• 한국발생생물학회:학술대회논문집
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• 한국발생생물학회 2003년도 제3회 국제심포지움 및 학술대회
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• pp.70-70
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• 2003

The uterus undergoes dynamic changes during the cycle and displays many features typical of developmental process. In order to be prepared for implantation, endometrium undergoes predictable, sequential phases of proliferation and secretory changes. The uterus during estrus cycle synthesize a complex of signaling molecules with specific spatial and temporal modes of expression and which are critical for cell proliferation and differentiation. The purpose of this investigation was to use cDNA microarrays to evaluate the expression of genes of rat uterus in estrus cycle. Animals were sacrificed on proestrus, estrus, metestrus, diestrus. Differential gene expression profiles were revealed(growth-related c-myc reponsive protein RCL, heat shock 47-kDa protein (HSP47), cytochrome c oxidase polypeptide Vlc2 (COX6C2), calreticulin (CALR)). Reverse transcription polymerase chain reaction (RT-PCR) was used to validate the relative expression pattern. Using this approach, we found several genes whose expression in rat uterus was altered with estrus cycle. Our long-term goal is to determine the role of these differentially expressed genes during estrus cycle. This study was supported by through the Biohealth Products Research Center(BPRC), Inje University.

• 한국수정란이식학회지
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• 제33권2호
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• pp.61-68
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• 2018

The major focus of this study is to analyze the expression of bovine MMPs and to monitor their activity during the estrus cycle and pregnancy. During pregnancy, MMP-2 expression was detectable around 30 days but became insignificant by 60 days, then started to increase again around 90 days and reached the maximum at 250 days. The activity of MMP-2 protein changed in accordance with its expression level. As expected, the level of TIMP-2 exhibited a reverse pattern. About MMP-9, high level expression was observed as early as 30 days and gradually increase until 90 days. Then started to decrease after 250 days. Again, the sites of MMP-9 expression were similar to those of MMP-2. On the other hand, expression of TIMP-3 remained low until 90 days but showed a small and temporal increase around 250 days. In summary, expression of different MMPs were differentially regulated during estrus cycle and pregnancy. While the expression of MMP-2 was high in estrus cycle, MMP-9 slowly takes over with the progression of pregnancy. These results indicated that the luteal tissue perform distinct functions during pregnancy and estrus. Perhaps the activity of MMP-2 is required for the structural remodeling of luteum, resulting the suppression of P4 inflow from blood. On the other hand, steady maintenance of MMP-9 throughout luteal development is important for the activation of cell proliferation, maturation and angiogenesis.

• 한국동물생명공학회지
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• 제38권2호
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• pp.62-69
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• 2023

Background: Research on the reproductive physiology of Water and Sika deer, an endemic in Korea, still needs to be completed. This study analyzed the ovarian development and morphological characteristics of wild Water deer and Sika deer. Methods: Water deer and Sika deer ovaries were collected from the Korean Peninsula and Russia-Korean Peninsula border during the estrus and pregnancy seasons, respectively. And, morphological and physiological analysis and immunohistochemistry were conducted to confirm the detection of Ca²⁺ and assess the morphological changes in the ovaries. Results: The results of morphological analysis of ovaries during pregnancy and estrus, the development of the corpus luteum and follicles of Water deer showed similar patterns to other mammals. In contrast, the corpus luteum of Sika deer differed in tissue morphology and composition from Water deer. Ca²⁺ related to tissue metabolism was detected in the theca cells zone of Water deer on the estrus and was highly detected in the luteum cells zone during pregnancy. The hormone receptor protein expression patterns were generally higher in the ovaries of Water deer on the estrus and the pregnancy than in Sika deer. The expression of LH receptor was relatively low in the lutein cell zone, unlikely that of Water deer. The expression of VEGF was also different from Water deer, and the response in Sika deer was relatively very low compared to Water deer in expressing all proteins-related development. Conclusions: Therefore, the results of the study were shown that the composition of the corpus luteum of Sika deer is not clear compared to Water deer, and there are many differences in the functional and morphological formation of the corpus luteum.

