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Article ; Online: Prenatal exposure to Di(2-ethylhexyl) phthalate and high-fat diet synergistically disrupts gonadal function in male mice†.

Barakat, Radwa / Lin, Po-Ching Patrick / Bunnell, Mary / Oh, Ji-Eun / Rattan, Saniya / Arnieri, Cyrus / Flaws, Jodi A / Ko, CheMyong J

Biology of reproduction

2024 Volume 110, Issue 5, Page(s) 1025–1037

Abstract: Prenatal exposure to Di (2-ethylhexyl) phthalate (DEHP) impairs the reproductive system and causes fertility defects in male offspring. Additionally, high-fat (HF) diet is a risk factor for reproductive disorders in males. In this study, we tested the ... ...

Abstract	Prenatal exposure to Di (2-ethylhexyl) phthalate (DEHP) impairs the reproductive system and causes fertility defects in male offspring. Additionally, high-fat (HF) diet is a risk factor for reproductive disorders in males. In this study, we tested the hypothesis that prenatal exposure to a physiologically relevant dose of DEHP in conjunction with HF diet synergistically impacts reproductive function and fertility in male offspring. Female mice were fed a control or HF diet 7 days prior to mating and until their litters were weaned on postnatal day 21. Pregnant dams were exposed to DEHP or vehicle from gestational day 10.5 until birth. The male offspring's gross phenotype, sperm quality, serum hormonal levels, testicular histopathology, and testicular gene expression pattern were analyzed. Male mice born to dams exposed to DEHP + HF had smaller testes, epididymides, and shorter anogenital distance compared with those exposed to HF or DEHP alone. DEHP + HF mice had lower sperm concentration and motility compared with DEHP mice. Moreover, DEHP + HF mice had more apoptotic germ cells, fewer Leydig cells, and lower serum testosterone levels than DEHP mice. Furthermore, testicular mRNA expression of Dnmt1 and Dnmt3a was two to eight-fold higher than in DEHP mice by qPCR, suggesting that maternal HF diet and prenatal DEHP exposure additively impact gonadal function by altering the degree of DNA methylation in the testis. These results suggest that the combined exposure to DEHP and high-fat synergistically impairs reproductive function in male offspring, greater than exposure to DEHP or HF diet alone.
MeSH term(s)	Animals ; Female ; Male ; Diethylhexyl Phthalate/toxicity ; Mice ; Pregnancy ; Prenatal Exposure Delayed Effects/chemically induced ; Diet, High-Fat/adverse effects ; Testis/drug effects ; Testis/pathology ; Spermatozoa/drug effects
Language	English
Publishing date	2024-05-16
Publishing country	United States
Document type	Journal Article
ZDB-ID	1118-6
ISSN	1529-7268 ; 0006-3363
ISSN (online)	1529-7268
ISSN	0006-3363
DOI	10.1093/biolre/ioae029
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Article ; Online: Single neonatal estrogen implant sterilizes female animals by decreasing hypothalamic KISS1 expression.

Park, Chan Jin / Minabe, Shiori / Hess, Rex A / Lin, Po-Ching Patrick / Zhou, Sherry / Bashir, Shah Tauseef / Barakat, Radwa / Gal, Arnon / Ko, CheMyong Jay

Scientific reports

2023 Volume 13, Issue 1, Page(s) 9627

Abstract: Reproductive sterilization by surgical gonadectomy is strongly advocated to help manage animal populations, especially domesticated pets, and to prevent reproductive behaviors and diseases. This study explored the use of a single-injection method to ... ...

