• Title/Summary/Keyword: mature cell

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Establishment of the expression system of human HtrA2 in the zebrafish (Zebrafish 동물모델에서 human HtrA2의 expression system 정립에 관한 연구)

  • Cho, Sung-Won;Park, Hyo-Jin;Kim, Goo-Young;Nam, Min-Kyung;Kim, Ho-Young;Ko, In-Ho;Kim, Cheol-Hee;Rhim, Hyang-Shuk
    • Journal of Life Science
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    • v.16 no.4
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    • pp.571-578
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    • 2006
  • HtrA2/Omi, a mitochondrial trypsin-like serine protease, is pivotal in regulating apoptotic cell death. Several lines of recent evidence suggest that HtrA2 is associated with the pathogenesis of neurodegenerative disorders; however, the physiological function of HtrA2 still remains elusive. For studying physiological function of HtrA2 in depth, it is necessary to develop a suitable expression system in the model animal. We therefore utilized the zebrafish as a model animal to establish expression of human HtrA2 (hHtrA2) in vivo. For expression of mature HtrA2 as GFP fusion in zebrafish embryos, the HtrA2 (WT) or (S306A) cDNAs with the C-terminal GFP tag were inserted into the pCS2+ plasmid. Expression patterns of HtrA2 in HEK293 cells were first monitored by immunofluorescence staining and immunoblot assays, showing approximately 64 kDa of the HtrA2-GFP fusion proteins. Subsequently, the hHtrA2 plasmid DNA or in vitro transcribed mRNA was microinjected into zebrafish embryos. The expression patterns of HtrA2 in Zebrafish embryos were monitored by GFP fluorescence in 24 hours-post-fertilization (hpf). Although expression patterns of HtrA2-GFP in developing embryos were different between the injected DNA and mRNA, both nucleic acids revealed good expression levels to further study the physiological role of HtrA2 in vivo. This study provides a suitable condition for expressing hHtrA2 in the zebrafish embryos as well as a method for generating useful system to investigate physiological properties of the specific human genes.

Structural Features of Various Trichomes in Vitex negundo during Development (방향성 좀목형(Vitex negundo)모용의 구조적 분화발달)

  • Lee, Seung-Hee;Kim, In-Sun
    • Applied Microscopy
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    • v.36 no.1
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    • pp.35-45
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    • 2006
  • Plants of Vitex negundo are known to develop numerous trichomes throughout their body, where certain trichome types have been believed to be one of the plausible structures for the unique scents. In the current study. structural aspects of the trichomes have been examined in leaves and stems of Vitex negundo using TEM and SEM. Trichome types as well as structural changes that occurred in certain trichomes during secretion have been mainly focused. Three type of glandular trichomes and two types of non-glandular trichomes were developed in the epidermis of young and mature Vitex negundo plants. The glandular trichomes included the peltate type (Type 1), the capitate type (Type 2), and degraded capitate type (Type 3), whereas the non-glandular warty trichomes contained the multicellular (Types 4) and unicellular type (Type 5). Type 1 and 2 consisted of head and stalk cells, but their number and size were different. One secretory cavity was formed from the four head cells in the former, but only two head cells were involved in the latter. The cytoplasmic density in the head cell was quite high and in particular, sER and Golgi bodies were well developed. At initiation of their development, the cuticle layer of the head cells separated from the outer tangential wall to form a secretory cavity. Subsequently the cavity expanded acropetally and a large number of secretory vesicles continuously produced from the head cells until they filled the entire cavity. The cavity contained materials that would be soon discharged into intercellular spaces and/or into the air. The cavity began to decrease the volume by contracting at initial secretion but degrade rapidly within short time. It has been suggested that the mode of secretion in V. negundo is probably the eccrine secretion, since no break or rupture of the cavity has been observed during examination. Contrastingly Type 3 exhibited deterioration of the head cell at early stage. Type 4 was about $110{\sim}190{\mu}m$ long, consisting of $2{\sim}3$ cells, and distributed more in the adaxial epidermis compared to the abaxial surface. However, $20{\sim}30{\mu}m$ long Type 5 was extremely dense in both epidermis. Among several trichome types, Type 1 and 2 probably play an important role in discharging unique aromatic scents in plants of V. negundo.