• 한국발생생물학회지:발생과생식
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• 제7권1호
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• pp.41-48
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• 2003

본 연구는 생식주기 중의 흰쥐 자궁에서 발현되는 유전자들 중 adhesion수용체 유전자들의 발현이 프로게스테론(P$_4$)에 의하여 차별적으로 조절되는 실험을 하였다. 첫째 실험군은 난소절제 흰쥐와 배란기 흰쥐를 사용하였고(OVX/estrus), 둘째 실험군은 난소절제 흰쥐와 난소절제 후 P4를 주사한 흰쥐를 사용하였다(OVX/OVX+P$_4$). 적출한 자궁조직에서 total RNA를 추출, [$\alpha$$^{32}$P]-dATP로 probe를 제작한 후 Rat Atlas away 1.2 II(Clontech)을 이용하여 발현되는 유전자들을 선별하였으며, 그 중 adhesion 수용체 유전자들의 발현양상을 RT-PCR 방법으로 확인하였다. OVX/estrus 자궁의 유전자 발현을 비교한 경우, 전체 1176개의 유전자들 중 P4에 의해 발현이 증가되는 adhesion 수용체 유전자들은 embigin protein, activated leukocyte cell adhesion molecule, afadin, neuroligin 2, semaphorin Z, osteonectin 등 이었다. OVX/OVX+P$_4$자궁의 유전자 발현을 비교한 경우, P$_4$에 의해 발현이 증가되는 adhesion수용체 유전자들은 osteonectin, afadin, neuroligin 2, semaphorin Z 등 이었다. 그리고 afadin, neuroligin 2, semaphorin Z은 두 실험 군에 서 모두 유전자 발현이 증가되었다. 이러한 결과로 보아 이 유전자들은 P$_4$에 의하여 발현이 조절되어 배란 후 착상 준비에 관여할 것으로 추측된다.

• 한국수정란이식학회지
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• 제27권2호
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• pp.93-100
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• 2012

To investigate the biochemical nature of changes in vaginal physiology during estrus and pregnancy, we examined the cytology and viscosity, and monitored the protein expression profile in vaginal mucus during estrus and pregnancy. The viscosity progressively decreased from estrus to pregnancy. Cell type analysis revealed that white blood cells progressively increased from estrus to pregnancy, while red blood cells progressively decreased during pregnancy. The cornification index (CI) was higher in estrus than in pregnancy. Protein mass spectrumetry identified the presence of ribosome-binding protein 1, GRIP 1 (Glutamate receptor-interacting protein 1)-associated protein 1, DUF729 (Domain of unknown function729) domain-containing protein 1, prolactin precursor, dihydrofolatereductase, and MMP (Matrix metalloprotease)-9 in vaginal mucus. MMP-2 and MMP-9 proteins in the vaginal mucus were active throughout estrus and gestation, as measured by a gelatinase assay, but most abundant in the vaginal mucus on day 0 of estrus. Results from ELISA of MMP-2 and MMP-9 were in accordance with the gelatinase assay. In light of the crucial role of metalloproteinases in extracellular matrix remodeling, the level of MMP-9 in vaginal mucus might be useful as an indicator of estrus and pregnancy to increase the efficiency of reproduction.

• 한국수정란이식학회지
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• 제23권4호
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• pp.251-255
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• 2008