Abstract	Reproductive sterilization by surgical gonadectomy is strongly advocated to help manage animal populations, especially domesticated pets, and to prevent reproductive behaviors and diseases. This study explored the use of a single-injection method to induce sterility in female animals as an alternative to surgical ovariohysterectomy. The idea was based on our recent finding that repetitive daily injection of estrogen into neonatal rats disrupted hypothalamic expression of Kisspeptin (KISS1), the neuropeptide that triggers and regulates pulsatile secretion of GnRH. Neonatal female rats were dosed with estradiol benzoate (EB) either by daily injections for 11 days or by subcutaneous implantation of an EB-containing silicone capsule designed to release EB over 2-3 weeks. Rats treated by either method did not exhibit estrous cyclicity, were anovulatory, and became infertile. The EB-treated rats had fewer hypothalamic Kisspeptin neurons, but the GnRH-LH axis remained responsive to Kisspeptin stimulation. Because it would be desirable to use a biodegradable carrier that is also easier to handle, an injectable EB carrier was developed from PLGA microspheres to provide pharmacokinetics comparable to the EB-containing silicone capsule. A single neonatal injection of EB-microspheres at an equivalent dosage resulted in sterility in the female rat. In neonatal female Beagle dogs, implantation of an EB-containing silicone capsule also reduced ovarian follicle development and significantly inhibited KISS1 expression in the hypothalamus. None of the treatments produced any concerning health effects, other than infertility. Therefore, further development of this technology for sterilization in domestic female animals, such as dogs and cats is worthy of investigation.
MeSH term(s)	Female ; Animals ; Cats ; Dogs ; Rats ; Kisspeptins/pharmacology ; Cat Diseases ; Dog Diseases ; Hypothalamus ; Gonadotropin-Releasing Hormone ; Animals, Domestic ; Sterilization ; Infertility ; Estrogens/pharmacology
Chemical Substances	Kisspeptins ; Gonadotropin-Releasing Hormone (33515-09-2) ; Estrogens
Language	English
Publishing date	2023-06-14
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-023-36727-8
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Article ; Online: Prenatal exposure to an environmentally relevant phthalate mixture disrupts testicular steroidogenesis in adult male mice.

Barakat, Radwa / Seymore, Talia / Lin, Po-Ching Patrick / Park, Chan Jin / Ko, CheMyong Jay

Environmental research

2019 Volume 172, Page(s) 194–201

Abstract: Endocrine disrupting chemicals (EDCs) in the environment are considered to be a contributing factor to the decline in the sperm quality. With growing evidence of the harmful effects of EDCs on the male reproductive system, we tested the hypothesis that ... ...

Abstract	Endocrine disrupting chemicals (EDCs) in the environment are considered to be a contributing factor to the decline in the sperm quality. With growing evidence of the harmful effects of EDCs on the male reproductive system, we tested the hypothesis that prenatal exposure to an environmentally relevant phthalate mixture adversely affects reproductive outcomes and androgen synthesis. In this study, an environmentally relevant composition of phthalates (15% DiNP, 21% DEHP, 36% DEP, 15% DBP, 8% DiBP, and 5% BBzP) that were detected in urine samples of pregnant women in Illinois, United States, was used. Pregnant CD-1 mice (F0) were orally dosed with a vehicle or the phthalate mixtures (20 µg/kg/day, 200 µg/kg/day, 200 mg/kg/day, or 500 mg/kg/day) from gestational day 10.5 to the day of birth. Then, the indices of the reproductive function of the F1 males born to these dams were assessed. Those male mice prenatally exposed to the phthalate mixture had smaller gonads, prostates and seminal vesicles, especially in the 20 µg/kg/day and 500 mg/kg/day phthalate mixture groups, compared to the controls. Importantly, at the age of 12 months, those prenatally exposed mice had significantly lower serum testosterone concentrations accompanied by the decreased mRNA expression of testicular steroidogenic genes (StAR, Cyp11, and Cyp17) and impaired spermatogenesis. Taken together, this study found that prenatal exposure to environmentally relevant doses of a phthalate mixture caused a life-long impact on the reproduction in male mice.
MeSH term(s)	Animals ; Diethylhexyl Phthalate ; Environmental Pollutants/toxicity ; Environmental Pollutants/urine ; Female ; Gene Expression Regulation/drug effects ; Genitalia, Male/drug effects ; Humans ; Illinois ; Male ; Mice ; Phthalic Acids/toxicity ; Phthalic Acids/urine ; Pregnancy ; Prenatal Exposure Delayed Effects ; Testosterone/blood ; Testosterone/metabolism
Chemical Substances	Environmental Pollutants ; Phthalic Acids ; Testosterone (3XMK78S47O) ; Diethylhexyl Phthalate (C42K0PH13C)
Language	English
Publishing date	2019-02-13
Publishing country	Netherlands
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	205699-9
ISSN	1096-0953 ; 0013-9351
ISSN (online)	1096-0953
ISSN	0013-9351
DOI	10.1016/j.envres.2019.02.017
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Article: Prenatal exposure to an environmentally relevant phthalate mixture disrupts testicular steroidogenesis in adult male mice

