Our previous study showed that transgenic (TG) pigs harboring human EPO (hEPO) gene have been shown to have reproductive disorders, including low pregnancy rates, irregular estrus cycle and low little size. To investigate these reasons, we assessed estrus behavior (standing response) and plasma $17{\beta}$-estradiol ($E_2$) level, which partly reflect reproductive function, during the estrus cycles after synchronization and superovulation by hormone treatments. Then, we analysed blood composition and expression of hEPO gene in TG pigs. Pigs were injected with PG600. After 10 days, pigs were fed with Regumate porcine for 6 days. Blood samples were collected from jugular vein. Analysis of blood composition and $E_2$ level were measured by Hemavet 950 and $E_2$ ELISA kit, respectively. And, the expression of hEPO gene in reproductive organs was quantitated by real-time RT-PCR. The percentage of estrus behavior in TG was significantly decreased. Hematocrit (HCT), hemoglobin (Hb) concentration and red blood cell (RBC) number were significantly higher in TG than wild type (WT). On the other hand, high expression of hEPO gene in TG was observed in the mammary gland as well as in the uterus. Moreover, plasma $E_2$ level was significantly higher in TG than WT. These results suggest that nonspecific expression of hEPO gene in the other organs of TG may affect blood composition and plasma $E_2$ level, thereby causing reproductive disorders.
Lee, Su-Yeon;Kang, Youn-Jung;Kwon, Jinie;Nishi, Yoshihiro;Yanase, Toshihiko;Lee, Kyung-Ah;Koong, Mi Kyoung
Clinical and Experimental Reproductive Medicine
/
v.47
no.3
/
pp.194-206
/
2020
Objective: The aim of this study was to investigate microRNAs (miRNAs) related to follicle-stimulating hormone (FSH) responsiveness using miRNA microarrays and to identify their target genes to determine the molecular regulatory pathways involved in FSH signaling in KGN cells. Methods: To change the cellular responsiveness to FSH, KGN cells were treated with FSH receptor (FSHR)-specific small interfering RNA (siRNA) followed by FSH. miRNA expression profiles were determined through miRNA microarray analysis. Potential target genes of selected miRNAs were predicted using bioinformatics tools, and their regulatory function was confirmed in KGN cells. Results: We found that six miRNAs (miR-1261, miR-130a-3p, miR-329-3p, miR-185-5p, miR-144-5p and miR-4463) were differentially expressed after FSHR siRNA treatment in KGN cells. Through a bioinformatics analysis, we showed that these miRNAs were predicted to regulate a large number of genes, which we narrowed down to cytochrome P450 family 19 subfamily A member 1 (CYP19A1) and estrogen receptor alpha (ESR1) as the main targets for miR-4463. Functional analysis revealed that miR-4463 is a regulatory factor for aromatase expression and function in KGN cells. Conclusion: In this study, we identified differentially expressed miRNAs related to FSH responsiveness. In particular, upregulation of miR-4463 expression by FSHR deficiency in human granulosa cells impaired 17β-estradiol synthesis by targeting CYP19A1 and ESR1. Therefore, our data might provide novel candidates for molecular biomarkers for use in research into poor responders.
Golden hamsters show the reproductive activity that is determined by the photoperiod (length of light per day). Photoperiod is an environmental factor that is predictable through an entire year. The hamsters are sexually active in summer during which day length exceeds night time. The critical length is at least 12.5 hours of light in a day where reproductive function is maintained. The information of photoperiod is mediated by the pineal gland because removal of pineal gland blocks the influence of photoperiod on reproductive activity. The hamsters without pineal gland maintain sexual activity and promote it in a situation that suppresses gonadal activity. The pineal gland secretes melatonin that reflects the photoperiod. The appropriate administrations of melatonin into both pineal intact and pinealectomized hamsters lead to a gonadal reression. The results suggest that melatonin constitutes a part of control mechanism whereby environmental information is transduced to neuroendocrine signal respensible for the functional integrity of the reproductive system. Despite of the intense studies, the action site of melatonin is on the whole unknown. It is mainly due to the lack of acute efffct of melatonin on the secretion of reproductive hormones. However, sexually regressed animals display the low levelsof gonadotropins and the augmentation of the hypothalamic gonadotropin-releasing hormone (GnRH) content, implying that the antigonadotropic effects either by photoperiod and/or by the treatment of melatonin are mediated by the GnRH neuronal system. The action mechanism by which melatonin exerts its effect on GnRH neuron needs to be investigated. Recent cloning of melatonin receptor will contribute to examine various and putative potencies of melatonin via its anatomical identification and the action mechanism of melatonin on target tissues at the molecular level.
