Studies of the Isolated Perfused Ovary: Methods and Examples of Application

Romanoff, Elijah B.; Pincus, Gregory; Burnett, Foster; Dyer, W. Wallace; O'Farrell, A. Marie

doi:10.1210/endo-71-5-752

Cited by 21 publications

(3 citation statements)

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“…In ruminants, the in vitro model of an isolated perfused ovary (IPO) was introduced several decades ago (Romanoff & Pincus 1962, Mills & Morrissette 1970, Stahler & Huch 1971, Sturm & Stahler 1971, Janson et al 1978, mainly to investigate ovarian steroid biosynthesis and energy metabolism. For studying mechanisms of LPS action on the bovine CL, with special consideration of the impact of uterine-derived PGF 2a , this model has important advantages.…”

Section: Introductionmentioning

confidence: 99%

Lipopolysaccharide enhances apoptosis of corpus luteum in isolated perfused bovine ovaries in vitro

Lüttgenau¹,

Möller²,

Kradolfer³

et al. 2016

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Lipopolysaccharide (LPS), the endotoxin of Gram-negative bacteria, has detrimental effects on the structure and function of bovine corpus luteum (CL) in vivo. The objective was to investigate whether these effects were mediated directly by LPS or via LPS-induced release of PGF 2a . Bovine ovaries with a mid-cycle CL were collected immediately after slaughter and isolated perfused for 240 min. After 60 min of equilibration, LPS (0.5 mg/ml) was added to the medium of five ovaries, whereas an additional six ovaries were not treated with LPS (control). After 210 min of perfusion, all ovaries were treated with 500 iu of hCG. In the effluent perfusate, concentrations of progesterone (P 4 ) and PGF 2a were measured every 10 and 30 min, respectively. Punch biopsies of the CL were collected every 60 min and used for RT-qPCR to evaluate mRNA expression of receptors for LPS (TLR2, -4) and LH (LHCGR); the cytokine TNFA; steroidogenic (STAR, HSD3B), angiogenic (VEGFA 121 , FGF2), and vasoactive (EDN1) factors; and factors of prostaglandin synthesis (PGES, PGFS, PTGFR) and apoptosis (CASP3, -8, -9). Treatment with LPS abolished the hCG-induced increase in P 4 (P%0.05); however, there was a tendency (PZ0.10) for increased release of PGF 2a at 70 min after LPS challenge. Furthermore, mRNA abundance of TLR2, TNFA, CASP3, CASP8, PGES, PGFS, and VEGFA 121 increased (P%0.05) after LPS treatment, whereas all other factors remained unchanged (PO0.05).In conclusion, reduced P 4 responsiveness to hCG in LPS-treated ovaries in vitro was not due to reduced steroidogenesis, but was attributed to enhanced apoptosis. However, an impact of luteal PGF 2a could not be excluded.

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Section: Introductionmentioning

confidence: 99%

Lipopolysaccharide enhances apoptosis of corpus luteum in isolated perfused bovine ovaries in vitro

Lüttgenau¹,

Möller²,

Kradolfer³

et al. 2016

Reproduction

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“…The method of perfusion of the ovary outside the body has been utilized for many years by a number of investigators to study steroidogenesis in ovarian tissue of the pig (Werthessen et al, 1953) and the cow (Romanoff and Pincus, 1962), and various related metabolic and morphologic events (Carrel and Lindbergh, 1935). These earlier techniques have been modified and perfected for in vitro perfusion of ovaries of the rabbit (Ah& et al, 1975), the sheep (Janson e t al., 1978), the cow (Stahler and Huch, 1971), and the human (Stahler et al, 1971; Kennedy et al, 1975).…”

Section: Introductionmentioning

confidence: 99%

The Study of Ovulation in the Isolated Perfused Rabbit Ovary. I. Methodology and Pattern of Steroidogenesis

Janson

LeMaire

Källfelt

et al. 1982

Biology of Reproduction

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A previously described technique for perfusion of the isolated rabbit ovary in a recirculating system (Ahrén et al., 1975) was modified for studies of preovulatory follicular development and ovulation. Follicular ruptures were induced either by injection of human chorionic gonadotropin (hCG) to the animal 5-6 h prior to perfusion or by adding luteinizing hormone (LH) directly to the medium during perfusion. Ovulation occurred 9-14 h after hCG injection in vivo and 7-13 h after the addition of LH in vitro. Ovulations did not occur unless the ovaries were stimulated by gonadotropin in vivo or in vitro. The patterns of release of estradiol-17 beta and progesterone into the medium are consistent with the measurements reported by others on these steroids in ovarian venous blood in vivo. This experimental model is likely to be useful in further exploration of the physiology, biochemistry and anatomy of ovulation.

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“…The techniques for the collection, flushing and perfusion of bovine ovaries have been described by Romanoff & Pincus (1962) required to fill a 1-ml section taken from a 5-ml pipette. Citrated bovine blood was used as the perfusate and perfusion pressure was maintained at 100 mm Hg.…”

Section: Methodsmentioning

confidence: 99%

Progesterone Synthesis by Perfused Bovine Ovaries of Early and Late Pregnancy

Mills¹,

Morrissette²

1970

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Summary. Fourteen bovine ovaries of early pregnancy and nineteen of late pregnancy were perfused with citrated bovine blood. During perfusion, the venous blood was collected from each ovary before and after addition of lh and was analysed for progesterone content. Average progesterone synthesis rates were 4-37+1-12 and 4-57 + 0-99 µg/min before addition of lh, for ovaries of early and late pregnancy respectively, and increased to 7-43+1-98 and 7-72+1-33 µg/min, respectively, after addition of lh. There was no significant difference in the rates of progesterone synthesis between ovaries of early and late pregnancy at the 5% level and their response to lh was highly significant (P<0-01). These data indicate that a decrease in progesterone synthesis during late pregnancy in the bovine is not due to a reduction in the ability of the corpus luteum to respond to gonadotrophins.

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Studies of the Isolated Perfused Ovary: Methods and Examples of Application

Cited by 21 publications

References 0 publications

Lipopolysaccharide enhances apoptosis of corpus luteum in isolated perfused bovine ovaries in vitro

Lipopolysaccharide enhances apoptosis of corpus luteum in isolated perfused bovine ovaries in vitro

The Study of Ovulation in the Isolated Perfused Rabbit Ovary. I. Methodology and Pattern of Steroidogenesis

Progesterone Synthesis by Perfused Bovine Ovaries of Early and Late Pregnancy

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