Neutral ceramidase gene: role in regulating ceramide-induced apoptosis.
The sphingolipid, ceramide, is a natural dietary constituent and a potent mediator of apoptosis. If left undegraded, it may induce apoptosis and cause disruption of cellular integrity. A potential mechanism to prevent ceramide-induced apoptosis in various organs may involve ceramidases that facilitate the degradation of ceramide. In this study, we first isolated and characterized the murine neutral ceramidase (N-CDase) gene, mapped its chromosomal location and determined its developmental and organ-specific expression. Then we used cultured mesangial cells as our in vitro model and mouse gastrointestinal (GI) tract as the in vivo model to determine the effects of an inhibitor of N-CDase, D-erythro-MAPP, to delineate whether N-CDase plays a role in preventing ceramide-induced apoptosis. Our results show that: (i) the structure of the murine neutral ceramidase gene is virtually identical to that of the human gene; (ii) it is localized on chromosome 19 at bands C2-C3 that is syntenic to human chromosome 10q24-26; (iii) N-CDase expression is developmentally regulated and it is expressed at high levels in cultured mesangial cells and in specific regions of the mouse small intestine; (iv) inhibition of N-CDase by D-erythro-MAPP leads to increased ceramide levels and consequent apoptosis in cultured mesangial cells; (v) mice treated with D-erythro-MAPP alone also caused apoptosis in the small intestine; and (vi) mice treated with D-erythro-MAPP prior to feeding C2 ceramide manifest markedly elevated levels of apoptosis in the GI tract raising the possibility that neutral ceramidase plays a detoxifying role against inadvertent stimulation of ceramide-induced apoptosis in organs that come in contact with this sphingolipid. We propose that N-CDase is an essential component of an innate detoxifying mechanism to prevent ceramide-induced apoptosis.
Choi
Moonsuk S
MS
Section on Developmental Genetics, Heritable Disorders Branch, The National Institute of Child Health and Human Development, The National Institutes of Health, Room 9S241, Building 10, Bethesda, MD 20892-1830, USA.
Anderson
Mary A
MA
Zhang
Zhongjian
Z
Zimonjic
Drazen B
DB
Popescu
Nicolae
N
Mukherjee
Anil B
AB
D000581:Amidohydrolases / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000502:physiology; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D002460:Cell Line; D055572:Ceramidases; D002518:Ceramides / Q000378:metabolism / Q000494:pharmacology*; D002874:Chromosome Mapping; D004305:Dose-Response Relationship, Drug; D005091:Exons; D015870:Gene Expression; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D005796:Genes / Q000235:genetics; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D020287:In Situ Nick-End Labeling; D007438:Introns; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008928:Mitochondria / Q000201:enzymology; D008969:Molecular Sequence Data; D009226:Myristates / Q000494:pharmacology; D055576:Neutral Ceramidase; D011412:Propanolamines / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA
D002518:Ceramides; D009226:Myristates; D011412:Propanolamines; D012333:RNA, Messenger; C099657:2-(N-myristoylamino)-1-phenyl-1-propanol; D000581:Amidohydrolases; C526744:ASAH2 protein, human; C526747:Asah2 protein, mouse; D055572:Ceramidases; D055576:Neutral Ceramidase