Examination of Granulosa Cell Glycoconjugates Which Change During Follicular Atresia in the Pig Ovary

Abstract

Abstract. Changes of glycoconjugates in granulosa cells during follicular atresia were examined by lectin histochemical and biochemical techniques. Twenty-four lectins were used to visualize the different glycoconjugates of granulosa cells in frozen sections of the pig ovary. Four lectins showed stronger staining of the granulosa cells in atretic than in healthy follicles. Very strong and/or strong staining with Sambucus sieboldiana agglutinin (SSA) was seen in scattered granulosa cells of atretic follicles, but those of healthy follicles were not stained with this lectin. In atretic follicles, very strong-moderate diffuse staining was seen with Aleuria aurantia lectin (AAL), Pisum sativum agglutinin (PSA) and Lens culinaris agglutinin (LCA) in granulosa cells, in contrast with very weak staining in those of healthy follicles. Furthermore, lectin blot analysis with three lectins (SSA, AAL, and PSA) indicated several glycoproteins with increased staining in atretic relative to healthy follicles for these three lectins. Especially, two SSA-positive glycoproteins (62 and 64 kD) of granulosa cells in atretic follicles were specific and characteristic which were not detected in those in healthy follicles. These results suggest that changes of some glycoconjugates in granulosa cells may be involved in granulosa cell apoptosis and follicular atresia.

Species-specific Differences in Apoptotic Cell Localization in Granulosa and Theca Interna Cells during Follicular Atresia in Porcine and Bovine Ovaries.

“…Unfortunately, no natural ligands that bind with PFG-1 and mediate granulosa cell apoptosis in atretic follicles have not been reported. In porcine follicles, the cell-surface glyco-structure of granulosa cells specifically changes during follicular atresia [22,23], and such changes are highly correlated with PFG-1 kinetics. In murine ovaries, Fas, which mediates apoptosis in a variety of lymphoid and tumor cells, induces apoptotic cell death in follicular granulosa cells and luteal cells, indicating that the Fas ligand-Fas system plays a key role in follicle selection and luteolysis in murine ovaries [24], but not in porcine ovaries [20].…”

Characteristic Changes in Carbohydrate Profile in the Kidneys of Hereditary Nephrotic Mice (ICGN Strain)

“…Carbohydrate binding specificities of the 24 lectins are summarized in Table 1. The localization of specific carbohydrate chains in the kidney sections was visualized using the avidin-biotin peroxidase complex (ABC) method as previously described [4,5,12,13]. Briefly, after deparaffinization and rehydration, the sections were incubated with avidin and biotin blocking solution (Vector Laboratories, Burlingame, CA, U.S.A.) at room temperature (RT; 23-25°C) for 15 min to block endogenous binding sites, and then washed with PBS.…”

“…As characteristic staining of Bandeiraea simplicifolia lectin-I (BSL-I) was demonstrated histochemically, lectin blot analysis was performed as previously described [4,5,12]. Briefly, frozen kidney sections (at least 10 mg) were mixed with 0.0625 M Tris-HCl (pH 6.8) containing 2% Nonidet-P40, 2% sodium dodecyl sulfate (SDS) and 5% 2-mercaptoethanol (Sigma Chemical, St. Louis, MO, U.S.A.), heated for 3 min at 100˚C, and then urea was added at a final concentration of 8 M. The protein concentration of each sample was determined by a modification of the method of Bradford [1].…”

“…For instance, tumor cells with malignant phenotypes show neoplastic transformation-related changes in oligosaccharide biosynthesis [2]. Cell-surface carbohydrates are reliable markers for histopathological diagnosis, and lectins recognizing these macromolecules or cell-surface carbohydrates have been extensively used as biochemical and histopathological probes in cell biology and pathology [4,5,7,12,13].The ICR-derived glomerulonephritis (ICGN) mice, a novel inbred strain of mice with a hereditary nephrotic syndrome of unknown etiology, are considered to be a good model for human idiopathic nephrotic syndrome. They were established at the National Institute of Infectious Diseases [21][22][23][24][25][26][27].…”

“…For instance, tumor cells with malignant phenotypes show neoplastic transformation-related changes in oligosaccharide biosynthesis [2]. Cell-surface carbohydrates are reliable markers for histopathological diagnosis, and lectins recognizing these macromolecules or cell-surface carbohydrates have been extensively used as biochemical and histopathological probes in cell biology and pathology [4,5,7,12,13].…”