Glycine receptor β–subunit gene mutation in spastic mouse associated with LINE–1 element insertion

Abstract

Congenital myoclonus is a widespread neurologic disorder characterized by hyperexcitability, muscular spasticity and myoclonus associated with marked reduction in neural glycine binding sites. The recessive mouse mutation spastic (spa) is a prototype of inherited myoclonus. Here we show that defects in the gene encoding the β–subunit of the glycine receptor (Glrb) underlie spa: Glrb maps to the same region of mouse chromosome 3 as spa, and Glrb mRNA is markedly reduced throughout brains of spa mice, most likely as a result of an insertional mutation of a 7.1 kilobase LINE–1 element within intron 6 of Glrb. These results provide evidence that Glrb is necessary for postsynaptic expression of glycine receptor complexes, and suggest Glrb as a candidate gene for inherited myoclonus in other species.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1

    Betz, H. Structure and function of inhibitory glycine receptors. Quart. Rev. Biophys. 25, 381–394 (1992).

  2. 2

    Betz, H. Ligand-gated ion channels in the brain: the amino acid receptor family. Neuron 5, 383–392 (1990).

  3. 3

    Grenningloh, G. et al. The strychnine-binding subunit of the glycine receptor shows homology with nicotinic acetylcholine receptors. Nature 328, 215–220 (1987).

  4. 4

    Kuhse, J., Schmieden, V. & Betz, H. Identification and functional expression of a novel ligand binding subunit of the inhibitory glycine receptor. J. biol. Chem. 265, 22317–22320 (1990).

  5. 5

    Kuhse, J., Schmieden, V. & Betz, H. A single amino acid exchange alters the pharmacology of neonatal rat glycine receptor subunit. Neuron 5, 867–873 (1990).

  6. 6

    Grenningloh, G. et al. Cloning and expression of the 58 kd β subunit of the inhibitory glycine receptor. Neuron 4, 963–970 (1990).

  7. 7

    Schmieden, V., Kuhse, J. & Betz, H. Agonist pharmacology of neonatal and adult glycine receptor α subunits: identification of amino acid residues involved in taurine activation. EMBO J. 11, 2025–2032 (1992).

  8. 8

    Schmieden, V., Grenningloh, G., Schofield, P.R. & Betz, H. Functional expression in Xenopus oocytes of the strychnine binding 48 kd subunit of the glycine receptor. EMBO J. 3, 695–700 (1989).

  9. 9

    Sontheimer, H. et al. Functional chloride channels by mammalian cell expression of rat glycine receptor subunit. Neuron 2, 1491–1497 (1989).

  10. 10

    Pribilla, I., Takagi, T., Langosch, D., Bormann, J. & Betz, H. The atypical M2 segment of the β subunit confers picrotoxinin resistance to inhibitory glycine receptor channels. EMBO J. 11, 4305–4311 (1992).

  11. 11

    White, W.F. & Heller, A.H. Glycine receptor alteration in the mutant mouse spastic. Nature 298, 655–657 (1982).

  12. 12

    Gundlach, A.L. et al. Deficit of glycine/strychnine receptors in inherited myoclonus of Poll Hereford calves. Science 241, 1807–1810 (1988).

  13. 13

    Gundlach, A.L. Disorder of the inhibitory glycine receptor: inherited myoclonus in Poll Hereford calves. FASEB J. 4, 2761–2766 (1990).

  14. 14

    Gundlach, A.L., Kortz, G., Burazin, T.C.D., Madigan, J. & Higgins, R.J. Deficit of inhibitory glycine receptors in spinal cord from Peruvian Pasos: evidence for an equine form of inherited myoclonus. Brain Res. 628, 263–270 (1993).

  15. 15

    Shiang, R. et al. Mutations in the α1 subunit of the inhibitory glycine receptor cause the dominant neurologic disorder, hyperekplexia. Nature Genet. 5, 351–358 (1993).

  16. 16

    Ryan, S.G. et al. A missense mutation in the gene encoding the α1 subunit of the inhibitory glycine receptor causes the spasmodic mouse phenotype. Nature Genet. 7, 131–135 (1994).

  17. 17

    Heller, A.H. & Hallett, M. Electrophysiological studies with the spastic mutant mouse. Brain Res. 234, 299–308 (1982).

