Abstract
The dorsal root ganglion (DRG) and dorsal horn of the spinal cord are areas through which primary afferent information passes enroute to the brain. Previous studies have reported that, during normal neuronal activity, the regional distribution of a second messenger, diacylglycerol (DG), which is derived from phosphoinositide turnover, is diverse in these areas. However, the way that DG is regulated in these organs remains unknown. The present study was performed to investigate mRNA expression and protein localization of DG kinase (DGK) isozymes, which play a central role in DG metabolism. Gene expression for DGK isozymes was detected with variable regional distributions and intensities in the spinal cord. Among the isozymes, most intense signals were found for DGKζ and DGKι in the DRG. By immunohistochemical analysis, DGKζ immunoreactivity was detected heterogeneously in the nucleus and cytoplasm of small DRG neurons with variable levels of distribution, whereas it was detected exclusively in the cytoplasm of large neurons. On the other hand, DGKι immunoreactivity was distributed solely in the cytoplasm of most of the DRG neurons. Double-immunofluorescent imaging of these isozymes showed that they coexisted in a large population of DRG neurons at distinct subcellular sites, i.e., DGKζ in the nucleus and DGKι in the cytoplasm. Thus, DGK isozymes may have different functional roles at distinct subcellular sites. Furthermore, the heterogeneous subcellular localization of DGKζ between the nucleus and cytoplasm implies the possible translocation of this isozyme in small DRG neurons under various conditions.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Akopian AN, Sivilotti L, Wood JN (1996) A tetrodotoxin-resistant voltage-gated sodium channel expressed by sensory neurons. Nature 379:257–262
Ali H, Nakano T, Saino-Saito S, Hozumi Y, Katagiri Y, Kamii H, Sato S, Kayama T, Kondo H, Goto K (2004) Selective translocation of diacylglycerol kinase ζ in hippocampal neurons under transient forebrain ischemia. Neurosci Lett 372:190–195
Altman J (1982) Morphological development of the rat cerebellum and some of its mechanisms. Exp Brain Res Suppl 6:8–49
Blitterswijk WJ van, Houssa B (2000) Properties and functions of diacylglycerol kinases. Cell Signal 12:595–605
Craner MJ, Klein JP, Renganathan M, black JA, Waxman SG (2002) Changes of sodium channel expression in experimental painful diabetic neuropathy. Ann Neurol 52:786–792
Dib-Hajj S, Black JA, Felts P, Waxman SG (1996) Down-regulation of transcripts for Na channel alpha-SNS in spinal sensory neurons following axotomy. Proc Natl Acad Sci USA 93:14950–14954
Donnerer J, Schuligoi R, Stein C (1992) Increased content and transport of substance P and calcitonin gene-related peptide in sensory nerves innervating inflamed tissue: evidence for a regulatory function of nerve growth factor in vivo. Neuroscience 49:693–698
Goto K, Kondo H (1993) Molecular cloning and expression of a 90-kDa diacylglycerol kinase that predominantly localizes in neurons. Proc Natl Acad Sci USA 90:7598–7602
Goto K, Kondo H (1996) A 104-kDa diacylglycerol kinase containing ankyrin-like repeats localizes in the cell nucleus. Proc Natl Acad Sci USA 93:11196–11201
Goto K, Kondo H (1999) Diacylglycerol kinase in the central nervous system—molecular heterogeneity and gene expression. Chem Phys Lipids 98:109–117
Goto K, Watanabe M, Kondo H, Yuasa H, Sakane F, Kanoh H (1992) Gene cloning, sequence expression and in situ localization of 80 kDa diacylglycerol kinase specific to oligodendrocyte of rat brain. Mol Brain Res 16:75–87
Goto K, Funayama M, Kondo H (1994) Cloning and expression of a cytoskeleton-associated diacylglycerol kinase that is dominantly expressed in the cerebellum. Proc Natl Acad Sci USA 91:13042–13046
Hökfelt T, Wiesenfeld-Hallin Z, Villar M, Melander T (1987) Increase of galanin-like immunoreactivity in rat dorsal root ganglion cells after peripheral axotomy. Neurosci Lett 83:217–220
Hozumi Y, Ito T, Nakano T, Nakagawa T, Aoyagi M, Kondo H, Goto K (2003) Nuclear localization of diacylglycerol kinase ζ in neurons. Eur J Neurosci 18:1448–1457
Ito T, Hozumi Y, Sakane F, Saino-Saito S, Kanoh H, Aoyagi M, Kondo H, Goto K (2004) Cloning and characterization of diacylglycerol kinase ι splice variants in rat brain. J Biol Chem 279:23317–23326
Jones GA, Carpenter G (1993) The regulation of phospholipase C-γ1 by phosphatidic acid. Assessment of kinetic parameters. J Biol Chem 268:20845–20850
