J Vet Sci.  2010 Dec;11(4):299-304. 10.4142/jvs.2010.11.4.299.

Expression of KA1 kainate receptor subunit in the substantia gelatinosa of the trigeminal subnucleus caudalis in mice

• Park SA
• Park SJ
• Han SK

• KMID: 1072178
• DOI: http://doi.org/10.4142/jvs.2010.11.4.299

Abstract

The KA1 kainate receptor (KAR) subunit in the substantia gelatinosa (SG) of the trigeminal subnucleus caudalis (Vc) has been implicated in the processing of nociceptive information from the orofacial region. This study compared the expression of the KA1 KAR subunit in the SG of the Vc in juvenile, prepubescent and adult mice. RT-PCR, Western blot and immunohistochemistry analyses were used to examine the expression level in SG area. The expression levels of the KA1 KAR subunit mRNA and protein were higher in juvenile mice than in prepubescent or adult mice. Quantitative data revealed that the KA1 KAR subunit mRNA and protein were expressed at levels approximately two and three times higher, respectively, in juvenile mice than in adult mice. A similar expression pattern of the KA1 KAR subunit was observed in an immunohistochemical study that showed higher expression in the juvenile (59%) than those of adult (35%) mice. These results show that the KA1 KAR subunits are expressed in the SG of the Vc in mice and that the expression level of the KA1 KAR subunit decreases gradually with postnatal development. These findings suggest that age-dependent KA1 KAR subunit expression can be a potential mechanism of age-dependent pain perception.

Keyword

immunohistochemistry; KA1; RT-PCR; substantia gelatinosa; trigeminal subnucleus caudalis; Western blot

MeSH Terms

Age Factors
Animals
Gene Expression Profiling
*Gene Expression Regulation, Developmental
Mice
Receptors, Kainic Acid/*metabolism
Substantia Gelatinosa/*metabolism

Figure

• 

  Fig. 1 (A) Expression of the KA1 subunit mRNA in the substantia gelatinosa (SG) of the trigeminal subnucleus caudalis (Vc). The KA1 subunit is more strongly expressed in juvenile than prepubescent or adult mice. (B) The relative expression of normalized KA1 mRNA in juvenile, prepubescent and adult mice. *p<0.05. NS: not significant, GAPDH: glyceraldehyde-3-phosphate dehydrogenase.

• 

  Fig. 2 KA1 subunit protein expressed in the punched lamina II containing SG neurons. Expression of the KA1 subunit decreased gradually with postnatal development.

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  Fig. 3 The KA1 subunit was expressed differently in juvenile and adult mice. (A) Immunoreactivities for the KA1 subunit containing kainate receptors were observed in the SG of the Vc in juvenile (Aa) and adult (Ab) mice. The closed and open arrowheads indicate intense and weak expression of KA1 subunits, respectively. (B) The percentage of neurons expressing the KA1 subunit in the juveniles and adults. *p<0.05. Immunohistochemical staining and couterstain with hematoxylin, ×400.

Reference

1. Bureau I, Dieudonne S, Coussen F, Mulle C. Kainate receptor-mediated synaptic currents in cerebellar Golgi cells are not shaped by diffusion of glutamate. Proc Natl Acad Sci USA. 2000. 97:6838–6843.

2. Chen ZL, Yu H, Yu WM, Pawlak R, Strickland S. Proteolytic fragments of laminin promote excitotoxic neurodegeneration by up-regulation of the KA1 subunit of the kainate receptor. J Cell Biol. 2008. 183:1299–1313.

3. Chiang CY, Hu JW, Sessle BJ. NMDA receptor involvement in neuroplastic changes induced by neonatal capsaicin treatment in trigeminal nociceptive neurons. J Neurophysiol. 1997. 78:2799–2803.

4. Contractor A, Sailer AW, Darstein M, Maron C, Xu J, Swanson GT, Heinemann SF. Loss of kainate receptor-mediated heterosynaptic facilitation of mossy-fiber synapses in KA2-/- mice. J Neurosci. 2003. 23:422–429.

5. Cui C, Mayer ML. Heteromeric kainate receptors formed by the coassembly of GluR5, GluR6, and GluR7. J Neurosci. 1999. 19:8281–8291.

6. Fernandes HB, Catches JS, Petralia RS, Copits BA, Xu J, Russell TA, Swanson GT, Contractor A. High-affinity kainate receptor subunits are necessary for ionotropic but not metabotropic signaling. Neuron. 2009. 63:818–829.

7. Furuyama T, Kiyama H, Sato K, Park HT, Maeno H, Takagi H, Tohyama M. Region-specific expression of subunits of ionotropic glutamate receptors (AMPA-type, KA-type and NMDA receptors) in the rat spinal cord with special reference to nociception. Brain Res Mol Brain Res. 1993. 18:141–151.

