title: Lipid interference with fluorometric assay of DNA in adipose tissues under various conditions.
abstract: Interference by lipids with fluorometric assay of DNA in adipose tissues using Hoechst 33258 was investigated. Mixed glycerides shifted the emission maximum of standard DNA and induced a dose-dependent increase in fluorescence intensity. Glycerides in the samples containing a known concentration of DNA yielded erroneously higher DNA concentrations. The DNA concentrations obtained from acetone-defatted white and brown adipose tissues (WAT and BAT) were lower than those of non-defatted ones, while DNA content did not differ in low lipid-containing skeletal muscle between defatted and non-defatted samples, indicating that large amounts of lipids interfere with DNA measurement using Hoechst 33258 and that acetone defatting is a simple method to avoid this interference. Using this defatting method, the cellularity of WAT and BAT was estimated in rats under various experimental conditions. Cold-acclimation and repetitive immobilization stress decreased the body weight gain and the epididymal WAT weight. Sucrose overfeeding increased WAT weight but not body weight. These treatments of 4 weeks' duration did not induce any significant difference in WAT cell number from controls, while cold-acclimation increased the tissue cell number as well as the BAT weight.
authors.affiliation: Department of Physiology I, Asahikawa Medical College, Japan.
authors.forename: S K
authors.initials: SK
authors.lastname: Saha
authors.forename: M
authors.initials: M
authors.lastname: Moriya
authors.forename: H
authors.initials: H
authors.lastname: Ohinata
authors.forename: A
authors.initials: A
authors.lastname: Kuroshima
mesh_terms: D000096:Acetone / Q000737:chemistry; D000273:Adipose Tissue / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D002001:Adipose Tissue, Brown / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D000818:Animals; D006690:Bisbenzimidazole / Q000737:chemistry; D001835:Body Weight / Q000187:drug effects / Q000502:physiology; D002725:Chloroform / Q000737:chemistry; D003080:Cold Temperature; D004247:DNA / Q000032:analysis / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D002149:Energy Intake / Q000502:physiology; D000431:Ethanol / Q000737:chemistry; D004986:Ether / Q000737:chemistry; D005989:Glycerides / Q000737:chemistry / Q000494:pharmacology*; D007103:Immobilization; D008297:Male; D000432:Methanol / Q000737:chemistry; D018482:Muscle, Skeletal / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D000069463:Olive Oil; D010938:Plant Oils / Q000737:chemistry / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D013050:Spectrometry, Fluorescence; D013395:Sucrose / Q000008:administration & dosage
chemical_list: D005989:Glycerides; D000069463:Olive Oil; D010938:Plant Oils; D004986:Ether; D000096:Acetone; D000431:Ethanol; D013395:Sucrose; D002725:Chloroform; D004247:DNA; D006690:Bisbenzimidazole; D000432:Methanol