Ferredoxin reductase levels in the ovaries of pigs and superovulated rats during follicular cell growth and luteinization.

The concentration of ferredoxin reductase, a component of the mitochondrial steroidogenic electron transport chain, was measured in the ovaries of pigs and superovulated rats by a protein blotting procedure using polyclonal antibodies to the purified protein. The concentration of ferredoxin reductase in porcine granulosa cells doubled during growth of follicles from small (1-2 mm diameter) to large (6-12 mm diameter). The concentration doubled again during the period of luteinization. This is in contrast to the rate of cholesterol side-chain cleavage, which showed little change during follicular growth but increased by more than tenfold during luteinization. A similar large increase in cholesterol side-chain cleavage occurs during the period of luteinization in the ovaries of superovulated rats, but as for the pig, only a small increase in ferredoxin reductase was observed. A threefold increase in the yield of mitochondrial protein from tissue homogenates was found between the granulosa cells of small-medium follicles and the cells of the corpora luteum. The increase in ferredoxin reductase during follicular development and luteinization, therefore, correlates well with the increase in mitochondria in the cells, but does not correlate with the dramatic increase in cholesterol side-chain cleavage activity which occurs during luteinization. Based on these results, it is unlikely that the level of ferredoxin reductase limits the expression of the full steroidogenic activity of the granulosa cells of the ovary.

[1]  R. Tuckey,et al.  The composition and distribution of lipid granules in the rat ovary , 1984, Molecular and Cellular Endocrinology.

[2]  P. Stevenson,et al.  Steroidogenesis despite a variant metabolism in primary cultures of pig granulosa cells. , 1983, Biochimica et biophysica acta.

[3]  G. S. Boyd,et al.  The role of cholesterol and cytochrome P-450 in the cholesterol side chain cleavage reaction in adrenal cortex and corpora lutea. , 1975, Journal of steroid biochemistry.

[4]  W. N. Burnette,et al.  "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. , 1981, Analytical biochemistry.

[5]  M. J. Dimino Differences in mitochondrial steroidogenesis between follicular and luteal tissues of porcine ovaries. , 1977, Endocrinology.

[6]  T. Kimura,et al.  Studies on adrenal steroid hydroxylases. Molecular and catalytic properties of adrenodoxin reductase (a flavoprotein). , 1973, The Journal of biological chemistry.

[7]  D. Stone,et al.  Studies on ACTH action in perfused bovine adrenals: the site of action of ACTH in corticosteroidogenesis. , 1954, Archives of biochemistry and biophysics.

[8]  Nils Björkman A study of the ultrastructure of the granulosa cells of the rat ovary. , 1962 .

[9]  E. Work,et al.  Laboratory techniques in biochemistry and molecular biology , 1969 .

[10]  S. Klinken,et al.  Changes in enzyme activities during the artificially stimulated transition from follicular to luteal cell types in rat ovary. , 1977, European journal of biochemistry.

[11]  R. Tuckey,et al.  Purification and analysis of phospholipids in the inner mitochondrial membrane fraction of bovine corpus luteum, and properties of cytochrome P-450scc incorporated into vesicles prepared from these phospholipids. , 1985, European journal of biochemistry.

[12]  Y. Ichikawa,et al.  Purification and biochemical characterization of hepatic ferredoxin (hepatoredoxin) from bovine liver mitochondria , 1986, FEBS letters.

[13]  E. Simpson Cholesterol side-chain cleavage, cytochrome P450, and the control of steroidogenesis , 1979, Molecular and Cellular Endocrinology.

[14]  M. Waterman,et al.  Evidence for the presence of cholesterol side chain cleavage cytochrome P-450 and adrenodoxin in fresh granulosa cells. Effects of follicle-stimulating hormone and cyclic AMP on cholesterol side chain cleavage cytochrome P-450 synthesis and activity. , 1983, The Journal of biological chemistry.

[15]  T. Yamano,et al.  Crystallization and properties of reduced nicotinamide adenine dinucleotide phosphate‐adrenodoxin reductase of pig adrenocortical mitochondria , 1977, FEBS letters.

[16]  R. Tuckey,et al.  Properties of ferredoxin reductase and ferredoxin from the bovine corpus luteum. , 1984, The International journal of biochemistry.

[17]  Y. Kang,et al.  The porcine ovarian follicle. II. Electron microscopic study of surface features of granulosa cells at different stages of development. , 1977, Biology of reproduction.

[18]  Y. Nakamura,et al.  Requirement of a new flavoprotein and a non-heme iron-containing protein in the steroid 11-beta- and 18-hydroxylase system. , 1966, Biochimica et biophysica acta.

[19]  M. Waterman,et al.  Induction of synthesis of cholesterol side chain cleavage cytochrome P-450 and adrenodoxin by follicle-stimulating hormone, 8-bromo-cyclic AMP, and low density lipoprotein in cultured bovine granulosa cells. , 1984, The Journal of biological chemistry.

[20]  P. Stevenson,et al.  Electron flow and cholesterol-side-chain cleavage in ovarian mitochondria. , 1971, European journal of biochemistry.

[21]  P. Stevenson,et al.  The differential development of mitochondrial cytochrome P-450 and the respiratory cytochromes in rat ovary. , 1981, Biochimica et Biophysica Acta.

[22]  R. Estabrook,et al.  Isolation from adrenal cortex of a nonheme iron protein and a flavoprotein functional as a reduced triphosphopyridine nucleotide-cytochrome P-450 reductase , 1966 .

[23]  M. Pesce,et al.  A new micromethod for determination of protein in cerebrospinal fluid and urine. , 1973, Clinical chemistry.