• Title/Summary/Keyword: IGF1

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Methylation Pattern of H19 Gene at Various Preimplantation Development Stages of In Vitro Fertilized and Cloned Porcine Embryos

  • Im, Young-Bin;Han, Dong-Wook;Gupta, Mukesh Kumar;Uhm, Sang-Jun;Heo, Young-Tae;Kim, Jin-Hoi;Park, Chan-Kyu;Lee, Hoon-Taek
    • Reproductive and Developmental Biology
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    • v.31 no.2
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    • pp.83-90
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    • 2007
  • Insulin-like growth factor II (IGF2) and H19 genes are mutually imprinted genes which may be responsible for abnormalities in the cloned fetuses and offspring. This study was performed to identify putative differentially methylated regions (DMRs) of porcine H19 locus and to explore its genomic imprinting in in vitro fertilized (IVF) and somatic cell nuclear transferred (SCNT) embryos. Based on mice genomic data, we identified DMRs on H19 and found porcine H19 DMRs that included three CTCF binding sites. Methylation patterns in IVF and SCNT embryos at the 2-, 4-, $8{\sim}16$-cells and blastocyst stages were analyzed by BS (Bisulfite Sequencing)-PCR. The CpGs in CTCF1 was significantly unmethylated in the 2-cell stage IVF embryos. However, the 4- (29.1%) and $8{\sim}16$-cell (68.2%) and blastocyst (48.2%) stages showed higher methylation levels (p<0.01). On the other hand, SCNT embryos were unmethylayted ($0{\sim}2%$) at all stages of development. The CpGs in CTCF2 showed almost unmethylation levels at the 2-,4- and $8{\sim}16$-cell and blastocyst stages of development in both IVF ($0{\sim}14.1%$) and SCNT ($0{\sim}6.4%$) embryos. At all stages of development, CTCF3 was unmethylated in IVF ($0{\sim}17.3%$) and SCNT ($0{\sim}1.2%$) embryos except at the blastocyst stage (54.5%) of IVF embryos. In conclusion, porcine SCNT embryos showed an aberrant methylation pattern comprised to IVF embryos. Therefore, we suggest that the aberrant methylation pattern of H19 loci may be a reason for increased abnormal fetus after embryo transfer of porcine SCNT embryos.

Monitoring of Chicken RNA Integrity as a Function of Prolonged Postmortem Duration

  • Malila, Yuwares;Srimarut, Yanee;U-chupaj, Juthawut;Strasburg, Gale;Visessanguan, Wonnop
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.11
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    • pp.1649-1656
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    • 2015
  • Gene expression profiling has offered new insights into postmortem molecular changes associated with meat quality. To acquire reliable transcript quantification, high quality RNA is required. The objective of this study was to analyze integrity of RNA isolated from chicken skeletal muscle (pectoralis major) and its capability of serving as the template in quantitative real-time polymerase chain reaction (qPCR) as a function of postmortem intervals representing the end-points of evisceration, carcass chilling and aging stages in chicken abattoirs. Chicken breast muscle was dissected from the carcasses (n = 6) immediately after evisceration, and one-third of each sample was instantly snap-frozen and labeled as 20 min postmortem. The remaining muscle was stored on ice until the next rounds of sample collection (1.5 h and 6 h postmortem). The delayed postmortem duration did not significantly affect $A_{260}/A_{280}$ and $A_{260}/A_{230}$ ($p{\geq}0.05$), suggesting no altered purity of total RNA. Apart from a slight decrease in the 28s:18s ribosomal RNA ratio in 1.5 h samples (p<0.05), the value was not statistically different between 20 min and 6 h samples ($p{\geq}0.05$), indicating intact total RNA up to 6 h. Abundance of reference genes encoding beta-actin (ACTB), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), hypoxanthine-guanine phosphoribosyltransferase (HPRT), peptidylprolylisomerase A (PPIA) and TATA box-binding protein (TBP) as well as meat-quality associated genes (insulin-like growth factor 1 (IGF1), pyruvate dehydrogenase kinase isozyme 4 (PDK4), and peroxisome proliferator-activated receptor delta (PPARD) were investigated using qPCR. Transcript abundances of ACTB, GAPDH, HPRT, and PPIA were significantly different among all postmortem time points (p<0.05). Transcript levels of PDK4 and PPARD were significantly reduced in the 6 h samples (p<0.05). The findings suggest an adverse effect of a prolonged postmortem duration on reliability of transcript quantification in chicken skeletal muscle. For the best RNA quality, chicken skeletal muscle should be immediately collected after evisceration or within 20 min postmortem, and rapidly preserved by deep freezing.

