Abstract
During a variety of insults to the brain adenine nucleotides are released in large quantities from damaged cells, triggering multiple cellular responses to injury. Here, we evaluated changes in extracellular ATP, ADP and AMP hydrolysis at different times (0–24 hours) after unilateral cortical stab injury (CSI) in adult rats. Results demonstrated that 24 hours following CSI, ATP and ADP hydrolyzing activities were not significantly altered in injured cortex. Based on calculated V ATP/V ADP ratio it was concluded that ATP/ADP hydrolysis was primarily catalyzed by NTPDase1 enzyme form. In contrast, AMP hydrolysis, catalyzed by 5’-nucleotidase, was significantly reduced at least 4 hours following CSI. Kinetic analysis and Lineweaver-Burk transformation of the enzyme velocities obtained over the range of AMP concentrations (0.05–1.50 mM) revealed that inhibition of 5’-nucleotidase activity after CSI was of the uncompetitive type. Taken together our data suggest that injured tissue has reduced potential for extracellular metabolism of adenine nucleotides in early stages after CSI.
Similar content being viewed by others
References
Burnstock G (2007) Physiology and pathophysiology of purinergic neurotransmission. Physiol Rev 87:659–797
Abbracchio MP, Burnstock G (1994) Purinoceptors: are there families of P2X and P2Y purinoceptors? Pharmacol Ther 64:445–475
Burnstock G (1972) Purinergic nerves. Pharmac Rev 24:509–581
Rathbone MP, Middlemiss PJ, Gysbers JW et al (1999) Trophic effects of purines in neurons and glial cells. Prog Neurobiol 59:663–690
Vizi ES, Liang SD, Sperlagh B et al (1997) Studies on the release and extracellular metabolism of endogenous ATP in rat superior cervical ganglion: support for neurotransmitter role of ATP. Neuroscience 79:893–903
Zimmermann H, Braun N (1999) Ecto-nucleotidases–molecular structures, catalytic properties, and functional roles in the nervous system. Prog Brain Res 120:371–385
Kegel B, Braun N, Heine P et al (1997) An ecto-ATPase and an ecto-ATP diphosphohydrolase are expressed in rat brain. Neuropharmacology 36:1189–1200
Smith TM, Kirley TL (1998) Cloning, sequencing and expression of a human brain ecto-apyrase. Biochim Biophys Acta 1386:65–78
Kukulski F, Komoszynski M (2003) Purification and characterization of NTPDase 1 (ecto-apyrase) and NTPDase 2 (ecto-ATPase) from porcine brain cortex synaptosomes. Eur J Biochem 270:3447–3454
Zimmermann H (1996) Biochemistry, localization and functional roles of ecto-nucleotidases in the nervous system. Prog Neurobiol 49:589–618
Fredholm BB, Abbracchio MP, Burnstock G et al (1994) Nomenclature and classification of purinoceptors. Pharmacol Rev 46:143–156
Melani A, Turchi D, Vannucchi MG et al (2005) ATP extracellular concentrations are increased in the rat striatum during in vivo ischemia. Neurochem Int 47:442–448
Zimmermann H (1994) Signaling via ATP in the nervous system. Trends Neurosci 17:420–426
Fields RD, Burnstock G (2006) Purinergic signaling in neuron-glia interactions. Nat Rev Neurosci 7:423–436
Rathbone MP, Middlemiss PJ, Kim JK et al (1992) Adenosine and its nucleotides stimulate proliferation of chick astrocytes and human astrocytoma cells. Neurosci Res 13:1–17
Middlemiss PJ, Gysbers JW, Rathbone MP (1995) Extracellular guanosine and guanosine-5’-triphosphate increase: NGF synthesis and release from cultured mouse neopallial astrocytes. Brain Res 677:152–156
Ferrari D, Chiozzi P, Falzoni S et al (1997) ATP-mediated cytotoxicity in microglial cells. Neuropharmacol 36:1295–1301
Neary JT, Kang Y, Shi YF et al (2006) P2 receptor signalling, proliferation of astrocytes, expression of molecules involved in cell-cell interactions. Novarstis Found Symp 276:131–143
Sorimachi M, Yamagami K, Wakomori M (2002) Activation of ATP receptor increases the cytosolic Ca(2+) concentration in ventral tegmental area neurons of rat brain. Brain Res 935:129–133
Inoue K, Koizumi S, Nakazawa K (1995) Glutamate-evoked release of adenosine 5’-triphosphate causing an increase in intracellular calcium in hippocampal neurons. Neuroreport 6:437–440
Nedeljkovic N, Bjelobaba I, Subasic S et al (2006) Up-regulation of ectonucleotidase activity after cortical stab injury in rats. Cell Biol Int 30:541–546
Nagy AK, Walton NY, Treiman DM (1997) Reduced cortical ecto-ATPase activity in rat brains during prolonged status epilepticus induced by sequential administration of lithium and pilocarpine. Mol Chem Neuropathol 31:135–147
Bonan CD, Walz R, Pereira GS et al (2000) Changes in synaptosomal ectonucleotidase activities in two rat models of temporal lobe epilepsy. Epilepsy Res 39:229–238
Bonan CD, Amaral OB, Rockenbach IC et al (2000) Altered ATP hydrolysis induced by pentylentetrazol kindling in rat brain synaptosomes. Neurochem Res 25:775–779
