All studies were approved by the IACUC of Texas Tech University Health Sciences Center, Lubbock, Texas (IACUC protocol# 20026). Experiments were performed in accordance with relevant guidelines and regulations. Female CD1 pregnant mice (Charles River Laboratories, Inc., Wilmington, MA; Cat# CRL: 22, RRID: IMSR_CRL:22) and after delivery their offspring were kept under standardized light and dark conditions (12 h), humidity (70%), and temperature (22 °C). Pregnant mice were singly housed. Offspring were separated into male and female after weaning (postnatal day 21–22) and housed in a group of 2–5. They were given ad libitum access to food and water. Animal behavior was monitored daily to minimize animal suffering. We applied the following exclusion criteria to our experiments: severe weight loss, infections, or significant behavioral deficits (decreased mobility, seizures, lethargy). No animal was excluded from this study. The research design is depicted in Fig. 1 . A total number of 176 (n = 16 for mother and n = 160 for offspring) mice were used to perform this study. All experiments were conducted in compliance with the ARRIVE guidelines.![]()
Study design. Pregnant CD1 were exposed to Blu e-cigarette from gestational day 5 (E5) to postnatal day 7 (PD7). At the end of the exposure, plasma nicotine and cotinine level were measured by LCMS/MS, and body weight was measured at PD7, PD23, PD45 and PD90. Mice were sacrificed and brain was extracted at every time point to evaluate blood-brain barrier (BBB) integrity by western blot and immunofluorescence. Open field test, novel object recognition test and morris water maze test were conducted at adolescent and adult time point to evaluate hyperactivity and learning-memory function
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