Ultrastructural and morphometric features of the acetylcholine innervation in adult rat parietal cortex: An electron microscopic study in serial sections

This study was aimed at characterizing the ultrastructural morphology of the normal acetylcholine (ACh) innervation in adult rat parietal cortex. After immunostaining with a monoclonal antibody against purified rat brain choline acetyltransferase (ChAT), more than 100 immunoreactive axonal varicosities (terminals) from each layer of the Par 1 area were photographed and examined in serial thin sections across their entire volume. These varicosities were relatively small, averaging 0.6 μm in diameter, 1.6 μM2 in surface, and 0.12 μM3 in volume. In every layer, a relatively low proportion exhibited a synaptic membrane differentiation (10% in layer I, 14% in II–III, 11% in IV, 21% in V, 14% in VI), for a I–VI average of 14%. These synaptic junctions were usually single, symmetrical (>99%), and occupied a small portion of the surface of varicosities (<3%). A majority were found on dendritic branches (76%), some on spines (24%), and none on cell bodies. On the whole, the ACh junctional varicosities were significantly larger than their nonjunctional counterparts, and both synaptic and nonsynaptic varicosities could be observed on the same fiber.

[1]  P. Clarke Nicotinic receptors in mammalian brain: localization and relation to cholinergic innervation. , 1993, Progress in brain research.

[2]  P S Goldman-Rakic,et al.  Association of m1 and m2 muscarinic receptor proteins with asymmetric synapses in the primate cerebral cortex: morphological evidence for cholinergic modulation of excitatory neurotransmission. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[3]  KM Harris,et al.  Dendritic spines of CA 1 pyramidal cells in the rat hippocampus: serial electron microscopy with reference to their biophysical characteristics , 1989, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[4]  L. Descarries,et al.  Serotonin nerve terminals in adult rat neocortex , 1975, Brain Research.

[5]  J. Changeux,et al.  Immunocytochemical localization of a neuronal nicotinic receptor: the beta 2-subunit , 1993, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[6]  A. Levey,et al.  Monoclonal antibodies to choline acetyltransferase: production, specificity, and immunohistochemistry , 1983, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[7]  D. McCormick Neurotransmitter actions in the thalamus and cerebral cortex and their role in neuromodulation of thalamocortical activity , 1992, Progress in Neurobiology.

[8]  F. Eckenstein,et al.  An anatomical study of cholinergic innervation in rat cerebral cortex , 1988, Neuroscience.

[9]  P. Novikoff,et al.  DIFFUSION ARTIFACTS IN 3,3'-DIAMINOBENZIDINE CYTOCHEMISTRY , 1972, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[10]  C. Saper Organization of cerebral cortical afferent systems in the rat. II. Magnocellular basal nucleus , 1984, The Journal of comparative neurology.

[11]  P. Goldman-Rakic,et al.  Heterogeneous targets of dopamine synapses in monkey prefrontal cortex demonstrated by serial section electron microscopy: a laminar analysis using the silver-enhanced diaminobenzidine sulfide (SEDS) immunolabeling technique. , 1993, Cerebral cortex.

[12]  M. Hasselmo,et al.  Cholinergic suppression specific to intrinsic not afferent fiber synapses in rat piriform (olfactory) cortex. , 1992, Journal of neurophysiology.

[13]  H. Thoenen,et al.  Cholinergic neurons in the rat cerebral cortex demonstrated by immunohistochemical localization of choline acetyltransferase , 1983, Neuroscience Letters.

[14]  H. Fibiger,et al.  Organization of central cholinergic systems. , 1989, Progress in brain research.

[15]  L. Hersh,et al.  An atlas of the regional and laminar distribution of choline acetyltransferase immunoreactivity in rat cerebral cortex , 1989, Neuroscience.

[16]  M. Mesulam,et al.  Central cholinergic pathways in the rat: An overview based on an alternative nomenclature (Ch1–Ch6) , 1983, Neuroscience.

