Research article| Volume 119, ISSUE 1, P79-84, October 1993

Download started.


Swelling and death of neuronal cells by lactic acid

      This paper is only available as a PDF. To read, Please Download here.


      Lactacidosis occurring in cerebral ischemia or trauma is a major mechanism of cytotoxic brain edema and brain damage. Respective effects of lactacidosis were currently analyzed in vitro by employment of the murine neuronal cell line, Neuro-2A, in order to obtain a better understanding of specific mechanisms underlying cell swelling and cell death in comparison with glial cells. The cells were suspended in a physiological medium in the presence of lactic acid at increasing concentrations. Levels of acidosis reaching from pH 6.8-5.6 were obtained while other parameters, such as osmolarity and electrolyte concentrations, were maintained in the physiological range. Assessment of cell swelling and cell viability using exclusion of propidium iodide was made by flow cytometry with employment of an advanced Coulter system. Swelling of Neuro-2A cells commenced once the pH in the medium was lowered to 6.8 or below. From this level downward, cell swelling was a function of the severity of acidosis and duration of exposure. For example, lactacidosis of pH 6.8 or 5.6 lasting 90 min led to an increase in cell volume to 109.5% or 159.6% of normal, respectively. Viability of the neuronal cells was 85% under control conditions. It remained in this range down to pH 6.2. At pH 5.6, however, cell viability decreased in a time-dependent fashion. At 90 min, only 48.9% of the neuronal cells were viable at pH 5.6. The swelling response and impairment of viability of the neuronal cells was compared with that of C6 glioma cells. A 60 min exposure of the glial cells to either pH 6.2 or pH 5.6 led to swelling of only 55% or 65%, respectively, of the cell volume increase observed in the Neuro-2A cells. In addition, the glial cells were less vulnerable to lactacidosis as demonstrated by better maintenance of cell viability. After suspension for 1 h at pH 5.6, only 53.9% of the neuronal cells were alive, in comparison to 74.1% of the C6 glioma cells. Taken together, the present findings demonstrate, as former observations on glial cells, that lactacidosis is a powerful mechanism of cell swelling and cell death in a neuronal cell line. As in the glial cells, different pH thresholds could be identified, associated either with cell swelling or a decrease in cell viability. While cell swelling occurred already at relatively mild levels of acidosis (pH 6.8), viability of the Neuro-2A cells was decreasing only at pH 5.6, confirming different susceptibilities of cell swelling and cell death to acidosis. The level of acidosis found to destroy nerve cells in vitro has been observed in severe forms of cerebral ischemia in vivo, for example in hyperglycemia.


      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'


      Subscribe to Journal of the Neurological Sciences
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect


