Earticle

An uterus is female reproductive tract organ that affected estrus cycle. During a various changes occur at uterus in estrus cycle, one of them is body fluids secretion be called uterine fluid. Therefore, the objective of this study was to investigate the changes of protein patterns using two-dimensional gel electrophoresis in uterus fluids during the follicular and luteal phases in estrus cycle of pigs. In changes of protein spots were confirmed during the follicular and luteal phases. The 136 spots were expressed in follicular phase, the 57 spots of them showed reproducibility. On the other hand, the 140 spots were expressed in luteal phase, the 73 spots of them showed reproducibility. Also, spots expressed in follicular phase were number 69 and 94 spots and spots expressed in luteal phase only were number 156, 157, 184～187, 190 and 191 spots. The spots which of higher expression levels in the luteal phase than in follicular phase were number 76 and 79 spots. In conclusion, the spots expressed in follicular and luteal phases were confirmed with difference levels and these differences are function of RNA resolving, protein synthesis and cytoskeletal architecture.

The objective of this study was to investigate the relationship between fertility and protein pattern change using in vitro fertilization, analysis of sperm characteristics and two-dimensional gel electrophoresis in different pig types. In results, the viability and mitochondria integrity of sperm were higher significantly (p<0.05) but the portions of acrosome reaction was lower significantly (p<0.05) in Duroc and F1 (potbellied × PWG miniature pig) than PWG miniature. On in vitro fertilization to investigate fertility, the fertility of F1 semen war higher significantly (p<0.05) than in Duroc and PWG miniature pig. On the other hand, protein patterns showed similar function among the different boar semen. Especially, the heat shock 70 kDa 1-like and G patch domain-containing protein 4 were significantly (p<0.05) higher expressed in F1 than in Duroc and PWG miniature pig. The proteins associated with mitochondria in Duroc were significantly (p<0.05) higher expressed than in F1 and PWG miniature pig. The developmental rates to blastocyst stage of oocytes fertilized with sperm of F1 pig were significantly (p<0.05) higher than in PWG miniature pig. However, phosphoglycerate kinase 2 and zinc finger protein 431 were significantly (p<0.05) higher expressed in PWG miniature pig than in F1 and Duroc pigs. In conclusion, the results of the present study indicate that different proteins were expressed in different pig types, and were associated with a sperm functions and embryo development.

Luteal cells produce progesterone that supports pregnancy. Steroidogenesis requires coordination of the anabolic and catabolic pathways of lipid metabolism. In the present study, the corpus luteum (CL) in early pregnancy established from luteal phase and pregnant phase was analyzed. The first study determined progesterone changes in the bovine CL at day 19 (early maternal recognition period) and day 90 in mid-pregnancy and compared them to the CL from day 12 of the estrous cycle. CL alternation was tested using two-dimensional polyacrylamide gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI- TOF). Comparing CL from luteal phase to those from pregnant phase counterparts, significant changes in expression level were found in 23 proteins. Of these proteins 17 were not expressed in pregnant phase CL but expressed in luteal phase counterpart, whereas, the expression of the other 6 proteins was limited only in pregnant phase CL. Among these proteins, vimentin is considered to be involved in regulation of post-implantation development. In particular, vimentin may be used as marker for CL development during pregnancy because the expression level changed considerably in pregnant phase CL tissue compared with its luteal phase counterpart. Data from 2-DE suggest that protein expression was disorientated in mid pregnancy from luteal phase, but these changes was regulated with progression of pregnancy. These findings demonstrate CL development during mid-pregnancy from luteal phase and suggest that alternations of specific CL protein expression may be involved in maintenance of pregnancy.

