Kuvaus

A missense variant in the autophagy-related ATG4D-gene has been associated with a progressive degenerative neurological disease in Lagotto Romagnolo (LR) dogs. In addition to neural lesions, affected dogs show an extraneural histopathological phenotype characterized by severe cytoplasmic vacuolization, a finding not previously linked with disturbed autophagy in animals. Here we aimed at testing the hypothesis that autophagy is altered in the affected dogs, at reporting the histopathology of extraneural tissues and at excluding lysosomal storage diseases. Basal and starvation-induced autophagy were monitored by Western blotting and immunofluorescence of microtubule associated protein 1A/B light chain3 (LC3) in fibroblasts from 2 affected dogs. The extraneural findings of 9 euthanized LRs and skin biopsies from 4 living affected LRs were examined by light microscopy, electron microscopy, and immunohistochemistry (IHC), using antibodies against autophagosomal membranes (LC3), autophagic cargo (p62), and lysosomal membranes (LAMP2). Biochemical screening of urine and fibroblasts of 2 affected dogs was performed. Under basal conditions, the affected fibroblasts contained significantly more LC3-II and LC3-positive vesicles than did the controls. Morphologically, several cells, including serous secretory epithelium, endothelial cells, pericytes, plasma cells, and macrophages, contained cytoplasmic vacuoles with an ultrastructure resembling enlarged amphisomes, endosomes, or multivesicular bodies. IHC showed strong membranous LAMP2 positivity only in sweat glands. The results show that basal but not induced autophagy is altered in affected fibroblasts. The ultrastructure of affected cells is compatible with altered autophagic and endo-lysosomal vesicular traffic. The findings in this spontaneous disease provide insight into possible tissue-specific roles of basal autophagy.
Alkuperäiskielienglanti
LehtiVeterinary Pathology
Vuosikerta54
Numero6
Sivut953-963
Sivumäärä11
ISSN0300-9858
DOI - pysyväislinkit
TilaJulkaistu - marraskuuta 2017
OKM-julkaisutyyppiA1 Alkuperäisartikkeli tieteellisessä aikakauslehdessä, vertaisarvioitu