Barakat, Radwa / Seymore, Talia / Lin, Po-Ching Patrick / Park, Chan Jin / Ko, CheMyong Jay

Environmental research. 2019 May, v. 172

2019

Abstract	Endocrine disrupting chemicals (EDCs) in the environment are considered to be a contributing factor to the decline in the sperm quality. With growing evidence of the harmful effects of EDCs on the male reproductive system, we tested the hypothesis that prenatal exposure to an environmentally relevant phthalate mixture adversely affects reproductive outcomes and androgen synthesis. In this study, an environmentally relevant composition of phthalates (15% DiNP, 21% DEHP, 36% DEP, 15% DBP, 8% DiBP, and 5% BBzP) that were detected in urine samples of pregnant women in Illinois, United States, was used. Pregnant CD-1 mice (F0) were orally dosed with a vehicle or the phthalate mixtures (20 μg/kg/day, 200 μg/kg/day, 200 mg/kg/day, or 500 mg/kg/day) from gestational day 10.5 to the day of birth. Then, the indices of the reproductive function of the F1 males born to these dams were assessed. Those male mice prenatally exposed to the phthalate mixture had smaller gonads, prostates and seminal vesicles, especially in the 20 μg/kg/day and 500 mg/kg/day phthalate mixture groups, compared to the controls. Importantly, at the age of 12 months, those prenatally exposed mice had significantly lower serum testosterone concentrations accompanied by the decreased mRNA expression of testicular steroidogenic genes (StAR, Cyp11, and Cyp17) and impaired spermatogenesis. Taken together, this study found that prenatal exposure to environmentally relevant doses of a phthalate mixture caused a life-long impact on the reproduction in male mice.
Keywords	adults ; blood serum ; endocrine-disrupting chemicals ; gene expression ; genes ; males ; maternal exposure ; messenger RNA ; mice ; phthalates ; pregnant women ; prostate gland ; seminal vesicles ; sperm quality ; spermatogenesis ; steroidogenesis ; testes ; testosterone ; urine ; Illinois
Language	English
Dates of publication	2019-05
Size	p. 194-201.
Publishing place	Elsevier Inc.
Document type	Article
ZDB-ID	205699-9
ISSN	1096-0953 ; 0013-9351
ISSN (online)	1096-0953
ISSN	0013-9351
DOI	10.1016/j.envres.2019.02.017
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Article ; Online: Generation and characterization of an endothelin-2 iCre mouse.

Cacioppo, Joseph A / Koo, Yongbum / Lin, Po-Ching Patrick / Gal, Arnon / Ko, CheMyong

Genesis (New York, N.Y. : 2000)

2015 Volume 53, Issue 2, Page(s) 245–256

Abstract: A novel transgenic mouse line that expresses codon-improved Cre recombinase (iCre) under regulation of the Endothelin-2 gene (edn2) promoter was developed for the conditional deletion of genes in Endothelin-2 lineage cells and for the spatial and ... ...

