title: Visualization of fibrillar amyloid deposits in living, transgenic Caenorhabditis elegans animals using the sensitive amyloid dye, X-34.
abstract: Transgenic Caenorhabditis elegans animals can be engineered to express high levels of the human beta amyloid peptide (Abeta). Histochemistry of fixed tissue from these animals reveals deposits reactive with the amyloid-specific dyes Congo Red and thioflavin S (Fay et al., J. Neurochem 71:1616, 1998). Here we show by immuno-electron microscopy that these animals contain intracellular immunoreactive deposits with classic amyloid fibrillar ultrastructure. These deposits can be visualized in living animals using the newly developed, intensively fluorescent, amyloid-specific dye X-34. This in vivo staining allows monitoring of amyloid deposition in individual animals over time. The specificity of this staining is demonstrated by examining transgenic animals expressing high levels of a non-fibrillar beta peptide variant, the beta single-chain dimer. These animals have deposits immunoreactive with anti-beta antibodies, but do not have X-34 deposits or deposits with a fibrillar ultrastructure. X-34 can also be used in vivo to visualize putative amyloid deposits resulting from accumulation of human transthyretin, another amyloidic protein. In vivo amyloid staining with X-34 may be a useful tool for monitoring anti-amyloidic treatments in real time or screening for genetic alterations that affect amyloid formation.
authors.affiliation: Institute for Behavioral Genetics, University of Colorado, Campus Box 447, Boulder, CO 80309, USA. linkc@colorado.edu
authors.forename: C D
authors.initials: CD
authors.lastname: Link
authors.forename: C J
authors.initials: CJ
authors.lastname: Johnson
authors.forename: V
authors.initials: V
authors.lastname: Fonte
authors.forename: M
authors.initials: M
authors.lastname: Paupard
authors.forename: D H
authors.initials: DH
authors.lastname: Hall
authors.forename: S
authors.initials: S
authors.lastname: Styren
authors.forename: C A
authors.initials: CA
authors.lastname: Mathis
authors.forename: W E
authors.initials: WE
authors.lastname: Klunk
mesh_terms: D000475:Alkenes; D016229:Amyloid beta-Peptides / Q000235:genetics*; D000686:Amyloidosis / Q000473:pathology*; D000818:Animals; D030801:Animals, Genetically Modified; D001565:Benzoates; D017173:Caenorhabditis elegans; D004195:Disease Models, Animal; D005456:Fluorescent Dyes; D016253:Microscopy, Immunoelectron; D009474:Neurons / Q000473:pathology* / Q000648:ultrastructure; D011228:Prealbumin / Q000235:genetics*; D013194:Staining and Labeling / Q000379:methods*
chemical_list: C413183:1,4-bis(3-carboxy-4-hydroxyphenylethenyl)-benzene; D000475:Alkenes; D016229:Amyloid beta-Peptides; D001565:Benzoates; D005456:Fluorescent Dyes; D011228:Prealbumin