The Effect of Insulin-like Growth Factor-1 on the Quantitative and Qualitative Composition of Phosphoinositide Cycle Components During the Damage and Regeneration of Somatic Nerves
, Anastasia Zavarykina, Egor Popkov and Victor Revin
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsThis study aimed to investigate the impact of Insulin-like Growth Factor-1 (IGF-1) on the quantitative and qualitative changes in phosphoinositide cycle components during the damage and regeneration of somatic nerves. To achieve this, the researchers focused on three key objectives: Effect of IGF-1 on Phosphoinositide Levels, Changes in Fatty Acid Composition, Role of Phosphoinositide Cycle in Nerve Regeneration.
General concept comments
The manuscript appears to be written in a rather careless and disorganized way, making it difficult to follow and understand. Key ideas and concepts are not clearly communicated, and the structure lacks clarity. This confusion detracts from the overall quality of the work and makes it challenging for readers to grasp the significance of the findings. More attention to detail and clearer, more precise writing would greatly improve the presentation and comprehensibility of the manuscript. The authors should underline the novelty of the work. The abstract is too long. It should be summarized. Some self-references have been detected. 25% of them are old, a literature backup is needed. Conclusions section is too long. Data Availability Statement is correctly reported, and the figures have a good resolution.
Specific comments
The abstract is too long. It should be summarized.
Line 99: is it not clear which discussion is referred to.
Line 111: is it not clear which study is referred to.
Comments on the Quality of English LanguageMinor editing of English language required
Author Response
Comments 1: The abstract is too long. It should be summarized.
Response 1: Thank you for pointing this out. We agree with this comment. Therefore, we have shortened the abstract. We have updated the text of the manuscript. This change can be found – page 1. We mark our revisions in red.
Comments 2: Line 99: is it not clear which discussion is referred to.
Response 2: Agree. We have changed this point: Results and Discussion – line 103.
Comments 3: Line 111: is it not clear which study is referred to.
Response 3: Agree. We have changed this point. We have updated the text of the manuscript. This change can be found – line 115.
Comments 4: Minor editing of English language required
Response 4: Agree. We will use a professional, certified editing MDPI Author Services to improve English
Comments 5: Some self-references have been detected. 25% of them are old, a literature backup is needed.
Response 5: Agree. We have changed this point – 4, 21, 24 old sources of literature have been replaced with more modern ones. We used such literature sources as 8, 9, 11 to describe the methods, since they are primary sources and reliable results were obtained using them.
Comments 6: Conclusions section is too long.
Response 6: Agree. The conclusions have been shortened. This change can be found – page 11.
Reviewer 2 Report
Comments and Suggestions for AuthorsParchaykina et al. examine a role of insulin-like growth factor-1 (IGF-1) in regulating regenerative processes in injured nerve fibers by focusing on phosphoinositide cycle components. The experiments were performed in the rat sciatic nerve by using biochemical and extracellular recording techniques. As a result, it was revealed that IGF-1 has an ability to activate several signaling pathways related to the formation of lipid metabolites. Figures 1-8 demonstrate the results obtained of monophospoinositide (MPI), diphosphoinositide (DPI), triphosphoinositide (TPI), diacylglycerol (DAG) and free fatty acids (FFAs) in the proximal and distal end of the sciatic nerve without and with IGF-1 treatment. The compound action potentials of sciatic nerve without and with IGF-1 treatment are shown in Fig. 9. This manuscript is not carefully written in English and from a scientific point of view. There are many points that should be addressed and could help to improve this manuscript, as follows:
1. Abstract: it is not right that the most pronounced effect of the drug is observed at a IGF-1 concentration of 75 ng/kg, mentioned in lines 20 and 21, because only two concentrations are used in the present study (see the following comment 14). The conclusion of this study should be stated at the end of this section.
2. Technical abbreviations are used in some places, but not in others after their definition (for example, see lines 14 and 27). Their definitions are repeatedly given (for example, see lines 58 and 92). Mistakes like these can be found throughout this manuscript.
3. Line 48: please use superscript in “Ca2+” throughout this manuscript.
4. Lines 53 and 72: once “FFAs” is defined, this should be used thereafter.
5. Lines 71-79: these three sentences should be written together.
6. Materials and Methods: the names of the companies from which the drugs and experimental equipments (amplifier etc.) used in the present study were purchased, or where those companies are located, are not mentioned. It is not clear why the stimulus parameters (1.5 V, 0.3 ms and 100 impulses/s) given in line 98 were used. Which part of the sciatic nerve was electrically stimulated? Was gas added to Ringer’s solution during recording of bioelectrical activity? No statistical analysis is described.
