Investigation of mammalian ovulation with an in vitro perfused rabbit ovary preparation

doi:10.1016/S0002-9378(78)80007-6

American Journal of Obstetrics and Gynecology

Volume 132, Issue 7, 1978, Pages 728-738

https://doi.org/10.1016/S0002-9378(78)80007-6 Get rights and content

An in vitro perfused rabbit ovary preparation has been developed to study local factors responsible for the ovulatory process. This preparation enables: (1) isolation of the ovary from systemic influences, (2) direct serial observations of follicle development and ovulation, (3) addition to the perfusing medium of substances that may influence ovulation acting at the level of the ovary, and (4) correlation of the times of individual follicle rupture with ovarian contractile patterns. With this model the temporal requirements for human chorionic gonadotropin in the process of ovulation have been studied. The direct effects on ovulation of prostaglandin F_2α(PGF_2α), the antihistamine chlorpheniramine, and calcium deprivation were also investigated. Chlorpheniramine and EDTA each led to inhibition of ovulation and depressed ovarian contractility. The use of the isolated in vitro ovary preparation permits characterization of the local requirements for ovulation.

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Prostaglandin E₂ (PGE₂) is the most common prostanoid and has a variety of bioactivities including a crucial role in urogenital function. Multiple enzymes are involved in its biosynthesis. Among 3 PGE₂ terminal synthetic enzymes, membrane-associated PGE₂ synthase-2 (mPGES-2) is the most recently identified, and its role remains uncharacterized. In previous studies, membrane-associated PGE₂ synthase-1 (mPGES-1) and cytosolic PGE₂ synthase (cPGES) were reported to be expressed along the urogenital tracts. Here we report the genomic structure and tissue distribution of mPGES-2 in the urogenital system. Analysis of several bioinformatic databases demonstrated that mouse mPGES-2 spans 7 kb and consists of 7 exons. The mPGES-2 promoter contains multiple Sp1 sites and a GC box without a TATA box motif. Real-time quantitative PCR revealed that constitutive mPGES-2 mRNA was most abundant in the heart, brain, kidney and small intestine. In the urogenital system, mPGES-2 was highly expressed in the renal cortex, followed by the renal medulla and ovary, with lower levels in the ureter, bladder and uterus. Immunohistochemistry studies indicated that mPGES-2 was ubiquitously expressed along the nephron, with much lower levels in the glomeruli. In the ureter and bladder, mPGES-2 was mainly localized to the urothelium. In the reproductive system, mPGES-2 was restricted to the epithelial cells of the testis, epididymis, vas deferens and seminal vesicle in males, and oocytes, stroma cells and corpus luteum of the ovary and epithelial cells of the oviduct and uterus in females. This expression pattern is consistent with an important role for mPGES-2-mediated PGE₂ in urogenital function.
Genomic structure and genitourinary expression of mouse cytosolic prostaglandin E<inf>2</inf> synthase gene
2003, Biochimica et Biophysica Acta - Molecular and Cell Biology of Lipids
Prostaglandin E₂ (PGE₂) plays an important role in genitourinary function. Multiple enzymes are involved in its biosynthesis. Here we report the genomic structure and tissue-selective expression of cytosolic PGE₂ synthase (cPGES) in genitourinary tissues. Full-length mouse cPGES cDNA was cloned by reverse transcript-polymerase chain reaction (RT-PCR) and 5′- and 3′-rapid amplification of cDNA ends (RACE). Analysis of a cPGES cDNA with partially sequenced cPGES genomic clones and bioinformatic databases demonstrates that the murine cPGES gene spans ∼22 kb and consists of eight exons. The cPGES gene promoter is GC-rich and contains many SP1 sites but lacks an obvious TATA box motif. RNase protection assay revealed constitutive expression of cPGES was greatest in the testis with lower levels in the ovary, kidney, bladder and uterus. In situ hybridization studies demonstrated that cPGES mRNA was most highly expressed in the epithelial cells of seminiferous tubules in the testis. In the female reproductive tissues, cPGES was mainly localized in ovarian primary and secondary follicles and oviductal epithelial cells with less expression in uterine endometrium. In the kidney cPGES expression was diffusely expressed. In urinary bladder, cPGES expression was restricted to the transitional epithelial cells. This expression pattern is consistent with an important role for cPGES-mediated PGE₂ in urogenital tissue function.
Leukocyte networks and ovulation
2002, Journal of Reproductive Immunology
