Decoding the Synaptic Proteome with Long-Term Exposure to Midazolam during Early Development
Round 1
Reviewer 1 Report
Authors Nguyen et.al. in their paper " Decoding the Synaptic Proteome With Long-Term Exposure to Midazolam During Early Development" studied long-term exposure to MDZ during very early stages of development (in neonates) induces changes at the synaptic level by quantitative mass spectrometry-based proteomics. This study is done for the first time to analyze the alterations in the synaptic proteome associated with long-term MDZ in a rodent model. Proper controls (saline, unexposed, and exposed to MDZ) were used to study the differential expression profiles of rat pups synaptosomes with MDZ in an escalating dose from postnatal 62 days (P)3 to P21. Synaptosomes from P21 rat pups were extracted and used mass-spectrometry to examine changes in the synaptic proteome.
Potential up-and down-regulated proteins involved in various molecular pathways like protein depolymerization, tricarboxylic cycle, central nervous system, neuron development, synaptogenesis signaling, protein kinase A signaling, and oxidative phosphorylation were seen with long-term exposure to MDZ. Altogether, these findings provide new insights pertaining to long-term exposure of MDZ during the early stages of development that can impact neurodevelopmental outcomes, in neonates.
The study is conducted well, written, and analyzed in a scientific and logical way.
Author Response
Authors Nguyen et.al. in their paper " Decoding the Synaptic Proteome With Long-Term Exposure to Midazolam During Early Development" studied long-term exposure to MDZ during very early stages of development (in neonates) induces changes at the synaptic level by quantitative mass spectrometry-based proteomics. This study is done for the first time to analyze the alterations in the synaptic proteome associated with long-term MDZ in a rodent model. Proper controls (saline, unexposed, and exposed to MDZ) were used to study the differential expression profiles of rat pups synaptosomes with MDZ in an escalating dose from postnatal 62 days (P)3 to P21. Synaptosomes from P21 rat pups were extracted and used mass-spectrometry to examine changes in the synaptic proteome. Potential up-and down-regulated proteins involved in various molecular pathways like protein depolymerization, tricarboxylic cycle, central nervous system, neuron development, synaptogenesis signaling, protein kinase A signaling, and oxidative phosphorylation were seen with long-term exposure to MDZ. Altogether, these findings provide new insights pertaining to long-term exposure of MDZ during the early stages of development that can impact neurodevelopmental outcomes, in neonates.
The study is conducted well, written, and analyzed in a scientific and logical way.
Thank you very much for your time and effort to provide us with positive reviews and comments.
Reviewer 2 Report
The manuscript by Nghi M. Nguyen et al. presents proteomic data on synaptosomes following exposure to midazolam. the topic is of great interest, given the possible effects of the drug use in NICU and the experiments are well designed. there are few issues that could be addressed for improving the manuscript:
- Figure n.3 (and to some extent, figure n.2) are difficult to read, especially the text part.
- It could be useful to compare the dosage used in the animal models to that normally used in babies.
Author Response
The manuscript by Nghi M. Nguyen et al. presents proteomic data on synaptosomes following exposure to midazolam. the topic is of great interest, given the possible effects of the drug use in NICU and the experiments are well designed. there are few issues that could be addressed for improving the manuscript:
- Figure n.3 (and to some extent, figure n.2) are difficult to read, especially the text part.
We have incorporated changes in the text size and enhanced the clarity in Figures 2 and 3 as recommended.
- It could be useful to compare the dosage used in the animal models to that normally used in babies.
Great critique. Generally, in a NICU setting, as a common practice, neonates are often given a dose ramping mode of sedatives starting with a low dose and slowing increasing over time. Also, such an exposure to sedatives tend to last for a few days to weeks. The cumulative doses over such long-term usage tend to fall in the ranges used in the manuscript. Notably, the usage of such dosage regimens has been well documented in rodent models, we accordingly have mimicked the dose ramping scenario in our current study. The relevant text and references have now been included in the revised manuscript. Please see line 261-264 on page 7.
