Abstract
The axon initial segment of cortical principal neurones contains an organelle consisting of two to four stacks of flat, membrane-delineated cisternae alternating with electron-dense, fibrillar material. These cisternal organelles are situated predominantly close to the synaptic junctions of GABAergic axo-axonic cell terminals. To examine the possibility that the cisternal organelle is involved in Ca2+ sequestration, we tested for the presence of Ca2+-ATPase in the cisternal organelles of pyramidal cell axons in the CA1 and CA3 regions of the hippocampus. Electron microscopic immunocyto-chemistry using antibodies to muscle sarcoplasmic reticulum ATPase revealed immunoreactivity associated with cisternal organelle membranes. The localisation of Ca2+-ATPase in cisternal organelles was also confirmed by enzyme cytochemistry, which produced reaction product in the lumen of the cisternae. These experiments provide evidence for the presence of a Ca2+ pump in the cisternal organelle membrane, which may play a role in the sequestration and release of Ca2+. Cisternal organelles are very closely aligned to the axolemma and the outermost cisternal membrane is connected to the plasma membrane by periodic electron-dense bridges as detected in electron micrographs. It is suggested that the interface acts as a voltage sensor, releasing Ca2+ from cisternal organelles upon depolarisation of the axon initial segment, in a manner similar to the sarcoplasmic reticulum of skeletal muscle. The increase in intra-axonal Ca2+ may regulate the GABAA receptors associated with the axo-axonic cell synapses, and could affect the excitability of pyramidal cells.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Ando T, Fujimoto K, Mayahara H, Miyajima H, Ogawa K (1981) A new one-step method for the histochemistry and cytochemistry of Ca2+-ATPase activity. Acta Histochem Cytochem 14:705–726
Andrews SB, Leapman RD, Landis DMD, Reese TS (1988) Activity-dependent accumulation of calcium in Purkinje cell dendritic spines. Proc Nat Acad Sci USA 85:1682–1685
Audigier SMP, Wang JKT, Greengard P (1988) Membrane depolarization and carbamoylcholine stimulate phosphatidylinositol turnover in intact nerve terminals. Proc Natl Acad Sci USA 85:2859–2863
Berridge MJ (1993) Inositol trisphosphate and calcium signalling. Nature 361:315–325
Berridge MJ, Irvine RF (1989) Inositol phosphates and cell signalling. Nature 341:197–205
Buhl EH, Halasy K, Somogyi P (1993) Hippocampal unitary IP-SPs: identified sources and number of release sites. Eur J Neurosci [Suppl 6]:225
Buhl EH, Halasy K, Somogyi P (1994a) Hippocampal unitary inhibitory postsynaptic potentials: diverse sources and number of release sites. Nature 368:823–828
Buhl EH, Han Z.-S., Lorinczi Z, Stezhka VV, Karnup SV, Somogyi P (1994b) Physiological properties of anatomically identified axo-axonic cells in the rat hippocampus. J Neurophysiol 71:1289–1307
Cohen RS, Kriho V (1991) Localization of ATPase activity in dendritic spines of the cerebral cortex. J Neurocytol 20:703–715
Conradi S (dy1969) Observations on the ultrastructure of the axon hillock and intial axon segment of lumbosacral motoneurones in the cat. Acta Physiol Scand [Supp. 332]: 65–84
de Zeeuw CI, Ruigrok TJH, Holstege JC, Schalekamp MPA, Voogd, J (1990) Intracellular labeling of neurons in the medial accessory olive of the cat: III. Ultrastructure of axon hillock and initial segment and their GABAergic innervation. J Comp Neurol 300:495–510
Freund TF, Martin KAC, Soltesz I, Somogyi P, Whitteridge D (1989) Arborisation pattern and postsynaptic targets of physiologically identified thalamocortical afferents in the monkey striate cortex. J Comp Neurol 289:315–336
Galione A (1992) Ca2+-induced Ca2+ release and its modulation by cyclic ADP-ribose. Trends Pharmacol Sci 13:304–306
Grover AK, Khan I (1992) Calcium pump isoforms: diversity, selectivity and plasticity. Cell Calcium 13:9–17