[17]  M. Mesulam,et al.  Cortical projections arising from the basal forebrain: A study of cholinergic and noncholinergic components employing combined retrograde tracing and immunohistochemical localization of choline acetyltransferase , 1984, Neuroscience.

[18]  W. Singer,et al.  Quantitative analysis of the choline acetyltransferase‐immunoreactive axonal network in the cat primary visual cortex: I. Adult cats , 1987, The Journal of comparative neurology.

[19]  G. Halliday,et al.  Serotonin-like immunoreactive cells and fibres in the rat ventromedial mesencephalic tegmentum , 1989, Brain Research Bulletin.

[20]  D. Lewis Distribution of choline acetyltransferase-immunoreactive axons in monkey frontal cortex , 1991, Neuroscience.

[21]  H. Thoenen,et al.  Production of specific antisera and monoclonal antibodies to choline acetyltransferase: characterization and use for identification of cholinergic neurons. , 1982, The EMBO journal.

[22]  H H Jasper,et al.  Acetylcholine Liberation from Cerebral Cortex during Paradoxical (REM) Sleep , 1971, Science.

[23]  L. Squire,et al.  Neuroanatomy of memory. , 1993, Annual review of neuroscience.

[24]  C. D. Woody,et al.  Effects of acetylcholine and cyclic GMP on input resistance of cortical neurons in awake cats , 1978, Brain Research.

[25]  D. Armstrong Ultrastructural characterization of choline acetyltransferase‐containing neurons in the basal forebrain of rat: Evidence for a cholinergic innervation of intracerebral blood vessels , 1986, The Journal of comparative neurology.

[26]  P. Lea,et al.  Resolution limit of serial sections for 3D reconstruction of tubular cristae in rat liver mitochondria. , 1991, Journal of electron microscopy technique.

[27]  Karl Zilles,et al.  Immunohisto- and cytochemical localization of cortical nicotinic cholinoceptors in rat and man , 1989, Brain Research.

[28]  J. E. Vaughn,et al.  Small cholinergic neurons within fields of cholinergic axons characterize olfactory-related regions of rat telencephalon , 1992, Neuroscience.

[29]  K. Krnjević,et al.  Pharmacological properties of acetylcholine‐sensitive cells in the cerebral cortex , 1963, The Journal of physiology.

[30]  S. Palay,et al.  The Fine Structure of the Nervous System: Neurons and Their Supporting Cells , 1991 .

[31]  C. Aoki,et al.  Cholinergic terminals in the cat visual cortex: Ultrastructural basis for interaction with glutamate-immunoreactive neurons and other cells , 1992, Visual Neuroscience.

[32]  T. Sejnowski,et al.  Thalamocortical oscillations in the sleeping and aroused brain. , 1993, Science.

[33]  H. Fibiger,et al.  Chapter 49: Role of forebrain cholinergic systems in learning and memory: relevance to the cognitive deficits of aging and Alzheimer's dementia , 1993 .

[34]  L. Descarries,et al.  Ultrastructural features of the serotonin innervation in adult rat hippocampus: An immunocytochemical description in single and serial thin sections , 1991, Neuroscience.

[35]  A. Cuello,et al.  Nerve growth factor-induced synaptogenesis and hypertrophy of cortical cholinergic terminals. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[36]  Low-affinity nerve growth factor receptor (p75NGFR)- and choline acetyltransferase (ChAT)-immunoreactive axons in the cerebral cortex and hippocampus of adult macaque monkeys and humans. , 1993, Cerebral cortex.

[37]  M. Frotscher,et al.  Cholinergic innervation of hippocampal GAD‐ and somatostatin‐immunoreactive commissural neurons , 1987, The Journal of comparative neurology.

[38]  C. Rossi-Arnaud,et al.  Mechanical deafferentation of basal forebrain-cortical pathways and neurotoxic lesions of the nucleus basalis magnocellularis: comparative effect on spatial learning and cortical acetylcholine release in vivo , 1993, Behavioural Brain Research.