Effects of Kyungohkgo(Qióng yù gào) on Growth and Learning Ability in Growth Deficiency Rat with Insufficient Nutrition Diet (경옥고(瓊玉膏)가 영양소 결핍으로 유도한 성장장애 흰쥐의 성장 및 학습효과에 미치는 영향)

  • Han, Seok-Hoon;Cha, Yun-Yeop;Lee, Eun
    • Journal of Korean Medicine Rehabilitation
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    • v.18 no.2
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    • pp.97-109
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    • 2008
  • Objectives : Effects of Kyungohkgo($Qi{\acute{o}}ng\;y{\grave{u}}\;g{\grave{a}}o$) on intellectual development and learning ability were investigated growth and intellectual impairment rat with insufficient nutrition diet. Methods : We divided male Sprague-Dawley rats into 4 groups(A, B, C, D). They were normal group(A), growth deficiency rat with insufficient nutrition diet group(B), growth deficiency rat with 0.1% Kyungohkgo($Qi{\acute{o}}ng\;y{\grave{u}}\;g{\grave{a}}o$) group(C) and growth deficiency rat with 0.2% Kyungohkgo($Qi{\acute{o}}ng\;y{\grave{u}}\;g{\grave{a}}o$) group(D). They were administered for 5 weeks. We measured body weight, serum growth hormone, insulin-like growth factor and thyroid stimulating hormone, RBC, concentration of Hb and PCV ratio, total WBC and its composition, the values of plasma glutamic oxaloacetic transaminase(GOT) and glutamic pyruvic transaminase(GPT) activities and morris water maze test in escape distance, escape time and escape speed. Results : 1. Body weight showed a tendency to increase in the Kyungohkgo($Qi{\acute{o}}ng\;y{\grave{u}}\;g{\grave{a}}o$) groups and 0.2% Kyungohkgo($Qi{\acute{o}}ng\;y{\grave{u}}\;g{\grave{a}}o$) group showed significantly different than control groups. 2. Serum growth hormone, insulin-like growth factor and thyroid stimulating hormone showed a tendency to increase in Kyungohkgo($Qi{\acute{o}}ng\;y{\grave{u}}\;g{\grave{a}}o$) groups, however these values showed no significantly different. 3. About the counts of RBC, 0.1% and 0.2% Kyungohkgo($Qi{\acute{o}}ng\;y{\grave{u}}\;g{\grave{a}}o$) groups showed significantly different than control groups. Concentration of Hb was higher than control group in Kyungohkgo($Qi{\acute{o}}ng\;y{\grave{u}}\;g{\grave{a}}o$) groups. And 0.2% Kyungohkgo($Qi{\acute{o}}ng\;y{\grave{u}}\;g{\grave{a}}o$) group showed significantly different than control groups in PCV ratio. 4. The counts of total WBC and its composition showed no significantly different in all treatment groups. 5. The values of plasma glutamic oxaloacetic transaminase(GOT) and glutamic pyruvic transaminase(GPT) activities showed no significantly different in all treatment groups. 6. In the morris water maze test, in escape distance and escape speed, Kyungohkgo($Qi{\acute{o}}ng\;y{\grave{u}}\;g{\grave{a}}o$) groups showed no significantly different than control group. But Kyungohkgo($Qi{\acute{o}}ng\;y{\grave{u}}\;g{\grave{a}}o$) groups showed the increasing tendency. Conclusions : So Kyungohkgo($Qi{\acute{o}}ng\;y{\grave{u}}\;g{\grave{a}}o$) have an effect of promoting growth of rats and might be effect to treat various kinds of growth delay in children.