Abstract	A novel transgenic mouse line that expresses codon-improved Cre recombinase (iCre) under regulation of the Endothelin-2 gene (edn2) promoter was developed for the conditional deletion of genes in Endothelin-2 lineage cells and for the spatial and temporal localization of Endothelin-2 expression. Endothelin-2 (EDN2, ET-2, previously VIC) is a transcriptionally regulated 21 amino acid peptide implicated in vascular homeostasis, and more recently in female reproduction, gastrointestinal function, immunology, and cancer pathogenesis that acts through membrane receptors and G-protein signaling. A cassette (edn2-iCre) was constructed that contained iCre, a polyadenylation sequence, and a neomycin selection marker in front of the endogenous start codon of the edn2 gene in a mouse genome BAC clone. The cassette was introduced into the C57BL/6 genome by pronuclear injection, and two lines of edn2-iCre positive mice were produced. The edn2-iCre mice were bred with ROSA26-lacZ and Ai9 reporter mice to visualize areas of functional iCre expression. Strong expression was seen in the periovulatory ovary, stomach and small intestine, and colon. Uniquely, we report punctate expression in the corneal epithelium, the liver, the lung, the pituitary, the uterus, and the heart. In the embryo, expression is localized in developing hair follicles and the dermis. Therefore, edn2-iCre mice will serve as a novel line for conditional gene deletion in these tissues.
MeSH term(s)	Animals ; Endothelin-2/genetics ; Female ; Gene Deletion ; Gene Knockout Techniques ; Genetic Engineering ; Integrases/genetics ; Male ; Mice, Inbred C57BL ; Mice, Transgenic ; Organ Specificity
Chemical Substances	Endothelin-2 ; Cre recombinase (EC 2.7.7.-) ; Integrases (EC 2.7.7.-)
Language	English
Publishing date	2015-02
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2004544-X
ISSN	1526-968X ; 1526-954X
ISSN (online)	1526-968X
ISSN	1526-954X
DOI	10.1002/dvg.22845
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Article ; Online: Granulosa cell endothelin-2 expression is fundamental for ovulatory follicle rupture.

Cacioppo, Joseph A / Lin, Po-Ching Patrick / Hannon, Patrick R / McDougle, Daniel R / Gal, Arnon / Ko, CheMyong

Scientific reports

2017 Volume 7, Issue 1, Page(s) 817

Abstract: Ovulation is dependent upon numerous factors mediating follicular growth, vascularization, and ultimately oocyte release via follicle rupture. Endothelin-2 (EDN2) is a potent vasoconstrictor that is transiently produced prior to follicle rupture by ... ...

Abstract	Ovulation is dependent upon numerous factors mediating follicular growth, vascularization, and ultimately oocyte release via follicle rupture. Endothelin-2 (EDN2) is a potent vasoconstrictor that is transiently produced prior to follicle rupture by granulosa cells of periovulatory follicles and induces ovarian contraction. To determine the role of Edn2 expression, surgical transplant and novel conditional knockout mice were super-ovulated and analyzed. Conditional knockout mice utilized a new iCre driven by the Esr2 promoter to selectively remove Edn2. Follicle rupture and fertility were significantly impaired in the absence of ovarian Edn2 expression. When ovaries of Edn2KO mice were transplanted in wild type recipients, significantly more corpora lutea containing un-ovulated oocytes were present after hormonal stimulation (1.0 vs. 5.4, p = 0.010). Following selective ablation of Edn2 in granulosa cells, Esr2-Edn2KO dams had reduced oocytes ovulated (3.8 vs. 16.4 oocytes/ovary) and smaller litters (4.29 ± l.02 vs. 8.50 pups/dam). However, the number of pregnancies per pairing was not different and the reproductive axis remained intact. Esr2-Edn2KO ovaries had a higher percentage of antral follicles and fewer corpora lutea; follicles progressed to the antral stage but many were unable to rupture. Conditional loss of endothelin receptor A in granulosa cells also decreased ovulation but did not affect fecundity. These data demonstrate that EDN2-induced intraovarian contraction is a critical trigger of normal ovulation and subsequent fecundity.
MeSH term(s)	Animals ; Endothelin-2/genetics ; Endothelin-2/metabolism ; Female ; Fertility ; Granulosa Cells/metabolism ; Mice ; Mice, Inbred C57BL ; Ovarian Follicle/cytology ; Ovarian Follicle/metabolism ; Ovarian Follicle/physiology ; Ovulation ; Receptor, Endothelin A/metabolism
Chemical Substances	Endothelin-2 ; Receptor, Endothelin A
Language	English
Publishing date	2017-04-11
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-017-00943-w
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Article ; Online: Generation of an estrogen receptor beta-iCre knock-in mouse.

Cacioppo, Joseph A / Koo, Yongbum / Lin, Po-Ching Patrick / Osmulski, Sarah A / Ko, Chunjoo D / Ko, CheMyong

Genesis (New York, N.Y. : 2000)

2016 Volume 54, Issue 1, Page(s) 38–52

Abstract: A novel knock-in mouse that expresses codon-improved Cre recombinase (iCre) under regulation of the estrogen receptor beta (Esr2) promoter was developed for conditional deletion of genes and for the spatial and/or temporal localization of Esr2 expression. ...