[39]  P. Davies,et al.  SELECTIVE LOSS OF CENTRAL CHOLINERGIC NEURONS IN ALZHEIMER'S DISEASE , 1976, The Lancet.

[40]  K. Krnjević,et al.  The mechanism of excitation by acetylcholine in the cerebral cortex , 1971, The Journal of physiology.

[41]  J. Coyle,et al.  Topographic analysis of the innervation of the rat neocortex and hippocampus by the basal forebrain cholinergic system , 1983, The Journal of comparative neurology.

[42]  H. Fibiger,et al.  Substance P in the ascending cholinergic reticular system , 1983, Nature.

[43]  P. Sargent,et al.  The diversity of neuronal nicotinic acetylcholine receptors. , 1993, Annual review of neuroscience.

[44]  A. Levey,et al.  Cholinergic innervation of cortex by the basal forebrain: Cytochemistry and cortical connections of the septal area, diagonal band nuclei, nucleus basalis (Substantia innominata), and hypothalamus in the rhesus monkey , 1983, The Journal of comparative neurology.

[45]  M. Mesulam,et al.  Three-dimensional representation and cortical projection topography of the nucleus basalis (Ch4) in the macaque: concurrent demonstration of choline acetyltransferase and retrograde transport with a stabilized tetramethylbenzidine method for horseradish peroxidase , 1986, Brain Research.

[46]  A. Silver,et al.  Choline acetylase in the central nervous system of man and some other mammals , 1956, The Journal of physiology.

[47]  L. Hösli,et al.  Receptors for neurotransmitters on astrocytes in the mammalian central nervous system , 1993, Progress in Neurobiology.

[48]  KM Harris,et al.  Three-dimensional structure of dendritic spines and synapses in rat hippocampus (CA1) at postnatal day 15 and adult ages: implications for the maturation of synaptic physiology and long-term potentiation [published erratum appears in J Neurosci 1992 Aug;12(8):following table of contents] , 1992, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[49]  E. Jaffé,et al.  Release of [3H]dopamine from rat prefrontal cortex: Modulation through presynaptic cholinergic heteroreceptors , 1989, Neuroscience Letters.

[50]  J. Richard,et al.  Evidence for the existence of serotonin type-2 receptors on cholinergic terminals in rat cortex , 1985, Brain Research.

[51]  J. E. Vaughn,et al.  Immunocytochemical localization of choline acetyltransferase in rat cerebral cortex: A study of cholinergic neurons and synapses , 1985, The Journal of comparative neurology.

[52]  W. Singer,et al.  Cholinergic innervation of the cat striate cortex: A choline acetyltransferase immunocytochemical analysis , 1986, The Journal of comparative neurology.

[53]  M. Mesulam,et al.  Choline acetyltransferase-immunoreactive neurons intrinsic to rodent cortex and distinction from acetylcholinesterase-positive neurons , 1984, Neuroscience.

[54]  K. Harris,et al.  Dendritic spines of rat cerebellar Purkinje cells: serial electron microscopy with reference to their biophysical characteristics , 1988, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[55]  S. Dunnett,et al.  Ultrastructural organization of choline acetyltransferase‐immunoreactive fibres innervating the neocortex from embryonic ventral forebrain grafts , 1986, The Journal of comparative neurology.

[56]  T. Freund,et al.  Cholinergic synapses in the rat brain: a correlated light and electron microscopic immunohistochemical study employing a monoclonal antibody against choline acetyltransferase , 1984, Brain Research.

[57]  A. Beaudet,et al.  Synaptic remodeling of serotonin axon terminals in rat agranular cerebellum , 1981, Brain Research.

[58]  D M Bowen,et al.  Neurotransmitter-related enzymes and indices of hypoxia in senile dementia and other abiotrophies. , 1976, Brain : a journal of neurology.

