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Official websites use. Share sensitive information only on official, secure websites. The use, distribution or reproduction in other forums is permitted, provided the original author s and the copyright owner s are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. Increased use of cannabis and cannabinoids for recreational and medical purposes has led to a growth in research on their effects in animal models. The majority of this work has employed cannabinoid injections; however, smoking remains the most common route of cannabis consumption. Following smoke exposure, trunk blood and brains were collected at 6 time points 10— min. Samples were collected 10 and 40 min following exposure. Cannabis is the most widely used illicit drug in the United States, with Department of Health and Human Services, With changing attitudes toward cannabis and increasing access to medicinal and recreational markets, there is a critical need for scientific literature to assess the effects of cannabis on human health to inform medical decisions and legal policy making Hutchison et al. The increasing accessibility and use of cannabis have also generated interest in understanding its potential therapeutic properties, particularly within the aging population. Various formulations of medical cannabis and cannabis products are currently indicated for the treatment of chronic pain, nausea and vomiting in cancer patients, epilepsy and seizures, and post-traumatic stress disorder Belendiuk et al. Ease in breeding, relatively short life spans, and the ability to generate transgenic lines also make mice a practical model for studying pharmacological treatments for a variety of disease conditions. When conducting such studies, however, it is important to consider variations in genetic background among mouse strains, as they can have considerable behavioral and physiological differences that alter drug pharmacokinetics. Strain differences in respiratory and cardiac functioning Reinhard et al. Thus, it is important to consider such potential differences in studies of drug pharmacokinetics, particularly in mouse models of disease conditions that may utilize only one or a few background strains. Inhalation, particularly via smoking, remains the most common route of cannabis use Knapp et al. For those reasons, an increasing number of pharmacological studies of cannabis in rodent models are using an inhaled route of administration, which calls for a thorough assessment of the pharmacokinetic profile of inhaled cannabis in mice. Experiment 1 used young adult 3 mo. Experiment 2 used young adult 4—7 mo. These were non-transgenic mice obtained from in-house transgenic breeding colonies, in keeping with the 3Rs of humane animal practices Russell and Burch, Experiment 3 used young adult 3 mo. Demographics of the mice in each experiment are described in Table 1. All experiments took place during the light phase. All animal procedures were approved and performed in accordance with the University of Florida Institutional Animal Care and Use Committee and followed National Institutes of Health guidelines. Cannabis cigarettes were lit and puffed 35 cm 3 puff volume, 1 puff per min, 2 s per puff in the ignition chamber of the smoking machine, from which the smoke both mainstream and sidestream was pumped into the exposure chamber via a series of holes on one side of the chamber, and continuously vented out through a series of holes on the opposite side of the chamber and exhausted to the exterior of the building. Once the final cigarette was fully burnt, carbon monoxide CO and total suspended particulate matter TSP measurements were obtained. For this measurement, air was pumped out of the exposure chamber and into the monitor for 2 minutes. Mice were exposed to smoke generated from sequentially burning cannabis cigarettes NIDA Drug Supply Program, approximately mg each containing 5. In Experiment 1, mice were exposed to smoke from sequentially burning 5 cannabis cigarettes in 1 h approximately 12 min per cigarette , after which they were removed from the exposure chamber. Upon removal, mice of each sex were euthanized at 10, 20, 40, 60, , and min time points post-smoke exposure mice from different cages were used at each time point. In Experiment 3, mice were exposed to smoke from burning either 3 cannabis cigarettes in 36 min or 5 cannabis cigarettes in 1 h, in order to determine cannabinoid levels following different durations doses of cannabis smoke exposure. Mice were euthanized via rapid decapitation at specific time points following smoke exposure, with 0 min defined as the time at which the cages were removed from the exposure chamber. In Experiment 2, tail clippings were collected for genotyping and temporarily stored in ice. Plasma was pipetted from each sample and transferred to a polypropylene 0. Mice on the FVB background were genotyped for either the human tau transgene Santacruz et al. Mice on the background were genotyped by Transnetyx for the tTA transgene Santacruz et al. Mice on the SW background were genotyped for the human tau transgene Lewis et al. Mice on the B6 background were genotyped for the presence of a progranulin knock-out allele Ahmed et al. A gradient method was applied to achieve chromatographic separation using a mobile phase consisting of water containing 0. The mass spectral analysis was achieved by multiple reaction monitoring MRM , and the compound parameters for analytes and IS are described in Table 2. Prior to analysis, each brain was weighed and homogenized in triple distilled water at a ratio of one part brain to three parts water. The same process was used to generate the lower limit of quantification LLOQ, 2. After spiking, the CS and QC samples were vortex-mixed