A simple automated approach to measure mouse whole gut transit

Kacmaz, Halil; Alto, Alecia; Knutson, Kaitlyn R.; Linden, David R.; Gibbons, Simon J.; Farrugia, Gianrico; Beyder, Arthur

doi:10.1111/nmo.13994

Cited by 9 publications

(16 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…When we assessed the GI function in Hoxb8 Cre+/- ;Piezo2 fl/fl ( Piezo2 Hoxb8 ) mice, we observed accelerated GI transit, increased defecation frequency, increased water content and reduced dry-stool weight in comparison to the wild-type ( Piezo2 WT ) littermates (Figure 3B, Figure S2-E), phenocopying the Piezo2 SNS model. Additionally, the shorter transit time was observed in Piezo2 Hoxb8 mice using a videorecorder to detect the colored fecal pellets instead of an experimenter to avoid stress on mice 43 (Figure S2-E). These results suggest that Piezo2-expressing intestinal epithelial cells or spinal afferents, rather than enteric or nodose neurons, are responsible for the accelerated GI transit phenotype.…”

Section: Resultsmentioning

confidence: 99%

“…Our previous GI transit experiments and others provide information on the time required for intestinal contents to travel from the stomach to the evacuation point 43,[49][50][51] , but lack details about the transit throughout the intermediate regions of the gut. To investigate whether Piezo2 -expressing somatosensory neurons modulate motility along the entire GI tract or in discrete regions, we functionally evaluated gastric emptying, intestinal transit, and colonic transit.…”

Section: Neuronal Piezo2 Mediates Gastric Emptying Intestinal Transit...mentioning

confidence: 99%

See 1 more Smart Citation

PIEZO2 in somatosensory neurons controls gastrointestinal transit

Servin‐Vences

Lam

Koolen

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

The gastrointestinal tract is in a state of constant motion. These movements are tightly regulated by the presence of food and help digestion by mechanically breaking down and propelling gut content. Mechanical sensing in the gut is thought to be essential for regulating motility; however, the identity of the neuronal populations, the molecules involved, and the functional consequences of this sensation are unknown. Here, we show that humans lacking PIEZO2 exhibit impaired bowel sensation and motility. Piezo2 in mouse dorsal root but not nodose ganglia is required to sense gut content, and this activity slows down food transit rates in the stomach, small intestine, and colon. Indeed, Piezo2 is directly required to detect colon distension in vivo. Our study unveils the mechanosensory mechanisms that regulate the transit of luminal contents throughout the gut, which is a critical process to ensure proper digestion, nutrient absorption, and waste removal.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Neuronal Piezo2 Mediates Gastric Emptying Intestinal Transit...mentioning

confidence: 99%

PIEZO2 in somatosensory neurons controls gastrointestinal transit

Servin‐Vences

Lam

Koolen

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…Time‐lapse videos were generated by Video Velocity (CandyLabs). In the Light setting, the time of pellet appearance and pellet color were composed as timelapse videos and analyzed as previously described 6 (Figure 1H). For experiments performed in the Dark (red light), timelapse videos showed pellets but did not distinguish color, so at the end of the experiment a simple light image was collected to determine the distribution of pellets and was analyzed via a reverse play‐through of the video to determine pellet sequence (Figure 1I,J).…”

Section: Methodsmentioning

confidence: 99%

“…The automated whole gut transit system is by previously published works. 6 LEDs (Daybetter Flexible LED Strip Lights Kit, #FLSL-R5032-RGB-US) were installed to improve light with red light lumination. Red light can be used during the active phase since mice are dichromatic.…”

