is a mediator of microglial phagocytosis. Nature 446:1091–1095CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Inoue K (2007) UDP facilitates microglial phagocytosis through P2Y6 receptors. Cell Adh Migr 1:131–132CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Zhang Z, Wang Z, Ren H, Yue M, Huang K, Gu H, Liu M, Du B, Qian M (2011) P2Y(6) agonist uridine 5′-diphosphate promotes host defense against bacterial infection via monocyte chemoattractant protein-1-mediated monocytes/macrophages recruitment. J Immunol 186:5376–5387CrossRefPubMedGoogle Scholar
  11. 11.
    Staudinger J, Zhou J, Burgess R, Elledge SJ, Olson EN (1995) PICK1: a perinuclear binding protein and substrate for protein kinase C isolated by the yeast two-hybrid system. J Cell Biol 128:263–271CrossRefPubMedGoogle Scholar
  12. 12.
    Xu J, Xia J (2006) Structure and function of PICK1. Neurosignals 15:190–201CrossRefPubMedGoogle Scholar
  13. 13.
    Hanley JG (2008) PICK1: a multi-talented modulator of AMPA receptor trafficking. Pharmacol Ther 118:152–160CrossRefPubMedGoogle Scholar
  14. 14.
    Hanley JG, Henley JM (2005) PICK1 is a calcium-sensor for NMDA-induced AMPA receptor trafficking. EMBO J 24:3266–3278CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Steinberg JP, Takamiya K, Shen Y, Xia J, Rubio ME, Yu S, Jin W, Thomas GM, Linden DJ, Huganir RL (2006) Targeted in vivo mutations of the AMPA receptor subunit GluR2 and its interacting protein PICK1 eliminate cerebellar long-term depression. Neuron 49:845–860CrossRefPubMedGoogle Scholar
  16. 16.
    Terashima A, Pelkey KA, Rah JC, Suh YH, Roche KW, Collingridge GL, McBain CJ, Isaac JT (2008) An essential role for PICK1 in NMDA receptor-dependent bidirectional synaptic plasticity. Neuron 57:872–882CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Wang YN, Zhou L, Li YH, Wang Z, Li YC, Zhang YW, Wang Y, Liu G, Shen Y (2015) Protein interacting with C-Kinase 1 deficiency impairs glutathione synthesis and increases oxidative stress via reduction of surface excitatory amino acid carrier 1. J Neurosci 35:6429–6443CrossRefPubMedGoogle Scholar
  18. 18.
    Liu B, Wang K, Gao HM, Mandavilli B, Wang JY, Hong JS (2001) Molecular consequences of activated microglia in the brain: overactivation induces apoptosis. J Neurochem 77:182–189CrossRefPubMedGoogle Scholar
  19. 19.
    Anggono V, Clem RL, Huganir RL (2011) PICK1 loss of function occludes homeostatic synaptic scaling. J Neurosci 31:2188–2196CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Zhu J, Shao CY, Yang W, Zhang XM, Wu ZY, Zhou L, Wang XX, Li YH, Xia J, Luo JH, Shen Y (2012) Chronic zinc exposure decreases the surface expression of NR2A-containing NMDA receptors in cultured hippocampal neurons. PLoS One 7:e46012CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Xie YJ, Zhou L, Jiang N, Zhang N, Zou N, Zhou L, Wang Y, Cowell JK, Shen Y (2015) Essential roles of leucine-rich glioma inactivated 1 in the development of embryonic and postnatal cerebellum. Sci Rep 5:7827CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Nguyen TT, Kim YM, Kim TD, Le OT, Kim JJ, Kang HC, Hasegawa H, Kanaho Y, Jou I, Lee SY (2013) Phosphatidylinositol 4-phosphate 5-kinase α facilitates Toll-like receptor 4-mediated microglial inflammation through regulation of the Toll/interleukin-1 receptor domain-containing adaptor protein (TIRAP) location. J Biol Chem 288:5645–5659CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Neniskyte U, Neher JJ, Brown GC (2011) Neuronal death induced by nanomolar amyloid β is mediated by primary phagocytosis of neurons by microglia. J Biol Chem 286:39904–39913CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Wang DJ, Su LD, Wang YN, Yang D, Sun CL, Zhou L, Wang XX, Shen Y (2014) Long-term potentiation at cerebellar parallel fiber-Purkinje cell synapses requires pre- and postsynaptic signaling cascades. J Neurosci 34:2355–2364CrossRefPubMedGoogle Scholar
  25. 25.
    Wu ZY, Zhu LJ, Zou N, Bombek LK, Shao CY, Wang N, Wang XX, Liang L, Xia J, Rupnik M, Shen Y (2012) AMPA receptors regulate exocytosis and insulin release in pancreatic β cells. Traffic 13:1124–1139CrossRefPubMedGoogle Scholar
  26. 26.
    Mosbacher J, Maier R, Fakler B, Glatz A, Crespo J, Bilbe G (1998) P2Y receptor subtypes differentially couple to inwardly-rectifying potassium channels. FEBS Lett 436:104–110CrossRefPubMedGoogle Scholar
  27. 27.
    Liu GD, Ding JQ, Xiao Q, Chen SD (2009) P2Y6 receptor and immunoinflammation. Neurosci Bull 25:161–164CrossRefPubMedGoogle Scholar
  28. 28.
    Deken SL, Beckman ML, Quick MW (2001) PICKing on transporters. Trends Neurosci 24:623–625CrossRefPubMedGoogle Scholar
  29. 29.
    Cao M, Mao Z, Kam C, Xiao N, Cao X, Shen C, Cheng KKY, Xu A, Lee KM, Jiang L, Xia J (2013) PICK1 and ICA69 control insulin granule trafficking and their deficiencies lead to impaired glucose tolerance. PLoS Biol 11:e1001541CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Department of Microbiology and ImmunologyJiaxing University School of MedicineJiaxingChina
  2. 2.Department of Blood Transfusion, Institute of Molecular MedicineZhejiang Provincial People’s HospitalHangzhouChina
  3. 3.Key Laboratory of Craniocerebral Diseases of Ningxia Hui Autonomous RegionNingxia Medical UniversityYinchuanChina
  4. 4.Department of NeurobiologyZhejiang University School of MedicineHangzhouChina

Personalised recommendations