7. Line 82: how many rats were used in this study? Details of how the transection was performed should be given.
8. Line 85: the “proximal end (segment)” or “distal end (segment)” is a very vague scientific statement. This end (segment) should be quantitatively mentioned.
9. Line 86: there is no statement about the composition of “Ringer’s solution”.
10. Lines 87 and 88: no references are given about “Bligh-Dyer method” and “Prokhorova method”.
11. Line 99: not “Results and Their Discussion” but “Results and Discussion”?
12. Line 118: not “Figure 1, 2” but “Figures 1, 2”.
13. Line 126: not “trifos ..” but “triphos ..”
14. Line 148: it is unknown whether IGF-1 at 75 ng/kg exhibits a maximal effect. IGF-1 at concentrations higher than 75 ng/kg should be tested.
15. Line 168: please give the “literature review”. Is “diacylglycerin” not English but Deutsch?
16. Figure 1 and this legend: there is no statement about what the bar accompanied by column means. SE or SD? How many sciatic nerve these data were obtained from?
17. Figure 2 and this legend: there is no statement about what the bar accompanied by column means. SE or SD? How many sciatic nerve these data were obtained from?
18. Figure 3 and this legend: there is no statement about what the bar accompanied by column means. SE or SD? How many sciatic nerve these data were obtained from?
19. Figure 4 and this legend: there is no statement about what the bar accompanied by column means. SE or SD? How many sciatic nerve these data were obtained from? There is no * in this figure (see line 189).
20. Line 198: there seems to be no explanation about “OL”.
21. Line 201: how many data “51.76±2.44” was calculated from?
22. Figure 5 and this legend: there is no statement about what the bar accompanied by column means. SE or SD? How many sciatic nerve these data were obtained from? There is no * in this figure (see line 215).
23. Line 218: is “fatty acids (Fas)” OK?
24. Line 244: there seems to be no explanation about “saturation index”.
25. Figure 6 and this legend: there is no statement about what the bar accompanied by column means. SE or SD? How many sciatic nerve these data were obtained from?
26. Figure 7 and this legend: there is no statement about what the bar accompanied by column means. SE or SD? How many sciatic nerve these data were obtained from? The bars accompanying the data points are not clearly visible.
27. Figure 8 and this legend: there is no statement about what the bar accompanied by column means. SE or SD? How many sciatic nerve these data were obtained from? The bars accompanying the data points are not clearly visible.
28. Line 295: it seems to be unlikely that Ca2+ enters through Na+ channels. Please give “the literature” (in line 294) showing that Ca2+ enters through Na+ channels.
29. Line 305: is “proteins of the Ca2+ channels” OK?
30. Line 321: is “action potential conduction with a small amplitude” OK?
31. Figure 9: the voltage and time scale bars should be given in this figure. It is unclear where is a stimulus artifact in this potential recordings. Data given in this figure should be quantitatively analyzed.
32. Line 356: what is “PKD”?
33. Conclusions (line 334): this section is too long. Some of this content should be put in “3. Results and Their Discussion”. Where is section 4?
34. References: each letter in the title of the reference begins (Ref. 6) or does not begin with a capital letter (Ref. 5). All of the references given should be consistent in style.
35. There are much more mistakes than pointed above. Please check your manuscript very carefully from English and scientific points of view.
Comments on the Quality of English LanguageEnglish Language should be amended.
Author Response
Comments 1: Abstract: it is not right that the most pronounced effect of the drug is observed at a IGF-1 concentration of 75 ng/kg, mentioned in lines 20 and 21, because only two concentrations are used in the present study (see the following comment 14). The conclusion of this study should be stated at the end of this section.
Response 1: Thank you for pointing this out. We agree with this comment. We have shortened the abstract according to the comments of the first reviewer and deleted this point from the abstract. We have updated the text of the manuscript. This change can be found – page 1. We mark our revisions in red.
Comments 2: Technical abbreviations are used in some places, but not in others after their definition (for example, see lines 14 and 27). Their definitions are repeatedly given (for example, see lines 58 and 92). Mistakes like these can be found throughout this manuscript.
Response 2: Agree. We have changed this point. This change can be found – line 17; line 46; line 84; line 172; lines 229 and 232.
Comments 3: Line 48: please use superscript in “Ca2+” throughout this manuscript.
Response 3: Agree. We have done superscript in “Ca2+” throughout this manuscript.
Comments 4: Lines 53 and 72: once “FFA” is defined, this should be used thereafter.
