Hippocalcin-like 4, a neural calcium sensor, has a limited contribution to pain and itch processing

Christopher G. Alvaro; João M. Braz; Mollie Bernstein; Katherine A. Hamel; Veronica Craik; Hiroki Yamanaka; Allan I. Basbaum

doi:10.1371/journal.pone.0226289

Peer Review History

Original SubmissionOctober 18, 2019
6 Nov 2019 Decision Letter - Karl-Wilhelm Koch, Editor PONE-D-19-29157 Contribution of the neural calcium sensor hippocalcin-like 4 to pain and itch processing PLOS ONE Dear Dr. Basbaum, Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process. We would appreciate receiving your revised manuscript by Dec 21 2019 11:59PM. When you are ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file. If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. To enhance the reproducibility of your results, we recommend that if applicable you deposit your laboratory protocols in protocols.io, where a protocol can be assigned its own identifier (DOI) such that it can be cited independently in the future. For instructions see: http://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols Please include the following items when submitting your revised manuscript: A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). This letter should be uploaded as separate file and labeled 'Response to Reviewers'. A marked-up copy of your manuscript that highlights changes made to the original version. This file should be uploaded as separate file and labeled 'Revised Manuscript with Track Changes'. An unmarked version of your revised paper without tracked changes. This file should be uploaded as separate file and labeled 'Manuscript'. Please note while forming your response, if your article is accepted, you may have the opportunity to make the peer review history publicly available. The record will include editor decision letters (with reviews) and your responses to reviewer comments. If eligible, we will contact you to opt in or out. We look forward to receiving your revised manuscript. Kind regards, Karl-Wilhelm Koch, Ph.D. Academic Editor PLOS ONE Journal Requirements: When submitting your revision, we need you to address these additional requirements. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at http://www.journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and http://www.journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf 1. In your data availability statement you write, "All relevant data are within the paper and its Supporting Information files." Please ensure you have provided the individual data points used to create the figures and determine means, medians and variance measures presented in the results, tables and figures (http://journals.plos.org/plosone/s/data-availability#loc-faqs-for-data-policy). If these data cannot be publicly deposited or included in the supporting information, e.g. due to patient privacy or ownership by a third party, explain why and explain how researchers may access them. 2. Thank you for including the following funding information within the acknowledgements section of your manuscript; "This work was supported by NIH R35 NS097306 and NS14627. " We note that you have provided funding information that is not currently declared in your Funding Statement. However, funding information should not appear in the Acknowledgments section or other areas of your manuscript. We will only publish funding information present in the Funding Statement section of the online submission form. Please remove any funding-related text from the manuscript and let us know how you would like to update your Funding Statement. Currently, your Funding Statement reads as follows: "The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript." [Note: HTML markup is below. Please do not edit.] Reviewers' comments: Reviewer's Responses to Questions Comments to the Author 1. Is the manuscript technically sound, and do the data support the conclusions? The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. Reviewer #1: Yes Reviewer #2: Yes ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes Reviewer #2: Yes ****** 3. Have the authors made all data underlying the findings in their manuscript fully available? The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified. Reviewer #1: Yes Reviewer #2: Yes ****** 4. Is the manuscript presented in an intelligible fashion and written in standard English? PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here. Reviewer #1: Yes Reviewer #2: Yes ****** 5. Review Comments to the Author Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters) Reviewer #1: Neuronal Calcium Sensors (NCS) is a protein superfamily, itself, within the vast group of calcium binding proteins. At least two members of the NCS superfamily have been associated to pain perception: NCS-1 and DREAM/KChIP3/calsenilin. References to these studies are missing in the manuscript while the pain-related phenotype of calbindin-D deficient mice is cited, though this protein is only distantly related to NCS proteins. Please correct/complete the name of all the authors in the same style: Yamanaka H Reviewer #2: In the manuscript presented by Christopher GA et al, they investigated the behavioral effects of global deletion of the neural calcium sensor Hpcal4 in acute and persistent pain models. However, only a mild reduction in acute baseline thermal responses were observed in the mutant mice. And they show no major deficits in pain or itch responses in the setting of tissue or nerve injury. The conclusion that Hpcal4 makes a limited contribution to pain and itch processing was drawn. The following questions need to be considered. 