Tieteenalat

  • 413 Eläinlääketiede
  • 1182 Biokemia, solu- ja molekyylibiologia

Lainaa tätä

Syrjä, Pernilla ; Anwar, Tahira ; Pääkkönen, Tarja ; Kyöstilä, Kaisa ; Hultin Jäderlund, Karin ; Cozzi, Francesca ; Rohdin, Cecilia ; Hahn, Kerstin ; Wohlsein, Peter ; Baumgärtner, Wolfgang ; Henke, Diana ; Oevermann, Anna ; Sukura, Antti ; Leeb, Tosso ; Lohi, Hannes ; Eskelinen, Eeva-Liisa. / Basal Autophagy is altered in Lagotto Romagnolo Dogs with an ATG4D mutation. Julkaisussa: Veterinary Pathology. 2017 ; Vuosikerta 54, Nro 6. Sivut 953-963.
@article{c8140fd24cf14d299b0a6de47906e317,
title = "Basal Autophagy is altered in Lagotto Romagnolo Dogs with an ATG4D mutation",
abstract = "A missense variant in the autophagy-related ATG4D-gene has been associated with a progressive degenerative neurological disease in Lagotto Romagnolo (LR) dogs. In addition to neural lesions, affected dogs show an extraneural histopathological phenotype characterized by severe cytoplasmic vacuolization, a finding not previously linked with disturbed autophagy in animals. Here we aimed at testing the hypothesis that autophagy is altered in the affected dogs, at reporting the histopathology of extraneural tissues and at excluding lysosomal storage diseases. Basal and starvation-induced autophagy were monitored by Western blotting and immunofluorescence of microtubule associated protein 1A/B light chain3 (LC3) in fibroblasts from 2 affected dogs. The extraneural findings of 9 euthanized LRs and skin biopsies from 4 living affected LRs were examined by light microscopy, electron microscopy, and immunohistochemistry (IHC), using antibodies against autophagosomal membranes (LC3), autophagic cargo (p62), and lysosomal membranes (LAMP2). Biochemical screening of urine and fibroblasts of 2 affected dogs was performed. Under basal conditions, the affected fibroblasts contained significantly more LC3-II and LC3-positive vesicles than did the controls. Morphologically, several cells, including serous secretory epithelium, endothelial cells, pericytes, plasma cells, and macrophages, contained cytoplasmic vacuoles with an ultrastructure resembling enlarged amphisomes, endosomes, or multivesicular bodies. IHC showed strong membranous LAMP2 positivity only in sweat glands. The results show that basal but not induced autophagy is altered in affected fibroblasts. The ultrastructure of affected cells is compatible with altered autophagic and endo-lysosomal vesicular traffic. The findings in this spontaneous disease provide insight into possible tissue-specific roles of basal autophagy.",
keywords = "413 Veterinary science, 1182 Biochemistry, cell and molecular biology, basal au tophagy, Western blot , immunofluorescence , electron microscopy , cytoplasmic vacuolization , pathology , ATG4D , dog , LYSOSOMAL STORAGE DISEASES , CELLS , MUCOPOLYSACCHARIDOSES , DEGRADATION , ACTIVATION , MATURATION , CLEARANCE , PROTEIN , FLUX",
author = "Pernilla Syrj{\"a} and Tahira Anwar and Tarja P{\"a}{\"a}kk{\"o}nen and Kaisa Ky{\"o}stil{\"a} and {Hultin J{\"a}derlund}, Karin and Francesca Cozzi and Cecilia Rohdin and Kerstin Hahn and Peter Wohlsein and Wolfgang Baumg{\"a}rtner and Diana Henke and Anna Oevermann and Antti Sukura and Tosso Leeb and Hannes Lohi and Eeva-Liisa Eskelinen",
year = "2017",
month = "11",
doi = "10.1177/0300985817712793",
language = "English",
volume = "54",
pages = "953--963",
journal = "Veterinary Pathology",
issn = "0300-9858",
publisher = "Sage",
number = "6",

}

Syrjä, P, Anwar, T, Pääkkönen, T, Kyöstilä, K, Hultin Jäderlund, K, Cozzi, F, Rohdin, C, Hahn, K, Wohlsein, P, Baumgärtner, W, Henke, D, Oevermann, A, Sukura, A, Leeb, T, Lohi, H & Eskelinen, E-L 2017, 'Basal Autophagy is altered in Lagotto Romagnolo Dogs with an ATG4D mutation' Veterinary Pathology, Vuosikerta 54, Nro 6, Sivut 953-963. https://doi.org/10.1177/0300985817712793

Basal Autophagy is altered in Lagotto Romagnolo Dogs with an ATG4D mutation. / Syrjä, Pernilla; Anwar, Tahira; Pääkkönen, Tarja; Kyöstilä, Kaisa; Hultin Jäderlund, Karin; Cozzi, Francesca; Rohdin, Cecilia; Hahn, Kerstin; Wohlsein, Peter; Baumgärtner, Wolfgang; Henke, Diana; Oevermann, Anna; Sukura, Antti; Leeb, Tosso; Lohi, Hannes; Eskelinen, Eeva-Liisa.

julkaisussa: Veterinary Pathology, Vuosikerta 54, Nro 6, 11.2017, s. 953-963.