Response 4: Agree. We have done.
Comments 5: Lines 71-79: these three sentences should be written together.
Response 5: Agree. Тhree sentences were combined into one.
Comments 6: Materials and Methods: the names of the companies from which the drugs and experimental equipments (amplifier etc.) used in the present study were purchased, or where those companies are located, are not mentioned. It is not clear why the stimulus parameters (1.5 V, 0.3 ms and 100 impulses/s) given in line 98 were used. Which part of the sciatic nerve was electrically stimulated? Was gas added to Ringer’s solution during recording of bioelectrical activity? No statistical analysis is described.
Response 6: We have indicated the name of the company and the country of origin of insulin-like growth factor-1 (RD Systems, USA, line 76) and experimental equipments (a GW Instek GDS-71042 Oscilloscope (China) and Laboratory Electrical stimulator (ESL-2) (Russia) – lines 98, 99. We used the stimulus parameters (1.5 V, 0.3 ms and 100 impulses/s) because they were selected during the long-term joint work of scientists from Ogarev Mordovia State University and Lomonosov Moscow State University in the field of research on the conduction of excitation along a nerve fiber on various objects (giant axons of squids, somatic nerves of rabbits and rats) for several decades and the results of these investigations has been tested in scientific articles, dissertations and educational literature (Revin V.V., Maksimov G.V., Cols O.R.). The proximal section of the damaged nerve was subjected to electrical stimulation, since in its distal part there is a complete loss of the ability to conduct an action potential. The perfused rat sciatic nerves were placed in Ringer’s solution at 37°C with a continuous oxygen flow. This change can be found – 100 line. No statistical analysis is described: our task was to clearly demonstrate the change in the amplitude of the action potential, which is an unambiguous proof of the restoration of the functional activity of the nerve, nevertheless, statistical processing of the results was carried out and the values of permissible errors do not exceed 5%.
Comments 7: Line 82: how many rats were used in this study? Details of how the transection was performed should be given.
Response 7: We have added that in this study we used 10 rats in each series of experiments: This change can be found – Materials and Methods, line 68. Also we have added detailed description of transection: access to the nerve trunk was carried out by dissecting the skin and subcutaneous fat of the hindlimb using blunt branches. Fascia was dissected in layers, and the muscles of the hip region were dissected along the posterior surface of the limb. The transection of the nerve into proximal and distal sections, each of which averaged 1.4 cm, occurred in the lower third of the thigh to the place of its bifurcation into separate – common fibular and tibia nerves. This change can be found –lines 69-74.
Comments 8: Line 85: the “proximal end (segment)” or “distal end (segment)” is a very vague scientific statement. This end (segment) should be quantitatively mentioned.
Response 8: The proximal section of the nerve is a section that retains connection with the central nervous system, which is accompanied by more pronounced regeneration processes and the formation of growth cones, unlike the distal section, which completely loses central innervation and undergoes Wallerian degeneration. The transection of the nerve into proximal and distal sections, each of which averaged 1.4 cm, occurred in the lower third of the thigh to the place of its bifurcation into separate – common fibular and tibia nerves.
Comments 9: Line 86: there is no statement about the composition of “Ringer’s solution”.
Response 9: Agree. We have added the composition of “Ringer’s solution”, consisting of sodium chloride – 0.8 g, potassium chloride – 0.02 g, calcium chloride – 0.02 g, sodium bicarbonate - 0.02 g, glucose – 0.1 g in 100 ml of water. This change can be found –lines 78-80.
Comments 10: Lines 87 and 88: no references are given about “Bligh-Dyer method” and “Prokhorova method”.
Response 10: Agree. We have added references about “Bligh-Dyer method” (line 81) and “Prokhorova method” line 82.
Comments 11: Line 99: not “Results and Their Discussion” but “Results and Discussion”?
Response 11: Agree. We have done. This change can be found –line 103.
Comments 12: Line 118: not “Figure 1, 2” but “Figures 1, 2”.
Response 12: Agree. We have done. This change can be found –line 122.
Comments 13: Line 126: not “trifos ..” but “triphos ..”
Response 13: Agree. We have done. This change can be found –line 130.
Comments 14: Line 148: it is unknown whether IGF-1 at 75 ng/kg exhibits a maximal effect. IGF-1 at concentrations higher than 75 ng/kg should be tested.
Response 14: Agree. We have changed this point. This change can be found –152 line.
Comments 15: Line 168: please give the “literature review”. Is “diacylglycerin” not English but Deutsch?
Response 15: Agree. We have changed this point. This change can be found –line 172.
