FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / Metab Brain Dis / Effect of serum on intracellular calcium homeostasis and sur...
Metabolic brain disease1994; 9(4); 333-345; doi: 10.1007/BF02098880

Effect of serum on intracellular calcium homeostasis and survival of primary cortical and hippocampal CA1 neurons following brief glutamate treatment.

Abstract: Glutamate neurotoxicity was studied in primary neuronal cultures prepared from rat cerebral cortex and hippocampal CA1 sector. Neurons were cultivated with 5% native horse serum and then exposed to 0.1 or 1.0 mM glutamate for 5 min. Subsequently, neurons were allowed to recover for 24 hours either in the presence or in the absence of 5% native horse serum. In the absence of serum, neurons showed morphological signs of degeneration and exhibited marked loss of vitality as tested by vital staining and release of lactate dehydrogenase (LDH). In contrast, when neurons were cultivated in the presence of serum, no degenerative changes were seen and the neurons survived. Heat inactivated serum did not prevent neuronal death but addition of basic fibroblast growth factor (bFGF) or transforming growth factor-beta 1 (TGF-beta 1) had the same protective effect as native serum. Measurements of intracellular calcium activity ([Ca2+]i) with the indicator dye fura-2 revealed a sharp increase during glutamate exposure. In the absence of serum, [Ca2+]i returned to near control within 5 min but it secondarily increased after 1 hour to almost the same level as during glutamate exposure. This delayed increase was more pronounced in CA1 than in cortical neurons, it correlated linearly with the initial rise during glutamate exposure, and it was greatly reduced in the presence of serum. These observations suggest that glutamate neurotoxicity in vitro is a function of the delayed and not of the primary rise of intracellular calcium activity, and that trophic factors prevent neurotoxicity by attenuating this delayed response.
Get Full Text
Publication Date: 1994-12-01 PubMed ID: 7898400DOI: 10.1007/BF02098880Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research article studies the damaging effects of the neurotransmitter glutamate in brain neurons and how these effects may be alleviated with the presence of serum. It also discusses the potential neuroprotective qualities of certain growth factors.

Research Context and Method

  • The focus of the research was on glutamate neurotoxicity, which occurs when excess glutamate destroys neurons in the brain.
  • The researchers used primary neuronal cultures from rat cerebral cortex and hippocampal CA1 sector for the experiment.
  • The neural cells were cultured with 5% native horse serum and then exposed to certain concentrations of glutamate.
  • Following the exposure, the neurons were either allowed to recover in the presence or absence of serum for 24 hours.

Findings

  • When there was no serum present during recovery, the neurons exhibited signs of degeneration and a significant loss of vitality. This was tested by using vital staining and assessing the release of lactate dehydrogenase (LDH), a marker for cellular damage.
  • However, when serum was present, the neurons showed no signs of degeneration and survived.
  • Heat inactivating the serum did not prevent neuronal death, indicating the role of bioactive compounds in protection.

Role of Growth Factors

  • Adding fundamental growth factors like basic fibroblast growth factor (bFGF) or transforming growth factor-beta 1 (TGF-beta 1) replicated the protective effect of the native serum.
  • This suggests that the serum was providing trophic factors which helped prevent neurotoxic effects.

Role of Calcium

  • Measurements of intracellular calcium activity with the indicator dye fura-2 showed a sharp increase during glutamate exposure.
  • In the absence of serum, the calcium activity returned to near control levels but increased again after an hour.
  • This delayed increase was more noticeable in some neurons than others, and it was significantly reduced in the presence of serum.
  • The researchers concluded that the toxic effects of glutamate were correlated more with the delayed rise in intracellular calcium levels, rather than the immediate rise in calcium levels after glutamate exposure.

Implications

  • The findings suggest a potential therapeutic avenue for conditions involving glutamate neurotoxicity, such as stroke or Alzheimer’s disease.
  • Further research into the nature of the trophic factors and calcium activity could yield applicable clinical strategies.