for 5 min at rpm to ensure homogeneity. The quenched samples were vortex-mixed for 5 min at rpm and then filtered through a 0. There were no endogenous substances eluting at the retention time of any analyte or IS when analyzing blank plasma or brain homogenate Supplementary Figure S1. The calibration curve had a concentration range of 2. Intra- and inter-day accuracy and precision were performed during three different days at each QC level for both plasma and brain homogenate Supplementary Table S1. Phoenix Version 6. Statistical analyses were conducted using SPSS In Experiment 2, strain comparisons between young adult mice were analyzed using a two-factor ANOVA, with strain 4 levels and time 2 levels as between-subjects factors. Significant effects of strain were further explored using a Tukey post hoc analysis. Additionally, the effect of sex was evaluated across strains in young adult mice using a two-factor ANOVA, with sex 2 levels and time 2 levels as factors. Demographics of outliers are described in Table 1. The experiment was conducted in two cohorts for a total of 72 B6 mice half female in each cohort. Data are represented as the mean of individual mouse analyte concentrations at each time point. Values represent the average of all samples at each time point male and female mice. Of those six samples below the LLOQ, five were from males at the and min time points. Because limitations in availability resulted in mice of both sexes and all ages not being represented equally across all strains and post-exposure time points see Table 1 , separate analyses were used to a compare strains in young adults; b compare males and females in young adult and middle-aged mice; and c compare age groups in the B6 and FVB strains. One mouse was excluded from the analyses due to positive transgenic status female B6 mouse. Mice were exposed to smoke from burning either 3 or 5 cannabis cigarettes, followed by sample collection 40 min post-smoke exposure. To our knowledge, this is the first evaluation of both plasma and brain concentrations of all three analytes following cannabis smoke exposure in male and female mice. These values are significantly higher than the C max values obtained in Experiment 1 Similar to the present study, mice in the Fantauzzi et al. Moreover, while the freely-moving, whole-body exposure model avoids the restraint stress necessary to implement nose-only exposure e. Similarly, in the current study, Experiment 1 found that females achieved C max values approximately half of those in males, and at a later time point compared to males, whereas in Experiment 2 there were no sex differences at the min time point in young adult mice collapsed across strain. These contrasting trends may have contributed to the absence of overall sex differences when all mice were collapsed across strains. One possible explanation for this result is that FVB mice are more active than mice Bothe et al. Due to its rapid oxidation into the secondary metabolite, OH-THC does not accumulate at high levels in either plasma or brain Huestis, The results of Experiment 1 are consistent with these properties, as plasma and brain concentrations of OH-THC were below the LLOQ for almost all samples obtained after the min time point. Although there were no main effects or interactions involving strain or age on plasma OH-THC concentrations in Experiment 2, unequal distribution of mouse demographics prevented full comparisons across all strains and age groups. Despite the fact that sex differences were not observed in OH-THC plasma concentrations, all three experiments revealed robust sex differences in COOH-THC plasma concentrations, with females achieving higher concentrations than males. Inhaled cannabis pharmacokinetic studies in rats have shown the same pattern Baglot et al. Human pharmacokinetic studies have not found consistent sex differences in COOH-THC concentrations, with reports of either lower concentrations in females compared to males Matheson et al. One possible explanation for these results concerns differences in cytochrome P enzyme activity. Limitations of this study include starting sample collection 10 min following smoke exposure. Additionally, in Experiment 2, more balanced demographics of mice could have allowed for strain and age comparisons with greater statistical power. The raw data supporting the conclusion of this article will be made available by the authors, without undue reservation. The studies were conducted in accordance with the local legislation and institutional requirements. No potentially identifiable images or data are presented in this study. EG led investigation, performed statistical analyses, and wrote the first draft of the manuscript. All authors contributed to the article and approved the submitted version. The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher. This section collects any data citations, data availability statements, or supplementary materials included in this article. As a library, NLM provides access to scientific literature. Front Pharmacol. Find articles by Emely A Gazarov. Find articles by Sabrina Zequeira. Find articles by Alexandria S Senetra. Find articles by John Howard. Find articles by Abhisheak Sharma. Find articles by Christopher R McCurdy. Find articles by Jada Lewis. Find articles by Jennifer L Bizon. Find articles by Barry Setlow. Received May 22; Accepted Aug 8; Collection date Open in a new tab. Abbreviations: LLOQ, lower limit of quantification. Click here for additional data file. Similar articles. Add to Collections. Create a new collection. Add to an existing collection. Choose a collection Unable to load your collection due to an error Please try again. Add Cancel. Middle Aged 10—15 months. T max h.

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