Treating Mares with the Long-Acting Kisspeptin Analog C6 Increases Circulating Gonadotropins but Does Not Trigger Ovulation
, Vincent Hellier 1, Vincent Aucagne 4, Anne Duittoz 1 and Massimiliano Beltramo 1,*Round 1
Reviewer 1 Report
It is recommended to increase the number of experimental animals, set different C6 administration time and dosage, and verify whether the short duration of LH peak results in anovulation.
Line35: The Oc6 animals did not ovulate throughout the oestrous cycle. Right? or do not ovulate during specific periods of time？If the horse never ovulates in the cycle, is there something wrong with the horse? Please confirm in the article.
Materials and Methods
Line187：The unit of KP in the article suggested replacing "mmol" with "mg" to facilitate comparison with other literature.
Line198：Figure 1: The flowchart is somewhat tedious and messy, and it is recommended to simplify it.
Line 200：For "T±/Χh", please verify the "Χ" writing format.
Line211： “（”，please delete.
Results
Figures 3, 4, 5: The standard deviation is large, and Figure A、B、C is recommended to be combined into one figure.
If possible, it is recommended to reconstitute the test hormone concentration.
Line 483-484: “Should we conclude from these results that stimulation of the Kp system is not required to induce ovulation in the mare?” If the experimental design is more perfect, this conclusion maybe basically be drawn. However, based on the currently known literature and results, author should avoid such descriptions.
Conclusions
Please condense, part of the description can be integrated into the discussion section.
Author Response
Referee 1
It is recommended to increase the number of experimental animals, set different C6 administration time and dosage, and verify whether the short duration of LH peak results in anovulation.
We agree with the referee that it would be interesting to perform experiments with a larger number of animals to evaluate different C6 doses and times of administration. However, experiments with horses are extremely cumbersome and time consuming and it would not be possible to perform them within the deadline for answer to referee criticisms imposed by the journal.
Line35: The Oc6 animals did not ovulate throughout the oestrous cycle. Right? or do not ovulate during specific periods of time？If the horse never ovulates in the cycle, is there something wrong with the horse? Please confirm in the article.
We thanks the referee for this comment. The phrase led to possible misinterpretation. Actually, the animals treated with oC6 ovulated, as shown in supplementary figure 3F, but ovulation was not anticipated compared to the control group. We modified the phrase to clarify this point.
Materials and Methods
Line187：The unit of KP in the article suggested replacing "mmol" with "mg" to facilitate comparison with other literature.
To comply with referee suggestion at the first occurrence of the indication of the dose we indicated also the amount in mg to facilitate comparison with other literature data.
Line198：Figure 1: The flowchart is somewhat tedious and messy, and it is recommended to simplify it.
We agree that the scheme in figure one could be tedious. However, we think that would not be possible to simplify it without loosing essential information regarding the experiment. So we decided to keep it without modification.
Line 200：For "T±/Χh", please verify the "Χ" writing format.
Because there are different times (T) indicated in the figure the X (hours) is the generic indication for this different times that could be either before (-) or after (+) time zero (time of compound injection).
Line211： “（”，please delete.
We modified the manuscript as suggested by the referee.
Results
Figures 3, 4, 5: The standard deviation is large, and Figure A、B、C is recommended to be combined into one figure.
We agree with the referee that the standard deviation is large. However, the statistical analysis takes into account this large standard deviation giving us confidence that the conclusion we drawn is reasonable.
Combining the panels A, B and C into one it would make quite difficult to appreciate the profile of each treatment so we preferred not to change the figures.
If possible, it is recommended to reconstitute the test hormone concentration.
We are sorry but we did not understand this referee’s comment.
Line 483-484: “Should we conclude from these results that stimulation of the Kp system is not required to induce ovulation in the mare?” If the experimental design is more perfect, this conclusion maybe basically be drawn. However, based on the currently known literature and results, author should avoid such descriptions.
We disagree with the referee. We pose a legitimate question and we tried to explore the possible issues that could be responsible for a lack of effect of the stimulation of the Kp system in the horse. The interpretation is not only derived from our present results, but also from previous literature data that we cited. Hence, we did not modify the text of the manuscript.
Conclusions
Please condense, part of the description can be integrated into the discussion section.
We feel that the conclusiions is sufficiently concise and do not required further reduction.
Reviewer 2 Report
The manuscript is very well written except for occasional language errors. The experimental design is good and has been executed properly.
-Lines 25-26: “plasma gonadotropin concentrations” instead of “gonadotropins plasma concentrations”
-Line 35: “transrectal echography”….did the authors mean to write “transrectal ultrasonography” instead?
-Line 44: I suggest “mammalian reproduction”
-Lines 82-85: This portion should be in the discussion section instead of the introduction because it is based on the results of the presented study.
-The materials and methods have been discussed in enough detail for someone else to reproduce the study.
-The authors have not mentioned if they tested the data for normality and which data was normally distributed and which wasn’t. The rest of the statistical analysis section is well described.
-The results are well described, presented and discussed.
Author Response
Referee 2
The manuscript is very well written except for occasional language errors. The experimental design is good and has been executed properly.
-Lines 25-26: “plasma gonadotropin concentrations” instead of “gonadotropins plasma concentrations”
We modified as requested by the referee.
-Line 35: “transrectal echography”….did the authors mean to write “transrectal ultrasonography” instead?
We modified the sentence as requested.
-Line 44: I suggest “mammalian reproduction”
We change the text as requested by the referee.
-Lines 82-85: This portion should be in the discussion section instead of the introduction because it is based on the results of the presented study.
We agree that the last paragraph of the introduction illustrate the results of the present study. However, many scientific journal guidelines and books on scientific writing suggest that the final sentence of the introduction should anticipate the results obtained in order to catch the reader attention. Based on this consideration we decided to keep the sentence in its present place.
