Changes in extracellular matrix components and steroidogenic enzymes during growth and atresia of antral ovarian follicles in the sheep.
To investigate the involvement of extracellular matrix (ECM) in folliculogenesis in the sheep, parallel changes in ECM components and key steroidogenic enzymes were studied by quantitative immunohistochemistry and immunoblotting during follicular growth and atresia. Growth of ovarian follicles from 1 to 5 mm in diameter was characterized by a progressive increase in P450 cholesterol sidechain cleavage levels in both thecal (p < 0.001) and granulosa cells (p < 0.001), an increase in P450 aromatase levels in granulosa cells of follicles larger than 3.5 mm (p < 0.001), and an increase in levels of P450 17 alpha-hydroxylase C17,20 lyase (P450(17 alpha)) in the theca interna. In addition, during follicular growth, a change in localization of cells expressing P450(17 alpha) within the theca interna was observed, positive cells being sparse within the theca interna of small follicles and specifically located close to the basal laminae in large follicles. In parallel, follicular growth was associated with an increase in levels of type I collagen in granulosa cell layers (p < 0.01) and an increase in levels of fibronectin (p < 0.05), particularly the specific ED-A alternatively spliced variant of fibronectin, in the theca externa. Follicular atresia was characterized by a loss of P450 aromatase in granulosa cells (p < 0.001) and a decrease in levels of P450(17 alpha) in the theca interna (p < 0.05). Simultaneously, levels of fibronectin (p < 0.05), particularly the ED-A variant of fibronectin, decreased in the theca externa of atretic follicles. Within the wall of granulosa cells, levels of fibronectin (p < 0.05), laminin, type IV collagen, and heparan sulfate proteoglycans strongly increased during follicular atresia. Overall, these results show that follicular growth and atresia were associated with distinct changes in levels of ECM components, suggesting that ECM components may play a role in the regulation of proliferation, differentiation, and apoptosis of follicular cells.
Huet
C
C
INRA, Station de Physiologie de la Reproduction des Mammifères Domestiques, Nouzilly, France.
Monget
P
P
Pisselet
C
C
Monniaux
D
D
D017398:Alternative Splicing; D000818:Animals; D001141:Aromatase / Q000378:metabolism; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000378:metabolism; D003094:Collagen / Q000096:biosynthesis; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D005109:Extracellular Matrix / Q000502:physiology*; D016326:Extracellular Matrix Proteins / Q000096:biosynthesis*; D005260:Female; D005353:Fibronectins / Q000096:biosynthesis; D005496:Follicular Atresia; D006107:Granulosa Cells / Q000166:cytology / Q000201:enzymology / Q000502:physiology*; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate / Q000096:biosynthesis; D007797:Laminin / Q000096:biosynthesis; D006080:Ovarian Follicle / Q000166:cytology / Q000201:enzymology / Q000502:physiology*; D011509:Proteoglycans / Q000096:biosynthesis; D012756:Sheep; D013254:Steroid 17-alpha-Hydroxylase / Q000378:metabolism
D001141:Aromatase; D002786:Cholesterol Side-Chain Cleavage Enzyme; D003094:Collagen; D003577:Cytochrome P-450 Enzyme System; D016326:Extracellular Matrix Proteins; D005353:Fibronectins; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate; D007797:Laminin; D011509:Proteoglycans; D013254:Steroid 17-alpha-Hydroxylase