Comments 16: Figure 1 and this legend: there is no statement about what the bar accompanied by column means. SE or SD? How many sciatic nerve these data were obtained from?
Response 16: The bar accompanied by column means SE. We used 10 sciatic nerves for each series of experiments, i.e. 10 nerves for the series D 7 days, 10 nerves for the series D 7 days +IGF-1 etc.
Comments 17: Figure 2 and this legend: there is no statement about what the bar accompanied by column means. SE or SD? How many sciatic nerve these data were obtained from?
Response 17: The bar accompanied by column means SE. We used 10 sciatic nerves for each series of experiments, i.e. 10 nerves for the series D 7 days, 10 nerves for the series D 7 days +IGF-1 etc.
Comments 18: Figure 3 and this legend: there is no statement about what the bar accompanied by column means. SE or SD? How many sciatic nerve these data were obtained from?
Response 18: The bar accompanied by column means SE. We used 10 sciatic nerves for each series of experiments, i.e. 10 nerves for the series D 7 days, 10 nerves for the series D 7 days +IGF-1 etc.
Comments 19: Figure 4 and this legend: there is no statement about what the bar accompanied by column means. SE or SD? How many sciatic nerve these data were obtained from? There is no * in this figure (see line 189).
Response 19: The bar accompanied by column means SE. Agree. We used 10 sciatic nerves for each series of experiments, i.e. 10 nerves for the series D 7 days, 10 nerves for the series D 7 days +IGF-1 etc. We have replaced the correct version of the drawing with *. This change can be found – Figure 4.
Comments 20: Line 198: there seems to be no explanation about “OL”.
Response 20: Incorrectly translated, corrected to "total lipids". This change can be found – 204 line
Comments 21: Line 201: how many data “51.76±2.44” was calculated from?
Response 21: The data “51.76±2.44” was calculated from 10 proximal ends of the rat sciatic nerves in 10 repetitions.
Comments 22: Figure 5 and this legend: there is no statement about what the bar accompanied by column means. SE or SD? How many sciatic nerve these data were obtained from? There is no * in this figure (see line 215).
Response 22: The bar accompanied by column means SE. We used 10 sciatic nerves for each series of experiments, i.e. 10 nerves for the series D 7 days, 10 nerves for the series D 7 days +IGF-1 etc. We have replaced the correct version of the drawing with *. This change can be found – Figure 5.
Comments 23: Line 218: is “fatty acids (Fas)” OK?
Response 23: Agree. We have done. This change can be found –line 219.
Comments 24: Line 244: there seems to be no explanation about “saturation index”.
Response 24: «Saturation index» is the ratio of saturated fatty acids to unsaturated ones. We have added this definition. This change can be found –line 245-246.
Comments 25: Figure 6 and this legend: there is no statement about what the bar accompanied by column means. SE or SD? How many sciatic nerve these data were obtained from?
Response 25: The bar accompanied by column means SE. We used 10 sciatic nerves for each series of experiments, i.e. 10 nerves for the series D 7 days, 10 nerves for the series D 7 days +IGF-1 etc.
Comments 26: Figure 7 and this legend: there is no statement about what the bar accompanied by column means. SE or SD? How many sciatic nerve these data were obtained from? The bars accompanying the data points are not clearly visible.
Response 26: The bar accompanied by column means SE. We used 10 sciatic nerves for each series of experiments, i.e. 10 nerves for the series D 7 days, 10 nerves for the series D 7 days +IGF-1 etc. We have tried to improve the clarity of the data points. This change can be found – Figure 7.
Comments 27: Figure 8 and this legend: there is no statement about what the bar accompanied by column means. SE or SD? How many sciatic nerve these data were obtained from? The bars accompanying the data points are not clearly visible.
Response 27: The bar accompanied by column means SE. We used 10 sciatic nerves for each series of experiments, i.e. 10 nerves for the series D 7 days, 10 nerves for the series D 7 days +IGF-1 etc. We have tried to improve the clarity of the data points. This change can be found – Figure 8.
Comments 28: Line 295: it seems to be unlikely that Ca2+ enters through Na+ channels. Please give “the literature” (in line 294) showing that Ca2+ enters through Na+ channels.
Response 28: Agree. We have deleted this sentence. This change can be found –line 295.
Comments 29: Line 305: is “proteins of the Ca2+ channels” OK?
Response 29: This concept is correct, but we have added a hyphen (Ca2+-channels). This change can be found – 305 line.
Comments 30: Line 321: is “action potential conduction with a small amplitude” OK?
