Factors influencing the measurement of lysosomal enzymes activity in human cerebrospinal fluid

Emanuele Persichetti, Davide Chiasserini, Lucilla Parnetti, Paolo Eusebi, Silvia Paciotti, Claudia De Carlo, Michela Codini, Nicola Tambasco, Aroldo Rossi, Omar Ali El-Agnaf, Paolo Calabresi, Tommaso Beccari

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Measurements of the activities of lysosomal enzymes in cerebrospinal fluid have recently been proposed as putative biomarkers for Parkinson's disease and other synucleinopathies. To define the operating procedures useful for ensuring the reliability of these measurements, we analyzed several pre-analytical factors that may influence the activity of β-glucocerebrosidase, α-mannosidase, β-mannosidase, β-galactosidase, α-fucosidase, β-hexosaminidase, cathepsin D and cathepsin E in cerebrospinal fluid. Lysosomal enzyme activities were measured by well-established fluorimetric assays in a consecutive series of patients (n = 28) with different neurological conditions, including Parkinson's disease. The precision, pre-storage and storage conditions, and freeze/thaw cycles were evaluated. All of the assays showed within- and between-run variabilities below 10%. At - 20°C, only cathepsin D was stable up to 40 weeks. At -80°C, the cathepsin D, cathepsin E, and β-mannosidase activities did not change significantly up to 40 weeks, while β-glucocerebrosidase activity was stable up to 32 weeks. The β-galactosidase and α-fucosidase activities significantly increased (+54.9±38.08% after 4 weeks and + 88.94±36.19% after 16 weeks, respectively). Up to four freeze/thaw cycles did not significantly affect the activities of cathepsins D and E. The β-glucocerebrosidase activity showed a slight decrease (-14.6%) after two freeze/thaw cycles. The measurement of lysosomal enzyme activities in cerebrospinal fluid is reliable and reproducible if pre-analytical factors are accurately taken into consideration. Therefore, the analytical recommendations that ensue from this study may contribute to the establishment of actual values for the activities of cerebrospinal fluid lysosomal enzymes as putative biomarkers for Parkinson's disease and other neurodegenerative disorders.

Original languageEnglish
Article numbere101453
JournalPLoS One
Volume9
Issue number7
DOIs
Publication statusPublished - 1 Jul 2014
Externally publishedYes

Fingerprint

Cerebrospinal fluid
cathepsin E
cathepsin D
Cathepsin D
Cathepsin E
Mannosidases
mannosidases
Enzyme activity
cerebrospinal fluid
Glucosylceramidase
Human Activities
Cerebrospinal Fluid
freeze-thaw cycles
Parkinson disease
Galactosidases
alpha-L-Fucosidase
enzyme activity
galactosidases
Parkinson Disease
Biomarkers

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Persichetti, E., Chiasserini, D., Parnetti, L., Eusebi, P., Paciotti, S., De Carlo, C., ... Beccari, T. (2014). Factors influencing the measurement of lysosomal enzymes activity in human cerebrospinal fluid. PLoS One, 9(7), [e101453]. https://doi.org/10.1371/journal.pone.0101453

Factors influencing the measurement of lysosomal enzymes activity in human cerebrospinal fluid. / Persichetti, Emanuele; Chiasserini, Davide; Parnetti, Lucilla; Eusebi, Paolo; Paciotti, Silvia; De Carlo, Claudia; Codini, Michela; Tambasco, Nicola; Rossi, Aroldo; Ali El-Agnaf, Omar; Calabresi, Paolo; Beccari, Tommaso.

In: PLoS One, Vol. 9, No. 7, e101453, 01.07.2014.

Research output: Contribution to journalArticle

Persichetti, E, Chiasserini, D, Parnetti, L, Eusebi, P, Paciotti, S, De Carlo, C, Codini, M, Tambasco, N, Rossi, A, Ali El-Agnaf, O, Calabresi, P & Beccari, T 2014, 'Factors influencing the measurement of lysosomal enzymes activity in human cerebrospinal fluid', PLoS One, vol. 9, no. 7, e101453. https://doi.org/10.1371/journal.pone.0101453
Persichetti E, Chiasserini D, Parnetti L, Eusebi P, Paciotti S, De Carlo C et al. Factors influencing the measurement of lysosomal enzymes activity in human cerebrospinal fluid. PLoS One. 2014 Jul 1;9(7). e101453. https://doi.org/10.1371/journal.pone.0101453
Persichetti, Emanuele ; Chiasserini, Davide ; Parnetti, Lucilla ; Eusebi, Paolo ; Paciotti, Silvia ; De Carlo, Claudia ; Codini, Michela ; Tambasco, Nicola ; Rossi, Aroldo ; Ali El-Agnaf, Omar ; Calabresi, Paolo ; Beccari, Tommaso. / Factors influencing the measurement of lysosomal enzymes activity in human cerebrospinal fluid. In: PLoS One. 2014 ; Vol. 9, No. 7.
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