Reviewer ’s report  
 
Title: Histopathological features of the proper gastric glands in FVB/N -background mice 
carrying constitutively -active aryl -hydrocarbon receptor  
 
Version: 0 Date: 30 Oct 2017  
 
Reviewer: Robert Coffey  
 
Reviewer's report:  
 
Please include all comments for the authors in this box rather than uploading your report as an 
attachment. Please only upload as attachments annotated versions of manuscripts, graphs, 
supporting materials or other aspects of your report which ca nnot be included in a text format.  
Please overwrite this text when adding your comments to the authors.  
The authors report that transgenic mice expressing constitutively -active AhR (CA -AhR) develop 
gastric lesions mimicking gastritis cystica profunda. How ever, this finding has been previously 
reported. The histological findings are more consistent with spasmolytic polypeptide -expressing 
metaplasia (SPEM) than intestinal metaplasia.  
 
Major points  
1. The authors report that CA -AhR induces intramucosal (IM) a nd extramucosal (EM) cysts, 
foveolar hyperplasia, fused gastric pit, decreased number of parietal and chief cells, lymphocytic 
infiltration near the cysts, and downward migration of the proliferation center. All the findings 
were previously reported using a different strain of mice. Only differences are that EM cysts 
develop earlier in the current study using FVB -CA-AhR mice (8 weeks) compared to prior 
studies using B6/CBA -CA-AhR mice (10 weeks) and that EM cysts are confined in the stomach 
in FVB -CA-AhR mi ce, whereas EM cysts develop also in cecum and ileum in B6/CBA -CA-AhR 
mice. Phenotypic differences in different strains have been well -described. The authors are 
encouraged to identify the factors that can explain the phenotypic differences between FVB -CA-
AhR and B6/CBA -CA-AhR.  
2. The authors argue that CA -AhR induces intestinal metaplasia in the stomach solely based on 
alcian blue positivity. However, it is known that deep glandular cells of gastric antrum are 
positive for alcian blue in mice. Intestinal m etaplasia in mice requires both histologic goblet cell 
morphology and immunohistochemical Muc2 - or Tff3 -positivity. In this reviewer's opinion, 
phenotypes of CA -AhR mice resemble SPEM rather than intestinal metaplasia. Immunostaining 
with SPEM markers such  as Tff2 or CD44v9 should be performed.  
 
Minor points  
1. Terms of lamina muscularis and muscular layer are repeatedly used in the manuscript. Correct 
terms are (lamina) muscularis mucosae and muscularis propria.  
2. [Line 259] The manuscript reads that the epithelial cells lining cysts are squamous or 
columnar shaped and refers Fig. 5d. However, Fig. 5d only shows columnar epithelium.  
3. In discussion, the authors say that it would be interesting to study the functional crosstalk 
between AhR activation and L gr5+ stem cells. However, phenotypes of AhR activation is 
confined in the corpus and Lgr5+ stem cells are not present in the corpus but present only in the 
antrum and cardia.  
Are the methods appropriate and well described?  
If not, please specify what is required in your comments to the authors.  
 
Yes 
 
Does the work inc lude the necessary controls?  
If not, please specify which controls are required in your comments to the authors.  
 
Yes 
 
Are the conclusions drawn adequately supported by the data shown?  
If not, please explain in your comments to the authors.  
 
No 
 
Are you able to assess any statistics in the manuscript or would you recommend an  
additional statistical review?  
If an additional statistical review is recommended, please specify what aspects require further  
assessment in your comments to the editors.  
 
I am able to assess the statistics  
 
Quality of written English  
Please indicate the quality of language in the manuscript:  
 
Acceptable  
 
  
Declaration of competing interests  
Please complete a declaration of competing interests, considering the following questions:  
 
1. Have  you in the past five years received reimbursements, fees, funding, or salary from an  
organisation that may in any way gain or lose financially from the publication of this  
manuscript, either now or in the future?  
 
2. Do you hold any stocks or shares in an organisation that may in any way gain or lose  
financially from the publication of this manuscript, either now or in the future?  
 
3. Do you hold or are you currently applying for any patents relating to the content of the  
manuscript?  
 
4. Have you received reimbursements, fees, funding, or salary from an organization that 
holds  or has applied for patents relating to the content of the manuscript?  
 
5. Do you have any other financial competing interests?  
 
6. Do you have any non -financial com peting interests in relation to this paper?  
 
If you can answer no to all of the above, write 'I declare that I have no competing interests' 
below.  If your reply is yes to any, please give details below.  
 
I declare that I have no competing interests  
 
I agree to the open peer review policy of the jou rnal. I understand that my name will be included  
on my report to the authors and, if the manuscript is accepted for publication, my named report  
including any attachments I upload will be posted on the website along with the authors'  
responses. I agree for  my report to be made available under an Open Access Creative Commons  
CC-BY license ( http://creativecommons.org/licenses/by/4.0/ ). I understand that any comments  
which I do not wish to be included in my named report can be included as confidential comments  
to the editors, which will not be published.  
 
