Thyroid  in gut development during Atlantic salmon smoltification

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Modulation of intestinal growth and differentiation by photoperiod and dietary treatment during smoltification in Atlantic salmon (Salmo salar, L.)

Publication . Fernandes Duarte, Vilma Vanessa; Gaetano, Pasqualina; Striberny, Anja; Hazlerigg, David; Jørgensen, Even H.; Fuentes, Juan; Campinho, Marco António

Atlantic salmon undergo smoltification, a process that prepares the fish to enter and thrive in seawater (SW). Several physiological changes occur during smolting, especially in osmoregulatory tissues, the gill, the kidney, and the intestine. Here we characterized the effects on intestinal morphogenesis of two different, commonly used smoltification regimes during the end of the freshwater phase, photoperiod and/or the addition of salt and amino acid supplements in the diet. We focused on intestinal morphological differentiation, i.e., external perimeter, absorptive perimeter, tissue thickness, and villi density. In addition, we quantified cell proliferation (PCNA positive) and Na+, K+-ATPase (NKA) and Na+, K+,2Cl- (NKCCs) co-transporters expression and enterocyte apicobasal distribution by immunohistochemistry. These analyses show that the anterior and posterior intestines have different developmental dynamics during smoltification. In both intestinal regions, photoperiod and dietary treatment increased the absorptive perimeter. In addition, diet and photoperiod treatments differentially stimulated NKA protein expression in the anterior intestine. NKCC apical-basolateral expression in the enterocytes increased after SW entry in the anterior and posterior intestines. In conclusion, our results show that, as smoltification progresses, the anterior intestine responds more readily to experimental conditions than the posterior intestine. In our study, photoperiod and dietary treatment seem to enhance the development of the capacity to tolerate SW.

2023Journal article

Restricted

Photoperiod and dietary treatment in freshwater modulate the short-term intestinal response to seawater in atlantic salmon (salmo salar)

Publication . Gaetano, Pasqualina; Fernandes Duarte, Vilma Vanessa; Striberny, Anja; Hazlerigg, David; Jørgensen, Even H.; Campinho, Marco António; Fuentes, Juan

Stimulation and timing of smoltification are essential for successful Atlantic salmon (Salmo salar) aquaculture. This study investigated intestinal responses during dietary and photoperiod manipulation in freshwater (FW) and after a subsequent seven days residence in seawater (SW). "Small" and "large" Atlantic salmon parr (-40 g and -130 g respectively) were treated in FW for 12 weeks and thereafter transferred to SW for seven days. During the FW phase, fish underwent two different light conditions, 24 L:0D - 24 L ("LL-LL" groups) and 7 L:17D - 24 L ("SP-LL" groups) or fed with either regular feed ("LL-LL C" and "SP-LL C" groups) or feed enriched with a salt mix plus free tryptophan ("LL-LL + diet" and "SP-LL + diet" groups). We analyzed Na+/K+-ATPase (NKA) activity, tissue bioelectrical properties in Ussing chambers, and intestinal fluid composition. The NKA activity showed minor variations in relation to fish size, treatments, or intestinal region (anterior or posterior). Photoperiod modulated epithelial bioelectrical properties (Isc and Rt) of the anterior and posterior intestine, particularly transepithelial resistance (Rt). Pharmacological experiments, targeting apical Na+/K+/2Cl- (NKCC2) and Na+/ Cl (NCC) co-transporters revealed intestinal region-and water salinity-dependent effects. In addition, stimu-lation of the intracellular cAMP with forskolin and IBMX showed intestinal region-, water salinity, and treatment-dependence responses with clear functional specialization of the anterior and posterior intestine. The intestinal fluid composition reflected the ability to process ingested SW and showed little variation in large fish. In sum-mary, our data suggest a better pre-adaptation of the intestine during light-stimulated smoltification (SP-LL groups), and the combination of light and diet might give, in an industrial aquaculture setting, an advantage to smaller, but not larger smolts. Intestinal fluid composition in small fish can be used as an index of intestinal function and may act as a long-term performance proxy in SW Atlantic salmon.

2023Journal article

Open Access

Molecular responses in the intestine of Atlantic salmon (Salmo salar) following light and diet stimulation of smoltification: Potential molecular markers for a seawater-ready smolt

Publication . Gaetano, Pasqualina; Fernandes Duarte, Vilma Vanessa; Striberny, Anja; Hazlerigg, David G.; Jørgensen, Even H.; Campinho, Marco António; Fuentes, Juan