The main objectives of this study were to investigate the concept of family function from the perspective of the contemporary Korean family, and to construct model of change of family function with chronic illness. The hybrid model approach was applied in which three phases(theoretical phase, empirical phase, and analytic phase) of concept development were explored for family functioning. The study was conducted from 1997 to 1998. In empirical phase, two groups of purposive samples were drawn : normal family group composed of six families without ill family member, and ill family group composed of seven families of which wives have rheumatoid arthritis. Only families with child(or children) in primary or secondary schools were included in the study. The results were as follows : In theoretical phase, six dimensions of family concept were emerged : affective, structural, control, cognitive, financial, and reproductive dimension. In order to analyse the Korean normal family function in middle class with middle-aged women, financial and reproductive dimension were not included. In empirical phase, five dimensions(affective, structural, control, cognitive, and external relationship) were found from the normal family data. External relationship dimension is very important factor as a resource of the support, especially when their parents or siblings had no help or support to them. In the affective dimension, Korean family emphasized harmony and balance rather than affective expression between couples and between parents and children. They also showed common goals of the families to solve their problems to control the family members. The priority of the goals was getting into the higher education of their children or helping their unhealthy parents or family members. Six dimensions (affective, structural, control, cognitive, external relationship, and financial) of family functions were emerged from the ill family data. From the analysis of ill family data, types of restructuring house chore after wives illness were developed : (a) negociated, (b) accomodated, and (c) isolated, enduring types. Although the dimensions of family functioning identified in this study are similar to the conceptualizations that exist in the western literature, there were distinct differences in the nature of major themes and subconcepts under these family function dimensions.
Reproductive traits of sows are determined chiefly by the genetic background of the dam herself. Whether the breed of the serviced boars also contributes is not yet clear. A total of 8,946 litters of Landrace (L) sows from 68 farms were examined. Of these, 876, 1,260, and 6,810 litters were produced from natural mating with L, Yorkshire (Y), and Duroc (D) breeds, respectively. The birth litter size (BLS), weaning litter size (WLS), live born piglets (LBP), and survival rate % (SR), of every litter were recorded. A general linear-model procedure was followed to estimate the effects of boars' breed (B), parity (P) and B${\times}$P interaction on the reproductive traits of the sows. Results show that BLS, WLS, and LBP were all significantly (p<0.001) affected by B and P. SR% was significantly influenced by B but not by P. L sows crossbred with Y or purebred with L produced litters with higher BLS, WLS, and LBP values than those bred with D. Pure L breeding yielded litters with lower SR% than did crossbreeding with D, while the difference between SR% due to LY and that due to LD crossbreeding was not significant. The interaction of B with P was significant with respect to BLS, WLS, and LBP (p<0.001), but not SR%. No significant B effect on reproductive traits was measured in sows at their first parity; but at latter parities, LL or LY produced litters with similarly high BLS, WLS and LBP, which values were all significantly greater than those of LD litters. The breed of boar evidently affected the subsequent reproductive performance of L sows and this effect may be further manipulated by the parity effect. Breed differences in semen quality and the success of fetus development with different interactions of the genetic background with the uterus function of the sow that may contribute to these effects are discussed.
Studies of the red algal genus Bostrychia over the last 15 years have made it a model system for many evolutionary processes within red algal species. The combination of newly developed, or first employed methods, in red algal species studies has made Bostrychia a pioneer genus in intraspecific studies. Bostrychia was the first genus in which a mitochondrial marker was used for intraspecific red algal phylogeny, and the first for which a 3-genome phylogeny was undertaken. The genus was the first red alga used to genetically show maternal plastid and mitochondria inheritance, and also to show correlation between cryptic species (genetically divergent intraspecific lineages) and reproductive incompatibility. The chemotaxonomic use, and physiological function of osmolytes, has also been extensively studied in Bostrychia. Our continuous studies of Bostrychia also highlight important aspects in algal species studies. Our worldwide sampling, and resampling in certain areas, show that intensive sampling is needed to accurately assess the genetic diversity and therefore phylogeographic history of algal species, with increased sampling altering evolutionary hypotheses. Our studies have also shown that long-term morphological character stability (stasis) and character convergence can only be correctly assessed with wide geographic sampling of morphological species. While reproductive incompatibility of divergent lineages supports the biological species nature of these lineages, reproductive incompatibility is also seen between isolates with little genetic divergence. It seems that reproductive incompatibility may evolve quickly in red algae and the unique early stages of fertilization (e.g., gametes covered by walls, active movement of spermatium nuclei to the distant egg nucleus), also well investigated in Bostrychia,. may be key to our understanding of this process.