The aim of this study was to evaluate the changes of protein patterns in granulosa cells and corpus luteum during the estrus cycle in bovine ovary by proteomics ^techniques. Our study was devided into five steps for follicular, ovulatory, early-lteal, midluteal and late-luteal. The protein was extracted from glanulosa cell and corpus luteum proteins by using M-PER Mammalian Protein Extraction Reagent. Proteins were refined by clean-up kit and quantified by Bradford method until total protein was 700 μg. Immobilized pH gradient (IPG) strip was used 18 cm and 3 11 NL. SDS-PAGE was used 10% acrylamide gel. The protein spots were visualized by Coomassie Brilliant Blue (CBB) staining, analyzed by MALDI mass spectrometry and searched on NCIBlnr. As the result, 61 spots of total 85 spots were repeated on follicular stage and 51 spots of total 114 spots were repeated on ovulatory stage. 40 spots of total 129 were repeated on early-luteal and 49 spots of total 104 spots were repeated on mid-luteal stage. Also 41 spots of total 60 spots were repeated on last-luteal stage. There were differences in the ovulation (follicular∼ovultory stage) in which the spots of follicular stage 19 was only and in ovulation stage was 10 spots. The difference between the luteinization (ovultory∼mid-luteal stage) was the spots counted in each stage. The spots of ovulatory stage was 1, early-luteal stage was 1 and in mid-luteal stage was 2. Eleven spots were found in mid-luteal stage and 2 spots were found in last-luteal stage. In conclusion, we confirmed that there were 7 spots in ovulation, 4 spots in luteinization and 2 spots in luteolysis. Spot No. 89-93 from ovulation were transferrin, and spot No.94 and 95 were HSP60. Spot No. 103 were Dusty PK, spot No. 135 were OGDC-E2, and spot No. 175, 176 were Rab GDI beta from luteinization. Spot No. 178 and 179 from luteolysis were vimentin.

Pregnancy-associated plasma protein-A (PAPP-A) is known as an important biomarker for fetal abnormality during first trimester and has a pivotal role in follicle development and corpus luteum formation. And also, it is being revealed that an expression of PAPP-A in various cells and tissues such as cancer and lesion area. PAPP-A is the major IGF binding protein-4 (IGFBP-4) protease. Cleavage of IGFBP-4 results in loss of binding affinity for IGF, causing increased IGF bioavailability for proliferation, survival, and migration. Additionally, PAPP-A can be used as a promising therapeutic target for healthy longevity. Despite growing interest, almost nothing is known about how PAPP-A expression is regulated in any tissue. This review will focus on what is currently known about the zinc metalloproteinase, PAPP-A, and its role in cells and tissues. PAPP-A is expressed in proliferating cells such as fetus in uterus, granulosa cells in follicle, dermis in wound, cancer cells, and Sertoli cells in testis. They have common characteristics of proliferation faster than normal cells with stimulating IGFs action and inhibiting IGFBPs. The PAPP-A functions and expression studies in livestock have not yet been conducted much. Further studies are needed to use PAPP-A as a marker for healthy longevity in animal science.

Palatal development is one of the crucial events in craniofacial morphogenesis, according to the significant signaling pathway. In the fusion of palatal shelves, EMT is a fundamental process to achieve the proper morphogenesis of palate. Mechanisms of EMT have been reported as the processes of migration, apoptosis or general EMT through the modulations through various signalling molecules. Rgs19, known as a RGS family through GTPase activity, showed the interesting epithelial expression patterns in various organogenesis including the significant expression patterns of Rgs19 in palatal development. To evaluate the precise function of Rgs19 in palatogenesis, we employed the loss of function studies using AS-ODN treatments while in vitro palate organ cultivations. Knock-down of Rgs19 using treatments of AS-ODN showed the retarded palatal fusion with the decreased patterns of apoptosis in mesial epithelium edge (MEE). In addition, alteration patterns of related genes were examined with the qRT-PCR. And EMT process was delayed in MEE throught staining of pancytokeratin, which known as epithelial cell marker. These results show that expression of Rgs19 in MEE has crucial role of EMT, also Rgs19 affects to palatal fusion by regulation of apoptosis through the signalling modulations. Palatal development is one of the crucial events in craniofacial morphogenesis, according to the significant signaling pathway. In the fusion of palatal shelves, EMT is a fundamental process to achieve the proper morphogenesis of palate. Mechanisms of EMT have been reported as the processes of migration, apoptosis or general EMT through the modulations through various signalling molecules. Rgs19, known as a RGS family through GTPase activity, showed the interesting epithelial expression patterns in various organogenesis including the significant expression patterns of Rgs19 in palatal development. To evaluate the precise function of Rgs19 in palatogenesis, we employed the loss of function studies using AS-ODN treatments while in vitro palate organ cultivations. Knock-down of Rgs19 using treatments of AS-ODN showed the retarded palatal fusion with the decreased patterns of apoptosis in mesial epithelium edge (MEE). In addition, alteration patterns of related genes were examined with the qRT-PCR. And EMT process was delayed in MEE throught staining of pancytokeratin, which known as epithelial cell marker. These results show that expression of Rgs19 in MEE has crucial role of EMT, also Rgs19 affects to palatal fusion by regulation of apoptosis through the signalling modulations.