Response 30: Yes, this concept is correct, since the magnitude of the amplitude of the action potential can be judged on the restoration of the functional activity of the nerve conductor.
Comments 31: Figure 9: the voltage and time scale bars should be given in this figure. It is unclear where is a stimulus artifact in this potential recordings. Data given in this figure should be quantitatively analyzed.
Response 31: We have added voltage scale: 20 mV per cell and time scale: 1 ms per cell. This change can be found –lines 333-334. We do not represent an exciting stimulus that precedes the emergence of an action potential, but the action potential itself. Our task was to clearly demonstrate the change in the amplitude of the action potential, which is an unambiguous proof of the restoration of the functional activity of the nerve, nevertheless, statistical processing of the results was carried out and the values of permissible errors do not exceed 5%.
Comments 32: Line 356: what is “PKD”?
Response 32: The conclusions have been revised.
Comments 33: Conclusions (line 334): this section is too long. Some of this content should be put in “3. Results and Their Discussion”. Where is section 4?
Response 33: Agree. The conclusions have been revised. We have replaced the Conclusion section. This change can be found –lines 336-361.
Comments 34: References: each letter in the title of the reference begins (Ref. 6) or does not begin with a capital letter (Ref. 5). All of the references given should be consistent in style.
Response 34: We have done.
Comments 35: There are much more mistakes than pointed above. Please check your manuscript very carefully from English and scientific points of view.
Response 35: Dear reviewer, thank you for your valuable questions and comments, which will certainly improve our manuscript. We tried to carefully check the manuscript again and made all the corrections.
Comments on the Quality of English Language: English Language should be amended.
Response: Agree. We will use a professional, certified editing MDPI Author Services to improve English.
Reviewer 3 Report
Comments and Suggestions for AuthorsThe manuscript scipharm-3195840 reports that IGF-1 initiates several signaling pathways associated with forming lipid metabolites. The manuscript is well-written, and the results are important to the scientific community. My main concern is the references (standard) used in chromatography to identify the analytes. Additionally, the authors used gas chromatography to quantify the fatty acid content and claimed that methylated the lipids. This is another concern because methylation can occur in the sp2 carbon of the long chain of the fatty acid. Thus, I strongly recommend that the authors conduct the esterification reaction instead of methylation. Please, better explore it. This is important to validate the manuscript.
Author Response
Dear reviewer, thank you for your valuable comments, which will certainly improve our manuscript. We have added information about visualization and identification the analytes. This change can be found –lines 85-92.
We have also replaced the esterification reaction instead of methylation. This change can be found –line 95.
Round 2
Reviewer 1 Report
Comments and Suggestions for Authorsthe paper has been improved as requested
Author Response
Dear reviewer, thank you for your valuable comments. If you don't have any more comments, could you please choose «I would like to sign my review report» in the «Open Review».
Reviewer 2 Report
Comments and Suggestions for AuthorsThis manuscript has been amended in minor points mentioned by this reviewer, but it still has a number of problems, as mentioned in the following:
Major points:
1. Abstract: this section should be written in the following order: short introduction, the method used, the results obtained and the physiological significance of the results. This revised manuscript is not written as such. Alternatively, the result obtained in Fig. 9 is not reflected in this abstract. The authors only examined changes in the amount of lipid metabolites following sciatic nerve transection and the effect of IGF-1 on these; much of the content of this abstract is merely a suggestion. The present study does not examine an involvement of IGF-1 receptor, PDK1 and Akt (protein kinase B) in the authors’ findings.
2. In order to confirm the authors’ findings, more concentrations than just two should be investigated, as mentioned in my previous comment (14).
3. It is not necessary to repeat “insulin-like growth factor-1 (IGF-1)” in the legends of Figs. 1-5. Instead, each of figure legends should state how many experiments each point or bar represents and whether the variability of the data is given as SE or SD, as mentioned in my previous comments (16)-(19), (22) and (25)-(27).
4. Fig. 9: judging from the change in baseline in A, B, and D, doubts arise as to whether these recording were made correctly. How many experiments were performed? These recordings should be replaced by ones without baseline drift as shown in C, and then the results obtained should be quantitatively analyzed, as mentioned in my previous comment (31). The expression, “per cell”, is not usually used. Time and voltage scales should be shown in the figure as horizontal and vertical bars together with recordings.
5. The action potential shown in Fig. 9 originates from a summation of action currents occurring in many nerve fibers contained in the sciatic nerve. This point should be taken into consideration to account for the result shown in Fig. 9.