Section: Automated Whole Gut Transit Setupmentioning

confidence: 99%

Circadian rhythm and whole gut transit in mice

Laible,

Wegner,

Knutson

et al. 2024

Neurogastroenterology Motil

Self Cite

View full text Add to dashboard Cite

BackgroundIn preclinical studies whole gut transit (WGT) in mice is a gold‐standard “leading‐edge” approach that measures the time between orogastric gavage of carmine red and defecation of the first carmine red pellet. Transit studies in humans are performed during the active day because GI motility and transit are suppressed during the night. Since mice are nocturnal, WGT studies traditionally done during the day occur during their rest phase. How circadian rhythm affects WGT in mice is not known.MethodsWe used an automated approach for high temporal resolution uninterrupted testing of mouse WGT and activity. We housed wild‐type Bl6/C57 mice under the standard 12 h light–dark cycles. At 8 weeks, we performed carmine red orogastric gavage and assessed WGT during Light (rest) conditions. Then, we exposed mice to a reverse 12 h light–dark cycle for 2 weeks and tested them in the Dark (active) under red light conditions. Timelapse videos were analyzed to quantify activity and to timestamp all pellets, and multiple parameters were analyzed.Key ResultWhen complementary light cycle reversal experiments were performed, we found a significant increase in mouse activity when mice were tested during their Dark (active) phase, compared to their Light (rest) phase. In mice tested in the Active phase compared to the Rest phase, we found a significant acceleration in WGT, increased rate and total number of pellets produced, and more pellet clustering. These data show that the mice tested in the Active phase have important differences in activity that correlate with multiple alterations in gastrointestinal transit.Conclusion & InferencesDuring the Active phase mice have faster WGT, produce more pellets, and cluster their output compared to testing in the Rest phase. Like in humans, circadian rhythm is an important consideration for transit studies in mice, and a simple reverse light cycle approach facilitates further studies on the role of circadian rhythm in GI motility.

show abstract

“…Moreover, biological rhythms, food intake and stress significantly affect gastrointestinal motility, [22][23][24][25][26] affirming the importance of the timing of testing and considerations for experimental housing conditions. Although other optimizations of the whole-gut transit have been described by others, 27,28 our method is, as far as we know, the first to include the influence of housing conditions and social interaction in the experimental design to improve welfare.…”

Section: Re Sultsmentioning

confidence: 99%

An optimization and refinement of the whole‐gut transit assay in mice

Schonkeren

Seeldrayers

Thijssen

et al. 2023

Neurogastroenterology Motil

View full text Add to dashboard Cite

Background Gastrointestinal motility measurements in mice are currently performed under suboptimal conditions, as these nocturnal animals are measured during light conditions. In addition, other stressors, like individual housing, placement in a new cage during observation, and lack of bedding and cage enrichment cause animal discomfort and might contribute to higher variability. Here we aimed to develop a refined method of the widely‐used whole‐gut transit assay. Methods Wildtype mice (N = 24) were subjected to the standard or refined whole‐gut transit assay, either with or without a standardized slowing in gastrointestinal motility induced by loperamide. The standard assay consisted of a gavage with carmine red, observation during the light period and individual housing in a new cage without cage enrichment. For the refined whole‐gut transit assay, mice were gavaged with UV‐fluorescent DETEX®, observed during the dark period, while pairwise housed in their home cage with cage enrichment. Time until excretion of the first colored fecal pellet was assessed, and pellets were collected to assess number, weight, and water content. Key Results The DETEX®‐containing pellets were UV‐detectable, allowing to measure the mice in their active period in the dark. The refined method caused less variation (20.8% and 16.0%) compared to the standard method (29.0% and 21.7%). Fecal pellet number, weight, and water content was significantly different between the standard and refined method. Conclusions & Inferences This refined whole‐gut transit assay provides a reliable approach to measure whole‐gut transit time in mice in a more physiological context, with reduced variability compared to the standard method.

show abstract

A simple automated approach to measure mouse whole gut transit

Cited by 9 publications

References 18 publications

PIEZO2 in somatosensory neurons controls gastrointestinal transit

PIEZO2 in somatosensory neurons controls gastrointestinal transit

Circadian rhythm and whole gut transit in mice

An optimization and refinement of the whole‐gut transit assay in mice

Contact Info

Product

Resources

About