Palatal development is one of the crucial events in craniofacial morphogenesis, according to the significant signaling pathway including the out growth, elevation, and fusion of palatal shelves. In the fusion of palatal shelves, epithelial to mesenchymal transition (EMT) is a fundamental process to achieve the proper morphogenesis of palate. Mechanisms of EMT have been reported as the processes of migration, apoptosis or general EMT through the modulations through various signalling molecules. Rgs19, known as a regulator of G protein signaling (RGS) family through GTPase activity, showed the interesting epithelial expression patterns in various organogeneses including the significant expression patterns of Rgs19 in palatal development. To evaluate the precise function of Rgs19 in palatogenesis, we employed the gain and loss of function studies using ASODN treatments and gene electroporations while in vitro palate organ cultivations. Knockdown of Rgs19 using treatments of AS-ODN showed the retarded palatal fusion with the decreased patterns of apoptosis in mesial epithelium edge (MEE). In addition, alteration patterns of related genes were examined with the qRT-PCR. And epithelial mesenchyme transition (EMT) process was delayed in medial edge epithelium (MEE) throught immunohistochemistry of pancytokeratin, which known as epithelial cell marker. Morphological changes were observed with the three dimensional reconstruction method. These results show that expression of Rgs19 in MEE has crucial role of EMT, also Rgs19 affects to palatal fusion by regulation of apoptosis through the signalling modulations.

This study was conducted to improve the postharvest storage techniques of managing and storing seeds, totest qualities and viabilities of the seeds and to examine the germination rate and the protein expression of Achyranthesjaponica Nakai. The seeds collected from different areas of Je-Cheon and Gwang-Ju were stored with different temperaturesand durations. Two plant growth regulators and two seed priming were treated to investigate their effect on the germinationrates and the days required for germination. The weight of one hundred seed collected in Gwang-Ju was heavier than thosein Je-Cheon. Seed length collected in Gwang-Ju was also longer about 5.12 ㎜ than those in Je-Cheon about 4.90 ㎜ andseed width was longer in Gwang-Ju than those in Je-Cheon. The rates of seed germination in two different collection areaswere higher about 2.9 to 13.0% in Gwang-Ju compared to those in Je-Cheon. Comparing its rates with the storing temperaturesand durations, they were not clearly different in between 4℃ and 25℃ and they also were gradually decreased withgetting longer storing durations. The germination rates treated by plant growth regulators were higher with GA3 than thosewith Kinetin. The highest seed germination rate was appeared at 50 ppm of GA3. Comparing its rates with different seedpriming, they were relatively higher with KNO3 than those with PEG6000. In protein expression patterns between before thegerminating and after the germinating of seeds, more and clear bands were appeared in the seed after the germination comparedto those before the germination of seeds, especially 10 ~ 20 kDa. These results showing more and clear bands weremore clearly appeared in Gwang-Ju compared to Je-Cheon. Comparing the protein expression with plant growth regulatorsand seed primings, GA3 was better expression than those with Kinetin and KNO3 was better than those with PEG6000. Moreand clear bands were closely related to the germination rates of seeds and more detailed studies would be required.