Specific points:
1. Line 13: there is no definition of “IGF-1”.
2. Line 20: “PDK1” should be spelled out (see line 352).
3. Line 46: “DAG” should be defined here, because this is the first occurrence in the text.
4. Line 59: please use “DAG”.
5. Lines 64, 74, 112, 113, 177, 178, 265, 266, 309, 317, 327, 341, 348, 349, 355 and 356: please use “IGF-1”.
6. Lines 77-79: the composition of Ringer’s solution used should be given in mM.
7. Line 90: please spell out “Rf”.
8. Lines 92, 93, 97 and 98: it should be stated where in the country the company is located
9. Line 231: it is unnecessary to repeatedly define “IGF-1”.
10. Lines 291 and 292: the sentences on those lines should be connected.
11. Line 306: the physiological effects of voltage-gated Ca2+ channels expressed in nerve fibers contained in the sciatic nerve appear to be small, because the channels make little contribution to the conduction of action potentials.
12. Judging by simple many mistakes as mentioned above, it appears that this manuscript was not checked before been resubmitted.
Comments on the Quality of English Language
English of this manuscript should be amended.
Author Response
Основные моменты:
1. Аннотация: этот раздел должен быть написан в следующем порядке: краткое введение, использованный метод, полученные результаты и физиологическое значение результатов. Эта пересмотренная рукопись не написана как таковая. В качестве альтернативы, результат, полученный на рис. 9, не отражен в этом реферате. Авторы только исследовали изменения количества липидных метаболитов после перерезки седалищного нерва и влияние IGF-1 на них; большая часть содержания этого реферата является лишь предположением. Настоящее исследование не изучает участие рецептора IGF-1, PDK1 и Akt (протеинкиназа B) в выводах авторов.
1. Ответ: Спасибо, что указали на это. Мы согласны с этим комментарием. Поэтому аннотация была переписана в соответствии с комментариями. Мы обновили текст рукописи. Это изменение можно найти на странице 1. Мы отмечаем наши исправления красным цветом.
2. Чтобы подтвердить выводы авторов, следует исследовать больше концентраций, чем две, как упоминалось в моем предыдущем комментарии (14).
2. Ответ: Согласен. В этом исследовании было показано, что IGF-1 влияет на изменение состава липидного компонента соматических нервов и восстановление проводимости нервных волокон. В дальнейшем мы планируем расширить и углубить наши исследования, используя более высокие концентрации IGF-1, и представить эти результаты в другой рукописи.
3. Не обязательно повторять «инсулиноподобный фактор роста-1 (ИФР-1)» в подписях к рисункам 1-5. Вместо этого в каждой подписи к рисункам должно быть указано, сколько экспериментов представляет каждая точка или полоса, и представлена ли изменчивость данных как SE или SD, как упоминалось в моих предыдущих комментариях (16)-(19), (22) и (25)-(27).
3. Ответ: Согласен. Мы удалили «инсулиноподобный фактор роста-1» в подписях к рисункам. Мы добавили, что все полосы погрешности указывают стандартную ошибку среднего (n = 10).
4. Рис. 9: судя по изменению базовой линии в A, B и D, возникают сомнения относительно того, были ли эти записи сделаны правильно. Сколько экспериментов было проведено? Эти записи следует заменить на те, у которых нет дрейфа базовой линии, как показано в C, а затем полученные результаты следует количественно проанализировать, как упоминалось в моем предыдущем комментарии (31). Выражение «на ячейку» обычно не используется. Шкалы времени и напряжения следует показать на рисунке в виде горизонтальных и вертикальных полос вместе с записями.
4. Ответ: Мы заменили эти чертежи в соответствии с вышеуказанными замечаниями.
5. Потенциал действия, показанный на рис. 9, возникает из суммирования токов действия, возникающих во многих нервных волокнах, содержащихся в седалищном нерве. Этот момент следует учитывать, чтобы объяснить результат, показанный на рис. 9.
5. Ответ: Согласен. Мы вынесли этот комментарий на обсуждение.
Конкретные моменты:
1.Строка 13: определение «ИФР-1» отсутствует.
1.Ответ: Согласен. Аннотация была переписана.
2.Строка 20: следует прописать «PDK1» (см. строку 352).
2.Ответ: Согласен. Мы удалили «PDK1» из аннотации.
3.Строка 46: Здесь следует определить «DAG», поскольку это первое упоминание в тексте.
3.Ответ: Согласен. Первое упоминание теперь в аннотации.
4. Строка 59: используйте «DAG».
4.Ответ: Согласен. Мы внесли исправление.
