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Journal of Molecular Neuroscience

, Volume 68, Issue 1, pp 135–143 | Cite as

Tuftelin Is Required for NGF-Induced Differentiation of PC12 Cells

  • Dekel ShiloEmail author
  • Gadi Cohen
  • Anat Blumenfeld
  • Koby Goren
  • Salem Hanhan
  • Shay Sharon
  • Amir Haze
  • Dan Deutsch
  • Philip Lazarovici
Article
  • 127 Downloads

Abstract

Nerve growth factor (NGF) promotes pleiotropic gene transcription-dependent biological effects, in neuronal and non-neuronal cells, including survival, proliferation, differentiation, neuroprotection, pain, and angiogenesis. It is hypothesized that during odontogenesis, NGF may be implicated in morphogenetic and mineralization events by affecting proliferation and/or differentiation of dental cells. Tuftelin belongs to the enamel associated teeth proteins and is thought to play a role in enamel mineralization. We previously reported that tuftelin transcript and protein, which are ubiquitously expressed in various tissues of embryos, adults, and tumors, were significantly upregulated during NGF-induced PC12 differentiation. To further confirm the involvement of tuftelin in the differentiation process, we established a tuftelin-knockdown neuronal PC12 cell model, using a non-cytotoxic siRNA directed towards sequences at the 3′ UTR of the tuftelin gene. Using real-time PCR, we quantified tuftelin mRNA expression and found that tuftelin siRNA, but not scrambled siRNA or transfection reagents, efficiently depleted about 60% of NGF-induced tuftelin mRNA transcripts. The effect of tuftelin siRNA was quantified up to 6 days of NGF-induced differentiation. Using immunofluorescence and western blot analyses, we also found a direct correlation between reduction of 60–80% in tuftelin protein expression and inhibition of about 50–70% in NGF-induced differentiation of the cells, as was detected after 3–6 days of treatment. These results demonstrate an important role for tuftelin in NGF-induced differentiation of PC12 cells. Tuftelin could be a useful target for drug development in disease where neurotrophin therapy is required.

Keywords

Nerve growth factor NGF PC12 cells Cell differentiation Neuronal outgrowth Tuftelin Tuft1 

Notes

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Dental Research Laboratory, Faculty of Dental Medicine, Institute of Dental SciencesThe Hebrew University of JerusalemJerusalemIsrael
  2. 2.Rambam Health Care CampusHaifaIsrael
  3. 3.School of Pharmacy, Institute for Drug Research, Faculty of MedicineThe Hebrew University of JerusalemJerusalemIsrael

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