Cite This Article

APA
Uto A, Dux E, Hossmann KA. (1994). Effect of serum on intracellular calcium homeostasis and survival of primary cortical and hippocampal CA1 neurons following brief glutamate treatment. Metab Brain Dis, 9(4), 333-345. https://doi.org/10.1007/BF02098880

Publication

Metabolic brain disease
ISSN: 0885-7490
NlmUniqueID: 8610370
Country: United States
Language: English
Volume: 9
Issue: 4
Pages: 333-345

Researcher Affiliations

Uto, A
  • Max-Planck-Institute for Neurological Research, Department of Experimental Neurology, Cologne, Germany.
Dux, E
    Hossmann, K A

      MeSH Terms

      • Animals
      • Blood
      • Blood Proteins / physiology
      • Calcium / metabolism
      • Cell Death / drug effects
      • Cell Survival / drug effects
      • Cells, Cultured
      • Glutamic Acid / toxicity
      • Hippocampus / drug effects
      • Horses
      • L-Lactate Dehydrogenase
      • Nerve Degeneration
      • Neurons / drug effects
      • Neurons / metabolism
      • Rats
      • Rats, Wistar

      References

      This article includes 54 references
      1. Hahn JS, Aizenman E, Lipton SA. Central mammalian neurons normally resistant to glutamate toxicity are made sensitive by elevated extracellular Ca2+: toxicity is blocked by the N-methyl-D-aspartate antagonist MK-801.. Proc Natl Acad Sci U S A 1988 Sep;85(17):6556-60.
        pubmed: 2901101doi: 10.1073/pnas.85.17.6556google scholar: lookup
      2. Rothman SM, Olney JW. Glutamate and the pathophysiology of hypoxic--ischemic brain damage.. Ann Neurol 1986 Feb;19(2):105-11.
        pubmed: 2421636doi: 10.1002/ana.410190202google scholar: lookup
      3. Zhang Y, Tatsuno T, Carney JM, Mattson MP. Basic FGF, NGF, and IGFs protect hippocampal and cortical neurons against iron-induced degeneration.. J Cereb Blood Flow Metab 1993 May;13(3):378-88.
        pubmed: 8478396doi: 10.1038/jcbfm.1993.51google scholar: lookup
      4. Randall RD, Thayer SA. Glutamate-induced calcium transient triggers delayed calcium overload and neurotoxicity in rat hippocampal neurons.. J Neurosci 1992 May;12(5):1882-95.
        pubmed: 1349638doi: 10.1523/JNEUROSCI.12-05-01882.1992google scholar: lookup
      5. Miyamoto M, Murphy TH, Schnaar RL, Coyle JT. Antioxidants protect against glutamate-induced cytotoxicity in a neuronal cell line.. J Pharmacol Exp Ther 1989 Sep;250(3):1132-40.
        pubmed: 2778712
      6. Tymianski M, Charlton MP, Carlen PL, Tator CH. Secondary Ca2+ overload indicates early neuronal injury which precedes staining with viability indicators.. Brain Res 1993 Apr 2;607(1-2):319-23.
        pubmed: 7683241doi: 10.1016/0006-8993(93)91523-ugoogle scholar: lookup
      7. Séquier JM, Hunziker W, Andressen C, Celio MR. Calbindin D-28k Protein and mRNA Localization in the Rat Brain.. Eur J Neurosci 1990;2(12):1118-1126.
        pubmed: 12106072doi: 10.1111/j.1460-9568.1990.tb00023.xgoogle scholar: lookup
      8. Maiese K, Boniece I, DeMeo D, Wagner JA. Peptide growth factors protect against ischemia in culture by preventing nitric oxide toxicity.. J Neurosci 1993 Jul;13(7):3034-40.
        pubmed: 7687284doi: 10.1523/JNEUROSCI.13-07-03034.1993google scholar: lookup