        • Andersen B.J.
        • Unterberg A.W.
        • Clarke G.D.
        • Marmarou A.
        Effect of posttraumatic hypoventilation on cerebral energy metabolism.
        J. Neurosurg. 1988; 68: 601-607
        • Astrup J.
        • Symon L.
        Thresholds in cerebral ischemia - the ischemic penumbra.
        Stroke. 1981; 12: 723-725
        • Baethmann A.
        Pathophysiological and pathochemical aspects of cerebral edema.
        Neurosurg. Rev. 1978; 1: 85-100
        • Busa W.B.
        • Nuccitelli R.
        Metabolic regulation via intracellular pH.
        Am. J Physiol. 1984; 246: R409-R438
        • Chan P.H.
        • Chu L.
        • Chen S.
        Effects of MK-801 on glutamate-induced swelling of astrocytes in primary culture.
        J. Neurosci. Res. 1990; 25: 87-93
        • Chow S.Y.
        • Yen-Chow Y.C.
        • White H.S.
        • Woodbury D.M.
        pH Regulation after acid load in primary cultures of mouse astrocytes.
        Dev. Brain Res. 1991; 60: 69-78
        • De Laat S.W.
        • Van der Saag P.T.
        The plasma membrane as a regulatory site in growth and differentiation of neuroblastoma cells.
        Int. Rev. Cytol. 1982; 74: 1-54
        • Grinstein S.
        • Rothstein A.
        Mechanisms of regulation of the Math Eq exchanger.
        J Membr. Biol. 1986; 90: 1-12
        • Hossmann K.A.
        Development and resolution of ischemic brain edema.
        in: Pappius H.M. Feindel W. Dynamics of Brain Edema. Springer, Berlin1976: 319
        • Kachel V.
        Basic principles of electrical sizing of cells and particles and their realization in the new instrument ‘Metricell’.
        J. Histochem. Cytochem. 1976; 24: 211-230
        • Kachel V.
        • Glossner E.
        • Kordwig E.
        • Ruhenstroth Bauer G.
        Fluvo-metricell, a combined cell volume and cell fluorescence analyzer.
        J Histochem. Cytochem. 1977; 25: 804-812
        • Katsura K.
        • Ekholm A.
        • Asplund B.
        • Siesjö B.K.
        Extracellular pH in the brain during ischemia: relationship to the severity of lactic acidosis.
        J. Cereb. Blood Flow Metab. 1991; 11: 597-599
        • Kempski O.
        • Chaussy L.
        • Groβ U.
        • Zimmer M.
        • Baethmann A.
        Volume regulation and metabolism of suspended C6 glioma cells: an in vitro model to study cytotoxic brain edema.
        Brain Res. 1983; 279: 217-228
        • Kempski O.
        • Staub F.
        • Jansen M.
        • Schödel F.
        • Baethmann A.
        Glial swelling during extracellular acidosis in vitro.
        Stroke. 1988; 19: 385-392
        • Kempski O.
        • Rosen F.v.
        • Weigt H.
        • Staub F.
        • Peters J.
        • Baethmann A.
        Glial ion transport and volume control.
        Ann. NY Acad. Sci. 1991; 633: 306-317
        • Klatzo I.
        Presidential address - Neuropathological aspects of brain edema.
        J. Neuropathol. Exp. Neurol. 1967; 26: 1-14
        • Kraig R.P.
        • Pulsinelli W.A.
        • Plum F.
        Heterogeneous distribution of hydrogen and bicarbonate ions during complete brain ischemia.
        in: Progress in Brain Research. Vol. 63. Elsevier, Amsterdam1985: 155-166
        • Kraig R.P.
        • Petito C.K.
        • Plum F.
        • Pulsinelli W.A.
        Hydrogen ions kill brain at concentrations reached in ischemia.
        J. Cereb. Blood Flow Metab. 1987; 7: 379-386
        • Mellergård P.E.
        • Siesjö B.K.
        Astrocytes fail to regulate intracellular pH at moderately reduced extracellular pH.
        NeuroReport. 1991; 2: 695-698
        • Moolenaar W.H.
        Regulation of cytoplasmic pH by a Math Eq exchange.
        Trends Biol. Sci. 1986; 2: 141-143
        • Nedergaard M.
        • Kraig R.P.
        • Tanabe J.
        • Pulsinelli W.A.
        Dynamics of interstitial and intracellular pH in evolving brain infarct.
        Am. J Physiol. 1990; 260: R581-R588
        • Nedergaard M.
        • Goldman S.A.
        • Desai S.
        • Pulsinelli W.A.
        Acid-induced death in neurons and glia.
        J. Neurosci. 1991; 11: 2489-2497
        • Rosner M.J.
        • Becker D.P.
        Experimental brain injury: successful therapy with the weak base, thromethamine.
        J Neurosurg. 1984; 60: 961-971
        • Rothe G.
        • Valet G.
        Phagocytosis, intracellular pH, and cell volume in the multifunctional analysis of granulocytes by flow cytometry.
        Cytometry. 1988; 9: 316-324
        • Siesjö B.K.
        Acidosis and ischemic brain damage.
        Neurochem. Pathol. 1988; 9: 31-88
        • Siesjö B.K.
        • Bendek G.
        • Koide T.
        • Westerberg E.
        • Wieloch T.
        Influence of acidosis on lipid peroxidation in brain tissues in vitro.
        J. Cereb. Blood Flow Metab. 1985; 5: 253-258
        • Smith M.L.
        • Hanwehr R.v.
        • Siesjö B.K.
        Changes in extra-and intracellular pH in the brain during and following ischemia in hyperglycemic and in moderately hypoglycemic rats.
        J. Cereb. Blood Flow Metab. 1986; 6: 574-583
        • Spoerri P.E.
        • Glees P.
        • Dresp W.
        The time course of synapse formation of mouse neuroblastoma cells in monolayer cultures.
        Cell Tissue Res. 1980; 205: 411-421
        • Staub F.
        • Baethmann A.
        • Peters J.
        • Weigt H.
        • Kempski O.
        Effects of lactacidosis on glial cell volume and viability.
        J. Cereb. Blood Flow Metab. 1990; 10: 866-876
        • Tanford D.
        Protein denaturation.
        in: Advances in Protein Chemistry. Vol. 23. Academic Press, New York1968: 121-161
        • Theodorsson Norheim E.
        Kruskal-Wallis test: BASIC computer program to perform nonparametric one-way analysis of variance and multiple comparisons on ranks of several independent samples.
        Comput. Methods Programs Biomed. 1986; 23: 57-62
        • Thompson J.M.
        • London E.D.
        • Johnson J.E.J.
        Ultrastructural, functional and biochemical characteristics of mouse and human neuroblastoma cell lines.
        Neuroscience. 1982; 7: 1807-1815
        • Trivedi B.
        • Danforth W.H.
        Effects of pH on the kinetics of frog muscle phosphofructokinase.
        J Biol. Chem. 1966; 241: 4110-4114
        • Van der Valk J.B.
        • Vijverberg H.P.
        Glutamate-induced inward current in a clonal neuroblastoma cell line.
        Eur. J. Pharmacol. 1990; 185: 99-102
        • Yoshida K.
        • Marmarou A.
        Effects of tromethamine and hyperventilation on brain injury in the cat.
        J Neurosurg. 1991; 74: 87-96