Hashimoto J, Bruno B, Lew DP, Pozzan T, Volpe P, Meldolesi J (1988) Immunocytochemistry of calciosomes in liver and pancreas. J Cell Biol 107:2523–2531
Henkart M, Landis DMD, Reese TS (1976) Similarity of junctions between plasma membranes and endoplasmic reticulum in muscle and neurons. J Cell Biol 70:338–347
Henzi V, MacDermott AB (1992) Characteristics and function of Ca2+- and inositol 1,4,5-trisphosphate-releasable stores of Ca2+ in neurons. Neuroscience 46:251–273
Inoue M, Oomura Y, Yakushiji T, Akaike N (1986) Intracellular calcium ions decrease the affinity of the GABA receptor. Nature 324:156–158
Inui M, Saito A, Fleischer S (1987) Purification of the ryanodine receptor and identity with feet structures of junctional terminal cisternae of sarcoplasmic reticulum from fast skeletal muscle. J Biol Chem 262:1740–1747
Kortje KH, Freihofer D, Rahmann H (1990) Cytochemical localization of high-affinity Ca2+-ATPase activity in synaptic terminals. J Histochem Cytochem 38:895–900
Kosaka T (1980) The axon initial segment as a synaptic site: ultra-structure and synaptology of the initial segment of the pyramidal cell in the rat hippocampus (CA3 region). J Neurocytol 9:861–882
Krenács T, Molnár E, Dobó E, Dux L (1989) Fibre typing using sarcoplasmic reticulum Ca2+-ATPase and myoglobin immunohistochemistry in rat gastrocnemius muscle. Histochem J 21:145–155
Kuwajima G, Futatsugi A, Niinobe M, Nakanishi S, Mikoshiba K (1992) Two types of ryanodine receptors in mouse brain: skeletal muscle type exclusively in Purkinje cells and cardiac muscle type in various neurons. Neuron 9:1133–1142
Lai FA, Dent M, Wickenden C, Xu L, Kumari G, Misra M, Lee HB, Sar M, Meissner G (1992) Expression of a cardiac Ca2+-ATPase channel isoform in mammalian brain. Biochem J 288:553–564
Leidenheimer NJ, Browning MD, Harris RA (1991) GABAA receptor phosphorylation: multiple sites, actions and artifacts. Trends Pharmacol Sci 12:84–87
Lewis DA, Lund JS (1990) Heterogeneity of chandelier neurons in monkey neocortex: corticotropin-releasing factor- and parval- bumin-immunoreactive populations. J Comp Neurol 293:599–615
Linden DJ, Routtenberg A (1989) The role of protein kinase C in long-term potentiation: a testable model. Brain Res Rev 14:279–296
Llano I, Leresche N, Marty A (1991) Calcium entry increases the sensitivity of cerebellar Purkinje cells to applied GABA and decreases inhibitory synaptic currents. Neuron 6:565–574
Maeda N, Niinobe M, Inoue Y, Mikoshiba K (1989) Developmental expression and intracellular location of P400 protein, characteristic of Purkinje cells in the mouse cerebellum. Dev Biol 133:67–76
Maggio K, Watrin A, Keicher E, Nicaise G, Hernandez-Nicaise M-L (1991) Ca2+-ATPase and Mg2+-ATPase in Aplysia glial and interstitial cells: an EM cytochemical study. J Histochem Cytochem 39:1645–1658
Mata M, Fink DJ (1989) Ca++-ATPase in the central nervous system: an EM cytochemical study. J Histochem Cytochem 37:971–980
Maxwell WL, Watt C, Pediani JD, Graham DI, Adams JH, Gennarelli TA (1991) Localisation of calcium ions and calcium-ATPase activity within myelinated nerve fibres of the adult guinea-pig optic nerve. J Anat 176:71–79
McPherson PS, Campbell KP (1993) The ryanodine receptor/Ca2+ release channel. J Biol Chem 268:13765–13768
Mignery GA, Sudhof TC, Takei K, De Camilli P (1989) Putative receptor for inositol 1,4,5-trisphosphate similar to ryanodine receptor. Nature 342:192–195
Molnár E, Seidler NW, Jona I, Martonosi AN (1990) The binding of monoclonal and polyclonal antibodies to the Ca2+-ATPase of sarcoplasmic reticulum: effects on interactions between ATPase molecules. Biochem Biophys Acta 1023:147–167
Molnár E, Varga S, Jóna I, Seidler NW, Martonosi A (1992) Immunological relatedness of the sarcoplasmic reticulum Ca2+-ATPase and the Na+, K+-ATPase. Biochem Biophys Acta 1103:281–295