      9. Shigeno T, Mima T, Takakura K, Graham DI, Kato G, Hashimoto Y, Furukawa S. Amelioration of delayed neuronal death in the hippocampus by nerve growth factor.. J Neurosci 1991 Sep;11(9):2914-9.
        pubmed: 1880556doi: 10.1523/JNEUROSCI.11-09-02914.1991google scholar: lookup
      10. Yamamori T. Molecular mechanisms for generation of neural diversity and specificity: roles of polypeptide factors in development of postmitotic neurons.. Neurosci Res 1992 Jan;12(5):545-82.
        pubmed: 1313952doi: 10.1016/0168-0102(92)90064-jgoogle scholar: lookup
      11. Griffiths T, Evans MC, Meldrum BS. Status epilepticus: the reversibility of calcium loading and acute neuronal pathological changes in the rat hippocampus.. Neuroscience 1984 Jun;12(2):557-67.
        pubmed: 6462462doi: 10.1016/0306-4522(84)90073-3google scholar: lookup
      12. Dienel GA, Pulsinelli WA. Uptake of radiolabeled ions in normal and ischemia-damaged brain.. Ann Neurol 1986 May;19(5):465-72.
        pubmed: 3013076doi: 10.1002/ana.410190507google scholar: lookup
      13. Siesjö BK, Memezawa H, Smith ML. Neurocytotoxicity: pharmacological implications.. Fundam Clin Pharmacol 1991;5(9):755-67.
        pubmed: 1686604doi: 10.1111/j.1472-8206.1991.tb00765.xgoogle scholar: lookup
      14. Dux E, Oschlies U, Wiessner C, Hossmann KA. Glutamate-induced ribosomal disaggregation and ultrastructural changes in rat cortical neuronal culture: protective effect of horse serum.. Neurosci Lett 1992 Jul 20;141(2):173-6.
        pubmed: 1359467doi: 10.1016/0304-3940(92)90888-egoogle scholar: lookup
      15. Ullrich A, Schlessinger J. Signal transduction by receptors with tyrosine kinase activity.. Cell 1990 Apr 20;61(2):203-12.
        pubmed: 2158859doi: 10.1016/0092-8674(90)90801-kgoogle scholar: lookup
      16. Rothman SM. Synaptic activity mediates death of hypoxic neurons.. Science 1983 Apr 29;220(4596):536-7.
        pubmed: 6836300doi: 10.1126/science.6836300google scholar: lookup
      17. Lehmann A. Pharmacological protection against the toxicity of N-methyl-D-aspartate in immature rat cerebellar slices.. Neuropharmacology 1987 Dec;26(12):1751-61.
        pubmed: 3325847doi: 10.1016/0028-3908(87)90128-6google scholar: lookup
      18. Cheng B, McMahon DG, Mattson MP. Modulation of calcium current, intracellular calcium levels and cell survival by glucose deprivation and growth factors in hippocampal neurons.. Brain Res 1993 Apr 2;607(1-2):275-85.
        pubmed: 8386974doi: 10.1016/0006-8993(93)91517-vgoogle scholar: lookup
      19. Prehn JH, Peruche B, Unsicker K, Krieglstein J. Isoform-specific effects of transforming growth factors-beta on degeneration of primary neuronal cultures induced by cytotoxic hypoxia or glutamate.. J Neurochem 1993 May;60(5):1665-72.
        pubmed: 8097233doi: 10.1111/j.1471-4159.1993.tb13389.xgoogle scholar: lookup
      20. Mattson MP, Rychlik B, Chu C, Christakos S. Evidence for calcium-reducing and excito-protective roles for the calcium-binding protein calbindin-D28k in cultured hippocampal neurons.. Neuron 1991 Jan;6(1):41-51.
        pubmed: 1670921doi: 10.1016/0896-6273(91)90120-ogoogle scholar: lookup