This study was conducted to investigate the quality of seeds, the germination rates and the days required forgermination, to examine the patterns of protein expressions during the germination and to improve the techniques of managingand storing seeds and viability of the seeds of Lithospermum erythrorhizon Sieb. et Zucc. After collecting and harvestingseeds, they were classified to white and brown colors of seed coat through testing their seed size, weight, and quality. Thegermination rates, the days required for germination, and the protein expressions were examined with different colors ofseed coats, storing temperatures and durations by treating the different plant growth regulators and primings. One hundredseed weight of white color was heavier about 1.17 g than those of brown one about 0.81 g. The germination rates in whitecolor of seed coat was higher, 3.05 ~ 5.75%, than those in brown one. Its rates were decreased with getting longer in storagedurations. There was no big differences on germination rates between storage temperatures. The plant growth regulator ofGA3 and Kinetin was affected to improve the seed germination. GA3 increased the seed germination clearly at 25 ppm level,while kinetin increased it gradually from 25 to 100 ppm levels. In germination by seed primings, PEG6000 made higher germinationrate with increasing their levels, whereas KNO3 increased the germination until 100 mM level and then decreasedit with 200 mM unlike PEG6000. The protein expressed during the seed germination were appeared more and clearer bandsin the seed after germination, especially 20 ~ 30 kDa, compared to those in the seed before germination. These results showingmore and clearer bands were positively related to the germination rates which were different by seed colors, storage temperaturesand durations, and plant growth regulators and primings.

The aim of this study was to evaluate the changes of protein patterns in granulosa cells and corpus luteum in ovaries during the estrus cycle in cows. The estrus cycle was devided into five steps of follicular, ovulatory, early-luteal, mid-luteal and late-luteal phases. In results, 61 spots of total 85 spots were repeated on follicular phase and 51 spots of total 114 spots were repeated on ovulatory phase. The 40 spots of total 129 spots were repeated on early-luteal phase and 49 spots of total 104 spots were repeated on mid-luteal phase. Also 41 spots of total 60 spots were repeated on late-luteal phase. On the other hands, the 16 spots were indicated difference in follicular phase and ovulation phase had a difference 10 spots. It was showed difference No. 103 spot in ovulation phase, No. 135 spot in early-luteal phase and No. 175 and 176 spots in mid-luteal phase. Also, the 11 spots were expressed specifically in mid-luteal phase and No. 178 and 179 spots were difference of expression in late-luteal phase. We confirmed that there were 7 spots for ovulation, 4 spots for luteinization and 2 spots for luteolysis. Spot No. 89～93 in ovulation phase were transferrin, and spot No.94～98 were HSP60. Spot No. 103 was Dusty PK, spot No. 135 was OGDC- E2, and spot No. 175 and 176 were Rab GDI beta from luteinization. Spot No. 178 and 179 in luteolysis were vimentin. This results suggest that will be help to basic data about infertility.

It was conducted the experiment, divided into three groups as normal, poor and polycystic ovary syndrome, to detect the change of protein patterns in follicular fluid on ovarian response following controlled ovarian hyperstimulation for human IVF outcome. In the normal group, it was confirmed reproducible 57 spots in the detected total 81 spots. Then 1 spot was not found in the other groups. In the poor responder group, it was found reproducible 53 spots in the detected total 98 spots. 6 spots were down-regulation and 7 spots were up-regulation comparable with normal group. There were not 5 spots in poor responder group comparable with other groups. In the polycystic ovary syndrome group, it was expressed reproducible 53 spots in the detected total 80 spots and 3 spots were just expressed in this group. However, 4 spots were not found in polycystic ovary syndrome. 9 spots were up-regulation comparable with normal group. Significant up and down-regulation spots among the each groups were identified. The results were a cytosolic carboxypeptidase, a signal-induced proliferation-associated protein 1, a ceruloplasmin, a keratin(type Ⅱ cytoskeletal 1), a polypeptide N-acetylgalactosaminyltransferase 2, a serine/threonine-protein phosphatase 4 regulatory subunit 4. It was identified that 8 spots, 6 kinds of protein are corresponded with NCBInr database research, but 10 spots were failed in the identification. In conclusion, it has been confirmed change and expression of protein on the ovarian response following COH of human.