Sills, E. Scott;Obregon-Tito, Alexandra J.;Gao, Harry;McWilliams, Thomas K.;Gordon, Anthony T.;Adams, Catharine A.;Slim, Rima
Clinical and Experimental Reproductive Medicine
/
v.44
no.1
/
pp.40-46
/
2017
Objective: To describe in vitro development of human embryos derived from an individual with a homozygous pathogenic variant in NLRP7 (19q13.42) and recurrent hydatidiform mole (HM), an autosomal recessive condition thought to occur secondary to an oocyte defect. Methods: A patient with five consecutive HM pregnancies was genomically evaluated via next generation sequencing followed by controlled ovarian hyperstimulation, in vitro fertilization (IVF) with intracytoplasmic sperm injection, embryo culture, and preimplantation genetic screening. Findings in NLRP7 were recorded and embryo culture and biopsy data were tabulated as a function of parental origin for any identified ploidy error. Results: The patient was found to have a pathogenic variant in NLRP7 (c.2810+2T>G) in a homozygous state. Fifteen oocytes were retrieved and 10 embryos were available after fertilization via intracytoplasmic sperm injection. Developmental arrest was noted for all 10 embryos after 144 hours in culture, thus no transfer was possible. These non-viable embryos were evaluated by karyomapping and all were diploid biparental; two were euploid and eight had various aneuploidies all of maternal origin. Conclusion: This is the first report of early human embryo development from a patient with any NLRP7 mutation. The pathogenic variant identified here resulted in global developmental arrest at or before blastocyst stage. Standard IVF should therefore be discouraged for such patients, who instead need to consider oocyte (or embryo) donation with IVF as preferred clinical methods to treat infertility.
Seo, Hee-Won;Park, Kyung-Je;Lee, Hyung-Chul;Kim, Dae-Yong;Song, Yong-Sang;Lim, Jeong-Mook;Song, Gwon-Hwa;Han, Jae-Yong
Journal of Animal Science and Technology
/
v.51
no.6
/
pp.485-492
/
2009
Estrogen has dramatic effects on the development and function of the reproductive tract in mammals. Although diethylstilbestrol (DES) triggers the development of reproductive organs in immature animals, continued exposure to DES induces dysfunction of the female reproductive tract in mice. To investigate the effects of neonatal estrogen exposure on the reproductive tract of female chickens, we implanted DES pellets into the abdominal region of immature female chicks and then examined the effects of DES on the oviducts of both immature chicks and sexually mature chickens (30 weeks old). DES induced mass growth and differentiation of the oviduct in immature chicks. The chick oviduct increased by 2.7- and 29-fold in length and weight, respectively, following primary DES stimulation. In secondary DES stimulation, the length and weight of the chick oviduct increased by 4.5- and 74-fold, respectively. Additionally, DES treatments caused abnormal development of the infundibulum and magnum in hen oviducts. Furthermore, infundibulum abnormality gave rise to unusual ovulation of follicles and resulted in infertility and dysfunction of the magnum, such as less production of egg white proteins. Our results indicate that DES exposure during early developmental stages in chickens has detrimental effects on the development and maintenance of the female reproductive tract after sexual maturation.
Objective: Stress is known to be an inhibitor of the reproductive hypothalamic-pituitary-gonadal (HPG) axis. However, the neural and molecular connections between stress and reproduction are not yet understood. It is well established that in both humans and rodents, kisspeptin (encoded by the kiss1 gene) is a strong stimulator of the HPG axis. In the present study we hypothesized that endocannabinoids, an important neuromodulatory system in the brain, can act on the HPG axis at the level of kiss1 expression to inhibit reproductive function under stress. Methods: Adult male Wistar rats were unilaterally implanted with an intracerebroventricular cannula. Afterwards, the animals were exposed to immobilization stress, with or without the presence of the cannabinoid CB1 receptor antagonist AM251 (1 ${\mu}g/rat$). Blood samples were collected through a retro-orbital plexus puncture before and after stress. Five hours after the stress, brain tissue was collected for reverse transcriptase-quantitative polymerase chain reaction measurements of kiss1 mRNA. Results: Immobilization stress (1 hour) resulted in a decrease in the serum luteinizing hormone concentration. Additionally, kiss1 gene expression was decreased in key hypothalamic nuclei that regulate gonadotrophin secretion, the medial preoptic area (mPOA), and to some extent the arcuate nucleus (ARC). A single central administration of AM251 was effective in blocking these inhibitory responses. Conclusion: These findings suggest that endocannabinoids mediate, at least in part, immobilization stress-induced inhibition of the reproductive system. Our data suggest that the connection between immobilization stress and the HPG axis is kiss1 expression in the mPOA rather than the ARC.
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