Barnes maze: The apparatus and experimental design were described in detail. Measures were taken to eliminate odor cues (++ for methods). Video data were analyzed using software, which used a three-point tracking system. This mitigates some concern for blinding (+ for blinding).

Reverse Barnes maze test: Conducted on PND104. Few details were provided, but noted that it was similar to the initial testing and was to test the ability to learn a new escape hole.

Social Behavior testing: tested on PND180, well after the other tests. The apparatus and experimental design were described in detail. Measures were taken to eliminate odor cues (++ for methods). Video data were analyzed using software, which used a three-point tracking system. This mitigates some concern for blinding (+ for blinding).

Vocalization: Animals were evaluated after social testing. The test apparatus was adequately described, and vocalizations were recorded with an Avisoft Bioacoustics CM16/CMPA40-5V microphone (Avisoft Bioacoustics, Germany) and data was analyzed with computer software. The apparatus was cleaned with an ethanol solution between tests. Use of computer software to analyze data mitigates concern for a lack of blinding. Information on the custom software was limited but not expected to impact the results (+ for methods, + for blinding, + overall).

Elevated plus maze:Animals were tested approximately a week after Barnes Maze testing (PND 97). The test apparatus and protocol were described in sufficient detail. The apparatus was cleaned with an ethanol solution between uses. Trials were recorded by camera (Sony Handycam HDR-CDX440) and data were analyzed using Observer software v11.5 (Noldus). Additionally, this may be a stressful test and was appropriately conducted last to avoid influence on less stressful tests. (++ for methods, + for blinding, + overall)

Brain mRNA isolation and microRNA profiles and qPCR: Standard methods were used and described (++ for methods). Blinding was not reported, but the test is mainly automated with internal checks (+ for blinding).

Kaur et al. 2021

Risk of bias

HAWC