      21. Sloviter RS. Calcium-binding protein (calbindin-D28k) and parvalbumin immunocytochemistry: localization in the rat hippocampus with specific reference to the selective vulnerability of hippocampal neurons to seizure activity.. J Comp Neurol 1989 Feb 8;280(2):183-96.
        pubmed: 2925892doi: 10.1002/cne.902800203google scholar: lookup
      22. Hayes RL, Jenkins LW, Lyeth BG, Balster RL, Robinson SE, Clifton GL, Stubbins JF, Young HF. Pretreatment with phencyclidine, an N-methyl-D-aspartate antagonist, attenuates long-term behavioral deficits in the rat produced by traumatic brain injury.. J Neurotrauma 1988;5(4):259-74.
        pubmed: 2854855doi: 10.1089/neu.1988.5.259google scholar: lookup
      23. Hara H, Onodera H, Yoshidomi M, Matsuda Y, Kogure K. Staurosporine, a novel protein kinase C inhibitor, prevents postischemic neuronal damage in the gerbil and rat.. J Cereb Blood Flow Metab 1990 Sep;10(5):646-53.
        pubmed: 2384538doi: 10.1038/jcbfm.1990.117google scholar: lookup
      24. Widmer HR, Knüsel B, Hefti F. Stimulation of phosphatidylinositol hydrolysis by brain-derived neurotrophic factor and neurotrophin-3 in rat cerebral cortical neurons developing in culture.. J Neurochem 1992 Dec;59(6):2113-24.
        pubmed: 1431896doi: 10.1111/j.1471-4159.1992.tb10102.xgoogle scholar: lookup
      25. Prehn JH, Backhauss C, Krieglstein J. Transforming growth factor-beta 1 prevents glutamate neurotoxicity in rat neocortical cultures and protects mouse neocortex from ischemic injury in vivo.. J Cereb Blood Flow Metab 1993 May;13(3):521-5.
        pubmed: 8097519doi: 10.1038/jcbfm.1993.67google scholar: lookup
      26. Mattson MP, Guthrie PB, Kater SB. A role for Na+-dependent Ca2+ extrusion in protection against neuronal excitotoxicity.. FASEB J 1989 Nov;3(13):2519-26.
        pubmed: 2572500doi: 10.1096/fasebj.3.13.2572500google scholar: lookup
      27. Choi DW, Maulucci-Gedde M, Kriegstein AR. Glutamate neurotoxicity in cortical cell culture.. J Neurosci 1987 Feb;7(2):357-68.
        pubmed: 2880937doi: 10.1523/JNEUROSCI.07-02-00357.1987google scholar: lookup
      28. Tymianski M, Wallace MC, Spigelman I, Uno M, Carlen PL, Tator CH, Charlton MP. Cell-permeant Ca2+ chelators reduce early excitotoxic and ischemic neuronal injury in vitro and in vivo.. Neuron 1993 Aug;11(2):221-35.
        pubmed: 8102532doi: 10.1016/0896-6273(93)90180-ygoogle scholar: lookup
      29. Nicotera P, Bellomo G, Orrenius S. The role of Ca2+ in cell killing.. Chem Res Toxicol 1990 Nov-Dec;3(6):484-94.
        pubmed: 2103319doi: 10.1021/tx00018a001google scholar: lookup
      30. Iacopino A, Christakos S, German D, Sonsalla PK, Altar CA. Calbindin-D28K-containing neurons in animal models of neurodegeneration: possible protection from excitotoxicity.. Brain Res Mol Brain Res 1992 Apr;13(3):251-61.
        pubmed: 1317497doi: 10.1016/0169-328x(92)90033-8google scholar: lookup
      31. Baimbridge KG, Miller JJ. Immunohistochemical localization of calcium-binding protein in the cerebellum, hippocampal formation and olfactory bulb of the rat.. Brain Res 1982 Aug 12;245(2):223-9.
        pubmed: 6751467doi: 10.1016/0006-8993(82)90804-6google scholar: lookup
