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PONE-D-22-32310Generalization of contextual fear is sex-specifically affected by high salt intakePLOS ONE

Dear Dr. Gilman,

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PLOS ONE

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Funding for this work was provided by Kent State University, and the Applied Psychology Center in the Department of Psychological Sciences at Kent State University.  

  

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Funding for this work was provided by Kent State University, and the Applied Psychology Center in the Department of Psychological Sciences at Kent State University.  

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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: Partly

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2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: Yes

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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

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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

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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: This is an interesting study. The following issues should be addressed.

1. Fig. 2. Adjust the Y axis to reflect the low freezing levels seen. Also, it would be better to show these data as bar graphs so that group differences can be appreciated.

2. Only 5 seconds post the shock are analyzed. It seems better to analyze the largest ISI possible.

3. Females and males respond different to singly housing. Therefore, it is conceivable that in experiment 3 sex-dependent effects of singly housing might have contributed to the sex-dependent effects seen. This should be acknowledged.

4. It seems better to move figure panels not showing any significance from the main figure to supplementary figures. Some figure, like figure 6, could be a main or supplementary table as well. Fig. 4 is the main figure and receives less attention now.

5. There seems more of a sex difference in the training context in Experiments 1 and 2 in Fig. 4 than in Experiment 3. One wonder whether this might have contributed to what is seen in Experiment 3. In females, the pattern in the neutral context seems definitely similar in Experiments 2 and 3.

6. The salt effect in females in the neutral context seems subtle and not different between the two salt diet groups. The salt effect in males in the neutral context seems more pronounced.

7. It would be better to show the real cort data so that others can compare with other designs.

8. It would be better to remove p values from the figures, especially if they do not reach significance.

9. The text in the results section is written very stats heavy. While all correct, this makes it harder to read for those less versed in statistical interactions. The authors are encouraged to rephrase some of the text to make this manuscript more readable. The tables with the stats data are good and allow focusing less on stats result in the text.

Reviewer #2: This manuscript evaluates that possibility that a forced, high-salt diet affects contextual fear acquisition, recall and generalization in male and female C57BL/6J mice. I think there is some important data here, and it should be seen. Most of my concerns are with how the data, statistics, etc. are presented. Given that this is a straight-forward experiment, it would benefit to attempt to streamline what is being presented to the reader. I found myself paying too much attention to small details the were being presented/suggested rather than being able to focus on the main message (or two) as I read through the Results and Discussion presentation.

Specific comments:

Introduction: Lines 68-70 – I think it would be useful to provide some specific detail on how hippocampus/amygdala are affected by high-salt.

Consider providing a clear “N per group/sex” in the Methods.

Describe in more detail why 5x shock was used. 1 or two is sufficient to engender contextual fear.

Page 8, Line 158 – Much more thorough justification needs to be provided for why the authors are selectively presenting/analyzing data from minutes 2-6 out of the 10-minute test. Without this, it appears that the data has been “cherry picked” to show some effects.

Figure 2 – It is confusing to refer to a “post-shock period” when these experiments did not employ shock. Also, I find using the sex-symbols as data points to be distracting. Could you more simply use blue/red squares and circles?

Consider placing the tables in a data supplement. There are 10 of them, several with multiple parts, and they are essentially all ANOVA tables.

Page 26, Line 526 – The overall statistical analyses appear to be rigorous. The notion of a “trend” presented in the Discussion detracts from this.

Page 27, Line 548 – The above is also true for the notion of a “bimodal” distribution, which would be very difficult to show with that number of data points.

Page 28, Line 579 – Same for “non-significant trends”

How do the authors know that 0.8mA is not painful?

While I think the authors are likely correct about the lack of social simulation and light-phase testing in their experiment, bringing this up distracts from the overall message regarding the data at hand. For example, it begs the question why reverse-phase lighting was not used in the first place?

Page 28, Line 566 – Sentence beginning “Nonetheless…”. I would like the authors to expand more upon this. From a biological standpoint, I’m left hanging and wondering “how would this be possible?”. Perhaps the authors can draw on other work for examples outside of hypertension/anxiety.

Do the authors think their findings would generalize to more classic mouse tests of “anxiety”, such as EPM or open field?

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Reviewer #1: No

Reviewer #2: No

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Generalization of contextual fear is sex-specifically affected by high salt intake

PONE-D-22-32310R1

Dear Dr. Gilman,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication.

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If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- 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.

Kind regards,

Thiago Fernandes, PhD

Academic Editor

PLOS ONE

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #1: All comments have been addressed

Reviewer #2: All comments have been addressed

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2. 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

**********

3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: Yes

**********

4. 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

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5. 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

**********

6. 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: The authors did a fine job addressing the raised concerns. No additional edits are requested from this reviewer.

Reviewer #2: (No Response)

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7. 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: No

Reviewer #2: No

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