[59]  F. Eckenstein,et al.  Cholinergic neurons and fibres in the rat visual cortex , 1986, Journal of neurocytology.

[60]  D. Price,et al.  Cholinergic immunoreactive fibers in monkey anterior temporal cortex. , 1992, Cerebral Cortex.

[61]  G. Paxinos,et al.  The Rat Brain in Stereotaxic Coordinates , 1983 .

[62]  J. Lauterborn,et al.  In situ hybridization localization of choline acetyltransferase mRNA in adult rat brain and spinal cord. , 1993, Brain research. Molecular brain research.

[63]  L. Descarries,et al.  Ultrastructural analysis of the serotonin hyperinnervation in adult rat neostriatum following neonatal dopamine denervation with 6-hydroxydopamine , 1992, Brain Research.

[64]  F. Eckenstein,et al.  Cholinergic Innervation in Cerebral Cortex , 1987 .

[65]  B. K. Hartman,et al.  Light and electron microscopic immunocytochemical analysis of the neurovascular relationships of choline acetyltransferase and vasoactive intestinal polypeptide nerve terminals in the rat cerebral cortex , 1994, The Journal of comparative neurology.

[66]  H J Gundersen,et al.  The efficiency of systematic sampling in stereology and its prediction * , 1987, Journal of microscopy.

[67]  G. Koelle The histochemical localization of cholinesterases in the central nervous system of the rat , 1954, The Journal of comparative neurology.

[68]  C. Shute,et al.  The cholinergic limbic system: projections to hippocampal formation, medial cortex, nuclei of the ascending cholinergic reticular system, and the subfornical organ and supra-optic crest. , 1967, Brain : a journal of neurology.

[69]  L. Heimer,et al.  A correlated light and electron microscopic immunocytochemical study of cholinergic terminals and neurons in the rat amygdaloid body with special emphasis on the basolateral amygdaloid nucleus , 1986, The Journal of comparative neurology.

[70]  L. Descarries,et al.  Ultrastructural features of serotonin neurons grafted to adult rat hippocampus: An immunocytochemical analysis of their cell bodies and axon terminals , 1991, Neuroscience.

[71]  M. Frotscher,et al.  The cholinergic innervation of the rat fascia dentata: Identification of target structures on granule cells by combining choline acetyltransferase immunocytochemistry and Golgi impregnation , 1986, The Journal of comparative neurology.

[72]  L. Butcher,et al.  Cholinergic systems in the rat brain: I. Projections to the limbic telencephalon , 1984, Brain Research Bulletin.

[73]  L. Descarries,et al.  Recovery of choline acetyltransferase activity without sprouting of the residual acetylcholine innervation in adult rat cerebral cortex after lesion of the nucleus basalis , 1993, Brain Research.

[74]  D. Jones,et al.  Morphological characteristics of perforated synapses in the latter stages of synaptogenesis in rat neocortex: stereological and three-dimensional approaches , 1993, Journal of neurocytology.

[75]  J. E. Vaughn,et al.  Organization and morphological characteristics of cholonergic neurons: an immunocytochemical study with a monoclonal antibody to choline acetyltransferase , 1983, Brain Research.

[76]  C. Shute,et al.  The ascending cholinergic reticular system: neocortical, olfactory and subcortical projections. , 1967, Brain : a journal of neurology.

[77]  Michael Frotscher,et al.  Cholinergic innervation of the rat hippocampus as revealed by choline acetyltransferase immunocytochemistry: A combined light and electron microscopic study , 1985, The Journal of comparative neurology.

[78]  K. Krnjević,et al.  Acetylcholine‐sensitive cells in the cerebral cortex , 1963, The Journal of physiology.

[79]  C. Shute,et al.  Cholinesterase-Containing Systems of the Brain of the Rat , 1963, Nature.

[80]  P. Somogyi,et al.  Enrichment of cholinergic synaptic terminals on GABAergic neurons and coexistence of immunoreactive GABA and choline acetyltransferase in the same synaptic terminals in the striate cortex of the cat , 1991, The Journal of comparative neurology.

