Antisense ‘knockdowns’ of M1 receptors induces transient anterograde amnesia in mice

The effect on memory processes of inactivation of the M1 gene by an antisense oligodeoxyribonucleotide (aODN) was investigated in the mouse passive avoidance test. Mice received a single intracerebroventricular (i.c.v.) injection of M1 aODN (0.3, 1.0 or 2.0 nmol per injection), degenerated ODN (dODN) or vehicle on days 1, 4 and 7. An amnesic effect, comparable to that produced by antimuscarinic drugs, was observed 12, 24, 48 and 72 h after the last i.c.v. aODN injection, whereas dODN and vehicle, used as controls, did not produce any effect. Reduction in the entrance latency to the dark compartment induced by aODN disappeared 7 days after the end of aODN treatment, which indicates the absence of any irreversible damage or toxicity caused by aODN. Quantitative reverse transcription-polymerase chain reaction analysis demonstrated that a decrease in M1 mRNA levels occurred only in the aODN-treated group, being absent in all control groups. Furthermore, a reduction in M1 receptors was observed in the hippocampus of aODN-treated mice. Neither aODN, dODN nor vehicle produced any behavioral impairment of mice. These results indicate that the integrity and functionality of M1 receptors are fundamental in the modulation of memory processes.

[1]  T. Crook,et al.  Anti-sense phosphorothioate oligonucleotides have both specific and non-specific effects on cells containing human papillomavirus type 16. , 1991, Nucleic acids research.

[2]  S. Capaccioli,et al.  Reversion of the invasive phenotype of human fibroblasts by antimessenger oligonucleotide inhibiting the urokinase receptor gene expression , 1994 .

[3]  Y. Cheng,et al.  Phosphorothioate oligonucleotides are inhibitors of human DNA polymerases and RNase H: implications for antisense technology. , 1992, Molecular pharmacology.

[4]  M. Giovannini,et al.  Stereoselective Increase in Cholinergic Transmission by R-(+)-hyoscyamine , 1997, Neuropharmacology.

[5]  S. Capaccioli,et al.  Effect of K+ channel modulation on mouse feeding behaviour. , 1997, European journal of pharmacology.

[6]  L. Neckers,et al.  Stability, clearance, and disposition of intraventricularly administered oligodeoxynucleotides: implications for therapeutic application within the central nervous system. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[7]  M. Packard,et al.  Post-training injection of the acetylcholine M2 receptor antagonist AF-DX 116 improves memory , 1990, Brain Research.

[8]  C. F. Bennett,et al.  Progress in antisense oligonucleotide therapeutics. , 1996, Annual review of pharmacology and toxicology.

[9]  L. Papucci,et al.  Intracellular enhancement of intact antisense oligonucleotide steady-state levels by cationic lipids. , 1994, Anti-cancer drug design.

[10]  L. Papucci,et al.  An antisense oligonucleotide on the mouse Shaker-like potassium channel Kv1.1 gene prevents antinociception induced by morphine and baclofen. , 1997, The Journal of pharmacology and experimental therapeutics.

[11]  H. Ladinsky,et al.  Binding and functional profiles of the selective M1 muscarinic receptor antagonists trihexyphenidyl and dicyclomine , 1986, British journal of pharmacology.

[12]  L. Papucci,et al.  Inhibition of MDR1 gene expression by antimessenger oligonucleotides lowers multiple drug resistance. , 1994, Oncology research.

[13]  G. Higgins,et al.  Central administration of the muscarinic receptor subtype ‐ selective antagonist pirenzepine selectively impairs passive avoidance learning in the mouse , 1983, The Journal of pharmacy and pharmacology.

[14]  M. Jarvik,et al.  An Improved One-Trial Passive Avoidance Learning Situation , 1967, Psychological reports.

[15]  J. Palacios,et al.  Muscarinic cholinergic receptor subtypes in the human brain. II. Quantitative autoradiographic studies , 1986, Brain Research.

[16]  J. Eras,et al.  Phosphorothioate oligonucleotides cause degradation of secretory but not intracellular serglycin proteoglycan core protein in a sequence-independent manner in human megakaryocytic tumor cells. , 1995, Antisense research and development.

[17]  C. Wahlestedt,et al.  Antisense oligonucleotide strategies in neuropharmacology. , 1994, Trends in pharmacological sciences.

