Targeted disruption of the M(r) 46,000 mannose 6-phosphate receptor gene in mice results in misrouting of lysosomal proteins.

Köster, Anja; Säftig, Paul; Matzner, Ulrich; Figura, Kurt Von; Peters, C; Pohlmann, R

doi:10.1002/j.1460-2075.1993.tb06217.x

Cited by 89 publications

(68 citation statements)

References 17 publications

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“…The targetting vector was linearized with NotI and introduced into the ES cell line E14-1 (7). Electroporation, culture, and selection conditions have been described (7,8). Colonies selected with G418 and gancyclovir were screened by Southern blot analysis of DNA digested with EcoRI and hybridization with the external 3Ј probe (Fig.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Phenotype of arylsulfatase A-deficient mice: Relationship to human metachromatic leukodystrophy

Hess

Säftig

Hartmann

et al. 1996

Proc. Natl. Acad. Sci. U.S.A.

239

206

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Metachromatic leukodystrophy is a lysosomal sphingolipid storage disorder caused by the deficiency of arylsulfatase A. The disease is characterized by progressive demyelination, causing various neurologic symptoms. Since no naturally occurring animal model of the disease is available, we have generated arylsulfatase A-deficient mice. Deficient animals store the sphingolipid cerebroside-3-sulfate in various neuronal and nonneuronal tissues. The storage pattern is comparable to that of affected humans, but gross defects of white matter were not observed up to the age of 2 years. A reduction of axonal cross-sectional area and an astrogliosis were observed in 1-year-old mice; activation of microglia started at 1 year and was generalized at 2 years. Purkinje cell dendrites show an altered morphology. In the acoustic ganglion numbers of neurons and myelinated fibers are severely decreased, which is accompanied by a loss of brainstem auditory-evoked potentials. Neurologic examination reveals significant impairment of neuromotor coordination.

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Section: Methodsmentioning

confidence: 99%

“…Among 89 colonies examined, two targeted clones were identified and injected into blastocysts of C57BL͞6J mice as described (8). Chimeric male mice were bred with C57BL͞6J females and the offspring was analyzed by Southern blot analysis of tail DNA for the presence of the homologously recombined ASA gene.…”

Section: Methodsmentioning

confidence: 99%

Phenotype of arylsulfatase A-deficient mice: Relationship to human metachromatic leukodystrophy

Hess

Säftig

Hartmann

et al. 1996

Proc. Natl. Acad. Sci. U.S.A.

239

206

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“…After 6, 12, and 24 h, samples of the medium were removed and cells were washed, scraped, and homogenized in TBS/1% Triton X-100. Lysosomal enzyme activities were detected using fluorometric and colorometric assays as described (Kö ster et al, 1993). To avoid a possible effect of the medium change leading to nonspecific increase of lysosomal enzyme activities, the enzyme activity in the medium after 6-h culture was withdrawn from the specific activities measured in the medium after 12 and 24 h. In each experiment the enzyme activities were measured from two parallel culture dishes.…”

Section: Enzyme Activitiesmentioning

confidence: 99%

Role of LAMP-2 in Lysosome Biogenesis and Autophagy

Eskelinen

Illert²,

Tanaka

et al. 2002

MBoC

313

260

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In LAMP-2–deficient mice autophagic vacuoles accumulate in many tissues, including liver, pancreas, muscle, and heart. Here we extend the phenotype analysis using cultured hepatocytes. In LAMP-2–deficient hepatocytes the half-life of both early and late autophagic vacuoles was prolonged as evaluated by quantitative electron microscopy. However, an endocytic tracer reached the autophagic vacuoles, indicating delivery of endo/lysosomal constituents to autophagic vacuoles. Enzyme activity measurements showed that the trafficking of some lysosomal enzymes to lysosomes was impaired. Immunoprecipitation of metabolically labeled cathepsin D indicated reduced intracellular retention and processing in the knockout cells. The steady-state level of 300-kDa mannose 6-phosphate receptor was slightly lower in LAMP-2–deficient hepatocytes, whereas that of 46-kDa mannose 6-phosphate receptor was decreased to 30% of controls due to a shorter half-life. Less receptor was found in the Golgi region and in vesicles and tubules surrounding multivesicular endosomes, suggesting impaired recycling from endosomes to the Golgi. More receptor was found in autophagic vacuoles, which may explain its shorter half-life. Our data indicate that in hepatocytes LAMP-2 deficiency either directly or indirectly leads to impaired recycling of 46-kDa mannose 6-phosphate receptors and partial mistargeting of a subset of lysosomal enzymes. Autophagic vacuoles may accumulate due to impaired capacity for lysosomal degradation

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“…Mice deficient in MPR46 and/or M6P/IGF2R have been generated (Köster et al, 1993;Ludwig et al, 1993;Ludwig et al, 1994). Studies on MPR46-and/or M6P/IGF2R-negative fibroblasts have indicated that both receptors are necessary for efficient lysosomal targeting.…”

Section: Introductionmentioning

confidence: 99%

The 46-kDa mannose 6-phosphate receptor does not depend on endosomal acidification for delivery of hydrolases to lysosomes

Probst

Ton

Svoboda

et al. 2006

Journal of Cell Science

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In mammalian cells, the mannose 6-phosphate receptor pathway accounts for the transport of most soluble acid hydrolases to lysosomes. It is believed that dissociation of mannose 6-phosphate receptors and their ligands is entirely driven by the acidic environment in endosomal compartments. Indeed, pH-perturbing substances such as ammonium chloride and monensin have been shown to inhibit lysosomal enzyme targeting in cells that express both known mannose 6-phosphate receptors. We now demonstrate that ammonium chloride and monensin exert modest effects on the intracellular retention of lysosomal hydrolases in murine cells that synthesize only the 46-kDa mannose 6-phosphate receptor. Neither ammonium chloride nor monensin induces changes to the subcellular localization of lysosomal hydrolases and the 46-kDa mannose 6-phosphate receptor in these cells. This suggests that endosomal dissociation of the receptor and its ligands still occurs in the presence of these agents. We conclude that the murine 46-kDa mannose 6-phosphate receptor has the capacity to deliver its cargo proteins to lysosomes even in the absence of endosomal acidification.

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Targeted disruption of the M(r) 46,000 mannose 6-phosphate receptor gene in mice results in misrouting of lysosomal proteins.

Cited by 89 publications

References 17 publications

Phenotype of arylsulfatase A-deficient mice: Relationship to human metachromatic leukodystrophy

Phenotype of arylsulfatase A-deficient mice: Relationship to human metachromatic leukodystrophy

Role of LAMP-2 in Lysosome Biogenesis and Autophagy

The 46-kDa mannose 6-phosphate receptor does not depend on endosomal acidification for delivery of hydrolases to lysosomes

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