I agree to the open peer review policy of the journal  
Reviewer ’s report  
 
Title: Histopathological features of the proper gastric glands in FVB/N -background mice 
carrying constitutively -active aryl -hydrocarbon receptor  
 
Version: 0 Date: 30 Oct 2017  
 
Reviewer: Robert Coffey  
 
Reviewer's report:  
 
Please include all comments for the authors in this box rather than uploading your report as an 
attachment. Please only upload as attachments annotated versions of manuscripts, graphs, 
supporting materials or other aspects of your report which ca nnot be included in a text format.  
Please overwrite this text when adding your comments to the authors.  
The authors report that transgenic mice expressing constitutively -active AhR (CA -AhR) develop 
gastric lesions mimicking gastritis cystica profunda. How ever, this finding has been previously 
reported. The histological findings are more consistent with spasmolytic polypeptide -expressing 
metaplasia (SPEM) than intestinal metaplasia.  
 
Major points  
1. The authors report that CA -AhR induces intramucosal (IM) a nd extramucosal (EM) cysts, 
foveolar hyperplasia, fused gastric pit, decreased number of parietal and chief cells, lymphocytic 
infiltration near the cysts, and downward migration of the proliferation center. All the findings 
were previously reported using a different strain of mice. Only differences are that EM cysts 
develop earlier in the current study using FVB -CA-AhR mice (8 weeks) compared to prior 
studies using B6/CBA -CA-AhR mice (10 weeks) and that EM cysts are confined in the stomach 
in FVB -CA-AhR mi ce, whereas EM cysts develop also in cecum and ileum in B6/CBA -CA-AhR 
mice. Phenotypic differences in different strains have been well -described. The authors are 
encouraged to identify the factors that can explain the phenotypic differences between FVB -CA-
AhR and B6/CBA -CA-AhR.  
2. The authors argue that CA -AhR induces intestinal metaplasia in the stomach solely based on 
alcian blue positivity. However, it is known that deep glandular cells of gastric antrum are 
positive for alcian blue in mice. Intestinal m etaplasia in mice requires both histologic goblet cell 
morphology and immunohistochemical Muc2 - or Tff3 -positivity. In this reviewer's opinion, 
phenotypes of CA -AhR mice resemble SPEM rather than intestinal metaplasia. Immunostaining 
with SPEM markers such  as Tff2 or CD44v9 should be performed.  
 
Minor points  
1. Terms of lamina muscularis and muscular layer are repeatedly used in the manuscript. Correct 
terms are (lamina) muscularis mucosae and muscularis propria.  
2. [Line 259] The manuscript reads that the epithelial cells lining cysts are squamous or 
columnar shaped and refers Fig. 5d. However, Fig. 5d only shows columnar epithelium.  
3. In discussion, the authors say that it would be interesting to study the functional crosstalk 
between AhR activation and L gr5+ stem cells. However, phenotypes of AhR activation is 
confined in the corpus and Lgr5+ stem cells are not present in the corpus but present only in the 
antrum and cardia.  
Are the methods appropriate and well described?  
If not, please specify what is required in your comments to the authors.  
 
Yes 
 
Does the work inc lude the necessary controls?  
If not, please specify which controls are required in your comments to the authors.  
 
Yes 
 
Are the conclusions drawn adequately supported by the data shown?  
If not, please explain in your comments to the authors.  
 
No 
 
Are you able to assess any statistics in the manuscript or would you recommend an  
additional statistical review?  
If an additional statistical review is recommended, please specify what aspects require further  
assessment in your comments to the editors.  
 
I am able to assess the statistics  
 
Quality of written English  
Please indicate the quality of language in the manuscript:  
 
Acceptable  
 
  
Declaration of competing interests  
Please complete a declaration of competing interests, considering the following questions:  
 
1. Have  you in the past five years received reimbursements, fees, funding, or salary from an  
organisation that may in any way gain or lose financially from the publication of this  
manuscript, either now or in the future?  
 
2. Do you hold any stocks or shares in an organisation that may in any way gain or lose  
financially from the publication of this manuscript, either now or in the future?  
 
3. Do you hold or are you currently applying for any patents relating to the content of the  
manuscript?  
 
4. Have you received reimbursements, fees, funding, or salary from an organization that 
holds  or has applied for patents relating to the content of the manuscript?  
 
5. Do you have any other financial competing interests?  
 
6. Do you have any non -financial com peting interests in relation to this paper?  
 
If you can answer no to all of the above, write 'I declare that I have no competing interests' 
below.  If your reply is yes to any, please give details below.  
 
I declare that I have no competing interests  
 
I agree to the open peer review policy of the jou rnal. I understand that my name will be included  
on my report to the authors and, if the manuscript is accepted for publication, my named report  
including any attachments I upload will be posted on the website along with the authors'  
responses. I agree for  my report to be made available under an Open Access Creative Commons  
CC-BY license ( http://creativecommons.org/licenses/by/4.0/ ). I understand that any comments  
which I do not wish to be included in my named report can be included as confidential comments  
to the editors, which will not be published.  
 