8. Gobel S, Falls WM, Bennett GJ, Abdelmoumene M, Hayashi H, Humphrey E. An EM analysis of the synaptic connections of horseradish peroxidase-filled stalked cells and islet cells in the substantia gelatinosa of adult cat spinal cord. J Comp Neurol. 1980. 194:781–807.

9. Guo W, Zou S, Tal M, Ren K. Activation of spinal kainate receptors after inflammation: behavioral hyperalgesia and subunit gene expression. Eur J Pharmacol. 2002. 452:309–318.

10. Hantman AW, van den Pol AN, Perl ER. Morphological and physiological features of a set of spinal substantia gelatinosa neurons defined by green fluorescent protein expression. J Neurosci. 2004. 24:836–842.

11. Huettner JE. Glutamate receptor channels in rat DRG neurons: activation by kainate and quisqualate and blockade of desensitization by Con A. Neuron. 1990. 5:255–266.

12. Huettner JE. Kainate receptors and synaptic transmission. Prog Neurobiol. 2003. 70:387–407.

13. Hunter RG, Bellani R, Bloss E, Costa A, McCarthy K, McEwen BS. Regulation of kainate receptor subunit mRNA by stress and corticosteroids in the rat hippocampus. PLoS One. 2009. 4:e4328.

14. Hwang SJ, Pagliardini S, Rustioni A, Valtschanoff JG. Presynaptic kainate receptors in primary afferents to the superficial laminae of the rat spinal cord. J Comp Neurol. 2001. 436:275–289.

15. Kalb RG, Fox AJ. Synchronized overproduction of AMPA, kainate, and NMDA glutamate receptors during human spinal cord development. J Comp Neurol. 1997. 384:200–210.

16. Kerchner GA, Wang GD, Qiu CS, Huettner JE, Zhuo M. Direct presynaptic regulation of GABA/glycine release by kainate receptors in the dorsal horn: an ionotropic mechanism. Neuron. 2001. 32:477–488.

17. Kerchner GA, Wilding TJ, Li P, Zhuo M, Huettner JE. Presynaptic kainate receptors regulate spinal sensory transmission. J Neurosci. 2001. 21:59–66.

18. Kumazawa T, Perl ER. Excitation of marginal and substantia gelatinosa neurons in the primate spinal cord: indications of their place in dorsal horn functional organization. J Comp Neurol. 1978. 177:417–434.

19. Lerma J. Roles and rules of kainate receptors in synaptic transmission. Nat Rev Neurosci. 2003. 4:481–495.

20. Lerma J. Kainate receptor physiology. Curr Opin Pharmacol. 2006. 6:89–97.

21. Leszek P, Korewicki J, Klisiewicz A, Janas J, Biederman A, Browarek A, Charlemagne D, Trouvé P. Reduced myocardial expression of calcium handling protein in patients with severe chronic mitral regurgitation. Eur J Cardiothorac Surg. 2006. 30:737–743.

22. Li P, Wilding TJ, Kim SJ, Calejesan AA, Huettner JE, Zhuo M. Kainate-receptor-mediated sensory synaptic transmission in mammalian spinal cord. Nature. 1999. 397:161–164.

23. Light AR, Kavookjian AM. Morphology and ultrastructure of physiologically identified substantia gelatinosa (lamina II) neurons with axons that terminate in deeper dorsal horn laminae (III-V). J Comp Neurol. 1988. 267:172–189.

24. Light AR, Trevino DL, Perl ER. Morphological features of functionally defined neurons in the marginal zone and substantia gelatinosa of the spinal dorsal horn. J Comp Neurol. 1979. 186:151–171.

25. Lu CR, Willcockson HH, Phend KD, Lucifora S, Darstein M, Valtschanoff JG, Rustioni A. Ionotropic glutamate receptors are expressed in GABAergic terminals in the rat superficial dorsal horn. J Comp Neurol. 2005. 486:169–178.

26. Lucifora S, Willcockson HH, Lu CR, Darstein M, Phend KD, Valtschanoff JG, Rustioni A. Presynaptic low- and high-affinity kainate receptors in nociceptive spinal afferents. Pain. 2006. 120:97–105.

27. Monnerie H, Le Roux PD. Glutamate receptor agonist kainate enhances primary dendrite number and length from immature mouse cortical neurons in vitro. J Neurosci Res. 2006. 83:944–956.

28. Mulle C, Sailer A, Pérez-Otaño I, Dickinson-Anson H, Castillo PE, Bureau I, Maron C, Gage FH, Mann JR, Bettler B, Heinemann SF. Altered synaptic physiology and reduced susceptibility to kainate-induced seizures in GluR6-deficient mice. Nature. 1998. 392:601–605.