Molnár E, Baude A, Richmond SA, Patel PB, Somogyi P, McIlhinney RAJ (1993) Biochemical and immunocytochemical characterization of antipeptide antibodies to a cloned GluR1 glutamate receptor subunit: cellular and subcellular distribution in the rat forebrain. Neuroscience 53:307–326
Nakanishi S, Kuwajima G, Mikoshiba K (1992) Immunohisto-chemical localisation of ryanodine receptors in mouse central nervous system. Neurosci Res 15:130–142
Nasu F, Inomata K (1990) Ultracytochemical demonstration of Ca2+-ATPase activity in the rat saphenous artery and its innervated nerve terminal. J Electron Microsc (Tokyo) 39:487–491
Numann R, Catterall WA, Scheuer T (1991) Functional modulation of brain sodium channels by protein kinase C phosphorylation. Science 254:115–118
Padua RA, Yamamoto T, Fyda D, Sawchuk MA, Geiger JD, Nagy JI (1992) Autoradiographic analysis of [3H]ryanodine binding sites in rat brain: regional distribution and the effects of lesions on sites in the hippocampus. J Chem Neuroanat 5:63–73
Palay SL, Sotelo C, Peters A, Orkand PM (1968) The axon hillock and the initial segment. J Cell Biol 38:193–201
Peters A, Proskauer CC, Kaiserman-Abramof IR (1968) The small pyramidal neuron of the rat cerebral cortex. The axon hillock and initial segment. J Cell Biol 39:604–619
Peters A, Palay SL, Webster H (1991) The fine structure of the nervous system, 3rd edn. Oxford University Press, New York
Pitler TA, Alger BE (1992) Postsynaptic spike firing reduces synaptic GABAA responses in hippocampal pyramidal cells. J Neurosci 12:4122–4132
Plessers L, Eggermont JA, Wuytack F, Casteels R (1991) A study of the organellar Ca2+-transport ATPase isozymes in pig cerebellar Purkinje cells. J Neurosci 11:650–656
Raymond LA, Blackstone CD, Huganir RL (1993) Phosphorylation of amino acid neurotransmitter receptors in synaptic plasticity. Trends Neurosci 16:147–153
Rosenbluth J (1962) Subsurface cisterns and their relationship to the neuronal plasma membrane. J Cell Biol 13:405–421
Ross CA, Meldolesi J, Milner TA, Satoh T, Supattapone S, Snyder SH (1989) Inositol 1,4,5-trisphosphate receptor localized to endoplasmic reticulum in cerebellar Purkinje neurons. Nature 339:468–470
Rossier MF, Putney JW (1991) The identity of the calcium-storing, inositol 1,4,5-trisphosphate-sensitive organelle in non-muscle cells: calciosome, endoplasmic reticulum or both? Trends Neurosci 14:310–314
Rusakov DA, Podini P, Villa A, Meldolesi J (1993) Tridimensional organization of Purkinje neuron cisternal stacks, a specialized endoplasmic reticulum subcompartment rich in inositol 1,4,5- trisphosphate receptors. J Neurocytol 22:273–282
Saito A, Inui M, Radermacher M, Frank J, Fleischer S (1988) Ultrastructure of the calcium release channel of sarcoplasmic reticulum. J Cell Biol 107:211–219
Sarkadi B, Enyedi A, Penniston JT, Verma AK, Dux L, Molnár E, Gardos G (1988) Characterization of membrane calcium pumps by simultaneous immunoblotting and 32P radiography. Biochem Biophys Acta 939:40–46
Satoh T, Ross CA, Villa A, Supattapone S, Pozzan T, Snyder SH, Meldolesi J (1990) The inositol 1,4,5-trisphosphate receptor in cerebellar Purkinje cells: quantitative immunogold labeling reveals concentration in an ER subcompartment. J Cell Biol 111:615–624
Sharp AH, McPherson PS, Dawson TM, Aoki C, Campbell KP, Snyder SH (1993) Differential immunohistochemical localization of inositol 1,4,5-trisphosphate- and ryanodine-sensitive Ca2+ release channels in rat brain. J Neurosci 13:3051–3063
Sloper JJ, Powell TPS (1979) A study of the axon initial segment and proximal axon of neurons in the primate motor and somatic sensory cortices. Phil Trans R Soc Lond B 285:173–197
Soji T, Nishizono H, Yashiro T, Herbert DC (1991) Cytochemistry of Ca2+-dependent adenosine triphosphatase (Ca-ATPase) in rat anterior pituitary cells. Tissue Cell 23:1–6
Somogyi P (1977) A specific ‘axo-axonal’ interneuron in the visual cortex of the rat. Brain Res 136:345–350
