Abstract
The lysosomal hydrolase galactocerebrosidase (GALC) catalyzes the removal of galactose from galactosylceramide and from other sphingolipids. GALC deficiency is responsible for globoid cell leukodystrophy (GLD), or Krabbe's disease, an early lethal inherited neurodegenerative disorder characterized by the accumulation of the neurotoxic metabolite psychosine in the central nervous system (CNS). The poor outcome of current clinical treatments calls for novel model systems to investigate the biological impact of GALC down-regulation and for the search of novel therapeutic strategies in GLD. Zebrafish (Danio rerio) represents an attractive vertebrate model for human diseases. Here, lysosomal GALC activity was demonstrated in the brain of zebrafish adults and embryos. Accordingly, we identified two GALC co-orthologs (named galca and galcb) dynamically co-expressed in CNS during zebrafish development. Both genes encode for lysosomal enzymes endowed with GALC activity. Single down-regulation of galca or galcb by specific antisense morpholino oligonucleotides results in a partial decrease of GALC activity in zebrafish embryos that was abrogated in double galca/. galcb morphants. However, no psychosine accumulation was observed in galca/. galcb double morphants. Nevertheless, double galca/. galcb knockdown caused reduction and partial disorganization of the expression of the early neuronal marker neuroD and an increase of apoptotic events during CNS development. These observations provide new insights into the pathogenesis of GLD, indicating that GALC loss-of-function may have pathological consequences in developing CNS independent of psychosine accumulation. Also, they underscore the potentiality of the zebrafish system in studying the pathogenesis of lysosomal neurodegenerative diseases, including GLD.
| Original language | English |
|---|---|
| Pages (from-to) | 665-675 |
| Number of pages | 11 |
| Journal | Biochimica et Biophysica Acta - Molecular Basis of Disease |
| Volume | 1842 |
| Issue number | 4 |
| DOIs | |
| Publication status | Published - Apr 2014 |
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Keywords
- Embryonic development
- Galactosylceramidase
- Krabbe disease
- Sphingolipid
- Zebrafish
ASJC Scopus subject areas
- Molecular Biology
- Molecular Medicine
Cite this
Molecular cloning and knockdown of galactocerebrosidase in zebrafish : New insights into the pathogenesis of Krabbe's disease. / Zizioli, Daniela; Guarienti, Michela; Tobia, Chiara; Gariano, Giuseppina; Borsani, Giuseppe; Bresciani, Roberto; Ronca, Roberto; Giacopuzzi, Edoardo; Preti, Augusto; Gaudenzi, Germano; Belleri, Mirella; Di Salle, Emanuela; Fabrias, Gemma; Casas, Josefina; Ribatti, Domenico; Monti, Eugenio; Presta, Marco.
In: Biochimica et Biophysica Acta - Molecular Basis of Disease, Vol. 1842, No. 4, 04.2014, p. 665-675.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Molecular cloning and knockdown of galactocerebrosidase in zebrafish
T2 - New insights into the pathogenesis of Krabbe's disease
AU - Zizioli, Daniela
AU - Guarienti, Michela
AU - Tobia, Chiara
AU - Gariano, Giuseppina
AU - Borsani, Giuseppe
AU - Bresciani, Roberto
AU - Ronca, Roberto
AU - Giacopuzzi, Edoardo
AU - Preti, Augusto
AU - Gaudenzi, Germano
AU - Belleri, Mirella
AU - Di Salle, Emanuela
AU - Fabrias, Gemma
AU - Casas, Josefina
AU - Ribatti, Domenico
AU - Monti, Eugenio
AU - Presta, Marco
PY - 2014/4
Y1 - 2014/4
N2 - The lysosomal hydrolase galactocerebrosidase (GALC) catalyzes the removal of galactose from galactosylceramide and from other sphingolipids. GALC deficiency is responsible for globoid cell leukodystrophy (GLD), or Krabbe's disease, an early lethal inherited neurodegenerative disorder characterized by the accumulation of the neurotoxic metabolite psychosine in the central nervous system (CNS). The poor outcome of current clinical treatments calls for novel model systems to investigate the biological impact of GALC down-regulation and for the search of novel therapeutic strategies in GLD. Zebrafish (Danio rerio) represents an attractive vertebrate model for human diseases. Here, lysosomal GALC activity was demonstrated in the brain of zebrafish adults and embryos. Accordingly, we identified two GALC co-orthologs (named galca and galcb) dynamically co-expressed in CNS during zebrafish development. Both genes encode for lysosomal enzymes endowed with GALC activity. Single down-regulation of galca or galcb by specific antisense morpholino oligonucleotides results in a partial decrease of GALC activity in zebrafish embryos that was abrogated in double galca/. galcb morphants. However, no psychosine accumulation was observed in galca/. galcb double morphants. Nevertheless, double galca/. galcb knockdown caused reduction and partial disorganization of the expression of the early neuronal marker neuroD and an increase of apoptotic events during CNS development. These observations provide new insights into the pathogenesis of GLD, indicating that GALC loss-of-function may have pathological consequences in developing CNS independent of psychosine accumulation. Also, they underscore the potentiality of the zebrafish system in studying the pathogenesis of lysosomal neurodegenerative diseases, including GLD.
AB - The lysosomal hydrolase galactocerebrosidase (GALC) catalyzes the removal of galactose from galactosylceramide and from other sphingolipids. GALC deficiency is responsible for globoid cell leukodystrophy (GLD), or Krabbe's disease, an early lethal inherited neurodegenerative disorder characterized by the accumulation of the neurotoxic metabolite psychosine in the central nervous system (CNS). The poor outcome of current clinical treatments calls for novel model systems to investigate the biological impact of GALC down-regulation and for the search of novel therapeutic strategies in GLD. Zebrafish (Danio rerio) represents an attractive vertebrate model for human diseases. Here, lysosomal GALC activity was demonstrated in the brain of zebrafish adults and embryos. Accordingly, we identified two GALC co-orthologs (named galca and galcb) dynamically co-expressed in CNS during zebrafish development. Both genes encode for lysosomal enzymes endowed with GALC activity. Single down-regulation of galca or galcb by specific antisense morpholino oligonucleotides results in a partial decrease of GALC activity in zebrafish embryos that was abrogated in double galca/. galcb morphants. However, no psychosine accumulation was observed in galca/. galcb double morphants. Nevertheless, double galca/. galcb knockdown caused reduction and partial disorganization of the expression of the early neuronal marker neuroD and an increase of apoptotic events during CNS development. These observations provide new insights into the pathogenesis of GLD, indicating that GALC loss-of-function may have pathological consequences in developing CNS independent of psychosine accumulation. Also, they underscore the potentiality of the zebrafish system in studying the pathogenesis of lysosomal neurodegenerative diseases, including GLD.
KW - Embryonic development
KW - Galactosylceramidase
KW - Krabbe disease
KW - Sphingolipid
KW - Zebrafish
UR - http://www.scopus.com/inward/record.url?scp=84893839340&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84893839340&partnerID=8YFLogxK
U2 - 10.1016/j.bbadis.2014.01.008
DO - 10.1016/j.bbadis.2014.01.008
M3 - Article
C2 - 24463171
AN - SCOPUS:84893839340
VL - 1842
SP - 665
EP - 675
JO - Biochimica et Biophysica Acta - Molecular Basis of Disease
JF - Biochimica et Biophysica Acta - Molecular Basis of Disease
SN - 0925-4439
IS - 4
ER -