Presence of Arylsulfatase A and Sulfogalactosylglycerolipid in Mouse Ovaries: Localization to the Corpus Luteum

Abstract

Arylsulfatase A (AS-A) is a lysosomal enzyme, which catalyzes the desulfation of certain sulfogalactolipids, including sulfogalactosylglycerolipid (SGG), a molecule implicated in cell adhesion. In this report, immunocytochemistry revealed the selective presence of AS-A in the corpus luteum of mouse ovaries. Immunoblotting indicated that mouse corpus luteum AS-A had a molecular mass of 66 kDa, similar to AS-A of other tissues. Corpus luteum AS-A was active, capable of desulfating the artificial substrate, p-nitrocatechol sulfate, at the optimum pH of five. To understand further the role of AS-A in female reproduction, levels of AS-A were determined during corpus luteum development in pseudopregnant mice and during luteolysis after cessation of pseudopregnancy. Immunocytochemistry, immunoblotting and desulfation activity showed that AS-A expression was evident at the onset of pseudopregnancy in the newly formed corpora lutea, and its level increased steadily during gland development. The increase in the expression and activity of AS-A continued throughout luteolysis after the decrease in serum progesterone levels. We also observed the selective presence of SGG on the luteal cell surface in developed corpora lutea, as shown by immunofluorescence of mouse ovary sections as well as high-performance thin-layer chromatography of lipids isolated from mouse and pig corpora lutea. The identity of the "SGG" band on the thin layer silica plate was further validated by electrospray ionization mass spectrometry. Significantly, SGG disappeared in regressing corpora lutea. Therefore, lysosomal AS-A may be involved in cell-surface remodeling during luteolysis by desulfating SGG after its endocytosis and targeting to the lysosome.

Quantification of seminolipid by LC-ESI-MS/MS-multiple reaction monitoring: compensatory levels in Cgt mice

“…Azure A is also useful for staining sulfolipids on thin layer chromatography (TLC) plates. The limit of detection (LOD) of the colorimetric assay for sulfolipids is around 10 µg in the original method and, despite modifi cations and improvements, is still around 0.3 µg ( 13,17 ). The LOD of Azure A staining of chromatographed lipids on high-performance (HP)TLC plates is even higher: at least 1 µg of a sulfolipid is required for detection.…”

Arylsulfatase A deficiency causes seminolipid accumulation and a lysosomal storage disorder in Sertoli cells

“…Testis lipid extracts from wild-type and Arsa Ϫ / Ϫ mice at both 5 and 8 months of age were subjected to high-performance thin layer chromatography (HPTLC) followed by orcinol staining, as previously described ( 24 ). Glycolipids were stained purple, whereas phospholipids turned brown.…”

“…Glycolipids were stained purple, whereas phospholipids turned brown. In Arsa Ϫ / Ϫ mice, an extra glycolipid band chromatographed between SGG and phosphatidylcholine was observed; the area of the HPTLC plate around this band, starting underneath the SGG band, was scraped for lipid extraction by the Bligh/Dyer method ( 24 ). Extracted lipids were subjected to ESI-MS/MS as previously described ( 23,25 ).…”

Sexual dimorphism in the white matter of rodents

“…Different concentrations of progesterone (P2), estrogen (E2) and dihydrotestosterone (DHT), either alone or in various combinations, were added to enriched Olg cultures to determine their effects upon total cell numbers, proliferation, and cell death [46]. Female fluctuation of P2 concentration is well known and depends on the phase of estrous cycle [47] as well as pregnancy [48], supporting theoretical contribution of P2 to Olg sexual dimorphism. When hormones were added to Olgs culture, P2 significantly increased the number of Olgs in both sexes (but more so in female cultures), E2 had a minor effect on Olg number and DHT reduced Olg numbers in both sexes.…”