Amplification of Olfactory Signals by Anoctamin 9 is Essential for Mammalian Olfaction: a Risk Factor for the Covid-19-associated Anosmia

Article activity feed

1. 

   ScreenIT

   Mar 7, 2022

   SciScore for 10.1101/2022.03.02.482745: (What is this?)

   Please note, not all rigor criteria are appropriate for all manuscripts.

   Table 1: Rigor

   Ethics

   Consent: Due to the domestic law that cadavers must be donated and managed under informed consent and ethical process, the institutional review board (IRB) of The Catholic University of Korea has decreed that studies involving cadavers do not need to be reviewed. IRB: Due to the domestic law that cadavers must be donated and managed under informed consent and ethical process, the institutional review board (IRB) of The Catholic University of Korea has decreed that studies involving cadavers do not need to be reviewed. Euthanasia Agents: Briefly, male and female adult mice (9-12 weeks old) were anesthetized with intraperitoneal injection of Ketamine/Xylazine (90 mg/10 mg/kg).

   Sex as a biological …

   SciScore for 10.1101/2022.03.02.482745: (What is this?)

   Please note, not all rigor criteria are appropriate for all manuscripts.

   Table 1: Rigor

   Ethics	Consent: Due to the domestic law that cadavers must be donated and managed under informed consent and ethical process, the institutional review board (IRB) of The Catholic University of Korea has decreed that studies involving cadavers do not need to be reviewed. IRB: Due to the domestic law that cadavers must be donated and managed under informed consent and ethical process, the institutional review board (IRB) of The Catholic University of Korea has decreed that studies involving cadavers do not need to be reviewed. Euthanasia Agents: Briefly, male and female adult mice (9-12 weeks old) were anesthetized with intraperitoneal injection of Ketamine/Xylazine (90 mg/10 mg/kg).
   Sex as a biological variable	Human Samples: Two tissue samples of the human OE were acquired from two male cadavers (age: 68 years and 80 years).
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   KLH/BSA conjugation was performed for effective immunization and generation of anti-peptide antibodies.	anti-peptide suggested: None
   monoclonal anti-c-Fos antibody (1:1000, ab208942, Abcam), Tuj-1 (1:500, T5076, Sigma), Ac-Tub (1:500, T7451, Sigma), or olfactory marker protein (1:200, 019-22291, FUJIFILM Wako Chemicals) was diluted in 0.1% Triton X-100 in 1 x PBS containing 1% bovine serum albumin.	anti-c-Fos suggested: (Abcam Cat# ab208942, RRID:AB_2747772) Tuj-1 suggested: None Ac-Tub suggested: None
   The cell lysates were mixed with anti-ANO9 antibodies and incubated overnight in 4°C.	anti-ANO9 suggested: None
   Then, these immunoprecipitants were blotted with anti- phosphoserine antibodies Statistical analysis: All results are expressed as means ± standard errors (S.E.).	anti- phosphoserine suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   The transfected HEK 293T cells were incubated at 5% CO2 at 37◦C in the Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum, 10 units/ml penicillin, and 10 μg/ml streptomycin.	HEK 293T suggested: None
   Experimental Models: Organisms/Strains
   Sentences	Resources
   Adult male and female C57BL/6 background, transgenic, and Ano9 KO mice were used for electrophysiological recordings, in-vivo Ca2+ imaging, or behavioral tests.	C57BL/6 suggested: None
   To delete Ano9 in the whole body, the mice genotyped for the presence of the loxP sites (Ano9fl/fl) were crossed with B6.C-Tg(CMV-cre)1Cgn/J (CMV-cre) to produce CMV-cre;Ano9fl/- mice.	B6.C-Tg(CMV-cre)1Cgn/J suggested: RRID:IMSR_JAX:006054) CMV-cre;Ano9fl/- suggested: None
   The CMV-cre;Ano9fl/- mice were crossed with Ano9fl/fl mice to generate Ano9 deficient mice (CMV-cre;Ano9fl/fl).	Ano9fl/fl suggested: None CMV-cre;Ano9fl/fl suggested: None
   Recombinant DNA
   Sentences	Resources
   We subcloned chick Ano9 into pEGFP-N1 vector from the XM_015286257.2:1133-3649 PREDICTED: Gallus gallus anoctamin 9 (Ano9), transcript variant X3, mRNA.	pEGFP-N1 suggested: RRID:Addgene_172284)
   Software and Algorithms
   Sentences	Resources
   Imaging data were analyzed with NeuroPlex.	NeuroPlex suggested: (NeuroPlex, RRID:SCR_016193)
   The electrical signals were digitized at 500 Hz and analyzed using pClamp 10.	pClamp suggested: (pClamp, RRID:SCR_011323)
   Multiple sequence alignment was obtained using the CLUSTAL OMEGA program (https://www.ebi.ac.uk/Tools/msa/clustalo/) to compare the ANO9 sequences of various species.	CLUSTAL OMEGA suggested: (Clustal Omega, RRID:SCR_001591)
   The analysis was performed with tailored Python and R scripts utilizing the following packages: Scanpy (v.1.7.1) 82 was used to import data and SciPy (v.1.4.1) for t-test 83 (see Code availability)	Python suggested: (IPython, RRID:SCR_001658) SciPy suggested: (SciPy, RRID:SCR_008058)

   ---

   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.

   ---

   Read the original source
2. 

   Version published to 10.1101/2022.03.02.482745 on bioRxiv

   Mar 3, 2022