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Research article
Spatiotemporal expression of sonic hedgehog signaling molecules in the embryonic mesencephalic dopaminergic neurons
Doaa Mokhtar
assuit university
Corresponding Author
ORCiD: https://orcid.org/0000-0001-7975-3926
License:
This work is licensed under a CC BY 4.0 License. Read Full License
The most recent version of this article is available here.
Background
Midbrain dopaminergic neurons (mDA) play an important role in controlling the voluntary motor movement, reward behavior and emotion-based behavior. Differentiation of mDA neurons from progenitors depends on a number of secreted proteins, such as sonic hedgehog (SHH), and transcription factors. Different groups from mDA neurons arise from the varied patterns of SHH expression during development. The present study attempted to elucidate 1) the possible role(s) of some SHH signaling components (Ptch1, Gli1, Gli2 and Gli3) in the spatiotemporal development of mDA neurons along the rostrocaudal axis of the midbrain and their possible roles in differentiation (E12, E14) and survival of mDA neurons (E18); 2) the main role of Boc (bioregional Cdon-binding protein), and Gas1 (growth arrest-specific 1) as novel accessory receptors for the SHH in the development of mDA neurons 3) the significance of using the primary culture and/or the dopaminergic cell line (MN9D) for studying the development of mDA neurons.
Results
Mice embryos from embryonic day (E)12 were used for in situ hybridization and immunohistochemistry. The primary culture and cell line MN9D were used for in vitro investigation. At E12 and E14, but not E18, only Ptch1 and Gli1 were expressed in ventrolateral midbrain domains. All examined SHH signaling molecules were not detected in mDA area. Whereas, in MN9D cells, many SHH signaling molecules were expressed and co-localized with dopaminergic marker; tyrosine hydroxylase (TH), and their expression were upregulated with SHH treatment of the MN9D cells.
Conclusion
These results suggest that mDA neurons differentiation and survival are independent of SHH. In addition, MN9D cell line is not the ideal in vitro model for investigating the differentiation of mesencephalic dopaminergic neurons, and hence, the ventral midbrain primary culture might be favored over MN9D line.
Keywords
Boc, Gas1, mesencephalon, dopaminergic, MN9D, SHH, ISH
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Enrique Estudillo
commented on 09 July, 2020
Interesting information. Ayala-Sarmiento and collaborators also found Gas1 expression in the choroid plexus of pups and adult rats, suggesting its secretion from this structure to the cerebrospinal fluid. These results may reinforce your findings. I encourage you to read this work: GAS1 is present in the cerebrospinal fluid and is expressed in the choroid plexus of the adult rat All the best, Enrique
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CURRENT STATUS: Under Review
No community comments so far
Editorial decision: Major revision
On 31 Jan, 2021
Review #2 received
Received 20 Jan, 2021
Reviewer #2 agreed
On 13 Jan, 2021
Reviewers invited
Invitations sent on 13 Jan, 2021
Reviewer #1 agreed
On 13 Jan, 2021
Review #1 received
Received 13 Jan, 2021
Editor assigned
On 19 Dec, 2020
Submission checks complete
On 19 Dec, 2020
Editor invited
On 19 Dec, 2020
Version 1
Posted 06 Jul, 2020
Community comments: 1
Editorial decision: Minor revision
On 05 Dec, 2020
Review #2 received
Received 27 Nov, 2020
Review #1 received
Received 11 Oct, 2020
Reviewer #2 agreed
On 24 Sep, 2020
Reviewer #1 agreed
On 23 Sep, 2020
Reviewers invited
Invitations sent on 22 Jul, 2020
Editor assigned
On 21 Jul, 2020
Submission checks complete
On 01 Jul, 2020
Editor invited
On 01 Jul, 2020
First submitted
On 24 Jun, 2020
Metrics
Comments: 1
PDF Downloads: ...
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This preprint is under consideration at BMC Veterinary Research. 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
Spatiotemporal expression of sonic hedgehog signaling molecules in the embryonic mesencephalic dopaminergic neurons
Doaa Mokhtar
assuit university
Corresponding Author
ORCiD: https://orcid.org/0000-0001-7975-3926
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: Under Review
No community comments so far
Editorial decision: Major revision
On 31 Jan, 2021
Review #2 received
Received 20 Jan, 2021
Reviewer #2 agreed
On 13 Jan, 2021
Reviewers invited
Invitations sent on 13 Jan, 2021
Reviewer #1 agreed
On 13 Jan, 2021
Review #1 received
Received 13 Jan, 2021
Editor assigned
On 19 Dec, 2020
Submission checks complete
On 19 Dec, 2020
Editor invited
On 19 Dec, 2020
Version 1
Posted 06 Jul, 2020
Community comments: 1
Editorial decision: Minor revision
On 05 Dec, 2020
Review #2 received
Received 27 Nov, 2020
Review #1 received
Received 11 Oct, 2020
Reviewer #2 agreed
On 24 Sep, 2020
Reviewer #1 agreed
On 23 Sep, 2020
Reviewers invited
Invitations sent on 22 Jul, 2020
Editor assigned
On 21 Jul, 2020
Submission checks complete
On 01 Jul, 2020
Editor invited
On 01 Jul, 2020
First submitted
On 24 Jun, 2020
Metrics
Comments: 1
PDF Downloads: ...
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
The most recent version of this article is available here.
Background
Midbrain dopaminergic neurons (mDA) play an important role in controlling the voluntary motor movement, reward behavior and emotion-based behavior. Differentiation of mDA neurons from progenitors depends on a number of secreted proteins, such as sonic hedgehog (SHH), and transcription factors. Different groups from mDA neurons arise from the varied patterns of SHH expression during development. The present study attempted to elucidate 1) the possible role(s) of some SHH signaling components (Ptch1, Gli1, Gli2 and Gli3) in the spatiotemporal development of mDA neurons along the rostrocaudal axis of the midbrain and their possible roles in differentiation (E12, E14) and survival of mDA neurons (E18); 2) the main role of Boc (bioregional Cdon-binding protein), and Gas1 (growth arrest-specific 1) as novel accessory receptors for the SHH in the development of mDA neurons 3) the significance of using the primary culture and/or the dopaminergic cell line (MN9D) for studying the development of mDA neurons.
Results
Mice embryos from embryonic day (E)12 were used for in situ hybridization and immunohistochemistry. The primary culture and cell line MN9D were used for in vitro investigation. At E12 and E14, but not E18, only Ptch1 and Gli1 were expressed in ventrolateral midbrain domains. All examined SHH signaling molecules were not detected in mDA area. Whereas, in MN9D cells, many SHH signaling molecules were expressed and co-localized with dopaminergic marker; tyrosine hydroxylase (TH), and their expression were upregulated with SHH treatment of the MN9D cells.
Conclusion
These results suggest that mDA neurons differentiation and survival are independent of SHH. In addition, MN9D cell line is not the ideal in vitro model for investigating the differentiation of mesencephalic dopaminergic neurons, and hence, the ventral midbrain primary culture might be favored over MN9D line.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Enrique Estudillo
commented on 09 July, 2020
Interesting information. Ayala-Sarmiento and collaborators also found Gas1 expression in the choroid plexus of pups and adult rats, suggesting its secretion from this structure to the cerebrospinal fluid. These results may reinforce your findings. I encourage you to read this work: GAS1 is present in the cerebrospinal fluid and is expressed in the choroid plexus of the adult rat All the best, Enrique