A Study on the Establishment of Acceptable Range for Internal Quality Control of Radioimmunoassay (핵의학 검체검사 내부정도관리 허용범위 설정에 관한 고찰)

  • Young Ji, LEE;So Young, LEE;Sun Ho, LEE
    • The Korean Journal of Nuclear Medicine Technology
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    • v.26 no.2
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    • pp.43-47
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    • 2022
  • Purpose Radioimmunoassay implement quality control by systematizing the internal quality control system for quality assurance of test results. This study aims to contribute to the quality assurance of radioimmunoassay results and to implement systematic quality control by measuring the average CV of internal quality control and external quality control by plenty of institutions for reference when setting the laboratory's own acceptable range. Materials and Methods We measured the average CV of internal quality control and the bounce rate of more than 10.0% for a total of 42 items from October 2020 to December 2021. According to the CV result, we classified and compared the upper group (5.0% or less), the middle group (5.0~10.0%) and the lower group (10.0% or more). The bounce rate of 10.0% or more was compared by classifying the item of five or more institutions into tumor markers, thyroid hormones and other hormones. The average CV was measured by the overall average and standard deviation of the external quality control results for 28 items from the first quarter to the fourth quarter of 2021. In addition, the average CV was measured by the overall average and standard deviation of the proficiency results between institutions for 13 items in the first half and the second half of 2021. The average CV of internal quality control and external quality control was compared by item so we compared and analyzed the items that implement well to quality control and the items that require attention to quality control. Results As a result of measuring the precision average of internal quality control for 42 items of six institutions, the top group (5.0% or less) are Ferritin, HGH, SHBG, and 25-OH-VitD, while the bottom group (≤10.0%) are cortisol, ATA, AMA, renin, and estradiol. When comparing more than 10.0% bounce rate of CV for tumor markers, CA-125 (6.7%), CA-19-9 (9.8%) implemented well, while SCC-Ag (24.3%), CA-15-3 (26.7%) were among the items that require attention to control. As a result of comparing the bounce rate of more than 10.0% of CV for thyroid hormones examination, free T4 (2.1%), T3 (9.3%) showed excellent performance and AMA (39.6%), ATA (51.6%) required attention to control. When comparing the bounce rate of 10.0% or more of CV for other hormones, IGF-1 (8.8%), FSH (9.1%), prolactin (9.2%) showed excellent performance, however estradiol (37.3%), testosterone (37.7%), cortisol (44.4%) required attention to control. As a result of measuring the average CV of the whole institutions participating at external quality control for 28 items, HGH and SCC-Ag were included in the top group (≤10.0%), however ATA, estradiol, TSI, and thyroglobulin included in bottom group (≥30.0%). Conclusion As a result of evaluating 42 items of six institutions, the average CV was 3.7~12.2% showing a 3.3 times difference between the upper group and the lower group. Cortisol, ATA, AMA, Renin and estradiol tests with high CV will require continuous improvement activities to improve precision. In addition, we measured and compared the overall average CV of the internal quality control, the external quality control and the proficiency between institutions participating of six institutions for 41 items excluding HBs-Ab. As a result, ATA, AMA, Renin and estradiol belong to the same subgroup so we require attention to control and consider setting a higher acceptable range. It is recommended to set and control the acceptable range standard of internal quality control CV in consideration of many things in the laboratory due to the different reagents and instruments, and the results vary depending on the test's proficiency and quality control materials. It is thought that the accuracy and reliability of radioimmunoassay results can be improved if systematic quality control is implemented based on the set acceptable range.