[18]  J. S. Cohen,et al.  Oligodeoxynucleotides as antisense inhibitors of gene expression. , 1992, Progress in nucleic acid research and molecular biology.

[19]  T. Haley,et al.  Pharmacological effects produced by intracerebral injection of drugs in the conscious mouse. , 1957, British journal of pharmacology and chemotherapy.

[20]  P. Whitehouse Neuronal Loss and Neurotransmitter Receptor Alterations in Alzheimer’s Disease , 1986 .

[21]  C. Ghelardini,et al.  S-(-)-ET 126: a potent and selective M1 antagonist in vitro and in vivo. , 1995, Life sciences.

[22]  D. Mash,et al.  Loss of M2 muscarine receptors in the cerebral cortex in Alzheimer's disease and experimental cholinergic denervation. , 1985, Science.

[23]  J. Goodchild Inhibition of Gene Expression by Oligonucleotides , 1989 .

[24]  R. Bartus,et al.  Short-term memory in the rhesus monkey: Disruption from the anti-cholinergic scopolamine , 1976, Pharmacology Biochemistry and Behavior.

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

[26]  D Rodbard,et al.  Ligand: a versatile computerized approach for characterization of ligand-binding systems. , 1980, Analytical biochemistry.

[27]  S. Capaccioli,et al.  Cationic lipids improve antisense oligonucleotide uptake and prevent degradation in cultured cells and in human serum. , 1993, Biochemical and biophysical research communications.

[28]  Y. Cheng,et al.  Antisense oligonucleotides as therapeutic agents--is the bullet really magical? , 1993, Science.

[29]  J A Deutsch,et al.  The Cholinergic Synapse and the Site of Memory , 1971, Science.

[30]  A. Fisher,et al.  (±)-cis-2-Methyl-spiro(1,3-oxathiolane-5,3′) quinuclidine (AF102B): A new M1 agonist attenuates cognitive dysfunctions in AF64A-treated rats , 1989, Neuroscience Letters.

[31]  D. Braida,et al.  Effect of centrally administered atropine and pirenzepine on radial arm maze performance in the rat. , 1991, European journal of pharmacology.

[32]  N. Birdsall,et al.  Pirenzepine distinguishes between different subclasses of muscarinic receptors , 1980, Nature.

[33]  L. Kedes,et al.  Evolution of the functional human beta-actin gene and its multi-pseudogene family: conservation of noncoding regions and chromosomal dispersion of pseudogenes , 1985, Molecular and cellular biology.

[34]  R. Bartus,et al.  The cholinergic hypothesis of geriatric memory dysfunction. , 1982, Science.

[35]  L. Costa,et al.  A comparison of the antinociceptive responses to the GABA-receptor agonists THIP and baclofen , 1985, Neuropharmacology.

[36]  S. Dei,et al.  The medicinal chemistry of Alzheimer's and Alzheimer-like diseases with emphasis on the cholinergic hypothesis. , 1995, Farmaco.

[37]  D. Birch,et al.  Prevention of pre-PCR mis-priming and primer dimerization improves low-copy-number amplifications. , 1992, Nucleic acids research.

[38]  F. Roberts,et al.  The effect of pirenzepine on spatial learning in the Morris Water Maze , 1988, Pharmacology Biochemistry and Behavior.

[39]  L. Nilvebrant,et al.  Dicyclomine, benzhexol and oxybutynine distinguish between subclasses of muscarinic binding sites. , 1986, European journal of pharmacology.

[40]  L. Papucci,et al.  Quantitation of bcl-2 oncogene in cultured lymphoma/leukemia cell lines and in primary leukemia B-cells by a highly sensitive RT-PCR method. , 1995, Haematologica.

[41]  Alzheimer's and Parkinson's diseases : strategies for research and development , 1986 .

[42]  M. M. Bradford A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[43]  L. Squire,et al.  Structure and function of declarative and nondeclarative memory systems. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[44]  B. Habecker,et al.  Isolation, sequence, and functional expression of the mouse M1 muscarinic acetylcholine receptor gene. , 1988, The Journal of biological chemistry.

[45]  I. Creese,et al.  Reduction in muscarinic receptors by antisense oligodeoxynucleotide. , 1994, Biochemical pharmacology.