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Dramatic neurological and biological effects by Botulinum Neurotoxin Type A on SH-SY5Y neuroblastoma cells, beyond the blockade of neurotransmitter release
Bal R. Singh
Corresponding Author
ORCiD: https://orcid.org/0000-0002-9069-1487
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This work is licensed under a CC BY 4.0 License. Read Full License
View the published version at BMC Pharmacology and Toxicology.
Background Gene expression profile analysis on mammalian cell lines and animal models after exposure to botulinum neurotoxin (BoNT) have been investigated in several studies in recent years. This provides a powerful tool to identify critical signaling pathways involved in the biological and inflammatory responses to BoNT and helps to determine the mechanism of the function of botulinum toxins.
One of the key clinical characteristics of BoNT is its prolonged on-site effects. The role of BoNT on the blockage of neurotransmitter acetylcholine release in the neuromuscular junction has been well established. However, the effects of the treatment time of BoNT on the human cellular model and its potential mechanism remain to be defined. This study is aimed to use the gene microarray technology to compare the two physiological critical time points of BoNT type A (BoNT/A) treatment of human neuroblastoma cells and to advance our understanding of the profound biological influences that toxin molecules play in the neuronal cellular system.
Results SH-SY5Y cells were treated with BoNT/A for 4 and 48 hours which represent the time needed for the entrance of toxin into the cells and the time necessary for the initial appearance of the on-site effects after BoNT application. A comparison of the two time points identified 122 functional groups that are significantly changed. The top five groups are the alternative splicing, phosphoprotein, nucleus, cytoplasm, and acetylation. Furthermore, after 48 hours there were 744 genes significantly up-regulated and 624 genes significantly down-regulated (p‹ 0.01). These genes fell into the following neurological and biological annotation groups: Nervous system development, proteinaceous extracellular matrix, signaling pathways regulating pluripotency of stem cells, cellular function and signal transduction, and apoptosis.
We have also noticed that the up-regulated groups contained neuronal cell development, nervous system development, and metabolic processes while the down-regulated groups contained many chromosomes and cell cycle categories.
Conclusions The effects of BoNT/A on neuronal cells extend beyond blocking the neurotransmitter release, and that BoNT/A is a multifunctional molecule that can evoke profound cellular responses which warrant a more in-depth understanding of the mechanism of the toxin’s effects after administration.
Keywords
Gene expression profile, Clostridium botulinum, Botulinum neurotoxin type A, host response, microarray analysis, Neuroblastoma SH-SY5Y cells, zinc, neurological signaling pathways, biological signaling
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CURRENT STATUS: Published
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Posted 18 Jun, 2020
Published
On 05 Sep, 2020
No community comments so far
Editorial decision: Minor revision
On 11 Aug, 2020
Review #2 received
Received 10 Aug, 2020
Reviewer #2 agreed
On 20 Jun, 2020
Reviewers invited
Invitations sent on 17 Jun, 2020
Reviewer #1 agreed
On 17 Jun, 2020
Review #1 received
Received 17 Jun, 2020
Editor assigned
On 16 Jun, 2020
Submission checks complete
On 15 Jun, 2020
Editor invited
On 15 Jun, 2020
No comments provided
Editorial decision: Major revision
On 15 May, 2020
Review #2 received
Received 25 Feb, 2020
Review #1 received
Received 25 Feb, 2020
Reviewer #2 agreed
On 11 Feb, 2020
Reviewer #1 agreed
On 05 Feb, 2020
Reviewers invited
Invitations sent on 03 Feb, 2020
Editor assigned
On 29 Jan, 2020
First submitted
On 28 Jan, 2020
Submission checks complete
On 28 Jan, 2020
Editor invited
On 28 Jan, 2020
Metrics
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Subject Areas
Toxicology
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This preprint is under consideration at BMC Pharmacology and Toxicology. A preprint is a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research article
Dramatic neurological and biological effects by Botulinum Neurotoxin Type A on SH-SY5Y neuroblastoma cells, beyond the blockade of neurotransmitter release
Bal R. Singh
Corresponding Author
ORCiD: https://orcid.org/0000-0002-9069-1487
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Published
Version 2
Posted 18 Jun, 2020
Published
On 05 Sep, 2020
No community comments so far
Editorial decision: Minor revision
On 11 Aug, 2020
Review #2 received
Received 10 Aug, 2020
Reviewer #2 agreed
On 20 Jun, 2020
Reviewers invited
Invitations sent on 17 Jun, 2020
Reviewer #1 agreed
On 17 Jun, 2020
Review #1 received
Received 17 Jun, 2020
Editor assigned
On 16 Jun, 2020
Submission checks complete
On 15 Jun, 2020
Editor invited
On 15 Jun, 2020
No comments provided
Editorial decision: Major revision
On 15 May, 2020
Review #2 received
Received 25 Feb, 2020
Review #1 received
Received 25 Feb, 2020
Reviewer #2 agreed
On 11 Feb, 2020
Reviewer #1 agreed
On 05 Feb, 2020
Reviewers invited
Invitations sent on 03 Feb, 2020
Editor assigned
On 29 Jan, 2020
First submitted
On 28 Jan, 2020
Submission checks complete
On 28 Jan, 2020
Editor invited
On 28 Jan, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Toxicology
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published version at BMC Pharmacology and Toxicology.
Background Gene expression profile analysis on mammalian cell lines and animal models after exposure to botulinum neurotoxin (BoNT) have been investigated in several studies in recent years. This provides a powerful tool to identify critical signaling pathways involved in the biological and inflammatory responses to BoNT and helps to determine the mechanism of the function of botulinum toxins.
One of the key clinical characteristics of BoNT is its prolonged on-site effects. The role of BoNT on the blockage of neurotransmitter acetylcholine release in the neuromuscular junction has been well established. However, the effects of the treatment time of BoNT on the human cellular model and its potential mechanism remain to be defined. This study is aimed to use the gene microarray technology to compare the two physiological critical time points of BoNT type A (BoNT/A) treatment of human neuroblastoma cells and to advance our understanding of the profound biological influences that toxin molecules play in the neuronal cellular system.
Results SH-SY5Y cells were treated with BoNT/A for 4 and 48 hours which represent the time needed for the entrance of toxin into the cells and the time necessary for the initial appearance of the on-site effects after BoNT application. A comparison of the two time points identified 122 functional groups that are significantly changed. The top five groups are the alternative splicing, phosphoprotein, nucleus, cytoplasm, and acetylation. Furthermore, after 48 hours there were 744 genes significantly up-regulated and 624 genes significantly down-regulated (p‹ 0.01). These genes fell into the following neurological and biological annotation groups: Nervous system development, proteinaceous extracellular matrix, signaling pathways regulating pluripotency of stem cells, cellular function and signal transduction, and apoptosis.
We have also noticed that the up-regulated groups contained neuronal cell development, nervous system development, and metabolic processes while the down-regulated groups contained many chromosomes and cell cycle categories.
Conclusions The effects of BoNT/A on neuronal cells extend beyond blocking the neurotransmitter release, and that BoNT/A is a multifunctional molecule that can evoke profound cellular responses which warrant a more in-depth understanding of the mechanism of the toxin’s effects after administration.
Figure 1
Figure 2
Figure 3