1. Usually, the SNI mouse model involves ligation of two of the three branches of the sciatic nerve, the tibial nerve and the common peroneal nerve, while the sural nerve is left intact. The authors used a different method, leaving the tibial nerve intact. What is the advantage of this approach? 2. Hpcal4-deficient mice showed lower heat sensitivity in the normal state and no alteration of mechanical hypersensitivity after CFA injection. How about the change of CFA-induced heat hypersensitivity? It is critical for the drawing the conclusion that Hpcal4-deficient mice exhibit no major deficits in inflammatory pain responses. 3. What is the functional relationship between TR4 and Hpcal4? The possible mechanisms leading to the reduced expression of Hpcal4 after TR4 gene deletion need to be discussed. In addition, the compensatory effect from Hpcal1 or other calcium binding proteins should be discussed as well. 4. Whether Hpcal4 is expressed in DRG neurons? Its expression pattern in DRG should be explained and discussed. 5. As demonstrated by the authors, Hpcal4 makes a limited contribution to pain and itch processing. Thus, the title of the manuscript should be more clear and more restricted. ****** 6. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review?** For information about this choice, including consent withdrawal, please see our Privacy Policy. Reviewer #1: Yes: Jose R Naranjo Reviewer #2: Yes: Ying Zhang, Neuroscience Research Institute, Peking University, Beijing, China [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files to be viewed.] While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email us at figures@plos.org. Please note that Supporting Information files do not need this step. https://doi.org/10.1371/journal.pone.0226289.r001
Revision 1
21 Nov 2019 Author Response Editorial requirements: 1. In your data availability statement you write, "All relevant data are within the paper and its Supporting Information files." Please ensure you have provided the individual data points used to create the figures and determine means, medians and variance measures presented in the results, tables and figures Response: We have uploaded an Excel file with the raw data. 2. Thank you for including the following funding information within the acknowledgements section of your manuscript; "This work was supported by NIH R35 NS097306 and NS14627. " We note that you have provided funding information that is not currently declared in your Funding Statement. However, funding information should not appear in the Acknowledgments section or other areas of your manuscript. We will only publish funding information present in the Funding Statement section of the online submission form. Please remove any funding-related text from the manuscript and let us know how you would like to update your Funding Statement. Currently, your Funding Statement reads as follows: "The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript." Response: We have removed funding-related text from the revised manuscript. Reviewer #1: Neuronal Calcium Sensors (NCS) is a protein superfamily, itself, within the vast group of calcium binding proteins. At least two members of the NCS superfamily have been associated to pain perception: NCS-1 and DREAM/KChIP3/calsenilin. References to these studies are missing in the manuscript while the pain-related phenotype of calbindin-D deficient mice is cited, though this protein is only distantly related to NCS proteins. Response: We thank the Reviewer for pointing this out. Indeed, we were remiss not to include these studies in our manuscript. Reference to these studies has now been included in the Introduction section. Please correct/complete the name of all the authors in the same style: Yamanaka H Response: The name has been corrected. Reviewer #2: In the manuscript presented by Christopher GA et al, they investigated the behavioral effects of global deletion of the neural calcium sensor Hpcal4 in acute and persistent pain models. However, only a mild reduction in acute baseline thermal responses was observed in the mutant mice. And they show no major deficits in pain or itch responses in the setting of tissue or nerve injury. The conclusion that Hpcal4 makes a limited contribution to pain and itch processing was drawn. The following questions need to be considered. 1. Usually, the SNI mouse model involves ligation of two of the three branches of the sciatic nerve, the tibial nerve and the common peroneal nerve, while the sural nerve is left intact. The authors used a different method, leaving the tibial nerve intact. What is the advantage of this approach? Response: There are, in fact, different methods to generate the spared nerve injury model. In our approach, we ligate and cut the sural and peroneal nerves but leave the tibial intact. This results in profound and long lasting mechanical allodynia, which we described in detail in our Shields et al (2003) manuscript that reported an adaptation of the DeCosterd et al rat model. There are some advantages to our approach. Namely, by leaving the tibial nerve intact, we have access to a much bigger area of the footpad to apply the von Frey filaments and there is considerably less motor impairment. 