TY - JOUR
T1 - Expression of hyaluronan synthase 1 and distribution of hyaluronan during follicular atresia in pig ovaries
AU - Miyake, Yuko
AU - Sakurai, Masahiro
AU - Tanaka, Sachi
AU - Tunjung, Woro Anindito Sri
AU - Yokoo, Masaki
AU - Matsumoto, Hiromichi
AU - Aso, Hisashi
AU - Yamaguchi, Takahiro
AU - Sato, Eimei
N1 - Copyright:
Copyright 2012 Elsevier B.V., All rights reserved.
PY - 2009/2
Y1 - 2009/2
N2 - CD44 on macrophages is recognized as a phagocytic receptor involved in the phagocytosis of apoptotic cells. Recently, we detected cd44 on macrophages in atretic follicles during atresia. In this study, we evaluated the distribution of the principal cd44 ligand hyaluronan(ha) and the expressions of ha synthases(has: Has1, has2, and has3) during atresia in pig ovaries. We determined the 2139-bp sequence of sus scrofa has1 and raised an anti-has1 polyclonal antibody. The s. Scrofa has1 sequence contained six putative ha-binding motifs and conserved amino acid residues crucial for glcnac transfer- Ase activity. Has1 mrna expression was upregulated during atresia; however, has2 and has3 mrna expression levels were low and very low to undetectable, respectively. Western blotting showed that has1 was markedly upregulated during atresia. Immunohistochemical analyses revealed has1 distribution in theca cells of healthy and early atretic(stages i and ii) follicles and in progressing atretic(stage iii) follicles. Hyaluronan was visualized with the ha-binding protein; it accumulated in the theca layer during all stages and in stage iii follicles. Hyaluronan assay showed a significantly increased ha concentration in follicular fluid at stage iii. Flow cytometry showed has1 expression in 55.7% of sirpa-positive macrophages in stage iii follicles. Our results suggest that the ha concentration in follicular fluids increased during atresia and that has1 may be the dominant has protein in theca cells to produce ha in pig ovaries.
AB - CD44 on macrophages is recognized as a phagocytic receptor involved in the phagocytosis of apoptotic cells. Recently, we detected cd44 on macrophages in atretic follicles during atresia. In this study, we evaluated the distribution of the principal cd44 ligand hyaluronan(ha) and the expressions of ha synthases(has: Has1, has2, and has3) during atresia in pig ovaries. We determined the 2139-bp sequence of sus scrofa has1 and raised an anti-has1 polyclonal antibody. The s. Scrofa has1 sequence contained six putative ha-binding motifs and conserved amino acid residues crucial for glcnac transfer- Ase activity. Has1 mrna expression was upregulated during atresia; however, has2 and has3 mrna expression levels were low and very low to undetectable, respectively. Western blotting showed that has1 was markedly upregulated during atresia. Immunohistochemical analyses revealed has1 distribution in theca cells of healthy and early atretic(stages i and ii) follicles and in progressing atretic(stage iii) follicles. Hyaluronan was visualized with the ha-binding protein; it accumulated in the theca layer during all stages and in stage iii follicles. Hyaluronan assay showed a significantly increased ha concentration in follicular fluid at stage iii. Flow cytometry showed has1 expression in 55.7% of sirpa-positive macrophages in stage iii follicles. Our results suggest that the ha concentration in follicular fluids increased during atresia and that has1 may be the dominant has protein in theca cells to produce ha in pig ovaries.
KW - Apoptosis
KW - Atresia
KW - Follicle
KW - Hyaluronan
KW - Ovary
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U2 - 10.1095/biolreprod.108.067694
DO - 10.1095/biolreprod.108.067694
M3 - Article
C2 - 18923162
AN - SCOPUS:59949084220
VL - 80
SP - 249
EP - 257
JO - Biology of Reproduction
JF - Biology of Reproduction
SN - 0006-3363
IS - 2
ER -