Developmental Changes of Monohexosylceramide and Free Ceramide in the Large Intestine of the Rat1

Abstract

Neutral glycosphingolipids were isolated from the colon of rats between birth and adulthood. The glycolipid concentration was stable during this period. Epithelial cells of the adult colon contained three times more glycolipids than the whole organ. The distribution pattern underwent only minor modifications during development. Free ceramide contributed for 23-27% of the total neutral sphingolipids at all ages. In 6-day-old rats, it was constituted of nonhydroxylated fatty acids linked to C18-sphingenine (57.3% of the bases), C18- and C20-4D-hydroxysphinganine (24.2 and 14.0% of the bases, respectively). This composition was essentially maintained during development. Glucosylceramide was the major glycolipid at all ages (40-50% of the total neutral sphingolipid content). At birth, 40% of its fatty acids were 2-hydroxylated and 93% of the bases were C18-4D-hydroxysphinganine. In adult epithelial cells, 75% of the fatty acids were 2-hydroxylated and C18- and C20-4D-hydroxysphinganine contributed for 66 and 25% of the bases, respectively. A transient increase of the contribution of nonhydroxylated fatty acids and C18-sphingenine was observed during the first week of life. C20-4D-hydroxysphinganine, which was characterized by gas-liquid chromatography of its aldehydes after periodate oxidation and of its N-acetyl O-trimethylsilyl derivatives, appeared after birth and reached 20% of the bases after two weeks. These findings are another example of the specificity of the lipidic part of glucosylceramide during the ontogenic differentiation.

Implications for invariant natural killer T cell ligands due to the restricted presence of isoglobotrihexosylceramide in mammals

“…When the tissue and species pattern of expression of isogloboseries GSLs was examined (see SI Table 1), it was clear that the rat utilizes this biosynthetic pathway in several tissues (17,(20)(21)(22)(23)(24)(25). In mouse, iGb3 and iGb4 were detected only in DRG, among the tissues screened, including the CNS.…”

“…All solvents were of analytical grade or higher (VWR, unless otherwise stated). HPLC standards were from Dextra Laboratories (Reading, U.K., iGb3 trisaccharide) or Alexis (Axxora U.K., Nottingham, U.K.; iGb3 lipid), prepared in-house (iGb4 (22), or kindly provided by Eric Samain (Centre de Recherches sur les Macromolécules Végétales-Centre Nationale de la Recherche Scientifique, Grenoble, France; Gb3). Tissue culture media were from GIBCO (Paisley, U.K.), unless otherwise stated.…”

“…The biosynthetic derivative of iGb3, iGb4 (Fig. 1a), has been identified in multiple tissues in the rat, including intestine (17), stomach (22), kidney (23), spleen, glomerular mesangial cells, lymphosarcoma, granuloma (24), and a rat-derived metastatic cell line (25). It has been reported that mouse and human kidney lack iGb4 (23) (SI Table 1).…”

“…For iGb4, no commercial standard was available. However, the major species in a neutral lipid preparation from rat stomach has been reported to be iGb4 (22). Therefore, neutral GSL-derived oligosaccharides from rat stomach were prepared and anthranilic acid (2AA) labeled.…”

Immunological Function of Sphingosine 1-Phosphate in the Intestine

“…Several lines of evidence demonstrate that intestinal tissues contain higher levels of sphingolipids, including S1P, than other tissues [17]. There is no evidence of intestinal uptake of sphingolipids from the blood, and germfree rats have comparable levels of sphingolipids in the intestine to conventional specific pathogen-free (SPF) rats [18].…”