Old) had been collected for 72 h. for 72 h. The image shows lipidupper lipid layers inside the extraction samples. fecal samples. weeks old) were collected The picture shows the upper the layers within 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 = 4, ten a four h-fasting period, h-fasting gavaged with 200 gavaged with 200 corn [3 containing two (n =female mice (nold). Afterweeks old). Soon 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 four h was measured 4 (h)post-gavage in (h) plasma and (i) isolated tissues by liquid scintillation Cysteinylglycine manufacturer 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). Information represent suggests + SD; p signifies + p 0.05 (), 0.01 (). 0.001 (). Student’s unpaired t-test.3.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 in the duodenum and WTD-fed LAL-KO mice accumulate lipids predominantly within the duodenum and jejunum, and the modest intestine is markedly shorter compared to manage mice (Figure 3a). jejunum, plus the tiny intestine is markedly shorter in comparison with manage mice (Figure 3a). We observed aa extreme intestinal accumulation neutral lipids in LAL-KO micemice (Figure observed serious 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 in the (Figure 3d), which can be constant with is consistent with preceding in the lamina propria lamina propria (Figure 3d), which prior reports describing reports models of LAL-D [12,42,43]. We’ve got not too long ago demonstrated the important role of in vivo describing in vivo models of LAL-D [12,42,43]. We’ve lately demonstrated the critical role of cytosolic lipases withinmetabolism of lipids derived in the basolateral cytosolic lipases inside enterocytes in the enterocytes within the metabolism of lipids derived from theside of the smaller intestine the smaller To ascertain no matter whether LAL-KO enterocytes (blood) basolateral (blood) side of [32,40]. intestine [32,40]. To ascertain regardless of whether LALKO enterocytes accumulate lipid species in the basolateral membrane of enterocytes,Cells 2021, ten,8 ofCells 2021, ten, x8 ofaccumulate lipid species in the basolateral membrane of enterocytes, we incorporated we incorporated [3H]oleate into an intralipid emulsion, injected it cis-4-Hydroxy-L-proline Epigenetics 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 diverse intestinal segments [32]. The incorporation of the incorporation of [.