Braun N, Zhu Y, Krieglstein J et al (1998) Upregulation of the enzyme chain hydrolyzing extracellular ATP after transient ferebrain ischemia in the rat. J Neurosci 18:4891–4900
Villa RF, Gorini A, Hoyer S (2002) ATPases of synaptic plasma membranes from hippocampus after ischemia and recovery during aging. Neurochem Res 27:861–870
Neary JT, Rathbone MP, Cattabeni F et al (1996) Trophic actions of extracellular nucleotides and nucleosides on glial and neuronal cells. Trends Neurosci 19:13–18
Chen S, Pickard JD, Harris NG (2003) Time course of cellular pathology after controlled cortical impact injury. Exp Neurol 182:87–102
Braun N, Lenz C, Gillardon F et al (1997) Focal cerebral ischemia enhances glial expression of ecto-5’-nucleotidase. Brain Res 766:213–226
Gray EG, Whittaker VP (1962) The isolation of nerve endings from brain: an electron microscopic study of cell fragments derived by homogenization and centrifugation. J Anat 96:79–88
Nedeljkovic N, Nikezic G, Horvat A et al (1998) Properties of Mg(2+)-ATPase rat brain synaptic plasma membranes. Gen Physiol Biophys 17:3–13
Markwell MA, Haas SA, Lieber L et al (1978) A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. Anal Biochem 87:206–210
Nedeljkovic N, Banjac A, Horvat A et al (2005) Developmental profile of NTPDase activity in synaptic plasma membranes isolated from rat cerebral cortex. Int J Dev Neurosci 23:45–51
Pennial R (1966) An improved method for determination of inorganic phosphate by the isobutanol-benzen extraction procedure. Anal Biochem 14:87–90
Nedeljkovic N, Banjac A, Horvat A et al (2003) Ecto-ATPase and ecto-ATP-diphosphohydrolase are co-localized in rat hippocampal and caudate nucleus synaptic plasma membranes. Physiol Res 52:797–804
Stone TW (2005) Adenosine, neurodegeneration and neuroprotection. Neurol Res 27:161–168
Davalos D, Grutzendler J, Yang G et al (2005) ATP mediates rapid microglial response to local brain injury in vivo. Nat Neurosci 8:752–758
Van der Toorn A, Sykova E, Dijkhuizen RM et al (1996) Dynamic changes in water ADC, energy metabolism, extracellular space volume and tortuosity in neonatal brain during global ischemia. Magn Reson Med 36:52–60
Wang Y, Roman R, Lidofsky SD et al (1996) Autocrine signaling through ATP release represents a novel mechanism for cell volume regulation. Proc Natl Acad Sci USA 93:12020–12025
Burger RM, Lowenstein JM (1975) 5’-Nucleotidase from smooth muscle of small intestine and from brain. Inhibition of nucleotides. Biochemistry 14:2362–2366
Naito Y, Lowenstein JM (1985) 5’-Nucleotidase from rat heart membranes. Inhibition by adenine nucleotides and related compounds. Biochem J 226:645–651
Gordon EL, Pearson JD, Slakey LL (1985) The hydrolysis of extracellular adenine nucleotides by cultured endothelial cells from pig aorta. Feed-forward inhibition of adenosine production at the cell surface. J Biol Chem 261:15496–15507
Navarro JM, Olmo N, Turnay J et al (1998) Ecto-5’-nucleotidase from a human colon adrenocarcinoma cell line. Correlation between enzyme activity and levels in intact cells. Mol Cell Biochem 187:121–131
James S, Richardson PJ (1993) The subcellular distribution of [3H]-CGS 21680 binding sites in the rat striatum: copurification with cholinergic nerve terminals. Neurochem Int 23:115–122
Delwing D, Delwing D, Sarkis JJ et al (2007) Proline induces alterations on nucleotide hydrolysis in synaptosomes from cerebral cortex of rats. Brain Res 1149:210–215
Dale N (1998) Delayed production of adenosine underlies temporal modulation of swimming in frog embryo. J Gen Physiol 511:265–272
During MJ, Spencer DD (1992) Adenosine: a potential mediator of seizure arrest and postictal refractoriness. Ann Neurol 32:618–624
Weigand MA, Michel A, Eckstein HH et al (1999) Adenosine: a sensitive indicator of cerebral ischemia during carotid endarterectomy. Anesthesiology 91:414–421
Clark RS, Carcillio JA, Kochanek PM et al (1997) Cerebrospinal fluid adenosine concentration and uncoupling of cerebral blood flow and oxidative metabolism after severe head injury in humans. Neurosurgery 41:1284–1292
Robertson CL, Bell MJ, Kochanek PM et al (2001) Increased adenosine in cerebrospinal fluid after severe traumatic brain injury in infants and children: association with severity of injury and excitotoxicity. Crit Care Med 29:2287–2293
Frenguelli BG, Wigmore G, Llaudet E et al (2007) Temporal and mechanistic dissociation of ATP and adenosine during ischemia in the mammalian hippocampus. J Neurochem 101:1400–1413
Acknowledgements
The study was supported by Serbian Ministry of Science, Grant No. 143005.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Nedeljkovic, N., Bjelobaba, I., Lavrnja, I. et al. Early Temporal Changes in Ecto-Nucleotidase Activity after Cortical Stab Injury in Rat. Neurochem Res 33, 873–879 (2008). https://doi.org/10.1007/s11064-007-9529-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11064-007-9529-0