The transfer to seawater (SW) represents a critical stage in the production of Atlantic salmon. The success of the transfer links with the optimal development of hypo-osmoregulatory capacities during smoltification. While various strategies are adopted in aquaculture to stimulate smoltification, considerable fish loss still occurs after transfer to sea cages. Therefore, we investigated the molecular responses in the anterior and posterior intestine of Atlantic salmon, following 1) a photoperiod treatment (24 h light (L):0 h dark (D) → 24 L:0D vs. 7 L:17D → 24 L:0D) and 2) dietary treatment (regular feed or feed enriched with a salt mix/tryptophan), combined with, or without a photoperiodic treatment in freshwater (FW), to evaluate how intestinal osmoregulatory mechanisms are modulated by these treatments, and to identify potential intestinal markers indicative of a SW-ready smolt. Using quantitative real-time PCR (qPCR), we investigated transcript levels of transporters and channels involved in ion movements through the enterocytes, tight junction components, and receptors (i.e., calcium-sensing re ceptor and prolactin receptor). The two intestinal regions showed different gene profiles and responsiveness towards the experimental treatments. In the anterior intestine, the exposure to short photoperiod (7 L:17D) upregulated Na+/K+ − ATPase subunit alpha 1c (nkaα1c), Na+/K+/2Cl− cotransporter 1 (nkcc1), Na+/K+/2Cl− cotransporter 2 (nkcc2), Cl− /HCO− 3 exchanger Slc26a6 (slc26a6), and cystic fibrosis transmembrane conductance regulator I (cftrI), in FW and SW. Also, Na+/K+ − ATPase subunit alpha 1b (nkaα1b), occludin (ocln), and prolactin receptor (prlr) were upregulated in FW and claudin 15 (cldn15) in SW groups exposed to this photo period. The posterior intestine was less responsive to the experimental treatments, although upregulation of nkcc1, nkcc2, slc26a6, and cftrI was observed in FW in the short photoperiod groups. Hence, our findings show that exposure to a winter signal in FW more effectively activates hypo-osmoregulatory mechanisms in the in testine of Atlantic salmon, where a coordinated and complementary role of the anterior and posterior intestine ensures optimal SW processing. Dietary treatment had a positive but more marginal effect on the regulation of the genes investigated, mainly enhancing the impact of short photoperiod when the two treatments were combined. Overall, we propose the apical Na+/K+/2Cl− cotransporter, nkcc2, and the apical Cl− / HCO− 3 exchanger, slc26a6, as potential FW molecular markers in the anterior intestine to assess “SW-readiness” in Atlantic salmon smolts.

2025-02Journal article

Open Access

Molecular responses in the intestine of Atlantic Salmon (Salmo salar) following light and diet stimulation of smoltification: Potential molecular markers for a seawater-ready smolt

Publication . Gaetano, Pasqualina; Fernandes Duarte, Vilma Vanessa; Striberny, Anja; Hazlerigg, David G.; Jørgensen, Even H.; Campinho, Marco António; Fuentes, Juan

The transfer to seawater (SW) represents a critical stage in the production of Atlantic salmon. The success of the transfer links with the optimal development of hypo-osmoregulatory capacities during smoltification. While various strategies are adopted in aquaculture to stimulate smoltification, considerable fish loss still occurs after transfer to sea cages. Therefore, we investigated the molecular responses in the anterior and posterior intestine of Atlantic salmon, following 1) a photoperiod treatment (24 h light (L):0 h dark (D) -* 24 L:0D vs. 7 L:17D -* 24 L:0D) and 2) dietary treatment (regular feed or feed enriched with a salt mix/tryptophan), combined with, or without a photoperiodic treatment in freshwater (FW), to evaluate how intestinal osmoregulatory mechanisms are modulated by these treatments, and to identify potential intestinal markers indicative of a SW-ready smolt. Using quantitative real-time PCR (qPCR), we investigated transcript levels of transporters and channels involved in ion movements through the enterocytes, tight junction components, and receptors (i.e., calcium-sensing receptor and prolactin receptor). The two intestinal regions showed different gene profiles and responsiveness towards the experimental treatments. In the anterior intestine, the exposure to short photoperiod (7 L:17D) upregulated Na+/K+ - ATPase subunit alpha 1c (nka alpha 1c), Na+/K+/2Cl- cotransporter 1 (nkcc1), Na+/K+/2Clcotransporter 2 (nkcc2), Cl- /HCO-3 exchanger Slc26a6 (slc26a6), and cystic fibrosis transmembrane conductance regulator I (cftrI), in FW and SW. Also, Na+/K+ - ATPase subunit alpha 1b (nka alpha 1b), occludin (ocln), and prolactin receptor (prlr) were upregulated in FW and claudin 15 (cldn15) in SW groups exposed to this photoperiod. The posterior intestine was less responsive to the experimental treatments, although upregulation of nkcc1, nkcc2, slc26a6, and cftrI was observed in FW in the short photoperiod groups. Hence, our findings show that exposure to a winter signal in FW more effectively activates hypo-osmoregulatory mechanisms in the intestine of Atlantic salmon, where a coordinated and complementary role of the anterior and posterior intestine ensures optimal SW processing. Dietary treatment had a positive but more marginal effect on the regulation of the genes investigated, mainly enhancing the impact of short photoperiod when the two treatments were combined. Overall, we propose the apical Na+/K+/2Cl- cotransporter, nkcc2, and the apical Cl- / HCO-3 exchanger, slc26a6, as potential FW molecular markers in the anterior intestine to assess "SW-readiness" in Atlantic salmon smolts.

2025-02Journal article

Open Access