title: Autophagy is disrupted in a knock-in mouse model of juvenile neuronal ceroid lipofuscinosis.
abstract: Juvenile neuronal ceroid lipofuscinosis is caused by mutation of a novel, endosomal/lysosomal membrane protein encoded by CLN3. The observation that the mitochondrial ATPase subunit c protein accumulates in this disease suggests that autophagy, a pathway that regulates mitochondrial turnover, may be disrupted. To test this hypothesis, we examined the autophagic pathway in Cln3(Deltaex7/8) knock-in mice and CbCln3(Deltaex7/8) cerebellar cells, accurate genetic models of juvenile neuronal ceroid lipofuscinosis. In homozygous knock-in mice, we found that the autophagy marker LC3-II was increased, and mammalian target of rapamycin was down-regulated. Moreover, isolated autophagic vacuoles and lysosomes from homozygous knock-in mice were less mature in their ultrastructural morphology than the wild-type organelles, and subunit c accumulated in autophagic vacuoles. Intriguingly, we also observed subunit c accumulation in autophagic vacuoles in normal aging mice. Upon further investigation of the autophagic pathway in homozygous knock-in cerebellar cells, we found that LC3-positive vesicles were altered and overlap of endocytic and lysosomal dyes was reduced when autophagy was stimulated, compared with wildtype cells. Surprisingly, however, stimulation of autophagy did not significantly impact cell survival, but inhibition of autophagy led to cell death. Together these observations suggest that autophagy is disrupted in juvenile neuronal ceroid lipofuscinosis, likely at the level of autophagic vacuolar maturation, and that activation of autophagy may be a prosurvival feedback response in the disease process.
authors.affiliation: Molecular Neurogenetics Unit and Center for Human Genetic Research, Massachusetts General Hospital, Boston, Massachusetts 02114, USA.
authors.forename: Yi
authors.initials: Y
authors.lastname: Cao
authors.forename: Janice A
authors.initials: JA
authors.lastname: Espinola
authors.forename: Elisa
authors.initials: E
authors.lastname: Fossale
authors.forename: Ashish C
authors.initials: AC
authors.lastname: Massey
authors.forename: Ana Maria
authors.initials: AM
authors.lastname: Cuervo
authors.forename: Marcy E
authors.initials: ME
authors.lastname: MacDonald
authors.forename: Susan L
authors.initials: SL
authors.lastname: Cotman
mesh_terms: D000818:Animals; D001343:Autophagy / Q000235:genetics*; D001921:Brain / Q000473:pathology; D004195:Disease Models, Animal; D006720:Homozygote; D006801:Humans; D008562:Membrane Glycoproteins / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D018832:Molecular Chaperones / Q000235:genetics; D009472:Neuronal Ceroid-Lipofuscinoses / Q000235:genetics* / Q000473:pathology; D011494:Protein Kinases / Q000235:genetics; D017384:Sequence Deletion; D058570:TOR Serine-Threonine Kinases
chemical_list: C105199:CLN3 protein, human; D008562:Membrane Glycoproteins; D018832:Molecular Chaperones; D011494:Protein Kinases; C546842:MTOR protein, human; C546843:mTOR protein, mouse; D058570:TOR Serine-Threonine Kinases