Abstract	A novel knock-in mouse that expresses codon-improved Cre recombinase (iCre) under regulation of the estrogen receptor beta (Esr2) promoter was developed for conditional deletion of genes and for the spatial and/or temporal localization of Esr2 expression. ESR2 is one of two classical nuclear estrogen receptors and displays a spatiotemporal expression pattern and functions that are different from the other estrogen receptor, ESR1. A cassette was constructed that contained iCre, a polyadenylation sequence, and a neomycin selection marker. This construct was used to insert iCre in front of the endogenous start codon of the Esr2 gene of a C57BL/6J embryonic stem cell line via homologous recombination. Resulting Esr2-iCre mice were bred with ROSA26-lacZ and Ai9-RFP reporter mice to visualize cells of functional iCre expression. Strong expression was observed in the ovary, the pituitary, the interstitium of the testes, the head and tail but not body of the epididymis, skeletal muscle, the coagulation gland (anterior prostate), the lung, and the preputial gland. Additional diffuse or patchy expression was observed in the cerebrum, the hypothalamus, the heart, the adrenal gland, the colon, the bladder, and the pads of the paws. Overall, Esr2-iCre mice will serve as a novel line for conditionally ablating genes in Esr2-expressing tissues, identifying novel Esr2-expressing cells, and differentiating the functions of ESR2 and ESR1.
MeSH term(s)	Animals ; Cell Line ; Estrogen Receptor beta/biosynthesis ; Estrogen Receptor beta/genetics ; Estrogen Receptor beta/metabolism ; Female ; Gene Transfer Techniques ; Integrases/chemistry ; Integrases/metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Ovary/metabolism ; Ovary/physiology ; Promoter Regions, Genetic ; Testis/metabolism ; Testis/physiology ; Transcriptome
Chemical Substances	Estrogen Receptor beta ; Cre recombinase (EC 2.7.7.-) ; Integrases (EC 2.7.7.-)
Language	English
Publishing date	2016-01
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2004544-X
ISSN	1526-968X ; 1526-954X
ISSN (online)	1526-968X
ISSN	1526-954X
DOI	10.1002/dvg.22911
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Article ; Online: Loss of function of endothelin-2 leads to reduced ovulation and CL formation.

Cacioppo, Joseph A / Oh, Sang Wook / Kim, Hey-young / Cho, Jongki / Lin, Po-Ching Patrick / Yanagisawa, Masashi / Ko, CheMyong

PloS one

2014 Volume 9, Issue 4, Page(s) e96115

Abstract: Endothelin-2 (EDN2), a potent vasoconstrictive peptide, is transiently produced by periovulatory follicles at the time of ovulation when corpus luteum (CL) formation begins. EDN2 induces contraction of ovarian smooth muscles ex vivo via an endothelin ... ...

Abstract	Endothelin-2 (EDN2), a potent vasoconstrictive peptide, is transiently produced by periovulatory follicles at the time of ovulation when corpus luteum (CL) formation begins. EDN2 induces contraction of ovarian smooth muscles ex vivo via an endothelin receptor A-mediated pathway. In this study, we aimed to determine if EDN2 is required for normal ovulation and subsequent CL formation in?vivo. In the ovaries of a mouse model that globally lacks the Edn2 gene (Edn2 knockout mouse; Edn2KO), histology showed that post-pubertal Edn2KO mice possess follicles of all developmental stages, but no corpora lutea. When exogenous gonadotropins were injected to induce super-ovulation, Edn2KO mice exhibited significantly impaired ovulation and CL formation compared to control littermates. Edn2KO ovaries that did ovulate in response to gonadotropins did not contain histologically and functionally identifiable CL. Intra-ovarian injection of EDN2 peptide results suggest partial induction of ovulation in Edn2KO mice. Endothelin receptor antagonism in wild type mice similarly disrupted ovulation, CL formation, and progesterone secretion. Overall, this study suggests that EDN2 is necessary for normal ovulation and CL formation.
MeSH term(s)	Animals ; Corpus Luteum/abnormalities ; Corpus Luteum/blood supply ; Corpus Luteum/physiology ; Endothelin Receptor Antagonists/pharmacology ; Endothelin-2/metabolism ; Female ; Gonadotropins/pharmacology ; Mice, Inbred C57BL ; Mice, Knockout ; Neovascularization, Physiologic ; Ovarian Follicle/cytology ; Ovulation/metabolism
Chemical Substances	Endothelin Receptor Antagonists ; Endothelin-2 ; Gonadotropins
Language	English
Publishing date	2014-04-24
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ISSN	1932-6203
ISSN (online)	1932-6203
DOI	10.1371/journal.pone.0096115
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Article ; Online: Prenatal Exposure to DEHP Induces Premature Reproductive Senescence in Male Mice.