  18. 18

    Chai, C.K. Hereditary spasticity in mice. J. Hered. 52, 241–243 (1961).

  19. 19

    Chai, C.K., Roberts, E. & Sidman, R.L. Influence of aminooxyacetic acid, a gamma-aminobutyrate transaminase inhibitor, on hereditary spastic defect in the mouse. Proc. Soc. exp. Biol. Med. 109, 491–495 (1962).

  20. 20

    Meier, H. & Chai, C.K. spastic, an hereditary neurological mutation in the mouse characterized by vertebral arthropathy and leptomeningeal cyst formation. Exp. Med. Surg. 28, 24–38 (1970).

  21. 21

    Ziv, I., Blackburn, N., Rang, M. & Koreska, J. Muscle growth in normal and spastic mice. Dev. Med. child Neurol. 26, 94–99 (1984).

  22. 22

    Lane, P.W. Two new mutations in linkage group XVI of the house mouse, flaky tail and varitint-waddler-J. J. Hered. 63, 135–140 (1972).

  23. 23

    Eicher, E.M. & Lane, P.W. Assignment of LG XVI to chromosome 3 in the mouse. J. Hered. 71, 315–318 (1980).

  24. 24

    White, W.F. The glycine receptor in the mutant mouse spastic (spa): strychnine binding characteristics and pharmacology Brain Res. 329, 1–6 (1985).

  25. 25

    Becker, C.M., Hermans-Borgmeyer, I., Schmitt, B. & Betz, H. The glycine receptor deficiency of the mutant mouse spastic: evidence for normal glycine receptor structure and localization. J. Neurosci. 6, 1358–1364 (1986).

  26. 26

    Becker, C.M., Schmieden, V., Tarroni, P., Strasser, U. & Betz, H. Isoform-selective deficit of glycine receptors in the mouse mutant spastic. Neuron 8, 283–289 (1992).

  27. 27

    White, W.F. & Heller, A.H. Glycine and GABA uptake in the mutant mouse spastic. Soc. Neurosci. (Abstr.) 8, 575 (1982).

  28. 28

    Biscoe, T.J., Fry, J.P., Martin, I.L. & Rickets, C. Binding of GABA and benzodiazepine receptor ligands in the spinal cord of the spastic mouse, (ab). J. Physiol. 317, 32–33 (1981).

  29. 29

    Biscoe, T.J. & Fry, J.P. GABA and benzodiazepine receptor in neurologically mutant mice. in Actions and Interactions of GABA and Benzodiazepines, N.G. Bowery, Ed. 217–237 (Raven Press, New York, 1984).

  30. 30

    Seldin, M.F. et al. Genetic analysis of autoimmune gld mice. I. Identification of a restriction fragment length polymorphism closely linked to the gld mutation within a conserved linkage group. J. exp. Med. 167, 688–693 (1988).

  31. 31

    Green, E.L. Linkage, recombination and mapping. in Genetics and Probability in Animal Breeding Experiments 77–113 (Macmillan, New York, 1981).

  32. 32

    Bishop, D.T. The information content of phase-known matings for ordering genetic loci. Genet. Epidemiol. 2, 349–361 (1985).

  33. 33

    Seldin, M.F., Prins, J-B., Rodrigues, N.R., Todd, J.A. & Meisler, M.H. Mouse chromosome 3. Mamm. Genome 4, S47–S57 (1993).

  34. 34

    El Mestikawy, S. et al. Characterization of an atypical member of the Na+/Cl- dependent transporter family: Chromosomal localization and distribution in GABAergic and glutamatergic neurons in the rat brain. J. Neurochem. 62, 445–455 (1994).

  35. 35

    Gregor, P. et al. Chromosomal localization of glutamate receptor genes: Relationships to familial amyotrophic lateral sclerosis and other neurologic disorders of mice and humans. Proc. natn. Acad. Sci. U.S.A. 90, 3053–3057 (1993).

  36. 36

    Moseley, W.S. & Seldin, M.F. Definition of mouse chromosome 1 and 3 gene linkage groups that are conserved on human chromosome 1: Evidence that a conserved linkage group spans the centromere of human chromosome 1. Genomics 5, 899–905 (1989).

