ACE2 expression in rat brain: Implications for COVID-19 associated neurological manifestations
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SciScore for 10.1101/2021.05.01.442293: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics Euthanasia Agents: For immunohistochemistry, rats were deeply anesthetized with pentobarbital 63 mg/kg; when eyelid reflex was abolished, the animals were perfused transcardially with 0.9% NaCl solution followed by 0.1 M phosphate buffer (PB, pH 7.4) containing 4% paraformaldehyde and 15% v/v of a saturated picric acid solution.
IRB: Experimental protocols were reviewed and approved by the local Research and Ethics Committee (CIEFM-062-2016).Sex as a biological variable Animals and brain section preparation: Four male Wistar rats from the local animal breeding facility were used in the present study. Randomization not detected. Blinding not detected. Power Analysis not detected. Table 2: Resources
… SciScore for 10.1101/2021.05.01.442293: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics Euthanasia Agents: For immunohistochemistry, rats were deeply anesthetized with pentobarbital 63 mg/kg; when eyelid reflex was abolished, the animals were perfused transcardially with 0.9% NaCl solution followed by 0.1 M phosphate buffer (PB, pH 7.4) containing 4% paraformaldehyde and 15% v/v of a saturated picric acid solution.
IRB: Experimental protocols were reviewed and approved by the local Research and Ethics Committee (CIEFM-062-2016).Sex as a biological variable Animals and brain section preparation: Four male Wistar rats from the local animal breeding facility were used in the present study. Randomization not detected. Blinding not detected. Power Analysis not detected. Table 2: Resources
Antibodies Sentences Resources Primary antibody rabbit anti-ACE2 (ab15348, 1:1000, Abcam, MA, USA, see also table 1 for antibody information) was used, diluted in TBST + 1% NDS during 48h at 4°C with gentle shaking. anti-ACE2suggested: (Abcam Cat# ab15348, RRID:AB_301861)ab15348suggested: (Millipore Cat# AB15348, RRID:AB_805291)After this blocking step, sections were incubated during 48hr at 4°C with cocktails of primary antibodies against ACE2 and combinations of the following markers: calretinin (CR, a calcium binding protein with a high expression in several nuclei of the brainstem and in the olfactory bulb); the glycine transporter 2 (GlyT2, a marker of glycinergic neurons, highly expressed in the pontomedullary region of the brain); tyrosine hydroxylase (TH, a marker for catecholaminergic neurons), tryptophan hydroxylase (TPH, a marker for serotoninergic neurons); AVP-neurophysin (a marker for vasopressinergic neurons); neuron-specific nuclear protein (NeuN, a pan-neuronal marker); Von Willebrand Factor (VWF, a marker for endothelial cells) and glial fibrillary acidic protein (GFAP, a marker for astrocytes). calretinin ( CRsuggested: Noneglycine transporter 2suggested: NoneGlyT2suggested: Nonetyrosine hydroxylasesuggested: Nonetryptophan hydroxylase ( TPHsuggested: (GeneTex Cat# GTX61516, RRID:AB_10621478)neuron-specific nuclear proteinsuggested: NoneNeuNsuggested: Noneglial fibrillary acidic proteinsuggested: NoneGFAPsuggested: NoneExperimental Models: Organisms/Strains Sentences Resources Animals and brain section preparation: Four male Wistar rats from the local animal breeding facility were used in the present study. Wistarsuggested: RRID:MGI:5657554)Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).
Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.Results from TrialIdentifier: No clinical trial numbers were referenced.
Results from Barzooka: We did not find any issues relating to the usage of bar graphs.
Results from JetFighter: We did not find any issues relating to colormaps.
Results from rtransparent:- Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
- Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
- No protocol registration statement was detected.
Results from scite Reference Check: We found no unreliable references.
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