THE EFFECT OF PRP ON THE BONE REGENERATION OF ${\beta}-TCP$ GRAFTED IN RABBIT CRANIAL BONE DEFECT (가토 두개골 결손부에 이식된 ${\beta}-TCP$의 골치유 과정에서 PRP의 효과에 관한 연구)

  • Lee, Soung-Hoon;Hwang, Kyung-Gyun;Park, Chang-Joo;Lim, Byung-Sup;Cho, Jung-Yeon;Paik, Seung-Sam;Shim, Kwang-Sup
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.28 no.5
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    • pp.417-433
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    • 2006
  • Purpose : Platelet rich plasma (PRP) is an autologous material with many growth factors, such as BMPs, PDGF, $TGF-{\beta}_1$, $TGF-{\beta}_2$, VEGF, and IGF, facilitating bone healing process. The prominent osteoconductive activity and the biodegradable nature of beta-tricalciumphosphate (${\beta}-TCP$) for bone grafts in animal experiments have been reported. The purpose of this study was to evaluate the effect of PRP on the osteogenesis of ${\beta}-TCP$. Materials & Methods : Two artificial calvarial bone defects were made in 32 rabbits which were divided into 2 groups. In one group of 16 rabbits, autogenous bone / ${\beta}-TCP$ was grafted on each side of cranial bone defect. In the other group of 16 rabbits, mixture of ${\beta}-TCP$ and PRP / PRP alone was grafted on each side of the cranial bone defect. The animals were sacrificed at 2, 4, 8, and 12 weeks after surgery. The specimens were harvested and examined histologically and immunohistochemically by the expression of BMP2/4/7, PDGF, VEGF and $TGF-{\beta}_1$. Results : The mean volume of new bone formation was significantly higher at 4, 8, 12 weeks in autogenous graft than that in ${\beta}-TCP$. The BMP2/4 expression was significantly higher at 4 weeks in autogenous bone graft and at 4 weeks in mixture of ${\beta}-TCP$ and PRP and at 12 weeks in ${\beta}-TCP$. The expression of BMP7, PDGF, VEGF and $TGF-{\beta}_1$ showed no significant difference in autogenous, ${\beta}-TCP$, mixture of ${\beta}-TCP$ and PRP, and PRP alone during grafted bone regeneration. Conclusion : The results showed that PRP had no additional value in promoting healing process of ${\beta}-TCP$ grafts.

Continuous Positive Airway Pressure Therapy in a Obstructive Sleep Apnea Syndrome associated with Active Acromegaly (지속적 양압공급치료로 임상적 호전을 보인 활동성 선단거대증에 동반된 폐쇄성 수면 무호흡증후군 1예)

  • Moon, Hwa-Sik;Choi, Young-Mee;Ahn, Seog-Ju;Kim, Chi-Hong;Kwon, Soon-Seog;Kim, Young-Kyoon;Kim, Kwan-Hyoung;Song, Jeong-Sup;Park, Sung-Hak
    • Tuberculosis and Respiratory Diseases
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    • v.42 no.4
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    • pp.610-617
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    • 1995
  • Sleep apnea occurs in approximately 50% of patients with acromegaly, and sleep apnea is associated with increased cardiovascular diseases and mortality. In view of these findings, sleep apnea may be a factor in the increased incidence of cardiovascular deaths in acromegaly patients. We experienced a case of active acromegaly patient, 54-yr-old man, associated with obstructive sleep apnea syndrome, congestive heart failure with dilated cardiomyopathy and serious cardiac arrhythmias. He was treated for obstructive sleep apnea syndrome, diagnosed by overnight polysomnography, with nasal continuous positive airway pressure(CPAP) for 4 months, which successfully controlled his loud snoring, sleep apnea, nocturnal hypoxemia and apnea-related symptoms. And also he was treated for underlying acromegaly and cardiac complications with bromocriptine and cardiogenic drugs for 4 months, but still had elevated growth hormone(GH) and insulin like growth factor-1(IGF-1) levels and serious cardiac arrhythmias. We describe our experience about the effect of CPAP treatment in an active acromegaly patient associated with obstructive sleep apnea syndrome and cardiac complications with review of literature.