Tutkimustuotos: ArtikkelijulkaisuArtikkeliTieteellinenvertaisarvioitu

TY - JOUR

T1 - Basal Autophagy is altered in Lagotto Romagnolo Dogs with an ATG4D mutation

AU - Syrjä, Pernilla

AU - Anwar, Tahira

AU - Pääkkönen, Tarja

AU - Kyöstilä, Kaisa

AU - Hultin Jäderlund, Karin

AU - Cozzi, Francesca

AU - Rohdin, Cecilia

AU - Hahn, Kerstin

AU - Wohlsein, Peter

AU - Baumgärtner, Wolfgang

AU - Henke, Diana

AU - Oevermann, Anna

AU - Sukura, Antti

AU - Leeb, Tosso

AU - Lohi, Hannes

AU - Eskelinen, Eeva-Liisa

PY - 2017/11

Y1 - 2017/11

N2 - A missense variant in the autophagy-related ATG4D-gene has been associated with a progressive degenerative neurological disease in Lagotto Romagnolo (LR) dogs. In addition to neural lesions, affected dogs show an extraneural histopathological phenotype characterized by severe cytoplasmic vacuolization, a finding not previously linked with disturbed autophagy in animals. Here we aimed at testing the hypothesis that autophagy is altered in the affected dogs, at reporting the histopathology of extraneural tissues and at excluding lysosomal storage diseases. Basal and starvation-induced autophagy were monitored by Western blotting and immunofluorescence of microtubule associated protein 1A/B light chain3 (LC3) in fibroblasts from 2 affected dogs. The extraneural findings of 9 euthanized LRs and skin biopsies from 4 living affected LRs were examined by light microscopy, electron microscopy, and immunohistochemistry (IHC), using antibodies against autophagosomal membranes (LC3), autophagic cargo (p62), and lysosomal membranes (LAMP2). Biochemical screening of urine and fibroblasts of 2 affected dogs was performed. Under basal conditions, the affected fibroblasts contained significantly more LC3-II and LC3-positive vesicles than did the controls. Morphologically, several cells, including serous secretory epithelium, endothelial cells, pericytes, plasma cells, and macrophages, contained cytoplasmic vacuoles with an ultrastructure resembling enlarged amphisomes, endosomes, or multivesicular bodies. IHC showed strong membranous LAMP2 positivity only in sweat glands. The results show that basal but not induced autophagy is altered in affected fibroblasts. The ultrastructure of affected cells is compatible with altered autophagic and endo-lysosomal vesicular traffic. The findings in this spontaneous disease provide insight into possible tissue-specific roles of basal autophagy.

AB - A missense variant in the autophagy-related ATG4D-gene has been associated with a progressive degenerative neurological disease in Lagotto Romagnolo (LR) dogs. In addition to neural lesions, affected dogs show an extraneural histopathological phenotype characterized by severe cytoplasmic vacuolization, a finding not previously linked with disturbed autophagy in animals. Here we aimed at testing the hypothesis that autophagy is altered in the affected dogs, at reporting the histopathology of extraneural tissues and at excluding lysosomal storage diseases. Basal and starvation-induced autophagy were monitored by Western blotting and immunofluorescence of microtubule associated protein 1A/B light chain3 (LC3) in fibroblasts from 2 affected dogs. The extraneural findings of 9 euthanized LRs and skin biopsies from 4 living affected LRs were examined by light microscopy, electron microscopy, and immunohistochemistry (IHC), using antibodies against autophagosomal membranes (LC3), autophagic cargo (p62), and lysosomal membranes (LAMP2). Biochemical screening of urine and fibroblasts of 2 affected dogs was performed. Under basal conditions, the affected fibroblasts contained significantly more LC3-II and LC3-positive vesicles than did the controls. Morphologically, several cells, including serous secretory epithelium, endothelial cells, pericytes, plasma cells, and macrophages, contained cytoplasmic vacuoles with an ultrastructure resembling enlarged amphisomes, endosomes, or multivesicular bodies. IHC showed strong membranous LAMP2 positivity only in sweat glands. The results show that basal but not induced autophagy is altered in affected fibroblasts. The ultrastructure of affected cells is compatible with altered autophagic and endo-lysosomal vesicular traffic. The findings in this spontaneous disease provide insight into possible tissue-specific roles of basal autophagy.

KW - 413 Veterinary science

KW - 1182 Biochemistry, cell and molecular biology

KW - basal au tophagy

KW - Western blot

KW - immunofluorescence

KW - electron microscopy

KW - cytoplasmic vacuolization

KW - pathology

KW - ATG4D

KW - dog

KW - LYSOSOMAL STORAGE DISEASES

KW - CELLS

KW - MUCOPOLYSACCHARIDOSES

KW - DEGRADATION

KW - ACTIVATION

KW - MATURATION

KW - CLEARANCE

KW - PROTEIN

KW - FLUX

U2 - 10.1177/0300985817712793

DO - 10.1177/0300985817712793

M3 - Article

VL - 54

SP - 953

EP - 963

JO - Veterinary Pathology

JF - Veterinary Pathology

SN - 0300-9858

IS - 6

ER -