Contributions of selective knockout studies to understanding cholinesterase disposition and function.
The complete knockout of the acetylcholinesterase gene (AChE) in the mouse yielded a surprising phenotype that could not have been predicted from deletion of the cholinesterase genes in Drosophila, that of a living, but functionally compromised animal. The phenotype of this animal showed a sufficient compromise in motor function that precluded precise characterization of central and peripheral nervous functional deficits. Since AChE in mammals is encoded by a single gene with alternative splicing, additional understanding of gene expression might be garnered from selected deletions of the alternatively spliced exons. To this end, transgenic strains were generated that deleted exon 5, exon 6, and the combination of exons 5 and 6. Deletion of exon 6 reduces brain AChE by 93% and muscle AChE by 72%. Deletion of exon 5 eliminates AChE from red cells and the platelet surface. These strains, as well as knockout strains that selectively eliminate the AChE anchoring protein subunits PRiMA or ColQ (which bind to sequences specified by exon 6) enabled us to examine the role of the alternatively spliced exons responsible for the tissue disposition and function of the enzyme. In addition, a knockout mouse was made with a deletion in an upstream intron that had been identified in differentiating cultures of muscle cells to control AChE expression. We found that deletion of the intronic regulatory region in the mouse essentially eliminated AChE in muscle and surprisingly from the surface of platelets. The studies generated by these knockout mouse strains have yielded valuable insights into the function and localization of AChE in mammalian systems that cannot be approached in cell culture or in vitro.
Camp
Shelley
S
Department Pharmacology, Skaggs School of Pharmacy & Pharmaceutical Sciences 0657, University of California-San Diego, La Jolla, CA 92093-0657, USA. scamp@ucsd.edu
Zhang
Limin
L
Krejci
Eric
E
Dobbertin
Alexandre
A
Bernard
Véronique
V
Girard
Emmanuelle
E
Duysen
Ellen G
EG
Lockridge
Oksana
O
De Jaco
Antonella
A
Taylor
Palmer
P
D000110:Acetylcholinesterase / Q000737:chemistry / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism*; D000818:Animals; D001921:Brain / Q000201:enzymology; D003094:Collagen / Q000172:deficiency / Q000235:genetics; D005091:Exons / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic; D055786:Gene Knockout Techniques; D007438:Introns / Q000235:genetics; D008565:Membrane Proteins / Q000172:deficiency / Q000235:genetics; D051379:Mice; D018345:Mice, Knockout; D009124:Muscle Proteins / Q000172:deficiency / Q000235:genetics; D009132:Muscles / Q000201:enzymology; D009419:Nerve Tissue Proteins / Q000172:deficiency / Q000235:genetics; D009928:Organ Specificity; D021122:Protein Subunits / Q000172:deficiency / Q000235:genetics; D012326:RNA Splicing / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D017384:Sequence Deletion; D013116:Spinal Cord / Q000201:enzymology
D008565:Membrane Proteins; D009124:Muscle Proteins; D009419:Nerve Tissue Proteins; D021122:Protein Subunits; D012333:RNA, Messenger; C486718:prima1 protein, mouse; D003094:Collagen; D000110:Acetylcholinesterase; C534672:Colq protein, mouse