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Growth performance, carcass traits, muscle fiber characteristics and skeletal muscle mRNA abundance in hair lambs supplemented with ferulic acid

  • Pena-Torres, Edgar Fernando;Castillo-Salas, Candelario;Jimenez-Estrada, Ismael;Muhlia-Almazan, Adriana;Pena-Ramos, Etna Aida;Pinelli-Saavedra, Araceli;Avendano-Reyes, Leonel;Hinojosa-Rodriguez, Cindy;Valenzuela-Melendres, Martin;Macias-Cruz, Ulises;Gonzalez-Rios, Humberto
    • Journal of Animal Science and Technology
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    • v.64 no.1
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    • pp.52-69
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    • 2022
  • Ferulic acid (FA) is a phytochemical with various bioactive properties. It has recently been proposed that due to its phytogenic action it can be used as an alternative growth promoter additive to synthetic compounds. The objective of the present study was to evaluate the growth performance, carcass traits, fiber characterization and skeletal muscle gene expression on hair-lambs supplemented with two doses of FA. Thirty-two male lambs (n = 8 per treatment) were individually housed during a 32 d feeding trial to evaluate the effect of FA (300 and 600 mg d-1) or zilpaterol hydrochloride (ZH; 6 mg d-1) on growth performance, and then slaughtered to evaluate the effects on carcass traits, and muscle fibers morphometry from Longissimus thoracis (LT) and mRNA abundance of β2-adrenergic receptor (β2-AR), MHC-I, MHC-IIX and IGF-I genes. FA increased final weight and average daily gain with respect to non-supplemented animals (p < 0.05). The ZH supplementation increased LT muscle area, with respect to FA doses and control (p < 0.05). Cross-sectional area (CSA) of oxidative fibers was larger with FA doses and ZH (p < 0.05). Feeding ZH increased mRNA abundance for β2-AR compared to FA and control (p < 0.05), and expression of MHC-I was affected by FA doses and ZH (p < 0.05). Overall, FA supplementation of male hair lambs enhanced productive variables due to skeletal muscle hypertrophy caused by MHC-I up-regulation. Results suggest that FA has the potential like a growth promoter in lambs.

EXPRESSION OF OSTEOGENESIS RELATED FACTORS ACCORDING TO DISTRACTION RATE IN THE DISTRACTION OSTEOGENESIS (신연 골형성술시 신연속도에 따른 골형성 관여 인자의 발현)

  • Jee, Yu-Jin;Kim, Yeo-Gab
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.34 no.3
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    • pp.250-265
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    • 2008
  • Distraction osteogenesis is a well-established clinical treatment for limb length discrepancy and skeletal deformities. Appropriate mechanical tension-stress is believed not to break the callus but rather to stimulate osteogenesis. In contrast to fracture healing, the mode of bone formation in distraction osteogenesis is primarily intramembranous ossification. Although the biomechanical, histological, and ultrastructural changes associated with distraction osteogenesis have been widely described, the basic biology of the process is still not well known. Moreover, the molecular mechanisms in distraction osteogenesis remain largely unclear. Recent studies have implicated the growth factor cascade is likely to play an important role in distraction. And current reserch suggested that mechanical tension-stress modulates cell shape and phenotype, and stimulates the expression of the mRNA for bone matrix proteins. The purpose of this study is to examine the pattern of expression of growth factors($TGF-{\beta}1$, IGF-I, bFGF) and extracellular matrix proteins(osteoclacin, osteonectin) related to osteogenesis by osteodistraction of the mandible in rabbits. 24 rabbits is used for this experiment. Experimental group are gradual distraction(0.7mm, twice/day), acute distraction(1.4mm, twice/day) and control group is only osteotomized. After 5 days latency, osteotomic site is distracted for each 7 days and 3.5 days. Consolidation period is 28 days. The animal is sacrificed at the 3th, 7th, 14th, 28th. The distracted bone is examined by immunohistochemical analysis and RT-PCR analysis. The results obtained from this study were as follow : No significant difference was found on clinical examination according to distraction rate, but gradual distraction was shown to improve regenerate bone formation on radiographic and histologic examination. Growth factors and extracelluar matrix proteins expression increased in distraction group than control group. From these results, it could be stated that graudal distraction is shown to improve and accelerate bone formation and mechanical stress like distraction has considerable effects on osteogenesis related factors. And rabbit is the most appropriate animal model for further reseach on the molecular mechanisms that mediate osteodistraction. It is believed that understanding the biomolecular mechanisms that mediate distraction osteogenesis may guide the development of targeted strategies designed to improve distraction osteogenesis and accelerate bone healing.