5. Строки 64, 74, 112, 113, 177, 178, 265, 266, 309, 317, 327, 341, 348, 349, 355 и 356: используйте «IGF-1».
5.Ответ: Согласен. Все правки внесены.
6. Строки 77-79: состав используемого раствора Рингера следует указать в мМ.
6.Ответ: Согласен. Мы сделали.
7. Строка 90: пожалуйста, напишите «Rf».
7.Ответ: Согласен. Мы сделали.
8. Строки 92, 93, 97 и 98: следует указать, в какой части страны находится компания.
8.Ответ: Согласен. Мы уточнили, в какой части страны находится компания.
9.Строка 231: нет необходимости повторно определять «ИФР-1».
9.Ответ: Согласен. Мы внесем исправления.
10.Строки 291 и 292: предложения в этих строках должны быть связаны.
10.Ответ: Согласен. Мы внесем исправления.
11. Строка 306: физиологические эффекты потенциалзависимых каналов Ca2+, экспрессируемых в нервных волокнах, содержащихся в седалищном нерве, по-видимому, невелики, поскольку каналы вносят небольшой вклад в проведение потенциалов действия.
11.Ответ: Согласен. Мы удалили это утверждение.
12. Судя по многочисленным ошибкам, упомянутым выше, эта рукопись, по-видимому, не была проверена перед повторной отправкой.
12.Ответ: Уважаемый рецензент, спасибо за ваши ценные комментарии. Мы постарались еще раз тщательно проверить рукопись и внесли все исправления.
Комментарии по качеству английского языка: Английский язык данной рукописи следует исправить.
Ответ: Текст этой рукописи на английском языке был изменен. Текст этой рукописи на английском языке был изменен.Изменения выделены зеленым цветом.
Author Response File: 
Author Response.docx
Round 3
Reviewer 2 Report
Comments and Suggestions for AuthorsSince the authors’ replies to my comments were written in Russian, I could not understand them. This re-revised manuscript has been partly amended according to my comments, but it is not enough.
Major points:
1. Abstract: this style has been amended as instructed by me, but it is somewhat difficult to understand the English written in this Abstract. Is “the accumulation the all forms” in line 14 OK in English? What is “the registration method of action potentials” in line 21? This Abstract should be amended in English and scientific writing.
2. Fig. 9: these recordings have been amended. However, it is difficult to understand “(all error bars indicate a standard error of mean (n = 10)” in line 330. How many experiments about rat sciatic nerve action potentials were performed? As pointed out in my previous major point (4), the peak amplitude of this action potential of the rat sciatic nerve should be quantitatively analyzed.
Specific points:
1. Lines 2 and 3: English should be amended (see lines 59 and 60).
2. Line 36: “IGF-1” should be defined here because of its first appearance in the text.
3. Line 45: “FFA” should be defined here because of its first appearance in the text.
4. Line 49: “DAG” should be defined here because of its first appearance in the text.
5. Line 60 and other lines: please use “PI” (see line 38).
6. Line 81: is “0.14 mM” of NaCl OK? This value is too low.
7. Line 86: not “4N” but “4 N”?
8. First paragraph on page 3: the order of the country and place names is reversed. For example, not “Switzerland, Muttenz” but “Muttenz, Switzerland”.
9. Line 101: not “China” but “Taiwan”?
10. Lines 120 and 138: not “Figure” but “Figures”.
11. Lines 179, 182 and so on: please use “DAG”.
12. Line 199: not “regulate” but “regulates”.
13. Lines 322 and 326: these values, written as “average”, should be shown with their variation (see the above-mentioned major point 2).
14. Line 344: not “phosphorylationof” but “phosphorylation of”.
15. References: all of the references given should be consistent in style. As can be seen by comparing titles in references 22 and 23, this is not the case.
16. The English in this manuscript is difficult to read in places. It should be corrected by a native English speaker who is familiar with scientific research.
Comments on the Quality of English Language
English Language should be amended.
Author Response
Major points:
1.Abstract: this style has been amended as instructed by me, but it is somewhat difficult to understand the English written in this Abstract. Is “the accumulation the all forms” in line 14 OK in English? What is “the registration method of action potentials” in line 21? This Abstract should be amended in English and scientific writing.
1.Response: We used the service of editing English by a native speaker in an academic style. The modified document is attached on the website.
2.Fig. 9: these recordings have been amended. However, it is difficult to understand “(all error bars indicate a standard error of mean (n = 10)” in line 330. How many experiments about rat sciatic nerve action potentials were performed? As pointed out in my previous major point (4), the peak amplitude of this action potential of the rat sciatic nerve should be quantitatively analyzed.
