• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of pnasPNASInfo for AuthorsSubscriptionsAboutThis Article
Proc Natl Acad Sci U S A. Dec 1, 1993; 90(23): 10989–10993.
PMCID: PMC47907

Induction of apoptosis in cerebellar granule neurons by low potassium: inhibition of death by insulin-like growth factor I and cAMP.

Abstract

High levels of extracellular K+ ensure proper development and prolong survival of cerebellar granule neurons in culture. We find that when switched from a culture medium containing high K+ (25 mM) to one containing a low but more physiological K+ concentration (5 mM), differentiated granule neurons degenerate and die. Death induced by low K+ is due to apoptosis (programmed cell death), a form of cell death observed extensively in the developing nervous system and believed to be necessary for proper neurogenesis. The death process is accompanied by cleavage of genomic DNA into internucleosome-sized fragments, a hallmark of apoptosis. Inhibitors of transcription and translation suppress apoptosis induced by low K+, suggesting the necessity for newly synthesized gene products for activation of the process. Death can be prevented by insulin-like growth factor I but not by several other growth/neurotrophic factors. cAMP but not the protein kinase C activator phorbol 12-myristate 13-acetate can also support survival in low K+. In view of the large numbers of granule neurons that can be homogeneously cultured, our results offer the prospect of an excellent model system to study the mechanisms underlying apoptosis in the central nervous system and the suppression of this process by survival factors such as insulin-like growth factor I.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.8M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Levi-Montalcini R. The nerve growth factor: thirty-five years later. EMBO J. 1987 May;6(5):1145–1154. [PMC free article] [PubMed]
  • Barde YA. Trophic factors and neuronal survival. Neuron. 1989 Jun;2(6):1525–1534. [PubMed]
  • Maisonpierre PC, Belluscio L, Squinto S, Ip NY, Furth ME, Lindsay RM, Yancopoulos GD. Neurotrophin-3: a neurotrophic factor related to NGF and BDNF. Science. 1990 Mar 23;247(4949 Pt 1):1446–1451. [PubMed]
  • Ferrari G, Minozzi MC, Toffano G, Leon A, Skaper SD. Basic fibroblast growth factor promotes the survival and development of mesencephalic neurons in culture. Dev Biol. 1989 May;133(1):140–147. [PubMed]
  • Knusel B, Michel PP, Schwaber JS, Hefti F. Selective and nonselective stimulation of central cholinergic and dopaminergic development in vitro by nerve growth factor, basic fibroblast growth factor, epidermal growth factor, insulin and the insulin-like growth factors I and II. J Neurosci. 1990 Feb;10(2):558–570. [PubMed]
  • Gallo V, Giovannini C, Levi G. Modulation of non-N-methyl-D-aspartate receptors in cultured cerebellar granule cells. J Neurochem. 1990 May;54(5):1619–1625. [PubMed]
  • 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):6421–6425. [PMC free article] [PubMed]
  • Franklin JL, Johnson EM., Jr Suppression of programmed neuronal death by sustained elevation of cytoplasmic calcium. Trends Neurosci. 1992 Dec;15(12):501–508. [PubMed]
  • Chalazonitis A, Fischbach GD. Elevated potassium induces morphological differentiation of dorsal root ganglionic neurons in dissociated cell culture. Dev Biol. 1980 Jul;78(1):173–183. [PubMed]
  • Ishida I, Deguchi T. Effect of depolarizing agents on choline acetyltransferase and acetylcholinesterase activities in primary cell cultures of spinal cord. J Neurosci. 1983 Sep;3(9):1818–1823. [PubMed]
  • Resink A, Boer GJ, Balázs R. Treatment with excitatory amino acids or high K+ and NMDA receptors in cerebellar granule cells. Neuroreport. 1992 Sep;3(9):757–760. [PubMed]
  • Lasher RS, Zagon IS. The effect of potassium on neuronal differentiation in cultures of dissociated newborn rat cerebellum. Brain Res. 1972 Jun 22;41(2):482–488. [PubMed]
  • Levi G, Aloisi F, Ciotti MT, Gallo V. Autoradiographic localization and depolarization-induced release of acidic amino acids in differentiating cerebellar granule cell cultures. Brain Res. 1984 Jan 2;290(1):77–86. [PubMed]
  • Hockberger PE, Tseng HY, Connor JA. Immunocytochemical and electrophysiological differentiation of rat cerebellar granule cells in explant cultures. J Neurosci. 1987 May;7(5):1370–1383. [PubMed]
  • Cull-Candy SG, Howe JR, Ogden DC. Noise and single channels activated by excitatory amino acids in rat cerebellar granule neurones. J Physiol. 1988 Jun;400:189–222. [PMC free article] [PubMed]
  • Andersson IK, Edwall D, Norstedt G, Rozell B, Skottner A, Hansson HA. Differing expression of insulin-like growth factor I in the developing and in the adult rat cerebellum. Acta Physiol Scand. 1988 Feb;132(2):167–173. [PubMed]
  • Bondy CA. Transient IGF-I gene expression during the maturation of functionally related central projection neurons. J Neurosci. 1991 Nov;11(11):3442–3455. [PubMed]