Increase in Neurogenesis of Neural Stem Cells Cultured from Postnatal Mouse Subventricular Zone by Nifedipine (L-type 칼슘 채널을 저해하는 저해제, nifedipine에 의한 쥐 뇌실하 영역 신경줄기세포의 신경세포로의 분화 촉진)

  • Park, Ki-Youb;Kim, Man Su
    • Journal of Life Science
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    • v.32 no.2
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    • pp.108-118
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    • 2022
  • The subventricular zone (SVZ) in the brain contains neural stem cells (NSCs) that generate new neurons throughout one's lifetime. Many extracellular and intracellular factors that affect cell proliferation and neuronal differentiation of NSCs are already well-known. Recently, L-type calcium channels have been reported to regulate neural development and are present in NSCs, differentiating neuroblasts, and mature neurons in the SVZ. Nifedipine, a blocker of L-type calcium channels, has been long used as a therapeutic drug for hypertension. However, studies on the use of nifedipine to inhibit L-type calcium channels of NSCs are lacking. Herein, we treated NSCs cultured from mouse postnatal SVZ with nifedipine during neuronal differentiation. Nifedipine increased the number of Tuj1-positive neurons but did not significantly change the number of Olig2-positive oligodendrocytes. Nifedipine increased cell division during early differentiation, which was detected using the 5-ethynyl-2'-deoxyuridine incorporation assay and immunocytochemistry assessment by staining the cells with phosphorylated histone H3, a mitosis marker. Nifedipine increased the transcription of Dlx2, a neurogenic transcription factor, and the level of Mash1, a marker for early neurogenesis. In addition to nifedipine, verapamil, which is also an L-type calcium channel blocker, showed a slight increase in neurogenesis, but its statistical significance was very low. In contrast, pimozide, a T-type calcium channel blocker, did not affect neurogenesis, although T-type calcium channel genes Cav3.1, Cav3.2, and Cav3.3 were expressed. In summary, nifedipine might promote the neuronal fate of NSCs during early differentiation and calcium signaling through L-type calcium channels might be involved in neuronal differentiation, especially during the early stages of differentiation.

Studies on the Life History of Bacciger harengulae (Bacciger harengulae의 생활사에 관한 연구)

  • KIM Young-Gill;CHUN Seh-Kyu
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.17 no.5
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    • pp.449-470
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    • 1984
  • The cercaria of Bacciger herengulae which is parasitized on the gonad of Solen strictus was investigated in order to reveal its entire life history. The area covered for the study was in the vicinity sea of Naechodo, the estuary of the Kum river in the western coast of Korea during the period of 1980-1983. Morphology and development as well as infection rates of sporocyst and cercaria within Solen strictus were examined. For accomplishing the objectives of this study, an artificial infection experiment and some investigations on the second intermediate host, the final host and the growing stages were also studied in both laboratory and natural habitat of Solen strictus. According to the study, it was revealed that the first intermediate hosts were Meretrix lusoria, Solen strictus, Tapes japonica and Laternula limicola, the second intermediate host was Palaemon (Exopalaemon) carinicauda and the final hosts were Konosirus punctatus and Harengula zunasi. A mature sporocyst which was found in the gonad of Solen strictus was $4.0-4.3{\times}0.2-0.21\;mm$ insize, and the cercaia with 27 pairs of setae, each seta consisting of 6 tufts, was $270{\times}147{\mu}m$ in body size and $550{\times}52{\mu}m$ in tail size. Oral sucker($52{\times}42{\mu}m$), pharynx, vental sucker and two testese were obviously seen within the cercaria. The excretory vesicles of cercaria were in V-shape and the flame cell were formula was expressed as 2[(3+3)+(3+3)]=24. The infection of cercaria in the first intermediate host, Solen strictus, was found throughout the year regardlless of the water temperature, and its mean infection rate was $9.67\%$ during the study period. The infection rate fluctuated with temperature, the highest being $28.0\%\;at\;28.0^{\circ}C$ water temperature in July and the lowest $2.4\%\;at\;19.5^{\circ}C$ in October, and it increased in proportion to the shell length on the host. But cercaria was not detected at below 4.0 cm in size of the host. Mature cercariae were found 6 months from May to October when water temperature was above $19.5^{\circ}C$. On the other hand, when water temperature was below $19.5^{\circ}C$, only immature cercariae and sporocysts were found. The cercariae were active for 35 hours and survived for 71 hours at $20^{\circ}C$, and 29 and 34 hours at $25^{\circ}C$ respectively, whereas the cercariae were inactive at less than $20^{\circ}C$ in water temperature. Cercaria, from Solen strictus, approached shrimp of 1-3 cm in body length as its second host. Then, it began to intrude in to the muscle of shrimp after 2-3 hours. The infected cercaria formed cyst after 7-8 hours, and became mature metacercaria. $420{\times}310{\mu}m$ in size, 15 days afer infection. The infection rate of metaceria to shrimp in the laboratory was highest, at $25^{\circ}C$ being $61\%$ and at $20^{\circ}C\;17%$. The infection rate of metacearia in shrimp was highest in the first abdominal segment, followed by cephalothorax, the second, and fifth abdominal segments, and in that order. Also, the infection rate of metacercaria in wild shrimp was high $9.6-11.1\%$ at $26.5^{\circ}C$ in June, and low $1.56-2.5\%$ at $28-29.5^{\circ}C$ from July to August. The infected shrimp with metacercaria was experimentally fed to Konosirus punctatus in the laboratory in order to know its final host. The metacercaria developed into the adult worm, $440-520{\times}310-360{\mu}m$ in size, within the intestine of Konosirus punctatus 20 days after infection. The adult worm was oval shape and $20-24{\times}11-20{\mu}m$ in size. The infection rate of adult worm to Konosirus punctatus and Harengula zunasi ranged 87.3 to $100\%$, the mean being $95.2\%$, regardless of the body length of their hosts. The infection rate was $100\%$ in June and July, but it decreased in September and October. The size and body structure of the trematode observed during the present study were well agreed with those ievestigated by Yamaguti(1938), thus, it may be concluded that the adult worm it identified as Bacciger harengulae.