  37. 37

    Malosio, M-L., Marqueze-Pouey, B., Kuhse, J. & Betz, H. Widespread expression of glycine receptor subunit mRNAs in the adult and developing rat brain. EMBO J. 10, 2401–2409 (1991).

  38. 38

    Betz, H. Glycine receptors: heterogeneous and widespread in the mammalian brain. Trends Neurosci. 14, 458–461 (1991).

  39. 39

    Sommer, B., Poustka, A., Spurr, N.K. & Seeburg, P.H. The murine GABAA receptor δ-subunit gene: structure and assignment to human chromosome 1. DNA Cell Biol. 9, 561–568 (1990).

  40. 40

    Kirkness, E.F., et al. Isolation, characterization, and localization of human genomic DNA encoding the β1 subunit of GABAA, receptor (GABRB1) Genomics 10, 985–995 (1991).

  41. 41

    Lasham, A., Vreugdenhil, E., Bateson, A.N., Barnard, E.A. & Darlison, M.G. Conserved organization of γ-aminobutyric acidA receptor genes: cloning and analysis of the chicken β4-subunit gene. J. Neurochem. 57, 352–355 (1991).

  42. 42

    Fanning, T.G. Size and structure of the highly repetitive BAM HI element in mice. Nuci. Acids Res. 11, 5073–5091 (1983).

  43. 43

    Loeb, D.D. et al. The sequence of a large L1 Md element reveals a tandemly repeated 5′ end and several features found in retrotransposons. Molec. cell. Biol. 6, 168–182 (1986).

  44. 44

    Kuhse, J., Laube, B., Magalei, D. & Betz, H. Assembly of the in hibitory glycine receptor: Identification of amino acid sequence motifs governing subunit stoichiometry. Neuron 11, 1049–1056 (1993).

  45. 45

    Hoch, W., Betz, H. & Becker, C.M. Primary cultures of mouse spinal cord express the neonatal isoform of the inhibitory glycine receptor. Neuron 3, 339–348 (1989).

  46. 46

    Eickbush, T.H. Transposing without ends: the non-LTR retrotransposable elements. New Biologist 4, 430–440 (1992).

  47. 47

    Kazazian, Jr H.H. et al. Hemophilia A resulting from de novo insertion of L1 sequences represents a novel mechanism for mutation in man. Nature 332, 164–166 (1988).

  48. 48

    Narita, N. et al. Insertion of a 5′ truncated L1 element into the 3′ end of exon 44 of the dystrophin gene resulted in skipping of the exon during splicing in a case of Duchenne muscular dystrophy. J. clin. Invest. 91, 1862–1867 (1993).

  49. 49

    Morse, B., Rotherg, P.G., South, V.J., Spandorfer, J.M. & Astrin, S.M. Insertional mutagenesis of the MYC locus by a LINE-1 sequence in a human breast carcinoma. Nature 333, 87–90 (1988).

  50. 50

    Steinmeyer, K. et al. Inactivation of muscle chloride channel by transposon Insertion in myotonic mice. Nature 354, 304–308 (1991).

  51. 51

    Adachi, M., Watanabe-Fukunaga, R. & Nagata, S. Aberrant transcription caused by the insertion of an early transposable element in an intron of the Fas antigen gene of Ipr mice. Proc. natn. Acad. Sci. U.S.A. 90, 1756–1760 (1993).

  52. 52

    Kobayashi, S., Hirano, T., Kakinuma, M. & Uede, T. Transcriptional repression and differential splicing of FAS mRNA by early transposon (ETn) insertion in autoimmune Ipr mice. Biochem. Biophys. res. Commun. 191, 617–624 (1993).

  53. 53

    Sambrook, J., Fritsch, E.F. & Maniatis, T. in Molecular Cloning: A Laboratory Manual 2nd edn (Cold Spring Harbor Laboratory Press, New York, 1989).

  54. 54

    Giros, B., El Mestikawy, S., Bertrand, L. & Caron, M.G. Cloning and functional characterization of a cocaine-sensitive dopamine transporter. FEBS Lett. 295, 149–154 (1991).

Download references

Author information

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Kingsmore, S., Giros, B., Suh, D. et al. Glycine receptor β–subunit gene mutation in spastic mouse associated with LINE–1 element insertion. Nat Genet 7, 136–142 (1994). https://doi.org/10.1038/ng0694-136

Download citation

Further reading