2. Hpcal4-deficient mice showed lower heat sensitivity in the normal state and no alteration of mechanical hypersensitivity after CFA injection. How about the change of CFA-induced heat hypersensitivity? It is critical for the drawing the conclusion that Hpcal4-deficient mice exhibit no major deficits in inflammatory pain responses. Response: As the reviewer notes, we did not analyze the consequences of Hpcal4 deletion on thermal thresholds after CFA and therefore we cannot rule out an effect on thermal hypersensitivity in the setting of inflammatory pain. As thermal hypersensitivity cannot reliably be generated in the SNI, partial peripheral nerve injury models (Shields et al., 2003), we decided to focus our analyses on the mechanical changes that occur after nerve and tissue injuries. We have included these considerations in the Discussion section of the revised manuscript. 3. What is the functional relationship between TR4 and Hpcal4? The possible mechanisms leading to the reduced expression of Hpcal4 after TR4 gene deletion need to be discussed. In addition, the compensatory effect from Hpcal1 or other calcium binding proteins should be discussed as well. Response: As we previously demonstrated, deletion of the orphan nuclear receptor TR4 results in profound defects in a wide variety of pain and itch responses that are associated with a developmental loss of excitatory interneurons in the superficial dorsal horn of the spinal cord. We presume that the Hpcal4 reduction observed in the TR4 deficient mouse results from the loss of this population of excitatory neuron, rather than the loss of a functional relationship between TR4 and Hpcal4. In fact, we have no evidence that TR4 and Hpcal4 are at all related. On the other hand, we have now discussed in the revised manuscript the possible compensatory effects provided by Hpcal1. 4. Whether Hpcal4 is expressed in DRG neurons? Its expression pattern in DRG should be explained and discussed. Response: To our knowledge, there are no neuroanatomical (immunocytochemical or in situ) studies that analyzed the expression pattern of Hpcal4 in sensory neurons. However, a recent transcriptome analysis did suggest that both Hpcal1 and 4 are expressed in subpopulations of DRG neurons. These considerations have been now added in the Discussion of the revised manuscript. Of course, we recognize that because this was a complete null, that there is always a possible contribution of the Hpcal4 loss in areas other than the dorsal horn. 5. As demonstrated by the authors, Hpcal4 makes a limited contribution to pain and itch processing. Thus, the title of the manuscript should be more clear and more restricted. Response: We have modified the title of the manuscript to better reflect our conclusions. Attachments Attachment Submitted filename: Response to Reviewers.doc https://doi.org/10.1371/journal.pone.0226289.r002
25 Nov 2019 Decision Letter - Karl-Wilhelm Koch, Editor Hippocalcin-like 4, a neural calcium sensor, has a limited contribution to pain and itch processing PONE-D-19-29157R1 Dear Dr. Basbaum, We are pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it complies with all outstanding technical requirements. Within one week, you will receive an e-mail containing information on the amendments required prior to publication. When all required modifications have been addressed, you will receive a formal acceptance letter and your manuscript will proceed to our production department and be scheduled for publication. Shortly after the formal acceptance letter is sent, an invoice for payment will follow. To ensure an efficient production and billing process, please log into Editorial Manager at https://www.editorialmanager.com/pone/, click the "Update My Information" link at the top of the page, and update your user information. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org. If your institution or institutions have a press office, please notify them about your upcoming paper to enable them to help maximize its impact. If they will be preparing press materials for this manuscript, you must inform our press team as soon as possible and no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org. With kind regards, Karl-Wilhelm Koch, Ph.D. Academic Editor PLOS ONE Additional Editor Comments (optional): Reviewers' comments: https://doi.org/10.1371/journal.pone.0226289.r003
Formally Accepted
23 Jan 2020 Acceptance Letter - Karl-Wilhelm Koch, Editor PONE-D-19-29157R1 Hippocalcin-like 4, a neural calcium sensor, has a limited contribution to pain and itch processing Dear Dr. Basbaum: I am pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department. If your institution or institutions have a press office, please notify them about your upcoming paper at this point, to enable them to help maximize its impact. If they will be preparing press materials for this manuscript, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information please contact onepress@plos.org. For any other questions or concerns, please email plosone@plos.org. Thank you for submitting your work to PLOS ONE. With kind regards, PLOS ONE Editorial Office Staff on behalf of Dr. Karl-Wilhelm Koch Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0226289.r004

Open letter on the publication of peer review reports

PLOS recognizes the benefits of transparency in the peer review process. Therefore, we enable the publication of all of the content of peer review and author responses alongside final, published articles. Reviewers remain anonymous, unless they choose to reveal their names.

We encourage other journals to join us in this initiative. We hope that our action inspires the community, including researchers, research funders, and research institutions, to recognize the benefits of published peer review reports for all parts of the research system.

Learn more at ASAPbio .