A single-day mouse mesenteric lymph surgery in mice: an updated approach to study dietary lipid absorption, chylomicron secretion, and lymphocyte dynamics

Dedousis, Nikolaos; Teng, Lihong; Kanshana, Jitendra S.; Kohan, Alison B.

doi:10.1016/j.jlr.2022.100284

Cited by 3 publications

(1 citation statement)

References 102 publications

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“…The difference in dietary lipid absorption profiles in mice kept for 6-h or 24-h post-surgery was recently tested 44 . We found that the 24-h procedure reduces the animal survival rates and the excursion of dietary lipids into the lymph.…”

Section: Notementioning

confidence: 99%

The Isolation of Flowing Mesenteric Lymph in Mice to Quantify <em>In Vivo</em> Kinetics of Dietary Lipid Absorption and Chylomicron Secretion

Dedousis

Teng

Kohan

2022

JoVE

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Intestinal lipoproteins, especially triglyceride-rich chylomicrons, are a major driver of metabolism, inflammation, and cardiovascular diseases. However, isolating intestinal lipoproteins is very difficult in vivo because they are first secreted from the small intestine into the mesenteric lymphatics. Chylomicron-containing lymph then empties into the subclavian vein from the thoracic duct to deliver components of the meal to the heart, lungs, and, ultimately, whole-body circulation. Isolating naïve chylomicrons is impossible from blood since chylomicron triglyceride undergoes hydrolysis immediately upon interaction with lipoprotein lipase and other lipoprotein receptors in circulation. Therefore, the original 2-day lymph fistula procedure, described by Bollman et al. in rats, has historically been used to isolate fresh mesenteric lymph before its entry into the thoracic vein. That protocol has been improved upon and professionalized by the laboratory of Patrick Tso for the last 45 years, allowing for the analysis of these critical lipoproteins and secretions from the gut. The Tso lymph fistula procedure has now been updated and is presented here visually for the first time. This revised procedure is a single-day surgical technique for installing a duodenal feeding tube, cannulating the mesenteric lymph duct, and collecting lymph after a meal in conscious mice. The major benefits of these new techniques include the ability to reproducibly collect lymph from mice (which harnesses the power of genetic mouse models); the reduced anesthesia time for mice during the implantation of the duodenal infusion tube and the lymph cannula; the ability to continuously sample lymph throughout the feeding and post-prandial period; the ability to quantitatively measure hormones and cytokines before their dilution and enzymatic hydrolysis in blood; and the ability to collect large quantities of lymph for isolating intestinal lipoproteins. This technique is a powerful tool for directly and

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Section: Notementioning

confidence: 99%

The Isolation of Flowing Mesenteric Lymph in Mice to Quantify <em>In Vivo</em> Kinetics of Dietary Lipid Absorption and Chylomicron Secretion

Dedousis

Teng

Kohan

2022

JoVE

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Impaired intestinal free fatty acid transport followed by chylomicron malformation, not pancreatic insufficiency, cause metabolic defects in cystic fibrosis

Teng,

Dedousis,

Adeshirlarijaney

et al. 2024

Journal of Lipid Research

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Characterization of genes and proteins involved in the absorption of long-chain fatty acids in the gastrointestinal tract of cattle

Hentz,

Batistel

2024

Front. Anim. Sci.

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IntroductionDietary fatty acids play a pivotal role in energy metabolism and various physical functions. However, the molecular mechanisms underlying the absorption of long-chain fatty acids (LCFA) in ruminant animals remain poorly understood. To address this gap, we investigated the occurrence and distribution of genes and proteins previously characterized in nonruminant models within the gastrointestinal tract of cattle.MethodsTissue samples from the rumen, descending duodenum, proximal jejunum, medial jejunum, distal jejunum, and distal ileum were collected at slaughter from eight Holstein steers. RT‒PCR and capillary electrophoretic-based chemiluminescence were used to determine the mRNA and protein levels of genes and proteins involved in the uptake, activation, intracellular trafficking, re-esterification, and lipoprotein assembly of LCFA.Results and discussionMeasurable mRNA levels were found for 38 genes involved in the intestinal absorption of LCFA in all examined sections of the bovine gastrointestinal tract. The mRNA abundances of all genes were greater in the intestinal sections than in the rumen sections (p < 0.05). At the protein level, the expression of nine key transporters and enzymes, namely, CD36, SLC27A4, ACSL1, FABP1, FABP2, DGAT1, MTP, Apo-AI, and ACAT2, was detected in most of the intestinal sections. The distribution profile of these proteins in the small intestine suggested a prominent role for protein-mediated absorption of LCFA in the proximal and distal intestine in cattle. The low expression levels of most proteins in the rumen suggest little absorption of LCFA at this location in cattle. Overall, the findings from this study support the presence, in cattle, of genes and proteins involved in the intestinal absorption of LCFA described in models of nonruminants. Identifying the proteins involved in the absorption of LCFA in the small intestine is the first step in understanding how the expression of these proteins influences the capacity of the small intestine to absorb fatty acids in cattle.

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A single-day mouse mesenteric lymph surgery in mice: an updated approach to study dietary lipid absorption, chylomicron secretion, and lymphocyte dynamics

Cited by 3 publications

References 102 publications

The Isolation of Flowing Mesenteric Lymph in Mice to Quantify <em>In Vivo</em> Kinetics of Dietary Lipid Absorption and Chylomicron Secretion

The Isolation of Flowing Mesenteric Lymph in Mice to Quantify <em>In Vivo</em> Kinetics of Dietary Lipid Absorption and Chylomicron Secretion

Impaired intestinal free fatty acid transport followed by chylomicron malformation, not pancreatic insufficiency, cause metabolic defects in cystic fibrosis

Characterization of genes and proteins involved in the absorption of long-chain fatty acids in the gastrointestinal tract of cattle

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