• Animal cells and systems
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• v.3 no.1
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• pp.1-7
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• 1999

Gonadotropin-releasing hormone (GnRH) was originally isolated as a hypothalamic peptide that regulates reproduction by stimulating the release of gonadotropins. Using comparative animal models has led to the discovery that GnRH has a more ancient evolutionary origin. Durinq evolution GnRH peptide underwent gene duplication and structural changes to give rise to multiple molecular forms of GnRHs. Mammalian GnRH initially considered to be the sole molecular form, is now grouped as a family of peptides along with GnRH variants determined from representatives in all classes of vertebrates. Vertebrate species including primates and humanshave more than one GnRH variant in individual brains; a unique GnRH form in the forebrain and chicken IIGnRH in the midbrain. Furthermore, several species of bony fish have three molecular variants of GnRH: salmon GnRH sea-bream GnRH and chicken II GnRH. Also, it has been shown that in addition to the olfactory placodes and the midbrain, there is a third embryonic source of GnRH neurons from the basal diencephalon in birds and fish, which might be true for other vertebrates. Therefore, comparative animal models like fish with discrete sites of expression of three molecular variants of GnRH in individual brains, could provide insight into novel functions of GnRH variants, conservation of gene regulation, and mechanisms governing reproduction in vertebrates.

• Clinical and Experimental Reproductive Medicine
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• v.47 no.2
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• pp.94-100
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• 2020

The inverse correlation between maternal age and pregnancy rate represents a major challenge for reproductive endocrinology. The high embryo ploidy error rate in failed in vitro fertilization (IVF) cycles reflects genetic misfires accumulated by older oocytes over time. Despite the application of different follicular recruitment protocols during IVF, gonadotropin modifications are generally futile in addressing such damage. Even when additional oocytes are retrieved, quality is frequently poor. Older oocytes with serious cytoplasmic and/or chromosomal errors are often harvested from poorly perfused follicles, and ovarian vascularity and follicular oxygenation impact embryonic chromosomal competency. Because stimulation regimens exert their effects briefly and immediately before ovulation, gonadotropins alone are an ineffective antidote to long-term hypoxic pathology. In contrast, the tissue repair properties (and particularly the angiogenic effects) of platelet-rich plasma (PRP) are well known, with applications in other clinical contexts. Injection of conventional PRP and/or its components (e.g., isolated platelet-derived growth factors as a cell-free substrate) into ovarian tissue prior to IVF has been reported to improve reproductive outcomes. Any derivative neovascularity may modulate oocyte competence by increasing cellular oxygenation and/or lowering concentrations of intraovarian reactive oxygen species. We propose a mechanism to support intrastromal angiogenesis, improved follicular perfusion, and, crucially, embryo ploidy rescue. This last effect may be explained by mRNA upregulation coordinated by PRP-associated molecular signaling, as in other tissue systems. Additionally, we outline an intraovarian injection technique for platelet-derived growth factors and present this method to help minimize reliance on donor oocytes and conventional hormone replacement therapy.

• Clinical and Experimental Reproductive Medicine
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• v.47 no.1
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• pp.12-19
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• 2020

Objective: The appropriate function of the hypothalamic-pituitary-gonadal axis is essential for maintaining proper reproductive function. In female mammals, the hypothalamic-pituitary-gonadal axis regulates reproductive changes that take place in the estrus cycle and are necessary for successful reproduction. This study was conducted to investigate the effect of thymectomy on the estrus cycle in neonatally thymectomized guinea pigs. Methods: In this study, 12 female guinea pigs, six thymectomized and six sham-operated, were studied. The effects of neonatal thymectomy at 5-7 days of age on parameters of the reproductive axis were examined in female guinea pigs. Gonadotropin and 17β-estradiol levels were assessed at regular intervals (days 0, 3, 6, 9, 12, and 15) of the estrus cycle, and the time of vaginal opening in the thymectomized and shamoperated guinea pigs was determined. Results: Significant reductions in gonadotropins and 17β-estradiol levels during estrus cycle were found in neonatally thymectomized female guinea pigs compared to sham-operated guinea pigs. Conclusion: The results of this study underscore the importance of the thymus in the neonatal period for normal female reproductive function.

• Development and Reproduction
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• v.26 no.2
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• pp.37-47
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• 2022

Photoperiod has well been established to regulate testicular activities in golden hamsters. These animals breed actively around summer but become infertile in winter. In males, testicles are full of multistep germ cells including spermatozoa in summer. But in winter only fundamental cells consisting of the testicles are detected. The testicular degeneration is accompanied by the reduced levels of blood gonadotropins and testosterone. In this study, the expressions of gonadotropin subunit genes were investigated in the reproductive active and inactive testicles. And parts of sequences of the gonadotropin subunits were identified and compared with those of other rodents. As results, common gonadotropin alpha (CGa), follicle-stimulating hormone (FSH) β, and luteinizing hormone (LH) β genes were equivalently detected in pituitaries of both sexually active and inactive animals. In considering low concentrations of gonadotropin hormones determined in pituitary, the present findings imply that the processes involved in translation and/or formation of functional hormones could be impeded in the sexually inactive hamsters. All the nucleotide sequences of gonadotropin subunits identified in this study were same as those reported previously except for one base in CGa. An unsure amino acid deduced from the CGa sequence was confirmed from mRNA sequencing. The outcomes mentioned above suggest that animals with regressed testes prepare for the sexually active period forthcoming in the future.

