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Alkaline Ceramidase 3 Deficiency Results in Purkinje Cell Degeneration and Cerebellar Ataxia Due to Dyshomeostasis of Sphingolipids in the Brain.

Wang K, Xu R, Schrandt J, Shah P, Gong YZ, Preston C, Wang L, Yi JK, Lin CL, Sun W, Spyropoulos DD, Rhee S, Li M, Zhou J, Ge S, Zhang G, Snider AJ, Hannun YA, Obeid LM, Mao C - PLoS Genet. (2015)

Bottom Line: However, mechanisms that maintain the homeostasis of these bioactive sphingolipids in the brain remain unclear.Acer3 knockout causes an age-dependent accumulation of various ceramides and C18:1-monohexosylceramide and abolishes the age-related increase in the levels of sphingosine and S1P in the brain; thereby resulting in Purkinje cell degeneration in the cerebellum and deficits in motor coordination and balance.Our results indicate that Acer3 plays critically protective roles in controlling the homeostasis of various sphingolipids, including ceramides, sphingosine, S1P, and certain complex sphingolipids in the brain and protects Purkinje cells from premature degeneration.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Stony Brook University, Stony Brook, New York, United States of America; Stony Brook Cancer Center, Stony Brook, New York, United States of America; Department of Hepatobiliary Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, China.

ABSTRACT
Dyshomeostasis of both ceramides and sphingosine-1-phosphate (S1P) in the brain has been implicated in aging-associated neurodegenerative disorders in humans. However, mechanisms that maintain the homeostasis of these bioactive sphingolipids in the brain remain unclear. Mouse alkaline ceramidase 3 (Acer3), which preferentially catalyzes the hydrolysis of C18:1-ceramide, a major unsaturated long-chain ceramide species in the brain, is upregulated with age in the mouse brain. Acer3 knockout causes an age-dependent accumulation of various ceramides and C18:1-monohexosylceramide and abolishes the age-related increase in the levels of sphingosine and S1P in the brain; thereby resulting in Purkinje cell degeneration in the cerebellum and deficits in motor coordination and balance. Our results indicate that Acer3 plays critically protective roles in controlling the homeostasis of various sphingolipids, including ceramides, sphingosine, S1P, and certain complex sphingolipids in the brain and protects Purkinje cells from premature degeneration.

No MeSH data available.


Related in: MedlinePlus

Generation of Acer3  mouse.A. Acer3 targeting strategy. The Acer3 gene consists of 11 exons (empty rectangles with the numerals inside). Exon 8 of the Acer3 gene was replaced by the Neo resistant gene cassette upon homologous recombination. B. Southern blot analyses of WT ES cells or ES cells from an Acer3-targeted ES clone. Genomic DNA was digested with EcoRV, resolved on a 0.8% agarose gel, transferred to a nitrocellulose membrane, which was labeled with a radioactive probe (P) corresponding to the region upstream of Exon 8 as shown in Panel A. C. PCR-based genotyping of Acer3+/+, Acer3+/-, and Acer3-/- mice. DNA was isolated from mouse tail biopsies and subjected to PCR analyses using the PCR primer pairs (F1 and B1 or F1 and B2) as shown in Panel A. The image in C represents the PCR product patterns of the three genotypes, Acer3+/+, Acer3+/-, and Acer3-/-.
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pgen.1005591.g002: Generation of Acer3 mouse.A. Acer3 targeting strategy. The Acer3 gene consists of 11 exons (empty rectangles with the numerals inside). Exon 8 of the Acer3 gene was replaced by the Neo resistant gene cassette upon homologous recombination. B. Southern blot analyses of WT ES cells or ES cells from an Acer3-targeted ES clone. Genomic DNA was digested with EcoRV, resolved on a 0.8% agarose gel, transferred to a nitrocellulose membrane, which was labeled with a radioactive probe (P) corresponding to the region upstream of Exon 8 as shown in Panel A. C. PCR-based genotyping of Acer3+/+, Acer3+/-, and Acer3-/- mice. DNA was isolated from mouse tail biopsies and subjected to PCR analyses using the PCR primer pairs (F1 and B1 or F1 and B2) as shown in Panel A. The image in C represents the PCR product patterns of the three genotypes, Acer3+/+, Acer3+/-, and Acer3-/-.

Mentions: To elucidate the physiological function of age-dependent upregulation of Acer3 in the brain, we generated a mouse model deficient in Acer3 as described in Materials and Methods (Fig 2). Interbreeding of mice heterozygous for an Acer3 allele produced WT (Acer3+/+), heterozygous (Acer3+/-), and homozygous (Acer3-/-) offspring at a Mendelian ratio (Acer3+/+: Acer3+/-: Acer3-/-; 151:319:143), suggesting the absence of significant embryonic lethality in Acer3 knockout mice. Intercrossing of Acer3-/- mice produced similar offspring numbers as with the intercrossing of Acer3+/+ mice (Fig 3A), suggesting that both male and female Acer3-/- mice have normal fertility. There was no significant difference in body weight between Acer3+/+ and Acer3-/- mice when measured at 6 weeks, 4, 6, 8, or 12 months of age (Fig 3B). The macroscopic and microscopic analyses found no abnormalities in the anatomy of major organs in young mice. These results suggest that Acer3 knockout does not appear to cause any major defect in mouse development at least prior to middle age.