The aim of the present recent study was to compare the protein patterns in the vaginal mucus of Hanwoo cattles during spontaneous and CIDR induced-estrus. Ten cattles, who had been observed in estrus, received no treatment and served as the group of cattles with normal spontaneous estrus. Thirteen cattles in the CIDR received an CIDR insert on day 14 were removed and cattles were injected GnRH on day 15. Vaginal mucus samples were collected from all cattles at the same time the single AI in cattles with spontaneous estrus and the AI in cattles with induced estrus. Spontaneous and CIDR-induced estrus vaginal mucus samples were analyzed on two different array surfaces: cation-exchange (CM10), anion-exchange (Q10). In addition, using the NaCl solution by which the proteins combined after washing are 0.5, 1 and 2 M, it was fractionated and a protein was collected successively. The results are summarized as follows: 1) Ionic surfaces chemistries (Q10 and CM10) gave the best results in terms of detectable protein peaks, with more than 100 protein peaks in the two fractions and under each condition. 2) Protein mass spectrometer using 11 different proteins in protein identification of 7 were able to determine the protein. List of identified proteins as follows; Ribosome-binding protein 1, GRIP 1-associated protein 1, Katanin p60 ATPase-containing subunit A-like 1, Protein FAM44A, DUF729 domain-containing protein 1, Prolactin precursor, Dihydrofolate erductase. Conclusively, on the basis of this study, protein expression in the vaginal mucus could be used as an indicator for time of estrus manifestation in order to increase conception rates by applying AI at an optional time.

• 한국동물생명공학회지
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• 제37권2호
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• pp.96-105
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• 2022

The ovary undergoes substantial physiological changes along with estrus phase to mediate negative/positive feedback to the upstream reproductive tissues and to play a role in producing a fertilizable oocyte in the developing follicles. However, the disorder of estrus cycle in female can lead to diseases, such as cystic ovary which is directly associated with decline of overall reproductive performance. In gene expression studies of ovaries, quantitative reverse transcription polymerase chain reaction (qPCR) assay has been widely applied. During this assay, although normalization of target genes against reference genes (RGs) has been indispensably conducted, the expression of RGs is also variable in each experimental condition which can result in false conclusion. Because the understanding for stable RG in porcine ovaries was still limited, we attempted to assess the stability of RGs from the pool of ten commonly used RGs (18S, B2M, PPIA, RPL4, SDHA, ACTB, GAPDH, HPRT1, YWHAZ, and TBP) in the porcine ovaries under different estrus phase (follicular and luteal phase) and cystic condition, using stable RG-finding programs (geNorm, Normfinder, and BestKeeper). The significant (p < 0.01) differences in Ct values of RGs in the porcine ovaries under different conditions were identified. In assessing the stability of RGs, three programs comprehensively agreed that TBP and YWHAZ were suitable RGs to study porcine ovaries under different conditions but ACTB and GAPDH were inappropriate RGs in this experimental condition. We hope that these results contribute to plan the experiment design in the field of reproductive physiology in pigs as reference data.

• Asian-Australasian Journal of Animal Sciences
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• 제20권3호
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• pp.340-350
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• 2007

The objective of this study was to determine whether peroxisome proliferator-activated receptor ${\gamma}$(PPAR${\gamma}$ is involved in the regulation of weaning to estrus of primiparous sows. Twelve sows composed of 6 groups of 2 full-sibs in a similar age (325.2 d), body weight (BW; 152.4 kg) and backfat thickness (BFT; 27.0 mm) at start of lactation, were allocated to accept 31 MJ (restricted group, R-group) or 53 MJ (control group, C-group) DE/d treatment, respectively. The experimental results indicated that the low energy intake resulted in excessive losses of BW and BFT during lactation in R-group sows, which may be related to decrease of serum 15-deoxy-${\Delta}^{12,14}$-prostaglandin $J_2$ (15d-$PGJ_2$), a ligand of PPAR${\gamma}$ The obvious peak and the frequency of LH, FSH and estradiol ($E_2$) were only observed in C-group sows. Except for $E_2$ at d 1 and 2, serum FSH, LH and $E_2$ concentrations in R-group were lower than those in C-group sows after weaning. However, the serum progesterone ($P_4$) level in R-group sows was always more than that in C-group. The expression abundances of PPAR${\gamma}$and GnRH receptor (GnRH-R) in pituitary, FSH receptor (FSH-R), LH receptor (LH-R), estrogen receptor (ES-R) and aromatase in ovary of anestrous sows were lower than those of estrous sows. Neither the BFT nor the BW was associated with the mRNA abundance of PPAR${\gamma}$in hypothalamus during lactation. Expressions of PPAR${\gamma}$in pituitary and ovary were affected evidently by the BFT changes and only by the loss of BW of sows during and after lactation. Furthermore, PPAR${\gamma}$mRNA level in ovary was significantly related to the expression abundances of GnRH-R, FSH-R, ES-R and aromatase, and GnRH-R was obviously associated with PPAR${\gamma}$expression in pituitary. However, PPAR${\gamma}$expression in hypothalamus likely has no effects on these genes expression and no obvious difference for all sows. Not serum $E_2$ or $P_4$ alone but the ratios of $E_2$ to $P_4$ and 15d-$PGJ_2$ to $P_4$, and serum FSH and LH were evidently related to PPAR${\gamma}$expression in pituitary and ovary. It is concluded that PPAR${\gamma}$is associated with body conditions, reproduction hormones and their receptor expression, which affected the functions of pituitary and ovary and ultimately the estrus after weaning of primiparous sows.

