Old) have been collected for 72 h. for 72 h. The picture shows lipidupper lipid layers inside the extraction samples. fecal samples. weeks old) have been collected The image shows the upper the layers in the extraction tubes of fecal tubes of Quantification of (f) total fecalof (f) total fecal lipids and (g) fecal neutral sterols. (h,i) Cholesterol absorptionin chow diet-fed female mice Quantification lipids and (g) fecal neutral sterols. (h,i) Cholesterol absorption was measured was measured in chow dietfed four, 10 weeks = four, 10 a 4 h-fasting period, h-fasting gavaged with 200 gavaged with 200 corn [3 containing 2 (n =female mice (nold). Afterweeks old). Right after a 4mice have been period, mice were corn oil containing 2 ioilH]cholesterol i 200 cholesterol. Radioactivity was Radioactivity post-gavage in h plasma and (i) isolated tissues by liquid and [3H]cholesterol and 200 cholesterol. measured 4 h was measured 4 (h)post-gavage in (h) plasma and (i) isolated tissues by liquid scintillation counting. (j) Enterocyte mRNA expression of cholesterol transporters relative to Gapdh as scintillation counting. (j) Enterocyte mRNA expression of cholesterol transporters relative to Gapdh as reference gene reference gene (n = five). Information represent 0.05 (),SD; p0.01 (), p p 0.001 (), p Student’s unpaired t-test. (n = 5). Data represent means + SD; p means + p 0.05 (), 0.01 (). 0.001 (). Student’s unpaired t-test.three.three. LAL-KO Intestines Accumulate Lipids from the Systemic Circulation 3.three. LAL-KO Intestines Accumulate Lipids in the Systemic Circulation WTD-fed LAL-KO mice accumulate lipids predominantly inside the duodenum and WTD-fed LAL-KO mice accumulate lipids predominantly in the duodenum and jejunum, along with the compact intestine is markedly shorter compared to manage mice (Figure 3a). jejunum, plus the compact intestine is markedly shorter compared to handle mice (Figure 3a). We observed aa severe intestinal accumulation neutral lipids in LAL-KO micemice (Figure observed extreme intestinal accumulation of of neutral lipids in LAL-KO (Figure 3b,c). We 3b,c). Electron microscopy confirmed the of lipid-filledof lipid-filled lysosomes Electron microscopy confirmed the abundance abundance lysosomes predominantly predominantly within the (Figure 3d), which can be consistent with is consistent with previous inside the lamina propria lamina propria (Figure 3d), which prior reports describing reports models of LAL-D [12,42,43]. We have not too long ago demonstrated the critical function of in vivo describing in vivo models of LAL-D [12,42,43]. We have recently demonstrated the essential part of cytosolic lipases withinmetabolism of lipids derived from the basolateral cytosolic lipases within AB928 manufacturer enterocytes inside the enterocytes in the metabolism of lipids derived from theside on the little intestine the smaller To determine whether or not LAL-KO enterocytes (blood) basolateral (blood) side of [32,40]. intestine [32,40]. To determine whether LALKO enterocytes accumulate lipid species in the basolateral membrane of enterocytes,Cells 2021, 10,8 ofCells 2021, ten, x8 ofaccumulate lipid species from the basolateral membrane of enterocytes, we incorporated we incorporated [3H]oleate into an Aztreonam In Vivo intralipid emulsion, injected it intraperitoneally, and [3 H]oleate into an intralipid emulsion, injected it intraperitoneally, and measured the tracer three measured the tracer in diverse intestinal segments [32]. [3 H]oleate as an alternative of cholesterol in unique intestinal segments [32]. The incorporation of the incorporation of [.