Alkaline Ceramidase 3 Deficiency Results in Purkinje Cell Degeneration and Cerebellar Ataxia Due to Dyshomeostasis of Sphingolipids in the Brain.

Wang K, Xu R, Schrandt J, Shah P, Gong YZ, Preston C, Wang L, Yi JK, Lin CL, Sun W, Spyropoulos DD, Rhee S, Li M, Zhou J, Ge S, Zhang G, Snider AJ, Hannun YA, Obeid LM, Mao C - PLoS Genet. (2015)

Generation of Acer3  mouse.A. Acer3 targeting strategy. The Acer3 gene consists of 11 exons (empty rectangles with the numerals inside). Exon 8 of the Acer3 gene was replaced by the Neo resistant gene cassette upon homologous recombination. B. Southern blot analyses of WT ES cells or ES cells from an Acer3-targeted ES clone. Genomic DNA was digested with EcoRV, resolved on a 0.8% agarose gel, transferred to a nitrocellulose membrane, which was labeled with a radioactive probe (P) corresponding to the region upstream of Exon 8 as shown in Panel A. C. PCR-based genotyping of Acer3+/+, Acer3+/-, and Acer3-/- mice. DNA was isolated from mouse tail biopsies and subjected to PCR analyses using the PCR primer pairs (F1 and B1 or F1 and B2) as shown in Panel A. The image in C represents the PCR product patterns of the three genotypes, Acer3+/+, Acer3+/-, and Acer3-/-.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4608763&req=5

pgen.1005591.g002: Generation of Acer3 mouse.A. Acer3 targeting strategy. The Acer3 gene consists of 11 exons (empty rectangles with the numerals inside). Exon 8 of the Acer3 gene was replaced by the Neo resistant gene cassette upon homologous recombination. B. Southern blot analyses of WT ES cells or ES cells from an Acer3-targeted ES clone. Genomic DNA was digested with EcoRV, resolved on a 0.8% agarose gel, transferred to a nitrocellulose membrane, which was labeled with a radioactive probe (P) corresponding to the region upstream of Exon 8 as shown in Panel A. C. PCR-based genotyping of Acer3+/+, Acer3+/-, and Acer3-/- mice. DNA was isolated from mouse tail biopsies and subjected to PCR analyses using the PCR primer pairs (F1 and B1 or F1 and B2) as shown in Panel A. The image in C represents the PCR product patterns of the three genotypes, Acer3+/+, Acer3+/-, and Acer3-/-.
Mentions: To elucidate the physiological function of age-dependent upregulation of Acer3 in the brain, we generated a mouse model deficient in Acer3 as described in Materials and Methods (Fig 2). Interbreeding of mice heterozygous for an Acer3 allele produced WT (Acer3+/+), heterozygous (Acer3+/-), and homozygous (Acer3-/-) offspring at a Mendelian ratio (Acer3+/+: Acer3+/-: Acer3-/-; 151:319:143), suggesting the absence of significant embryonic lethality in Acer3 knockout mice. Intercrossing of Acer3-/- mice produced similar offspring numbers as with the intercrossing of Acer3+/+ mice (Fig 3A), suggesting that both male and female Acer3-/- mice have normal fertility. There was no significant difference in body weight between Acer3+/+ and Acer3-/- mice when measured at 6 weeks, 4, 6, 8, or 12 months of age (Fig 3B). The macroscopic and microscopic analyses found no abnormalities in the anatomy of major organs in young mice. These results suggest that Acer3 knockout does not appear to cause any major defect in mouse development at least prior to middle age.

Bottom Line: However, mechanisms that maintain the homeostasis of these bioactive sphingolipids in the brain remain unclear.Acer3 knockout causes an age-dependent accumulation of various ceramides and C18:1-monohexosylceramide and abolishes the age-related increase in the levels of sphingosine and S1P in the brain; thereby resulting in Purkinje cell degeneration in the cerebellum and deficits in motor coordination and balance.Our results indicate that Acer3 plays critically protective roles in controlling the homeostasis of various sphingolipids, including ceramides, sphingosine, S1P, and certain complex sphingolipids in the brain and protects Purkinje cells from premature degeneration.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Stony Brook University, Stony Brook, New York, United States of America; Stony Brook Cancer Center, Stony Brook, New York, United States of America; Department of Hepatobiliary Surgery, Nanfang Hospital, Southern Medical University, Guangzhou, China.

ABSTRACT
Dyshomeostasis of both ceramides and sphingosine-1-phosphate (S1P) in the brain has been implicated in aging-associated neurodegenerative disorders in humans. However, mechanisms that maintain the homeostasis of these bioactive sphingolipids in the brain remain unclear. Mouse alkaline ceramidase 3 (Acer3), which preferentially catalyzes the hydrolysis of C18:1-ceramide, a major unsaturated long-chain ceramide species in the brain, is upregulated with age in the mouse brain. Acer3 knockout causes an age-dependent accumulation of various ceramides and C18:1-monohexosylceramide and abolishes the age-related increase in the levels of sphingosine and S1P in the brain; thereby resulting in Purkinje cell degeneration in the cerebellum and deficits in motor coordination and balance. Our results indicate that Acer3 plays critically protective roles in controlling the homeostasis of various sphingolipids, including ceramides, sphingosine, S1P, and certain complex sphingolipids in the brain and protects Purkinje cells from premature degeneration.

No MeSH data available.


Related in: MedlinePlus