• 한국수정란이식학회지
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• 제29권1호
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• pp.43-50
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• 2014

To describe the macroscopic anatomy and ovarian-physiological difference of the genital organs of the female Korean water deer, organs from captured animals in a wild area of Korea were dissected. The ovary of estrus group was about $1.10{\pm}0.02mm$ long and weighed about $0.50{\pm}0.02g$. And pregnant group was about $1.3{\pm}0.10mm$ long and weighed about $0.40{\pm}0.05g$. And the crowns of corpora lutea were found in the estrus group, but we couldn't find crowns at the pregnant group. Especially, the estrus ovaries tended (p=0.04) to be heavier than the ovaries during pregnancy. The MMP-9 activity was higher at the Graafian follicles of pregnant group than that in estrus group. However, with regard to follicles of estrus group, MMP-2 level was higher than that in pregnant group. Furthermore, apoptosis detection marker (Casp-3) was highly expressed in Graafian follicle of the pregnant group and the corpora lutea of estrus group. Thus, the differential expression of MMPs observed in this study suggests that the reflected the mechanisms underlying of monovulatory in estrus and/or pregnancy. Our results may be very useful as it provides with information that may be considered for the development of reproductive biotechnologies in endangered animals.

• 생명과학회지
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• 제12권3호
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• pp.317-324
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• 2002

흰쥐 발정주기와 난소절제에 따른 질의 GCs, ER-$\alpha$, c-fos 및 c-jun 변화를 조직화학적 및 면역조직화학적으로 관찰하였다. 질상피는 발정사이기와 발정전기로 이어 지는 점액세포화과정에서 현저한 GCs의 양적 증가를 관찰할 수 있으며 발정사이기의 SBA, 발정전기와 발정기의 Con A와 같이 발정주기에 따른 특이적 GCs가 관찰되었다. 난소절제시에는 매우 위축된 표면층 평평세포에서만 미량의 GCs가 관찰되었다. 질에서 ER-$\alpha$, c-fos, c-jun등은 주로 핵에서 반응을 나타내는데, ER- $\alpha$는 상피세포 중 바닥층에서 주로 관찰되며, 반응세포수로 보아 발정주기에 따른 변화는 없었으나 버팀질세포에서는 발정사이기부터 발정기사이에 가장 많이 관찰되었다. c-fos는 상피의 바닥층과 중간층세포 그리고 버팀질세포에서 발정전기와 발정기사이에 가장 많이 관찰되며 c-jun은 발정기의 상피 바닥층에서 가장 많이 관찰되나 버팀질세포에서는 발정기에만 관찰되었다. 난소절제시 ER-$\alpha$, c-fos, c-jun모두 상피의 적은 세포에서만 관찰되며 버팀질 세포에서는 관찰되지 않았다. 이상의 결과로 보아 발정주기와 난소절제에 따라 특이적인 GCs분포를 보일 뿐 아니라 ER-$\alpha$, c-fos, c-jun 같은 단백질의 상이한 분포를 보여 주고 있어 이들이 질상피세포의 증식과 분화에 관여함을 알 수 있다.