Old) were collected for 72 h. for 72 h. The image shows lipidupper lipid layers within the extraction samples. fecal samples. weeks old) have been collected The image shows the upper the layers inside 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). Just after a 4mice were period, mice were corn oil containing two 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 = 5). Information represent 0.05 (),SD; p0.01 (), p p 0.001 (), p Student’s unpaired t-test. (n = five). Data represent means + SD; p implies + p 0.05 (), 0.01 (). 0.001 (). Student’s unpaired t-test.three.three. LAL-KO Intestines Accumulate Lipids in the Systemic Circulation three.three. LAL-KO Intestines Accumulate Lipids in the Systemic Circulation WTD-fed LAL-KO mice accumulate lipids predominantly in the duodenum and WTD-fed LAL-KO mice accumulate lipids predominantly in the duodenum and jejunum, plus the compact intestine is markedly shorter compared to control mice (Figure 3a). jejunum, as well as the small intestine is markedly shorter in comparison to manage mice (Figure 3a). We observed aa serious 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 inside the (Figure 3d), which is consistent with is constant with earlier inside the lamina propria lamina propria (Figure 3d), which preceding reports describing reports models of LAL-D [12,42,43]. We’ve Ionomycin Purity lately demonstrated the critical function of in vivo describing in vivo models of LAL-D [12,42,43]. We’ve got not too long ago demonstrated the vital role of cytosolic lipases withinmetabolism of lipids derived from the basolateral cytosolic lipases within enterocytes in the enterocytes in the metabolism of lipids derived from theside of the compact intestine the small To establish irrespective of whether LAL-KO enterocytes (blood) basolateral (blood) side of [32,40]. intestine [32,40]. To identify whether or not LALKO enterocytes accumulate lipid species from the basolateral membrane of enterocytes,Cells 2021, 10,8 ofCells 2021, ten, x8 ofaccumulate lipid species in the basolateral membrane of enterocytes, we incorporated we incorporated [3H]Biotinyl tyramide custom synthesis oleate into an 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 distinct intestinal segments [32]. [3 H]oleate instead of cholesterol in different intestinal segments [32]. The incorporation on the incorporation of [.