Barakat, Radwa / Lin, Po-Ching Patrick / Rattan, Saniya / Brehm, Emily / Canisso, Igor F / Abosalum, Mohamed E / Flaws, Jodi A / Hess, Rex / Ko, CheMyong

Toxicological sciences : an official journal of the Society of Toxicology

2017 Volume 156, Issue 1, Page(s) 96–108

Abstract: Di-(2-ethylhexyl) phthalate (DEHP) is the most commonly used phthalate, and it is an endocrine-disrupting chemical. This study tested a hypothesis that prenatal exposure to DEHP lays the foundation for premature gonadal dysfunction and subsequent ... ...

Abstract	Di-(2-ethylhexyl) phthalate (DEHP) is the most commonly used phthalate, and it is an endocrine-disrupting chemical. This study tested a hypothesis that prenatal exposure to DEHP lays the foundation for premature gonadal dysfunction and subsequent reproductive senescence in male mice. Pregnant female CD-1 mice were orally dosed with vehicle control (tocopherol-stripped corn oil) or with 20 μg/kg/day, 200 μg/kg/day, 500 mg/kg/day, or 750 mg/kg/day of DEHP from gestational day 11 to birth. Overall, the prenatal DEHP exposure did not cause any overt physical health problems in male offspring, as no significant differences in their body nor gonadal weight were seen up to the age of 23 months. However, an age- and dose-dependent gonadal dysfunction was observed. As early as 7 months of age, the 750 mg/kg/day group of mice exhibited significantly reduced fertility. At 19 months of age, 86% of the 750 mg/kg/day mice became infertile, whereas only 25% of the control mice were infertile. At this age, all of the DEHP-exposed mice had lower serum testosterone levels, higher serum estradiol levels, and higher LH levels compared with control mice. Histological evaluations showed that mice prenatally exposed to DEHP displayed a wide array of gonadal and epididymal abnormalities such as increased germ cell apoptosis, degenerative seminiferous tubules, oligozoospermia, asthenozoospermia, and teratozoospermia in comparison to age-matching control mice. In summary, this study shows that prenatal exposure to DEHP induces premature reproductive senescence in male mice.
MeSH term(s)	Administration, Oral ; Aging, Premature/blood ; Aging, Premature/chemically induced ; Aging, Premature/pathology ; Animals ; Animals, Outbred Strains ; Apoptosis/drug effects ; Diethylhexyl Phthalate/administration & dosage ; Diethylhexyl Phthalate/toxicity ; Dose-Response Relationship, Drug ; Endocrine Disruptors/administration & dosage ; Endocrine Disruptors/toxicity ; Epididymis/drug effects ; Epididymis/pathology ; Estradiol/blood ; Female ; Infertility, Male/blood ; Infertility, Male/chemically induced ; Infertility, Male/pathology ; Luteinizing Hormone/blood ; Male ; Mice ; Organ Size/drug effects ; Plasticizers/administration & dosage ; Plasticizers/toxicity ; Pregnancy ; Prenatal Exposure Delayed Effects ; Survival Analysis ; Testis/drug effects ; Testis/pathology ; Testosterone/blood
Chemical Substances	Endocrine Disruptors ; Plasticizers ; Testosterone (3XMK78S47O) ; Estradiol (4TI98Z838E) ; Luteinizing Hormone (9002-67-9) ; Diethylhexyl Phthalate (C42K0PH13C)
Language	English
Publishing date	2017--01
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S. ; Research Support, Non-U.S. Gov't
ZDB-ID	1420885-4
ISSN	1096-0929 ; 1096-6080
ISSN (online)	1096-0929
ISSN	1096-6080
DOI	10.1093/toxsci/kfw248
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Article ; Online: Periovulatory leukocyte infiltration in the rat ovary.