Cytological analysis of pregnancy-associated plasma protein-A expression in porcine neonatal testis (미성숙 돼지 정소에서 pregnancy-associated plasma protein-A의 발현의 세포학적 분석)

  • Kim, Ji-youn;Oh, Keon Bong;Byun, Sung June;Ock, Sun-A;Lee, Hwi-Cheul;Hwang, Seong-Su;Park, SangHyun;Ha, Wootae;Woo, Jae-Seok;Song, Hyuk
    • Journal of Embryo Transfer
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    • v.33 no.3
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    • pp.177-183
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    • 2018
  • The identification of biomarkers of a living tissues is essentially required to understand specific functions of the cells. In previous study, we reported IGFBP 3 as one of the putative biomarkers, by showing specific expression at porcine spermatogonial stem cells (SSCs) of early stage of porcine testis. In this study, we analyzed the expression of seven members of IGFBP family (IGFBPs) in SSCs and histological expression pattern of pregnancy-associated plasma protein-A (PAPP-A), which plays a role on the growth promoting enzyme by cleavage of IGFBPs in testis of 5 days old pig. RT-PCR analysis showed that IGFBP 1, 2, 3, 4, and 6 were expressed at high level specifically in porcine SSCs compared with whole testis. We performed immunohisotochemical staining of testis sections with PAPP-A and protein gene product 9.5 (PGP9.5) which are the known biomarkers for SSCs. We were not able to find co-expression of PAPP-A and PGP9.5; PAPP-A was expressed only in Sertoli cells and PGP9.5 expression was confirmed in spermatogonium. Additionally, we were able to confirm the GATA4 expression in Sertoli and Leydig cells as a regulator of Sertoli cell function was not detected PGP9.5 expressing cells, indicating indirect evidence of that cytolocalization of PAPP-A expression is limited in Sertoli cells. These results suggested that the PAPP-A expressed in Sertoli cells may play role on regulation of development and differentiation of testicular cells through the IGF axis in neonatal porcine testis.

Microscopic Study of the Pig Peri-implantation Embryos (전자현미경에 의한 착상 전후 돼지수정란의 형태학적 변화에 관한 연구)

  • 김진회;백청순;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.18 no.2
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    • pp.141-150
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    • 1994
  • Morphological features of the interaction between the hatching blastocyst and implantation in pig were studied by electron microscopy. The observations extended from late blastocyst stage to the completion of trophoblastic erosion of the epithelium and early decidual transformation of the epithelium and early decidual transformation of the stromal cells. Between day 7 and 17 of pregnancy, blastocysts from 0.3 to 12 mm in diameter were flushed from the uterine horns of Dutch Landrace pigs. On the 7th of development in the pig blastocyst, the blastocyst shedded of the zona pellucida established the tips of microvilli and with bleb-like cytoplasmic protrusions of the epithelial cells. From day 11 on in pig embryo, the bilayered trophoblast undergoes a dramatic phase of elongation so that the initially spherical expanded blastocyst becomes tubular. In pig, close apposition to the uterine wall beg-ins at about 12 $^1$/$_2$ days and then attachment occurred during the afternoon of the 16th or 18th day post coitum. At this stage, embryonic loss compared with corpus luteum number is up to 40% of ovulated oocytes. Therefore, the implantation failture of these embryos may be mainly caused by morphological abnormality and failture of zona shedding.

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