Responses of rainbow trout intestinal epithelial cells to different kinds of nutritional deprivation

Pumputis, Patrick G.; Dayeh, Vivian R.; Lee, Lucy E. J.; Pham, Phuc H.; Liu, Zhenzhen; Viththiyapaskaran, Senthuri; Bols, Niels C.

doi:10.1007/s10695-018-0511-3

Cited by 11 publications

(4 citation statements)

References 51 publications

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“…To examine whether P. acidilactici could promote gut barrier integrity and modulate immune-related genes in the established RTgutGC in vitro model, a 48 h endpoint coincubation experiment was performed using differentiated RTgutGC cells ( Figure 1A ) ( 56 ). However, P. acidilactici did not cause a significant change in the expression of the tight junction genes assessed ( cldn3 , cldn12 , and zo-1 ) nor did it induce changes in expression of key proinflammatory cytokines ( Figure 1B ).…”

Section: Resultsmentioning

confidence: 99%

“…glyceraldehyde-3-phosphate dehydrogenase (gapdh), 18S ribosomal RNA (18S), and internal transcribed spacer 2 locus (its2)] were assessed in 50% of the samples (i.e., in 3 random samples per treatment) using cDNA representing 6.25 ng of input total RNA. Reference gene stability was assessed using geNorm, NormFinder, BestKeeper, and Delta Ct comparison, through the bioinformatic open-access portal RefFinder (51)(52)(53)(54)(55). Most stable genes assessed were ef1a and actb.…”

Section: Qpcr Analysis Of Chinook Salmon Hindgutmentioning

confidence: 99%

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The effects of Pediococcus acidilactici MA18/5M on growth performance, gut integrity, and immune response using in vitro and in vivo Pacific salmonid models

Soto-Dávila,

Langlois Fiorotto,

Heath

et al. 2024

Front. Immunol.

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Microbial management is central to aquaculture’s efficiency. Pediococcus acidilactici MA18/5M has shown promising results promoting growth, modulation of the immune response, and disease resistance in many fishes. However, the mechanisms through which this strain confers health benefits in fish are poorly understood, particularly in Pacific salmonid models. Briefly, the aims of this study were to i) assess the protective effects of P. acidilactici MA18/5M by examining gut barrier function and the expression of tight junction (TJ) and immune genes in vitro and in vivo, and ii) to determine the protective effects of this strain against a common saltwater pathogen, Vibrio anguillarum J382. An in vitro model of the salmonid gut was employed utilizing the cell line RTgutGC. Barrier formation and integrity assessed by TEER measurements in RTgutGC, showed a significant decrease in resistance in cells exposed only to V. anguillarum J382 for 24 h, but pre-treatment with P. acidilactici MA18/5M for 48 h mitigated these effects. While P. acidilactici MA18/5M did not significantly upregulate tight junction and immune molecules, pre-treatment with this strain protected against pathogen-induced insults to the gut barrier. In particular, the expression of ocldn was significantly induced by V. anguillarum J382, suggesting that this molecule might play a role in the host response against this pathogen. To corroborate these observations in live fish, the effects of P. acidilactici MA18/5M was evaluated in Chinook salmon reared in real aquaculture conditions. Supplementation with P. acidilactici MA18/5M had no effect on Chinook salmon growth parameters after 10 weeks. Interestingly, histopathological results did not show alterations associated with P. acidilactici MA18/5M supplementation, indicating that this strain is safe to be used in the industry. Finally, the expression pattern of transcripts encoding TJ and immune genes in all the treatments suggest that variation in expression is more likely to be due to developmental processes rather than P. acidilactici MA18/5M supplementation. Overall, our results showed that P. acidilactici MA18/5M is a safe strain for use in fish production, however, to assess the effects on growth and immune response previously observed in other salmonid species, an assessment in adult fish is needed.

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Section: Resultsmentioning

confidence: 99%

Section: Qpcr Analysis Of Chinook Salmon Hindgutmentioning

confidence: 99%

The effects of Pediococcus acidilactici MA18/5M on growth performance, gut integrity, and immune response using in vitro and in vivo Pacific salmonid models

Soto-Dávila,

Langlois Fiorotto,

Heath

et al. 2024

Front. Immunol.