• Clinical and Experimental Reproductive Medicine
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• v.44 no.4
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• pp.232-238
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• 2017

Objective: To determine whether reducing the cetrorelix dose in the antagonist protocol to 0.125 mg had any deleterious effects on follicular development, the number and quality of retrieved oocytes, or the number of embryos, and to characterize its effects on the affordability of assisted reproductive technology. Methods: This randomized controlled study was conducted at the Fertility Unit of Tanta Educational Hospital of Tanta University, the Egyptian Consultants' Fertility Center, and the Qurrat Aien Fertility Center, from January 1 to June 30, 2017. Patients' demographic data, stimulation protocol, costs, pregnancy rate, and complications were recorded. Patients were randomly allocated into two groups: group I (n = 61) received 0.125 mg of cetrorelix (the study group), and group II (n = 62) received 0.25 mg of cetrorelix (the control group). Results: The demographic data were comparable regarding age, parity, duration of infertility, and body mass index. The dose of recombinant follicle-stimulating hormone units required was $2,350.43{\pm}150.76$ IU in group I and $2,366.25{\pm}140.34$ IU in group II, which was not a significant difference (p= 0.548). The duration of stimulation, number of retrieved oocytes, and number of developed embryos were not significantly different between the groups. The clinical and ongoing pregnancy rates likewise did not significantly differ. The cost of intracytoplasmic sperm injection per cycle was significantly lower in group I than in group II (US $ $494.66{\pm}4.079$ vs. US $ $649.677{\pm}43.637$). Conclusion: Reduction of the cetrorelix dose in the antagonist protocol was not associated with any significant difference either in the number of oocytes retrieved or in the pregnancy rate. Moreover, it was more economically feasible for patients in a low-resource country.

• Journal of Veterinary Clinics
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• v.10 no.2
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• pp.227-235
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• 1993

The effectiveness of sodium carboxymethylcellulose(SCMC) and dextran 70 in the prevention of adhesion formation in abdominal cavity of rabbits following artificial injuries was elucidated and the effects -of these medicines on blood pictures were also examined. After abrasion treatment on jejunum in gonadotropins primed rabbits, 1, 2 and 3% of SCMC solutions, 6 and 10% dextran 70 solutions and a synthetic solution of 1% SCMC and loft dextran 70 in 0.9% saline solution were infused into the abdominal cavity. Four weeks later the abdominal cavities were reopened under general anesthesia. The synthetic solution showed the highest adhesion reduction rate(100%), while 1% SCMC, 6 and loft dextran solutions showed no significantly evident effect of adhesion prevention. The SCMC solutions showed better adhesion reduction effect than dextran 70 solutions. After infusion of these adhesion preventive medicines, the changes of total leucocytes, erythrocytes, lymphocytes, PCV, plasma fibrinogen and protein contents were examined. No remarkable difference in blood pictures was shown between the synthetic solution and the other medicines. Therefore, it can be suggested that the synthetic solution of 1% SCMC and 10% dextran 70 in 0.9% saline solution at dose of 5$m\ell$/kg of body weight is prominently effective in the prevention of postoperative adhesion formation and wolf be safe in animals and human.

• Development and Reproduction
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• v.21 no.3
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• pp.327-333
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• 2017

Gonadotropins are heterodimers consisting an alpha chain ($Cg{\alpha}$) and a beta chain. Interestingly, presence of complicated $LH-{\beta}$ transcripts in rat testis was accidently found; testicular $LH-{\beta}$ transcripts were confined in seminiferous tubules to spermatids, and the translated products were localized in the elongated spermatids. We hypothesized that mouse testis has potential to produce the tissue specific $LH-{\beta}$ with similar structure to the rat testicular forms. To verify our hypothesis, we examined the adult mouse (ICR) testis using RT-PCR and immunohistochemistry. The PCR revealed the presence of the identical products in the reactions for three LH subunit types. The expected product sizes for mouse $Cg{\alpha}$ and $LH-{\beta}$ known as pituitary type were 224 bp and 503 bp, respectively. The testicular type $LH-{\beta}$ products were produced by a primer set based on the rat sequences, with unexpected size of 800 bp. Sequencing revealed that the proximal and distal parts (2-82 and 661- 773 bp, respectively) were homologous to rat testicular $LH-{\beta}$ cDNA, and middle part (83-660 bp) was a unique mouse-specific region. Both $Cg{\alpha}$ and $LH-{\beta}$ positive signals were in the round and elongated spermatids and mature sperms, and the $LH-{\beta}$ signals were more intense. In conclusion, our study demonstrated that the presence and localization of the LH subunits in mouse testis. Further studies will be needed to understand the precise structure and function of mouse testicular LH.