Oakley, Oliver R / Kim, HeyYoung / El-Amouri, Ismail / Lin, Po-Ching Patrick / Cho, Jongki / Bani-Ahmad, Mohammad / Ko, Chemyong

Endocrinology

2010 Volume 151, Issue 9, Page(s) 4551–4559

Abstract: Ovulation is preceded by intraovarian inflammatory reactions that occur in response to the preovulatory gonadotropin surge. As a main inflammatory event, leukocytes infiltrate the ovary and release proteolytic enzymes that degrade the extracellular ... ...

Abstract	Ovulation is preceded by intraovarian inflammatory reactions that occur in response to the preovulatory gonadotropin surge. As a main inflammatory event, leukocytes infiltrate the ovary and release proteolytic enzymes that degrade the extracellular matrix weakening the follicular wall, a required step for follicle rupture. This study aimed to quantitatively measure the infiltrating leukocytes, determine their cell types, and localize infiltration sites in the periovulatory rat ovary. Cycling adult and gonadotropin-stimulated immature rats were used as animal models. Ovaries were collected at five different stages of estrous cycle in the adult rats (diestrus, 1700 h; proestrus, 1500 h; proestrus, 2400 h; estrus, 0600 h; and metestrus, 1700 h) and at five different time points after superovulation induction in the immature rats (pregnant mare's serum gonadotrophin, 0 h; pregnant mare's serum gonadotrophin, 48 h; human chorionic gonadotropin, 6 h; human chorionic gonadotropin, 12 h; and human chorionic gonadotropin, 24 h). The ovaries were either dissociated into a single cell suspension for flow cytometric analysis or fixed for immunohistochemical localization of the leukocytes. Similar numbers of leukocytes were seen throughout the estrous cycle (approximately 500,000/ovary), except proestrus 2400 when 2-fold higher numbers of leukocytes were found (approximately 1.1 million/ovary). A similar trend of periovulatory rise of leukocyte numbers was seen in the superovulation-induced immature rat model, recapitulating a dramatic increase in leukocyte numbers upon gonadotropin stimulation. Both macrophage/granulocytes and lymphocytes were among the infiltrating leukocytes and were localized in the theca and interstitial tissues, where platelet-endothelial cell adhesion molecule-1 and intercellular adhesion molecule-1 may play roles in the transmigration of leukocytes, because their expressions correlates spatiotemporally with the infiltrating leukocytes. In addition, a strong inverse relationship between leukocyte numbers in the ovary and spleen, as well as significant reduction of leukocyte infiltration in the splenectomized rats, were seen, indicating that the spleen may serve as an immediate supplier of leukocytes to the periovulatory ovary.
MeSH term(s)	Animals ; CD11b Antigen/analysis ; CD11c Antigen/analysis ; CD3 Complex/analysis ; Cell Movement/drug effects ; Estrous Cycle/physiology ; Female ; Flow Cytometry ; Gonadotropins/pharmacology ; Humans ; Immunohistochemistry ; Intercellular Adhesion Molecule-1/analysis ; Leukocyte Common Antigens/analysis ; Leukocyte Count ; Leukocytes/cytology ; Leukocytes/metabolism ; Ovary/drug effects ; Ovary/metabolism ; Ovary/physiology ; Ovulation/physiology ; Platelet Endothelial Cell Adhesion Molecule-1/metabolism ; Rats ; Rats, Sprague-Dawley ; Spleen/cytology ; Spleen/surgery ; Splenectomy ; Time Factors
Chemical Substances	CD11b Antigen ; CD11c Antigen ; CD3 Complex ; Gonadotropins ; Platelet Endothelial Cell Adhesion Molecule-1 ; Intercellular Adhesion Molecule-1 (126547-89-5) ; Leukocyte Common Antigens (EC 3.1.3.48)
Language	English
Publishing date	2010-06-30
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	427856-2
ISSN	1945-7170 ; 0013-7227
ISSN (online)	1945-7170
ISSN	0013-7227
DOI	10.1210/en.2009-1444
Database	MEDical Literature Analysis and Retrieval System OnLINE

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