Ovulation, the process whereby the oocyte is expelled from the interior of the follicle, is the final process of folliculogenesis. During the last decade, data have accumulated to suggest that tissue-bound leukocytes are major effector cells in several physiological processes within the reproductive tract. Some specific subclasses of leukocytes seem to be critically involved in the process of ovulation. The main components of this ovulatory process are degradation of the extracellular matrix (ECM) at the follicular apex and changes in the follicular vasculature. The leukocytes participate actively in these events by secretion of proteases and vasoactive substances. This review covers our current understanding of the mechanisms by which the leukocytes are attracted to the preovulatory follicle after the LH-surge and the roles that the activated leukocytes play in the follicle during the ovulatory period.
Immunological aspects of ovarian function: role of the cytokines
1995, European Journal of Obstetrics and Gynecology
Interaction between the immune system and reproduction is multiple. Either directly or indirectly through their products, immune cells are associated with the regulation of every level of the hypothalamus-pituitary-ovarian axis. Immune cells are present in the ovaries and their numbers increase during the cycle. During the follicular development cytokines assist granulosa cell growth while inhibiting their differentiation. During the LH peak, an influx of immune cells occurs and several cytokines are released. The rupture of the follicle is considered as an inflammatory reaction. IL-1, TNF-α are the main cytokines involved in this process. During the luteal phase, the installation of the corpus luteum needs the setting up of neovascularization. Cytokines are probable candidates for this function, but they also promote cellular differentiation resulting in steroid synthesis. In the absence of pregnancy T lymphocytes and eosinophils are involved in corpus luteum regression. Their products are directly cytotoxic for the luteal cells. They attract macrophages which are locally activated to phagocytose the damaged luteal cells. They can induce apoptosis of endothelial and luteal cells through gene expression. Cytokines are members of a larger regulatory network residing in the ovary and involving hormones and growth factors. The various stages of ovarian cycle will be shown from an immunological point of view. Understanding the role of the cytokines should enable us to go beyond a purely descriptive stage, and allow us to envisage new ovulation induction therapy and treatment in certain cases of premature menopause.
A review - Role of eicosanoids in vertebrate ovulation
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Toxicity testing using the isolated in vitro perfused ovary
1993, Reproductive Toxicology
This article discusses the use of in vitro perfusion techniques as a tool for toxicity testing in the ovary and how the rat ovary has been adapted for this purpose. A brief review of the development of in vitro ovarian perfusion is provided, focusing on steroidogensis and physiology of ovulation. Adaptation of this model for use as a toxicologic model is discussed in the context of other isolated organ models, (that is, liver, heart, lung). Surgical procedures, perfusate and criteria for viability are outlined. Advantages of this technique are highlighted including ability to administer high doses of drugs directly to intact organ devoid of other influences. Applications of this model are discussed and data from studies of glutathione depleted ovaries perfused with hexachlorobenzene (HCB) are presented. Increased oxygen consumption after addition of HCB is suggestive of a disordered respiratory metabolism and is an example of future markers of ovarian injury using this innovative technique.

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: Supported by National Institutes of Health Grant HD-05948 and The Mitchell and Lillian Duberstein Foundation.

: Presented by invitation at the One Hundred and First Annual Meeting of the American Gynecological Society, Coronado, California, April 26–29, 1978.

^*: Connelly Foundation Fellow in Reproductive Biology at Pennsylvania Hospital.

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Investigation of mammalian ovulation with an in vitro perfused rabbit ovary preparation

Prostaglandins

Prostaglandins

Prostaglandins

Life Sci.

Prostaglandins

Am. J. Obstet. Gynecol.

Fertil. Steril.

Fertil. Steril.

Fertil: Steril.

Prostaglandins

Prostaglandins

Prostaglandins

In vitro studies of ovulation in the perfused rabbit ovary

Fertil. Steril.