Somogyi P, Hamori J (1976) A quantitative electron microscopic study of the Purkinje cell axon initial segment. Neuroscience 1:361–366
Somogyi P, Cowey A (1981) Combined Golgi and electron microscopic study on the synapses formed by double bouquet cells in the visual cortex of the cat and monkey. J Comp Neurol 195:547–566
Somogyi P, Takagi H (1982) A note on the use of picric acid- paraformaldehyde-glutaraldehyde fixative for correlated light and electron microscopic immunocytochemistry. Neuroscience 7:1779–1783
Somogyi P, Nunzi MG, Gorio A, Smith AD (1983a) A new type of specific interneuron in the monkey hippocampus forming synapses exclusively with the axon initial segments of pyramidal cells. Brain Res 259:137–142
Somogyi P, Smith AD, Nunzi MG, Gorio A, Takagi H, Wu J-Y (1983b) Glutamate decarboxylase immunoreactivity in the hippocampus of the cat. Distribution of immunoreactive synaptic terminals with special reference to the axon initial segment of pyramidal neurons. J Neurosci 3:1450–1468
Somogyi P, Freund TF, Hodgson AJ, Somogyi J, Beroukas D, Chubb IW (1985) Identified axo-axonic cells are immunoreactive for GABA in the hippocampus and visual cortex of the cat. Brain Res 332:143–149
Stelzer A (1992) Intracellular regulation of GABAA-receptor function. In: Narahashi T (ed) Ion Channels, vol 3. Plenum Press, New York, pp 83–136
Stuart GJ, Sakmann B (1994) Active propagation of somatic action potentials into neocortical pyramidal cell dendrites. Nature 367:69–72
Takei K, Stukenbrok H, Metcalf A, Mignery GA, Sudhof TC, Volpe P, De Camilli P (1992) Ca2+ Stores in Purkinje neurons: endoplasmic reticulum subcompartments demonstrated by the heterogenous distribution of the InsP3 receptor, Ca2+- ATPase, and calsequestrin. J Neurosci 12:489–505
Takeshima H, Nishimura S, Matsumoto T, Ishida H, Kangawa K, Minamino N, Matsuo H, Ueda M, Hanaoka M, Hirose T, Numa S (1989) Primary structure and expression from complementary DNA of skeletal muscle ryanodine receptor. Nature 339:439–445
Theibert AB, Supattapone S, Worley PF, Baraban JM, Meek JL, Snyder SH (1987) Demonstration of inositol 1,3,4,5-tetrakisphosphate receptor binding. Biochem Biophys Res Commun 148:1283–1289
Treves S, De Mattei M, Landfredi M, Villa A, Green NM, MacLennan DH, Meldolesi J, Pozzan T (1990) Calreticulin is a candidate for a calsequestrin-like function in Ca2+ storage compartments (calciosomes) of liver and brain. Biochem J 271:473–480
Tsunoda Y (1993) Receptor-operated Ca2+ signaling and crosstalk in stimulus secretion coupling. Biochem Biophys Acta 1154:105–156
Volpe P, Krause K-H, Hashimoto S, Zorzato F, Pozzan T, Meldolesi J, Lew DP (1988) ‘Calciosome,’ a cytoplasmic organelle: The inositol 1,4,5-trisphosphate-sensitive Ca2+ store of non-muscle cells? Proc Natl Acad Sci USA 85:1091–1095
Westrum LE (1993) Axon hillocks and initial segments in spinal trigeminal nucleus with emphasis on synapses including axo-axo-axonic contacts. J Neurocytol 22:793–803
Whiting P, McKernan RM, Iversen LL (1990) Another mechanism for creating diversity in γ-aminobutyrate type A receptors: RNA splicing directs expression of two forms of γ2 sub-unit, one of which contains a protein kinase C phosphorylation site. Proc Natl Acad Sci USA 87:9966–9970
Yamamoto T, Hertzberg EL, Nagy JI (1990) Epitopes of gap junctional proteins localized to neuronal subsurface cisterns. Brain Res 527:135–139
Yamamoto T, Hertzberg EL, Nagy JI (1991) Subsurface cisterns in α-motoneurons of the rat and cat: immunohistochemical detection with antibodies against connexin 32. Synapse 8:119–136
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Benedeczky, I., Molnár, E. & Somogyi, P. The cisternal organelle as a Ca2+-storing compartment associated with GABAergic synapses in the axon initial segment of hippocampal pyramidal neurones. Exp Brain Res 101, 216–230 (1994). https://doi.org/10.1007/BF00228742
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF00228742