      32. Harafuji H, Ogawa Y. Re-examination of the apparent binding constant of ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid with calcium around neutral pH.. J Biochem 1980 May;87(5):1305-12.
        pubmed: 6771253doi: 10.1093/oxfordjournals.jbchem.a132868google scholar: lookup
      33. Crumrine RC, Dubyak G, LaManna JC. Decreased protein kinase C activity during cerebral ischemia and after reperfusion in the adult rat.. J Neurochem 1990 Dec;55(6):2001-7.
        pubmed: 2230806doi: 10.1111/j.1471-4159.1990.tb05788.xgoogle scholar: lookup
      34. Bonni A, Frank DA, Schindler C, Greenberg ME. Characterization of a pathway for ciliary neurotrophic factor signaling to the nucleus.. Science 1993 Dec 3;262(5139):1575-9.
        pubmed: 7504325doi: 10.1126/science.7504325google scholar: lookup
      35. Choi DW. Calcium-mediated neurotoxicity: relationship to specific channel types and role in ischemic damage.. Trends Neurosci 1988 Oct;11(10):465-9.
        pubmed: 2469166doi: 10.1016/0166-2236(88)90200-7google scholar: lookup
      36. Siesjö BK. Cell damage in the brain: a speculative synthesis.. J Cereb Blood Flow Metab 1981;1(2):155-85.
        pubmed: 6276420doi: 10.1038/jcbfm.1981.18google scholar: lookup
      37. Raley-Susman KM, Lipton P. In vitro ischemia and protein synthesis in the rat hippocampal slice: the role of calcium and NMDA receptor activation.. Brain Res 1990 May 7;515(1-2):27-38.
        pubmed: 2162718doi: 10.1016/0006-8993(90)90572-sgoogle scholar: lookup
      38. Choi DW. Glutamate neurotoxicity in cortical cell culture is calcium dependent.. Neurosci Lett 1985 Aug 5;58(3):293-7.
        pubmed: 2413399doi: 10.1016/0304-3940(85)90069-2google scholar: lookup
      39. Faden AI, Simon RP. A potential role for excitotoxins in the pathophysiology of spinal cord injury.. Ann Neurol 1988 Jun;23(6):623-6.
        pubmed: 2841902doi: 10.1002/ana.410230618google scholar: lookup
      40. Freese A, Finklestein SP, DiFiglia M. Basic fibroblast growth factor protects striatal neurons in vitro from NMDA-receptor mediated excitotoxicity.. Brain Res 1992 Mar 20;575(2):351-5.
        pubmed: 1349256doi: 10.1016/0006-8993(92)90104-hgoogle scholar: lookup
      41. Abe K, Iri Y, Takayanagi M, Saito H. Involvement of protein kinase activation in neurotrophic effects of basic fibroblast growth factor in cultured brain neurons.. Jpn J Pharmacol 1991 Aug;56(4):563-6.
        pubmed: 1744998doi: 10.1254/jjp.56.563google scholar: lookup
      42. Benveniste H, Drejer J, Schousboe A, Diemer NH. Elevation of the extracellular concentrations of glutamate and aspartate in rat hippocampus during transient cerebral ischemia monitored by intracerebral microdialysis.. J Neurochem 1984 Nov;43(5):1369-74.
        pubmed: 6149259doi: 10.1111/j.1471-4159.1984.tb05396.xgoogle scholar: lookup
      43. Ip NY, Li Y, Yancopoulos GD, Lindsay RM. Cultured hippocampal neurons show responses to BDNF, NT-3, and NT-4, but not NGF.. J Neurosci 1993 Aug;13(8):3394-405.
        pubmed: 7688038doi: 10.1523/JNEUROSCI.13-08-03394.1993google scholar: lookup
      44. Zivin JA, Kochhar A, Saitoh T. Protein phosphorylation during ischemia.. Stroke 1990 Nov;21(11 Suppl):III117-21.