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“…Expression of the zo -1 was confirmed in rainbow trout intestinal epithelial cell line by immunofluorescence (Pumputis et al . 2018 ). E-cadherin and claudin can interact with zo -1 to maintain the intestinal epithelial barrier and signal transduction between adjacent cells, and they were also expressed in the rainbow trout intestinal cells (RTpi-MI and RTdi-MI) that can form the intestinal epithelial barrier (Pasquariello et al .…”

Section: Discussionmentioning

confidence: 99%

Derivation and characterization of new cell line from intestine of turbot (Scophthalmus maximus)

Liu

et al. 2023

In Vitro Cell.Dev.Biol.-Animal

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A continuous intestine cell line from turbot (Scophthalmus maximus) designated as SMI was established utilizing the tissue explant technique. Primary SMI cell was cultured at 24 °C in a medium with 20% fetal bovine serum (FBS), then subcultured in 10% FBS after 10 passages. Impacts of medium or temperature on the growth of SMI were examined and the results indicated it grew well in DMEM supplemented with 10% FBS at 24 °C. The SMI cell line was subcultured more than 60 times. Karyotyping, chromosome number, and ribosomal RNA genotyping analysis revealed that SMI had a modal diploid chromosome number of 44 and originated from turbot. After being transfected with pEGFP-N1 and FAM-siRNA, a large number of green fluorescence signals were observed in SMI, indicating that SMI could be used as an ideal platform to explore gene function in vitro. In addition, the expression of epithelium-associated genes such as itga6, itgb4, gja1, claudin1, zo-1, and E-cadherin in SMI suggested the SMI had some characteristics of epidermal cells. The upregulation of immune-associated genes such as TNF-β, NF-κB, and IL-1β in SMI after stimulation with pathogen-associated molecular patterns suggested the SMI might exhibit immune functions similar to the intestinal epithelium in vivo.

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“…In general, enterocytes are responsible for assimilation of luminal FAs and also support the de novo synthesis of FAs and cholesterol and serve as a site for production of major lipid transport proteins such as apolipoproteins. Recently, the RTgutGC cell line has been used as a model to study nutrient uptake (Antony Jesu Prabhu et al, 2018;Kim et al, 2018;Pumputis et al, 2018), test functional ingredients and gut immune function (Wang et al, 2019;Holen et al, 2021) and toxicity (Langan et al, 2017;Schug et al, 2020). To our best knowledge, most studies of FA uptake and trafficking by intestinal epithelial cells were conducted in immortalized cell lines such as Caco2 and IEC-6 as enterocyte models.…”

Section: Introductionmentioning

confidence: 99%

Intracellular trafficking of fatty acids in the fish intestinal epithelial cell line RTgutGC

Selvam

Saito²,

Sissener³

et al. 2022

Front. Mar. Sci.

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The shift towards higher inclusion of vegetable oils (VOs) in aquafeeds has resulted in major changes in dietary fatty acid composition, especially increased amounts of monounsaturated fatty acids (MUFAs) and decreased polyunsaturated fatty acids (PUFAs) and saturated fatty acids (SFAs). However, little is known about how this change in fatty acid (FA) profile affects the intracellular fate of these fatty acids in the intestinal cells. To investigate this topic, we used the rainbow trout intestinal epithelial cell line (RTgutGC) as an in vitro model. The cells were incubated with either palmitic acid (16:0, PA), oleic acid (18:1n-9, OA), or arachidonic acid (20:4n-6, ARA), to represent the SFA, MUFA, and PUFA, respectively. In all experiments, the RTgutGC were incubated with either non-labeled or radiolabeled FA (PA, OA, or ARA) for 16 h at 190C. The cells were then analyzed for the occurrence of cytosolic lipid droplets (CLD) with confocal microscopy, transcriptomic analysis (non-labeled FA experiments) and lipid class composition in the cells and serosal media from the basolateral side of the cells (radiolabeled FA experiments). CLD accumulation was higher in RTgutGC exposed to OA compared to cells given PA or ARA. This was coupled with increased volume, diameter, and surface area of CLDs in OA treated cells than with other FAs (PA, ARA). The results from radiolabeled FAs performed on permeable transwell inserts showed that OA increased the triacylglycerides (TAG) synthesis and was primarily stored in the cells in CLDs; whereas a significant amount of ARA was transported as TAG to the basolateral compartment. A significant proportion of free FAs was found to be excreted to the serosal basolateral side by the cells, which was significantly higher for PA and OA than ARA. Although there were clear clusters in differentially expressed genes (DEGs) for each treatment group, results from transcriptomics did not correlate to lipid transport and CLD analysis. Overall, the accumulation of TAG in CLDs was higher for oleic acid (OA) compared to arachidonic acid (ARA) and palmitic acid (PA). To conclude, carbon chain length and saturation level of FA differently regulate their intracellular fate during fatty acid absorption.

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Responses of rainbow trout intestinal epithelial cells to different kinds of nutritional deprivation

Cited by 11 publications

References 51 publications

The effects of Pediococcus acidilactici MA18/5M on growth performance, gut integrity, and immune response using in vitro and in vivo Pacific salmonid models

The effects of Pediococcus acidilactici MA18/5M on growth performance, gut integrity, and immune response using in vitro and in vivo Pacific salmonid models

Derivation and characterization of new cell line from intestine of turbot (Scophthalmus maximus)

Intracellular trafficking of fatty acids in the fish intestinal epithelial cell line RTgutGC

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