I agree to the open peer review policy of the journal  
Please review this paper.
user
Reviewer ’s report  
 
Title: Histopathological features of the proper gastric glands in FVB/N -background mice 
carrying constitutively -active aryl -hydrocarbon receptor  
 
Version: 0 Date: 30 Oct 2017  
 
Reviewer: Robert Coffey  
 
Reviewer's report:  
 
Please include all comments for the authors in this box rather than uploading your report as an 
attachment. Please only upload as attachments annotated versions of manuscripts, graphs, 
supporting materials or other aspects of your report which ca nnot be included in a text format.  
Please overwrite this text when adding your comments to the authors.  
The authors report that transgenic mice expressing constitutively -active AhR (CA -AhR) develop 
gastric lesions mimicking gastritis cystica profunda. How ever, this finding has been previously 
reported. The histological findings are more consistent with spasmolytic polypeptide -expressing 
metaplasia (SPEM) than intestinal metaplasia.  
 
Major points  
1. The authors report that CA -AhR induces intramucosal (IM) a nd extramucosal (EM) cysts, 
foveolar hyperplasia, fused gastric pit, decreased number of parietal and chief cells, lymphocytic 
infiltration near the cysts, and downward migration of the proliferation center. All the findings 
were previously reported using a different strain of mice. Only differences are that EM cysts 
develop earlier in the current study using FVB -CA-AhR mice (8 weeks) compared to prior 
studies using B6/CBA -CA-AhR mice (10 weeks) and that EM cysts are confined in the stomach 
in FVB -CA-AhR mi ce, whereas EM cysts develop also in cecum and ileum in B6/CBA -CA-AhR 
mice. Phenotypic differences in different strains have been well -described. The authors are 
encouraged to identify the factors that can explain the phenotypic differences between FVB -CA-
AhR and B6/CBA -CA-AhR.  
2. The authors argue that CA -AhR induces intestinal metaplasia in the stomach solely based on 
alcian blue positivity. However, it is known that deep glandular cells of gastric antrum are 
positive for alcian blue in mice. Intestinal m etaplasia in mice requires both histologic goblet cell 
morphology and immunohistochemical Muc2 - or Tff3 -positivity. In this reviewer's opinion, 
phenotypes of CA -AhR mice resemble SPEM rather than intestinal metaplasia. Immunostaining 
with SPEM markers such  as Tff2 or CD44v9 should be performed.  
 
Minor points  
1. Terms of lamina muscularis and muscular layer are repeatedly used in the manuscript. Correct 
terms are (lamina) muscularis mucosae and muscularis propria.  
2. [Line 259] The manuscript reads that the epithelial cells lining cysts are squamous or 
columnar shaped and refers Fig. 5d. However, Fig. 5d only shows columnar epithelium.  
3. In discussion, the authors say that it would be interesting to study the functional crosstalk 
between AhR activation and L gr5+ stem cells. However, phenotypes of AhR activation is 
confined in the corpus and Lgr5+ stem cells are not present in the corpus but present only in the 
antrum and cardia.  
Are the methods appropriate and well described?  
If not, please specify what is required in your comments to the authors.  
 
Yes 
 
Does the work inc lude the necessary controls?  
If not, please specify which controls are required in your comments to the authors.  
 
Yes 
 
Are the conclusions drawn adequately supported by the data shown?  
If not, please explain in your comments to the authors.  
 
No 
 
Are you able to assess any statistics in the manuscript or would you recommend an  
additional statistical review?  
If an additional statistical review is recommended, please specify what aspects require further  
assessment in your comments to the editors.  
 
I am able to assess the statistics  
 
Quality of written English  
Please indicate the quality of language in the manuscript:  
 
Acceptable  
 
  
Declaration of competing interests  
Please complete a declaration of competing interests, considering the following questions:  
 
1. Have  you in the past five years received reimbursements, fees, funding, or salary from an  
organisation that may in any way gain or lose financially from the publication of this  
manuscript, either now or in the future?  
 
2. Do you hold any stocks or shares in an organisation that may in any way gain or lose  
financially from the publication of this manuscript, either now or in the future?  
 
3. Do you hold or are you currently applying for any patents relating to the content of the  
manuscript?  
 
4. Have you received reimbursements, fees, funding, or salary from an organization that 
holds  or has applied for patents relating to the content of the manuscript?  
 
5. Do you have any other financial competing interests?  
 
6. Do you have any non -financial com peting interests in relation to this paper?  
 
If you can answer no to all of the above, write 'I declare that I have no competing interests' 
below.  If your reply is yes to any, please give details below.  
 
I declare that I have no competing interests  
 
I agree to the open peer review policy of the jou rnal. I understand that my name will be included  
on my report to the authors and, if the manuscript is accepted for publication, my named report  
including any attachments I upload will be posted on the website along with the authors'  
responses. I agree for  my report to be made available under an Open Access Creative Commons  
CC-BY license ( http://creativecommons.org/licenses/by/4.0/ ). I understand that any comments  
which I do not wish to be included in my named report can be included as confidential comments  
to the editors, which will not be published.  
 
I agree to the open peer review policy of the journal  
assistant