• Development and Reproduction
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• v.24 no.1
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• pp.43-52
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• 2020

NUCB2/nesfatin-1 known to regulate appetite and energy homeostasis is expressed not only in the hypothalamus, but also in various organs and tissues. Our previous reports also demonstrated that NUCB2/nesfatin-1 was expressed in the reproductive organs, including the ovaries, uterus, and testes of mice. However, it is yet known whether NUCB2/nesfatin-1 is expressed in the oviduct and how its expression is regulated. Therefore, we investigated the expression of NUCB2/nesfatin-1 in the oviduct and its expression is regulated by gonadotropin. Immunohistochemical staining results showed that nesfatin-1 protein was localized in epithelial cells of the oviduct. As a result of quantitative real-time PCR (qRT-PCR) and Western blot, NUCB2/nesfatin-1 was detected strongly in the oviducts. During the estrus cycle, NUCB2/nesfatin-1 expression in the oviducts was markedly higher in the proestrus stage than in other estrus stages. In order to elucidate whether the expression of NUCB2 mRNA is controlled by the gonadotropins, we injected PMSG and hCG and measured NUCB2 mRNA level in the oviduct after injection. Its level was increased in the oviduct after PMSG injection, but no significant change after hCG injection. In addition, NUCB2 mRNA levels were markedly reduced after ovariectomy, while recovered after 17β-estradiol (E2) injection, but not by progesterone (P4). This study demonstrated that NUCB2/nesfatin-1 is highly expressed in the oviduct of mouse and its expression is regulated by E2 secreted by the ovaries. These results suggest that NUCB2/nesfatin-1 expressed by the oviduct may affect the function of the oviduct regulated by the ovaries.

• Clinical and Experimental Reproductive Medicine
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• v.25 no.3
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• pp.281-286
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• 1998

Premature ovarian failure is a condition causing amenorrhea, hypoestrogenism, and elevated gonadotropins in women younger than 40 years. Many causes of premature ovarian failure were reported, including genetic abnormalities, enzymatic defects, defects in gonadotropin secretion or action, autoimmune disorders, physical and idiopathic causes. Recently, Finnish group reported a point mutation in the follicle stimulating hormone (FSH) receptor gene in premature ovarian failure patients. But it was reported that the group from United States could not find any mutation in FSH receptor gene. So we analysed C566T point mutation of FSH receptor gene using restriction fragment length polymorphism (RFLP) and compared the result between premature ovarian failure patient with idiopathic and known causes. But we did not find 556C${\rightarrow}$T mutation in the FSH receptor gene in both groups. These findings suggest that the missense mutation in the human FSH receptor gene reported in Finnish women with premature ovarian failure is uncommon in Korean women with premature ovarian failure.

• Clinical and Experimental Reproductive Medicine
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• v.16 no.1
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• pp.23-34
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• 1989

Estradiol (E)은 난소내 과립세포 (granulosa cell, GC)를 증가시키고 생식소 자극호르몬과 협동으로 배란을 유도한다. Androgen은 E의 작용과 반대의 작용을 나타내며 여포의 폐쇄요인으로 알려지고 있다. Testosterone(T)이 폐쇄여포의 여포액내 다량 존재하는 것이 알려짐에 따라 난소내에도 자가조절분비(autocrine)또는 paracrine regulation에 의해 작용을 나타낼 것으로 가정되어 난소내 여포가 폐쇄됨에 따라 그 수용체의 변화하는 양상을 조사하고져 하였다. 흰쥐의 과립세포의 세포질에는 $51.3{\pm}6.1$fmol/mg protein의 Estradiol 수용체(ER) ; $153.1{\pm}25.3$의 Testosterone수용체(TR) ; 또한 $35.1{\pm}8.1$의 Progesterone수용체(PR)가 존재하였다. 과립세포내 ER용 세포질내 E를 제거한 후에 정량이 가능하였고 또한 과립세포내에도 TR이 사람에서는 $23.4{\pm}7.2$ fmol/mg protein, 돼지는 $98.5{\pm}23.1$로 상당량 존재함을 관찰하였다. Dihydrotestosterone Enanthate(DHTE)를 100ug/흰쥐의 농도로 처리한 결과 난소내 TR의 농도는 변화가 없이 ER의 농도만 현저히 저하되고 쥐의 난소무게 역시 감소하는 것을 발견하였다. 위의 결과로 보아 난소내에도 스테로이드 호르몬은 autocrine(자가조절)방법으로 작용하며 An-drogen이 난소의 무게를 감소시키는 것은 ER의 수를 감소시켜 E의 작용이 억제되고 여포들이 폐쇄를 일으켜 그 증식이 저하된 때문으로 사료된다.