  • Lesniak MA, Hill JM, Kiess W, Rojeski M, Pert CB, Roth J. Receptors for insulin-like growth factors I and II: autoradiographic localization in rat brain and comparison to receptors for insulin. Endocrinology. 1988 Oct;123(4):2089–2099. [PubMed]
  • Edwards SN, Buckmaster AE, Tolkovsky AM. The death programme in cultured sympathetic neurones can be suppressed at the posttranslational level by nerve growth factor, cyclic AMP, and depolarization. J Neurochem. 1991 Dec;57(6):2140–2143. [PubMed]
  • Martin DP, Schmidt RE, DiStefano PS, Lowry OH, Carter JG, Johnson EM., Jr Inhibitors of protein synthesis and RNA synthesis prevent neuronal death caused by nerve growth factor deprivation. J Cell Biol. 1988 Mar;106(3):829–844. [PMC free article] [PubMed]
  • Jones KH, Senft JA. An improved method to determine cell viability by simultaneous staining with fluorescein diacetate-propidium iodide. J Histochem Cytochem. 1985 Jan;33(1):77–79. [PubMed]
  • Murphy TH, Malouf AT, Sastre A, Schnaar RL, Coyle JT. Calcium-dependent glutamate cytotoxicity in a neuronal cell line. Brain Res. 1988 Mar 22;444(2):325–332. [PubMed]
  • Hockenbery D, Nuñez G, Milliman C, Schreiber RD, Korsmeyer SJ. Bcl-2 is an inner mitochondrial membrane protein that blocks programmed cell death. Nature. 1990 Nov 22;348(6299):334–336. [PubMed]
  • Schramm M, Eimerl S, Costa E. Serum and depolarizing agents cause acute neurotoxicity in cultured cerebellar granule cells: role of the glutamate receptor responsive to N-methyl-D-aspartate. Proc Natl Acad Sci U S A. 1990 Feb;87(3):1193–1197. [PMC free article] [PubMed]
  • Ellis RE, Yuan JY, Horvitz HR. Mechanisms and functions of cell death. Annu Rev Cell Biol. 1991;7:663–698. [PubMed]
  • Raff MC. Social controls on cell survival and cell death. Nature. 1992 Apr 2;356(6368):397–400. [PubMed]
  • Ernfors P, Lönnerberg P, Ayer-LeLievre C, Persson H. Developmental and regional expression of basic fibroblast growth factor mRNA in the rat central nervous system. J Neurosci Res. 1990 Sep;27(1):10–15. [PubMed]
  • Maisonpierre PC, Belluscio L, Friedman B, Alderson RF, Wiegand SJ, Furth ME, Lindsay RM, Yancopoulos GD. NT-3, BDNF, and NGF in the developing rat nervous system: parallel as well as reciprocal patterns of expression. Neuron. 1990 Oct;5(4):501–509. [PubMed]
  • Bartlett WP, Li XS, Williams M, Benkovic S. Localization of insulin-like growth factor-1 mRNA in murine central nervous system during postnatal development. Dev Biol. 1991 Sep;147(1):239–250. [PubMed]
  • Yeh HJ, Ruit KG, Wang YX, Parks WC, Snider WD, Deuel TF. PDGF A-chain gene is expressed by mammalian neurons during development and in maturity. Cell. 1991 Jan 11;64(1):209–216. [PubMed]
  • Wanaka A, Johnson EM, Jr, Milbrandt J. Localization of FGF receptor mRNA in the adult rat central nervous system by in situ hybridization. Neuron. 1990 Sep;5(3):267–281. [PubMed]
  • Lamballe F, Klein R, Barbacid M. trkC, a new member of the trk family of tyrosine protein kinases, is a receptor for neurotrophin-3. Cell. 1991 Sep 6;66(5):967–979. [PubMed]
  • McConkey DJ, Hartzell P, Orrenius S. Rapid turnover of endogenous endonuclease activity in thymocytes: effects of inhibitors of macromolecular synthesis. Arch Biochem Biophys. 1990 Apr;278(1):284–287. [PubMed]
  • Batistatou A, Greene LA. Aurintricarboxylic acid rescues PC12 cells and sympathetic neurons from cell death caused by nerve growth factor deprivation: correlation with suppression of endonuclease activity. J Cell Biol. 1991 Oct;115(2):461–471. [PMC free article] [PubMed]
  • Dessi F, Charriaut-Marlangue C, Khrestchatisky M, Ben-Ari Y. Glutamate-induced neuronal death is not a programmed cell death in cerebellar culture. J Neurochem. 1993 May;60(5):1953–1955. [PubMed]
  • D'Mello SR, Galli C. SGP2, ubiquitin, 14K lectin and RP8 mRNAs are not induced in neuronal apoptosis. Neuroreport. 1993 Apr;4(4):355–358. [PubMed]
  • Calissano P, Ciotti MT, Battistini L, Zona C, Angelini A, Merlo D, Mercanti D. Recombinant human insulin-like growth factor I exerts a trophic action and confers glutamate sensitivity on glutamate-resistant cerebellar granule cells. Proc Natl Acad Sci U S A. 1993 Sep 15;90(18):8752–8756. [PMC free article] [PubMed]
  • Segal RA, Takahashi H, McKay RD. Changes in neurotrophin responsiveness during the development of cerebellar granule neurons. Neuron. 1992 Dec;9(6):1041–1052. [PubMed]
  • Rydel RE, Greene LA. cAMP analogs promote survival and neurite outgrowth in cultures of rat sympathetic and sensory neurons independently of nerve growth factor. Proc Natl Acad Sci U S A. 1988 Feb;85(4):1257–1261. [PMC free article] [PubMed]
  • Rukenstein A, Rydel RE, Greene LA. Multiple agents rescue PC12 cells from serum-free cell death by translation- and transcription-independent mechanisms. J Neurosci. 1991 Aug;11(8):2552–2563. [PubMed]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • Compound
    Compound
    PubChem Compound links
  • MedGen
    MedGen
    Related information in MedGen
  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...