29. Mulle C, Sailer A, Swanson GT, Brana C, O'Gorman S, Bettler B, Heinemann SF. Subunit composition of kainate receptors in hippocampal interneurons. Neuron. 2000. 28:475–484.

30. Pan YZ, Pan HL. Primary afferent stimulation differentially potentiates excitatory and inhibitory inputs to spinal lamina II outer and inner neurons. J Neurophysiol. 2004. 91:2413–2421.

31. Partin KM, Patneau DK, Winters CA, Mayer ML, Buonanno A. Selective modulation of desensitization at AMPA versus kainate receptors by cyclothiazide and concanavalin A. Neuron. 1993. 11:1069–1082.

32. Petralia RS, Wang YX, Wenthold RJ. Histological and ultrastructural localization of the kainate receptor subunits, KA2 and GluR6/7, in the rat nervous system using selective antipeptide antibodies. J Comp Neurol. 1994. 349:85–110.

33. Pinheiro PS, Perrais D, Coussen F, Barhanin J, Bettler B, Mann JR, Malva JO, Heinemann SF, Mulle C. GluR7 is an essential subunit of presynaptic kainate autoreceptors at hippocampal mossy fiber synapses. Proc Natl Acad Sci USA. 2007. 104:12181–12186.

34. Ririe DG, Eisenach JC. Age-dependent responses to nerve injury-induced mechanical allodynia. Anesthesiology. 2006. 104:344–350.

35. Ritter LM, Vazquez DM, Meador-Woodruff JH. Ontogeny of ionotropic glutamate receptor subunit expression in the rat hippocampus. Brain Res Dev Brain Res. 2002. 139:227–236.

36. Rozen S, Skaletsky H. Misener S, Krawetz S, editors. Primer3 on the WWW for general users and for biologist programmers. Bioinformatics Methods and Protocols: Methods in Molecular Biology. 2000. Totowa: Humana Press;365–386.

37. Sato K, Kiyama H, Park HT, Tohyama M. AMPA, KA and NMDA receptors are expressed in the rat DRG neurones. Neuroreport. 1993. 4:1263–1265.

38. Sessle BJ. Acute and chronic craniofacial pain: brainstem mechanisms of nociceptive transmission and neuroplasticity, and their clinical correlates. Crit Rev Oral Biol Med. 2000. 11:57–91.

39. Simmons RM, Li DL, Hoo KH, Deverill M, Ornstein PL, Iyengar S. Kainate GluR5 receptor subtype mediates the nociceptive response to formalin in the rat. Neuropharmacology. 1998. 37:25–36.

40. Sommer B, Burnashev N, Verdoorn TA, Keinänen K, Sakmann B, Seeburg PH. A glutamate receptor channel with high affinity for domoate and kainate. EMBO J. 1992. 11:1651–1656.

41. Stegenga SL, Kalb RG. Developmental regulation of N-methyl-D-aspartate- and kainate-type glutamate receptor expression in the rat spinal cord. Neuroscience. 2001. 105:499–507.

42. Tölle TR, Berthele A, Zieglgänsberger W, Seeburg PH, Wisden W. The differential expression of 16 NMDA and non-NMDA receptor subunits in the rat spinal cord and in periaqueductal gray. J Neurosci. 1993. 13:5009–5028.

43. Ullal G, Fahnestock M, Racine R. Time-dependent effect of kainate-induced seizures on glutamate receptor GluR5, GluR6, and GluR7 mRNA and protein expression in rat hippocampus. Epilepsia. 2005. 46:616–623.

44. Walker SM, Howard RF, Keay KA, Fitzgerald M. Developmental age influences the effect of epidural dexmedetomidine on inflammatory hyperalgesia in rat pups. Anesthesiology. 2005. 102:1226–1234.

45. Wu LJ, Ko SW, Zhuo M. Kainate receptors and pain: from dorsal root ganglion to the anterior cingulate cortex. Curr Pharm Des. 2007. 13:1597–1605.

46. Xu H, Wu LJ, Zhao MG, Toyoda H, Vadakkan KI, Jia Y, Pinaud R, Zhuo M. Presynaptic regulation of the inhibitory transmission by GluR5-containing kainate receptors in spinal substantia gelatinosa. Mol Pain. 2006. 2:29.

47. Youn DH, Randic M. Modulation of excitatory synaptic transmission in the spinal substantia gelatinosa of mice deficient in the kainate receptor GluR5 and/or GluR6 subunit. J Physiol. 2004. 555:683–698.

48. Ziak D, Chvátal A, Syková E. Glutamate-, kainate- and NMDA-evoked membrane currents in identified glial cells in rat spinal cord slice. Physiol Res. 1998. 47:365–375.