The food-borne pathogen Listeria monocytogenes is commonly associated with meats and unpasteurized dairy products. To identify this pathogen in meats more efficiently than has been done in the past, we purchased meats from Korean markets and performed sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and serotyping analysis on Listeria organisms isolated from meat samples. Each Listeria species showed specific protein band patterns on SDS-PAGE. Whole-cell protein SDS-PAGE profiles indicated that the organisms isolated from meats sold in local Korean markets were L. monocytogenes with the serotypes 1/2a, 1/2b, 1/2c, and 4b. We suggest that it is possible to carry out molecular subtyping of L. monocytogenes using SDS-PAGE.

각종(各種) 어류(魚類)의 가열(加熱)된 어육단백(魚肉蛋白)을 acrylamide gel을 사용(使用)하여 전기영동(電氣泳動)한 결과(結果)로 나타나는 분획상(分劃像)의 특이성(特異性)으로서 어류품종(魚類品種)을 분류(分類)할 수 있었다. 즉 숭어 대구 삼치 고등어 참조기 방어 갈치 금태 아지 병어의 특징있는 전기영동분획상(電氣泳動分劃像) 10 개(個)를 만들었다. 이로써 acrylamide gel 을 이용(利用)한 disc-전기영동(電氣泳動)을 통하여 조리(調理)된 어육제품(魚肉製品) 소편(小片)으로 그 어류품종(魚類品種)을 감별(鑑別)해 낼 수 있는 표준(標準)이 확실(確實)하게 되었다. 그러나 아직 본(本) 실험결과(實驗結果)를 통조림된 어육(魚肉)에는 적용(適用)할 수가 없다.

This studies' objective methods of identifying fish species are based on the species-specific protein-separation patterns obtained on electrophoresis of watersoluble sarcoplasmic proteins of fish muscle. As the proteins must be in their native undenatured state, electrophoretic identification of fish species has, so far, been restricted to raw fish. An extention of the electrophoretic method to the identification of cooked fish is discribed. The protein fragments extractable in 10M urea from the denatured proteins of cooked muscle can also be separated by electrophoresis into species' characteristic patterns that could be used for species identification. The separation patterns obtained on polyacrylamide gel for the urea extracts of cooked Mugil cephalus, Gadus macrocephalus, Scomberomorus niphonius, Scomber japonicus, Pseudosciaena manchurica, Seriola quinqueradiata, Trichius lepturus, Duderleinia berycoides, Lophimus setigerus, Pampus argenteus are presented. In its present form the method does not apply to canned fish.

There is a pair of labial gland of hydropsychid caddisfly (Hydropsyche kozhantschikovi Martynov) larva. It is in ‘Z’ formation in the body and the total length is about 20mm. Hydropsychid caddishfly larvae that were raised under the lab conditions were able to form a nest-spining by connecting the small grains of sand provided. By repeatedly treating the extracted labial gland with methanol/D.W., the cell layer was removed. Accordingly, only the matrix within the labial gland that did not dissolve in water was obtained. The matrix inside the methanol/D.W. treated labial gland was dissolved with 5% acetic acid. Then the results of an acidic electrophoresis with a number of conditions indicated that 5% acetic acid/ 5M urea/ 8% PAGE was the most effective. Moreover, the result of 2-D PAGE on the labial gland of these hydropsychid caddisfly larva, the number of proteins in the labial gland including the cell layer was about 350 and the number of proteins in the labial gland treated with methanol/D.W. was about 80, showing a substantially small number of proteins.