[81]  J. Mitchell,et al.  The spontaneous and evoked release of acetylcholine from the cerebral cortex , 1963, The Journal of physiology.

[82]  Gastone G. Celesia,et al.  Acetylcholine released from cerebral cortex in relation to state of activation , 1966, Neurology.

[83]  O. Lindvall,et al.  Distribution of putative neurotransmitters in the neocortex , 1979, Neuroscience.

[84]  S. Tuček Short-term control of the synthesis of acetylcholine. , 1993, Progress in biophysics and molecular biology.

[85]  J. Changeux,et al.  Nicotinic and muscarinic modulations of excitatory synaptic transmission in the rat prefrontal cortexin vitro , 1993, Neuroscience.

[86]  K. Baskerville,et al.  Topography of cholinergic afferents from the nucleus basalis of meynert to representational areas of sensorimotor cortices in the rat , 1993, The Journal of comparative neurology.

[87]  Nancy J. Woolf,et al.  Cholinergic systems in mammalian brain and spinal cord , 1991, Progress in Neurobiology.

[88]  D. Jones,et al.  Perforated and non-perforated synapses in rat neocortex: three-dimensional reconstructions , 1991, Brain Research.

[89]  Changiz Geula,et al.  Differential cholinergic innervation within functional subdivisions of the human cerebral cortex: A choline acetyltransferase study , 1992, The Journal of comparative neurology.

[90]  H. Gundersen Stereology of arbitrary particles * , 1986, Journal of microscopy.

[91]  Karl J. Zilles,et al.  The Cortex of the Rat: A Stereotaxic Atlas , 1985 .

[92]  D. Clarke Cholinergic innervation of the rat dentate gyrus: an immunocytochemical and electron microscopical study , 1985, Brain Research.

[93]  TATSUYA KANAI,et al.  Mesencephalic Reticular Activating System and Cortical Acetylcholine Output , 1965, Nature.

[94]  Prof. Dr. Karl Zilles The Cortex of the Rat , 1985, Springer Berlin Heidelberg.

[95]  Laurent Descarries,et al.  Noradrenergic axon terminals in the cerebral cortex of rat. III. Topometric ultrastructural analysis , 1977, Brain Research.

[96]  W. Feldberg,et al.  Acetylcholine synthesis in different regions of the central nervous system , 1948, The Journal of physiology.

[97]  C. Houser Cholinergic synapses in the central nervous system: studies of the immunocytochemical localization of choline acetyltransferase. , 1990, Journal of electron microscopy technique.

[98]  H. Fibiger,et al.  The nucleus basalis magnocellularis: The origin of a cholinergic projection to the neocortex of the rat , 1980, Neuroscience.

[99]  J. H. Quastel,et al.  Choline ester formation in, and choline esterase activities of, tissues in vitro. , 1936, The Biochemical journal.

[100]  P. Salvaterra,et al.  Interaction of monoclonal antibodies with mammalian choline acetyltransferase. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[101]  M. Brann,et al.  Muscarinic acetylcholine receptor subtypes: localization and structure/function. , 1993, Progress in brain research.

[102]  S. Foote,et al.  Distribution of choline acetyltransferase‐, serotonin‐, dopamine‐β‐hydroxylase‐, tyrosine hydroxylase‐immunoreactive fibers in monkey primary auditory cortex , 1987, The Journal of comparative neurology.

[103]  F. James Rohlf,et al.  Biometry: The Principles and Practice of Statistics in Biological Research , 1969 .

[104]  K. Krnjević,et al.  A histochemical study of cholinergic fibres in the cerebral cortex. , 1965, Journal of anatomy.

[105]  L. Mendell,et al.  Quantitative ultrastructure of Ia boutons in the ventral horn: scaling and positional relationships , 1993, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[106]  James M. Bower,et al.  Acetylcholine and memory , 1993, Trends in Neurosciences.