Ju G, Hökfelt T, Brodin E, Fahrenkrug J, Fischer JA, Frey P, Elde RP, Brown JC (1987) Primary sensory neurons of the rat showing calcitonin gene-related peptide immunoreactivity and their relation to substance P-, somatostatin-, galanin-, vasoactive intestinal polypeptide- and cholecystokinin-immunoreactive ganglion cells. Cell Tissue Res 247:417–431
Katagiri Y, Ito T, Saino-Saito S, Hozumi Y, Suwabe A, Otake K, Sata M, Kondo H, Sakane F, Kanoh H, Kubota I, Goto K (2005) Expression and localization of diacylglycerol kinase isozymes and enzymatic features in rat lung. Am J Physiol Lung Cell Mol Physiol 288:L1171-L1178
Kawasaki H, Springett GM, Toki S, Canales JJ, Harlan P, Blumenstiel JP, Chen EJ, Bany IA, Mochizuki N, Ashbacher A, Matsuda M, Housman DE, Graybiel AM (1998) A Rap guanine nucleotide exchange factor enriched highly in the basal ganglia. Proc Natl Acad Sci USA 95:13278–13283
Kohyama-Koganeya A, Watanabe M, Hotta Y (1997) Molecular cloning of a diacylglycerol kinase isozyme predominantly expressed in rat retina. FEBS Lett 409:258–264
Levine JD, Fields HL, Basbaum AI (1993) Peptides and the primary afferent nociceptor. J Neurosci 13:2273–2286
Limatola C, Schaap D, Moolenaar WH, Blitterswijk WJ van (1994) Phosphatidic acid activation of protein kinase C-ζ overexpressed in COS cells: comparison with other protein kinase C isotypes and other acidic lipids. Biochem J 304:1001–1008
Lindsay RM (1992) The role of neurotrophic factors in functional maintenance of mature sensory neurons. In: Scott SA (ed) Sensory neurons. Diversity, development, and plasticity. Oxford University Press, Oxford, pp 404–420
Lindsay RM, Lockett C, Sternberg J, Winter J (1989) Neuropeptide expression in cultures of adult sensory neurons: modulation of substance P and calcitonin gene-related peptide levels by nerve growth factor. Neuroscience 33:53–65
Lukacova N, Marsala J (1997) Regional distribution of phospholipids and polyphosphatidyl inositides in the rabbit's spinal cord. Neurochem Res 22:687–692
Nishizuka Y (1984) The role of protein kinase C in cell surface signal transduction and tumor promotion. Nature 308:693–698
Noguchi K, Morita Y, Kiyama H, Ono K, Tohyama M (1988) A noxious stimulus induces the preprotachykinin-A gene expression in the rat dorsal root ganglion: a quantitative study using in situ hybridization histochemistry. Brain Res 464:31–35
Noguchi K, Senba E, Morita Y, Sato M, Tohyama M (1989) Prepro-VIP and preprotachykinin mRNAs in the rat dorsal root ganglion cells following peripheral axotomy. Mol Brain Res 6:327–330
Noguchi K, Senba E, Morita Y, Sato M, Tohyama M (1990) Alpha-CGRP and beta-CGRP mRNAs are differentially regulated in the rat spinal cord and dorsal root ganglion. Mol Brain Res 7:299–304
Rhee SG, Suh PG, Ryu SH, Lee SY (1989) Studies of inositol phospholipid-specific phospholipase C. Science 244:546–550
Ron D, Kazanietz MG (1999) New insights into the regulation of protein kinase C and novel phorbol ester receptors. FASEB J 13:1658–1676
Sakane F, Kanoh H (1997) Molecules in focus: diacylglycerol kinase. Int J Biochem Cell Biol 29:1139–1143
Takeda M, Kagaya Y, Takahashi J, Sugie T, Ohta J, Watanabe J, Shirato K, Kondo H, Goto K (2001) Gene expression and in situ localization of diacylglycerol kinase isozymes in normal and infarcted rat hearts. Circ Res 89:265–272
Toya M, Hozumi Y, Ito T, Takeda M, Sakane F, Kanoh H, Saito H, Hiroi M, Kurachi H, Kondo H, Goto K (2005) Gene expression, cellular localization and enzymatic activity of diacylglycerol kinase isozymes in rat ovary and placenta. Cell Tissue Res 320:525–533
Tsai MH, Yu CL, Stacey DW (1990) A cytoplasmic protein inhibits the GTPase activity of H-Ras in a phospholipid-dependent manner. Science 250:982–985
Watanabe M, Mishina M, Inoue Y (1994) Distinct spatiotemporal expressions of five NMDA receptor channel subunit mRNAs in the cerebellum. J Comp Neurol 343:513–519
Waxman SG, Dib-Hajj S, Cummins TR, Black JA (1999) Sodium channels and pain. Proc Natl Acad Sci USA 96:7635–7639
Author information
Authors and Affiliations
Corresponding author
Additional information
The work was supported by grants-in-aid from the Ministry of Education, Science, Culture, and Sports of Japan (M.T., K.G.) and from the Ono Medical Research Foundation, Kato Memorial Bioscience Foundation, and Janssen Pharmaceutical (K.G.) and by the 21st Century Center of Excellence Program of the Ministry of Education, Culture, Sports, Science and Technology of Japan.
Rights and permissions
About this article
Cite this article
Sasaki, H., Hozumi, Y., Hasegawa, H. et al. Gene expression and localization of diacylglycerol kinase isozymes in the rat spinal cord and dorsal root ganglia. Cell Tissue Res 326, 35–42 (2006). https://doi.org/10.1007/s00441-006-0219-z
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00441-006-0219-z