An acetylcholine receptor alpha-subunit promoter conferring preferential synaptic expression in muscle of transgenic mice.
We have obtained transgenic mice expressing nuclearly targeted beta-galactosidase (nls-beta-gal) under the control of a chicken acetylcholine receptor alpha-subunit promoter. The expression of the transgene was detected in early somites, starting before embryonic day 9.5. In 13-day embryos, the expression pattern of the transgene closely paralleled that of the endogenous mouse alpha-subunit gene, assessed by in situ hybridization. Our results illustrate, with single-cell resolution, the tissue specificity of this alpha-subunit promoter during embryogenesis. After birth, the overall beta-galactosidase activity rapidly decreased with age. However, in diaphragms of newborn animals, beta-galactosidase activity selectively persisted in nuclei underlying the motor endplates. The latter were revealed by an acetylcholinesterase stain. Nls-beta-gal was also visualized by indirect immunofluorescence, while endplates were labelled with fluorescent alpha-bungarotoxin. Confocal microscopy unambiguously identified the more intensely stained nuclei as synaptic 'fundamental nuclei', and allowed estimates of relative staining levels. Thus an 842 bp acetylcholine receptor gene promoter confers preferential synaptic expression to a reporter gene within myofibres in vivo.
Klarsfeld
A
A
Unité de Neurobiologie Moléculaire, URA CNRS D1284, Institut Pasteur, Paris, France.
Bessereau
J L
JL
Salmon
A M
AM
Triller
A
A
Babinet
C
C
Changeux
J P
JP
D000818:Animals; D015139:Blotting, Southern; D002642:Chick Embryo; D002645:Chickens; D004247:DNA / Q000032:analysis / Q000235:genetics; D003964:Diaphragm; D005314:Embryonic and Fetal Development; D046911:Macromolecular Substances; D051379:Mice; D008822:Mice, Transgenic; D009132:Muscles / Q000502:physiology*; D009693:Nucleic Acid Hybridization; D011401:Promoter Regions, Genetic; D011950:Receptors, Cholinergic / Q000235:genetics* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013569:Synapses / Q000502:physiology*; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D046911:Macromolecular Substances; D011950:Receptors, Cholinergic; D011993:Recombinant Fusion Proteins; D004247:DNA; D001616:beta-Galactosidase