title: Overexpression of tissue-nonspecific alkaline phosphatase increases the expression of neurogenic differentiation markers in the human SH-SY5Y neuroblastoma cell line.
abstract: Patients suffering from the rare hereditary disease hypophosphatasia (HPP), which is based on mutations in the ALPL gene, tend to develop central nervous system (CNS) related issues like epileptic seizures and neuropsychiatric illnesses such as anxiety and depression, in addition to well-known problems with the mineralization of bones and teeth. Analyses of the molecular role of tissue-nonspecific alkaline phosphatase (TNAP) in transgenic SH-SY5Y(TNAPhigh) neuroblastoma cells compared to SH-SY5Y(TNAPlow) cells indicate that the enzyme influences the expression levels of neuronal marker genes like RNA-binding protein, fox-1 homolog 3 (NEUN) and enolase 2, gamma neuronal (NSE) as well as microtubule-binding proteins like microtubule-associated protein 2 (MAP2) and microtubule-associated protein tau (TAU) during neurogenic differentiation. Fluorescence staining of SH-SY5Y(TNAPhigh) cells reveals TNAP localization throughout the whole length of the developed projection network and even synapsin Ι co-localization with strong TNAP signals at some spots at least at the early time points of differentiation. Additional immunocytochemical staining shows higher MAP2 expression in SH-SY5Y(TNAPhigh) cells and further a distinct up-regulation of tau and MAP2 in the course of neurogenic differentiation. Interestingly, transgenic SH-SY5Y(TNAPhigh) cells are able to develop longer cellular processes compared to control cells after stimulation with all-trans retinoic acid (RA). Current therapies for HPP prioritize improvement of the bone phenotype. Unraveling the molecular role of TNAP in extraosseous tissues, like in the CNS, will help to improve treatment strategies for HPP patients. Taking this rare disease as a model may also help to dissect TNAP's role in neurodegenerative diseases and even improve future treatment of common pathologies.
authors.affiliation: Orthopedic Department, Orthopedic Center for Musculoskeletal Research, University of Wuerzburg, Germany.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Graser
authors.affiliation: Orthopedic Department, Orthopedic Center for Musculoskeletal Research, University of Wuerzburg, Germany.
authors.forename: Birgit
authors.initials: B
authors.lastname: Mentrup
authors.affiliation: Orthopedic Department, Orthopedic Center for Musculoskeletal Research, University of Wuerzburg, Germany.
authors.forename: Doris
authors.initials: D
authors.lastname: Schneider
authors.affiliation: Institute of Cell Biology, Faculty of Medicine, University of Duisburg-Essen, Germany.
authors.forename: Ludger
authors.initials: L
authors.lastname: Klein-Hitpass
authors.affiliation: Orthopedic Department, Orthopedic Center for Musculoskeletal Research, University of Wuerzburg, Germany.
authors.forename: Franz
authors.initials: F
authors.lastname: Jakob
authors.affiliation: Children's Hospital, Section of Pediatric Rheumatology and Osteology, University of Wuerzburg, Germany. Electronic address: hofmann_c5@ukw.de.
authors.forename: Christine
authors.initials: C
authors.lastname: Hofmann
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism*; D015153:Blotting, Western; D002454:Cell Differentiation / Q000235:genetics*; D045744:Cell Line, Tumor; D006801:Humans; D007014:Hypophosphatasia / Q000235:genetics; D007150:Immunohistochemistry; D009474:Neurons / Q000166:cytology / Q000201:enzymology*; D020411:Oligonucleotide Array Sequence Analysis; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D019076:Transgenes
chemical_list: C548267:ALPL protein, human; D000469:Alkaline Phosphatase
keywords: Cellular processes; Hypophosphatasia; MAP2; Overexpression; Transgenic