        pubmed: 2237967
      45. Akaneya Y, Enokido Y, Takahashi M, Hatanaka H. In vitro model of hypoxia: basic fibroblast growth factor can rescue cultured CNS neurons from oxygen-deprived cell death.. J Cereb Blood Flow Metab 1993 Nov;13(6):1029-32.
        pubmed: 8408312doi: 10.1038/jcbfm.1993.130google scholar: lookup
      46. Snyder SH, Bredt DS. Nitric oxide as a neuronal messenger.. Trends Pharmacol Sci 1991 Apr;12(4):125-8.
        pubmed: 1712138doi: 10.1016/0165-6147(91)90526-xgoogle scholar: lookup
      47. Collazo D, Takahashi H, McKay RD. Cellular targets and trophic functions of neurotrophin-3 in the developing rat hippocampus.. Neuron 1992 Oct;9(4):643-56.
        pubmed: 1389181doi: 10.1016/0896-6273(92)90028-cgoogle scholar: lookup
      48. WROBLEWSKI F, LADUE JS. Lactic dehydrogenase activity in blood.. Proc Soc Exp Biol Med 1955 Oct;90(1):210-3.
        pubmed: 13273400doi: 10.3181/00379727-90-21985google scholar: lookup
      49. Cheng B, Barger SW, Mattson MP. Staurosporine, K-252a, and K-252b stabilize calcium homeostasis and promote survival of CNS neurons in the absence of glucose.. J Neurochem 1994 Apr;62(4):1319-29.
        pubmed: 7510777doi: 10.1046/j.1471-4159.1994.62041319.xgoogle scholar: lookup
      50. Gross CE, Bednar MM, Howard DB, Sporn MB. Transforming growth factor-beta 1 reduces infarct size after experimental cerebral ischemia in a rabbit model.. Stroke 1993 Apr;24(4):558-62.
        pubmed: 8465363doi: 10.1161/01.str.24.4.558google scholar: lookup
      51. Ben-Ari Y. Limbic seizure and brain damage produced by kainic acid: mechanisms and relevance to human temporal lobe epilepsy.. Neuroscience 1985 Feb;14(2):375-403.
        pubmed: 2859548doi: 10.1016/0306-4522(85)90299-4google scholar: lookup
      52. Jørgensen MB, Diemer NH. Selective neuron loss after cerebral ischemia in the rat: possible role of transmitter glutamate.. Acta Neurol Scand 1982 Nov;66(5):536-46.
        pubmed: 7148396doi: 10.1111/j.1600-0404.1982.tb03140.xgoogle scholar: lookup
      53. Koike T, Martin DP, Johnson EM Jr. Role of Ca2+ channels in the ability of membrane depolarization to prevent neuronal death induced by trophic-factor deprivation: evidence that levels of internal Ca2+ determine nerve growth factor dependence of sympathetic ganglion cells.. Proc Natl Acad Sci U S A 1989 Aug;86(16):6421-5.
        pubmed: 2548215doi: 10.1073/pnas.86.16.6421google scholar: lookup
      54. Nozaki K, Finklestein SP, Beal MF. Basic fibroblast growth factor protects against hypoxia-ischemia and NMDA neurotoxicity in neonatal rats.. J Cereb Blood Flow Metab 1993 Mar;13(2):221-8.
        pubmed: 8436614doi: 10.1038/jcbfm.1993.27google scholar: lookup

      Citations

      This article has been cited 1 times.
      1. Bostan S, Serdengeçti S, Bayat FK, Bay S, Sezer A, Ayşit N, Öztürk G. A Novel Method for Culturing Telencephalic Neurons in Axolotls. J Comp Neurol 2025 Jun;533(6):e70066.
        doi: 10.1002/cne.70066pubmed: 40533890google scholar: lookup
      Subscribe to our newsletter
      Join 99,357 horse owners like you who receive our email updates on horse health and nutrition.