Screening for presenilin inhibitors using the free-living nematode, Caenorhabditis elegans.
Caenorhabditis elegans contains 3 homologs of presenilin genes that are associated with Alzheimer s disease. Loss-of-function mutations in C. elegans genes cause a defect in egg laying. In humans, loss of presenilin-1 (PS1) function reduces amyloid-beta peptide processing from the amyloid protein precursor. Worms were screened for compounds that block egg laying, phenocopying presenilin loss of function. To accommodate even relatively high throughput screening, a semi-automated method to quantify egg laying was devised by measuring the chitinase released into the culture medium. Chitinase is released by hatching eggs, but little is shed into the medium from the body cavity of a hermaphrodite with an egg laying deficient (egl) phenotype. Assay validation involved measuring chitinase release from wild-type C. elegans (N2 strain), sel-12 presenilin loss-of-function mutants, and 2 strains of C. elegans with mutations in the egl-36 K(+) channel gene. Failure to find specific presenilin inhibitors in this collection likely reflects the small number of compounds tested, rather than a flaw in screening strategy. Absent defined biochemical pathways for presenilin, this screening method, which takes advantage of the genetic system available in C. elegans and its historical use for anthelminthic screening, permits an entry into mechanism-based discovery of drugs for Alzheimer's disease.
Ellerbrock
Brenda R
BR
Pfizer, Central Research, Groton, CT 06340, USA. brenda.r.ellerbrock@pfizer.com
Coscarelli
Eileen M
EM
Gurney
Mark E
ME
Geary
Timothy G
TG
D000818:Animals; D017173:Caenorhabditis elegans / Q000235:genetics*; D029742:Caenorhabditis elegans Proteins / Q000235:genetics; D002688:Chitinases / Q000032:analysis / Q000378:metabolism; D017238:Genes, Helminth; D008565:Membrane Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics; D053764:Presenilin-1
D029742:Caenorhabditis elegans Proteins; C109040:Hop-1 protein, C elegans; D008565:Membrane Proteins; D053764:Presenilin-1; C095907:SEL-12 protein, C elegans; C076714:spe-4 protein, C elegans; D002688:Chitinases