Transcriptomic and functional proteomics analyses to unveil the common and unique pathway(s) of neuritogenesis induced by Russell’s viper venom nerve growth factor in rat pheochromocytoma neuronal cells

ABSTRACT

Background: The snake venom nerve growth factor (NGF)-induced signal transduction mechanism has never been explored.

Research design and methods: Homology modeling and molecular dynamic studies of the interaction between Russell’s viper venom NGF (RVV-NGFa) and mammalian tropomyosin-receptor kinase A (TrkA) was done by computational analysis. Transcriptomic and quantitative tandem mass spectrometry analyses determined the expression of intracellular genes and proteins, respectively, in RVV-NGFa-treated PC-12 neuronal cells. Small synthetic inhibitors of the signal transduction pathways were used to validate the major signaling cascades of neuritogenesis by RVV-NGFa.

Results: A comparative computational analysis predicted the binding of RVV-NGFa, mouse 2.5S-NGF (conventional neurotrophin), and Nn-α-elapitoxin-1 (non-conventional neurotrophin) to different domains of the TrkA receptor in PC-12 cells. The transcriptomic and quantitative proteomic analyses in unison showed differential expressions of common and unique genes and intracellular proteins, respectively, in RVV-NGFa-treated cells compared to control (untreated) mouse 2.5S-NGF and Nn-α-elapitoxin-1-treated PC-12 cells. The RVV-NGFa primarily triggered the mitogen-activated protein kinase-1 (MAPK1) signaling pathway for inducing neuritogenesis; however, 36 pathways of neuritogenesis were uniquely expressed in RVV-NGFa-treated PC-12 cells compared to mouse 2.5S NGF or Nn-α-elapitoxin-1 treated cells.

Conclusion: The common and unique intracellular signaling pathways of neuritogenesis by classical and non-classical neurotrophins were identified.

KEYWORDS:

• PC-12 cell
• neuritogenesis
• tyrosine kinase receptor
• Russell’s viper venom
• mitogen-activated protein kinase-1 (MAPK) signaling pathway
• snake venom nerve growth factor
• signal transduction

Article highlights

• NGF is a minor component of snake venom, though its exact function in venom has not been elucidated.

• The signal transduction mechanism of snake venom NGF and its comparison to classical neurotrophins was unknown.

• The major and unique signaling pathways of neuritogenesis and the survival of PC-12 cells post treatment with RVV-NGFa are illustrated.

• The TrkA receptor-binding region of RVV-NGFa may be developed as a peptide-based drug prototype for preventing and/or treating neurodegenerative disorders.

• This study describes the variation in the receptor-binding moiety (in TrkA) with respect to different ligands (classical and non-classical neurotrophins).

Acknowledgments

The authors thank C-CAMP, NCBS Bengaluru for LC-MS/MS analysis; Molsys Pvt. Ltd., Bengaluru for transcriptomic analysis and Ms. M. Majumdar, Dept. of MBBT, T.U for cell culture assays. T.I received INSPIRE Senior Research Fellowships from DST, New Delhi. DM received JRF from DST project grant.

Declaration of interest

The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

Author’s contribution

A.K. Mukherjee conceived the idea, designed the study, and edited the final manuscript; T. Islam and D. Madhubala performed the experiments and analyzed the data; R. Mukhopadhyay analyzed the data and edited the manuscript.

Reviewer disclosures

Peer reviewers on this manuscript have no relevant financial or other relationships to disclose.

Supplementary material

Supplemental data for this article can be accessed here.

Additional information

Funding

This manuscript was funded by the Science and Engineering Research Board, New Delhi (EMR/2017/001829) to A.K. Mukherjee.