如何回复审稿人意见(Response to Reviews)

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Dear Editor,

We have studied the valuable comments from you, the assistant editor and reviewers carefully, and tried our best to revise the manuscript. The point to point responds to the reviewer’s comments are listed as following:

Responds to the reviewer’s comments:

Reviewer 1

Comment 1: in page 3, line 40, we fed rats..." changed to rats were fed with...

Response: According to the reviewer’s comment, we have corrected the sentence. Furthermore, we have had the manuscript polished with a professional assistance in writing.

Comment 2: page 25. The style of reference 40 is not right (using initials for the first names). Since this paper has been published, the volume and page Nos should be provided.

Response: Thank you for your careful work. We have added the volume and page numbers for reference 40.

Reviewer 2

Comment: I would like to thank the authors for their efforts in addressing the criticisms with additional experiments. The one criticism that they did not address was relating to energy expenditure as the reason that the animals on the low calcium diet gained more weight. While I understand that performing this experiment will not affect the conclusion of this manuscript, I do believe that this point could be discussed in the Discussion section.

Response: Thank you for your valuable advice. Based on the previous revision, we further address the relationship between low calcium diet and energy expenditure in the section of discussion according to your thoughtful comments.

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Reviewer 3

Comment 1: In the text you often write: “As previously described”. Unless that paper is from your lab or one of the method paper co-authors is on the present MS this is not quite proper since the statement infers method development from your lab. There are numerous instances like that in the methods section; these should all be changed “according to those described by…..”

Response: We are sorry for this language mistake. We have carefully corrected this phrase throughout the manuscript according to your comment.

Comment 2: There are still some wording, sentence structure and grammatical issues even in this basically well put together MS. For example, while authors may have been excited about the data you cannot start a sentence with “Excitedly” in line 418 or “Whatever” in line 395.

Response: Thank you very much to point out the sentence structure and grammatical issues in our manuscript. According to the comments from you and the editors, we polished the manuscript with a professional assistance in writing, conscientiously.

Comment 3: In my view a big omission in this work is ignoring the anabolic side of lipid metabolism as well as thermogenesis issues. For example all animals consumed the same amount of feed but we had extra fat storage in the low Ca diet groups. So where did the extra energy go? Zemel et al (citation 34) in similar work indicate that increased thermogenesis on the high Ca diet explains the dissipation of dietary energy. Further even though Zemel et al (#34) indicated lipogenesis was enhanced in the low Ca diets that was in 2000 and you should have monitored expression of FAS and UCP either as mRNA abundance or actual FAS/UCP changes via proteomics or blotting techniques. In any case these controls are missing here and not emphasized in the MS. Casual reading of this paper would lead to the conclusion that the dietary Ca effect on fat deposition is strictly a function of increased or decreased lipolysis. While lipolysis appears to be a major player, lipogenesis and thermogenesis cannot be ignored for completeness. In Fig 8 you also show a decline in cAMP for the low Ca diet. Well beta agonists or cAMP enhancers regulate transcription of adipose and liver FAS (in rats (J Biol Chem 271:2307, 1996) and recently with large animal models (Hausman et al J Animal Science 87:1218, 2009 and Halsey et al J Animal Science 89: 1011, 2011). In addition

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cAMP levels could have been monitored. I really do not like the last sentence in the Abstract line 47-50 where you state that “low calcium diet-induced increase in fat mass was due to enhanced lipogenesis mediated by an upregulated CaSR signaling pathway” Your results here show no such thing, this is a completely false statement based on data herein. Correct. You show that high Ca diets enhance lipolysis and low Ca diets are antilipolytic. You did not monitor lipid anabolism here at all. See also line 255-257 and lines 333-335 of your MS.

Response: Thank you for your valuable and thoughtful comments. As you suggested that the anabolic side of lipid metabolism as well as thermogenesis issues should be monitored. We really agree with your viewpoints. In the present study, we did find that low calcium diet increased the mRNA level of fatty acid synthase (FAS) in white adipose tissue. Furthermore, the FAS mRNA level were also increased in adipocytes after treatment with 1,25-(OH)2D3 in in-vitro experiments. However, the increased FAS mRNA levels were not affected by preventing either the nuclear vitamin D receptor (nVDR) or calcium-sensing receptor (CaSR), suggesting that FAS might not be involved in the CaSR pathway. In addition, we thought that FAS played its role in fatty acid synthesis mainly in liver previously. Besides, the manuscript was required to restrict number of total words and our previous focus was on the antilolytic role of CaSR in the process of fat accumulation. So we ignored to provide the data of FAS mRNA levels in the submitted manuscript. In the newly submitted manuscript, we have provided the mRNA levels according to your helpful suggestion.

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