2.Response: We have presented a quantitative analysis in the table.
Specific points:
1.Lines 2 and 3: English should be amended (see lines 59 and 60).
1.Response: We used the service of editing English by a native speaker in an academic style. The modified document is attached on the website.
2.Line 36: “IGF-1” should be defined here because of its first appearance in the text.
2.Response: Agree. We have made the correction and highlighted in red.
3.Line 45: “FFA” should be defined here because of its first appearance in the text.
3.Response: Agree. We have made the correction and highlighted in red.
4.Line 49: “DAG” should be defined here because of its first appearance in the text.
4.Response: Agree. We have made the correction and highlighted in red.
5.Line 60 and other lines: please use “PI” (see line 38).
5.Response: Agree. We use "PI" throughout the text
6.Line 81: is “0.14 mM” of NaCl OK? This value is too low.
6.Response: Agree. We have made the correction and highlighted in red.
- Line 86: not “4N” but “4 N”?
7.Response: Agree. We have made the correction and highlighted in red.
8.First paragraph on page 3: the order of the country and place names is reversed. For example, not “Switzerland, Muttenz” but “Muttenz, Switzerland”.
8.Response: Agree. We have made the correction and highlighted in red.
9.Line 101: not “China” but “Taiwan”?
9.Response: Agree. We have made the correction and highlighted in red.
10.Lines 120 and 138: not “Figure” but “Figures”.
10.Response: Agree. We have made the correction and highlighted in red.
11.Lines 179, 182 and so on: please use “DAG”.
11.Response: Agree. We have made the correction and highlighted in red.
12.Line 199: not “regulate” but “regulates”.
12.Response: After the transfer: "FFAs regulate".
13.Lines 322 and 326: these values, written as “average”, should be shown with their variation (see the above-mentioned major point 2).
13.Response: Agree. We have made the correction and highlighted in red.
14.Line 344: not “phosphorylationof” but “phosphorylation of”.
14.Response: Agree. We have made the correction and highlighted in red.
15.References: all of the references given should be consistent in style. As can be seen by comparing titles in references 22 and 23, this is not the case.
15.Response: Agree. We have made the correction.
16.The English in this manuscript is difficult to read in places. It should be corrected by a native English speaker who is familiar with scientific research.
16.Response: Agree. We have made the correction. We will use a professional, certified editing MDPI Author Services to improve English.
Author Response File: Author Response.docx
Round 4
Reviewer 2 Report
Comments and Suggestions for AuthorsThis re-revised manuscript addresses almost all of my comments. There are only minor points that should be taken into consideration before publication, as follows:
1. Line 171: “2” in “A2” should be subscript.
2. Line 251: “MPI” is defined here as “phosphatidylinositol” while being defined as “monophosphoinositide” in line 123. This may cause confusion. Please amend this point.
3. Line 315: not “1,5” but “1.5”.
4. Line 321: as stated in my previous major point (4), “(all error bars indicate a standard error of mean (n = 10))” should be deleted. Fig. 9 does not indicate “error bars” and is not obtained from data of “(n = 10)”. “(n = 10)” should be given in Table 1.
Although pointed out in my previous specific point (15), the titles in refs. 22 and 23 are different in style. In ref. 22 the paper title begin with a capital letter, whereas in ref. 23 it does not. Please amend this point. All references should be checked in style.
Comments on the Quality of English LanguageThe Quality of English Language is fine.
Author Response
1.Line 171: “2” in “A2” should be subscript.
1.Response: Agree. We have made the correction.
2.Line 251: “MPI” is defined here as “phosphatidylinositol” while being defined as “monophosphoinositide” in line 123. This may cause confusion. Please amend this point.
2.Response: Agree. We have made the correction.
3.Line 315: not “1,5” but “1.5”.
3.Response: Agree. We have made the correction.
4.Line 321: as stated in my previous major point (4), “(all error bars indicate a standard error of mean (n = 10))” should be deleted. Fig. 9 does not indicate “error bars” and is not obtained from data of “(n = 10)”. “(n = 10)” should be given in Table 1.
4.Response: Agree. We have made the correction.
Although pointed out in my previous specific point (15), the titles in refs. 22 and 23 are different in style. In ref. 22 the paper title begin with a capital letter, whereas in ref. 23 it does not. Please amend this point. All references should be checked in style.
Response: Agree. We have made the correction.
Dear reviewer, thank you for your valuable comments that help improve our manuscript.
Author Response File: Author Response.docx