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Effects of Activation Treatments and Culture Condition on In Vitro Development of Caprine In Vivo and In Vitro Oocytes (재래산양의 체내 및 체외유래 난자의 활성화 처리방법 및 배양조건이 단위발생란의 체외발달에 미치는 영향)

  • Park H. S.;Kim T. S.;Lee Y. H.;Jung S. Y.;Lee M. Y.;Jin J. I.;Park J. K.;Lee J. S.;Kim C. H.
    • Reproductive and Developmental Biology
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    • v.28 no.3
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    • pp.181-185
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    • 2004
  • This study was conducted to examine whether activation treatments, source of oocytes and culture conditions affect in vitro developmental ability of caprine oocytes. Mature Korean native goats were pretreated with intravaginal CIDR for 10 days. The goats were then treated with a single intramuscular injection of 1,000 IU PMSG on Day 8 or twice daily injection of a total of 70 mg FSH for 3 days from Day 8 of CIDR insertion for superovulation. All the goats were injected with 10 mg PGF/sub 2a/ on Day 8 and 400 IU hCG on Day 10 of CIDR. Oocytes were surgically collected by oviduct flushing(in vivo maturation) or direct follicle aspiration(in vitro maturation) through mid-ventral incision at 35 h after hCG injection. Fifteen to twenty oocytes were placed in TCM-199 medium containing 25 mM Hepes and hormones under mineral oil at 39℃ in a humudified atmosphere of 5% CO₂ in air for 22 to 24 h. After maturation, the oocytes were activated by electric stimulation or ionomycin + 6-DMAP. The activated oocytes were then cultured in M16, TCM-199 and mSOF media supplemented with proteins at 39℃ for 6 to 7 days. Activation treatments did not affect cleavage of the oocytes. The cleavage rates were 64.1% (41/64) in oocytes activated by electric stimulation and 76.5% (218/285) in oocytes activated by ionomycin + 6-DMAP. The proportion of development to blastocyst was 15.6% (34/218) in oocytes activated by ionomycin + 6-DMAP, but activation by electric stimulation did not support embryos developed beyond morula stage. There were no differences in the cleavage rates of activated oocytes experiencing in vivo (86.8%, 66/76) and in vitro maturation (69.0%, 127/184). However, the development rate to blastocyst stage was significantly (P<0.05) higher for oocytes matured in vivo (50.0%, 33/66) compared to in vitro (0.8%, 1/127). Culture conditions did not affect the cleavage of -activated oocytes. The cleavage rates were 51.6% (49/95) in M16, 64.3% (18/28) in TCM-199 and 81.0% (145/179) in mSOF, respectively. By contrast, the development rate of activated oocytes to stage was greater (P<0.05) for oocytes cultured in mSOF medium (23.4%, 34/145) than in M16 or TCM-199 (0.0%). Our results suggest that source of oocytes and culture conditions are major factors affecting in vitro development of caprine parthenogenetic oocytes.

Studies on the Viability of In Vitro-Matured Bovine Oocytes Vitrified by Microdrop and Straw Method (Microdrop과 Straw 방법으로 초자화 동결한 소 난자의 생존율에 관한 연구)

  • Yang, B.C.;Yang, B.S.;Sung, H.H.;Im, S.K.;Park, S.B.;Chang, W.K.;Lee, C.K.
    • Journal of Animal Science and Technology
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    • v.44 no.6
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    • pp.701-710
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    • 2002
  • Studies on the Viability of In Vitro-Matured Bovine Oocytes Vitrified by Microdrop and Straw Method To establish vitrification method for bovine oocytes, mature bovine oocytes were vitrified by microdrop (MD) or straw (Straw) method and the viability of vitrified oocytes with or without cumulus cells (CC) were examined by several methods; a) parthenogenetic activation; b) pronuclear formation after in vitro fertilization (IVF); and c) embryonic development after IVF. The survival rate of vitrified oocytes by MD was significantly higher than by Straw (92.50 vs. 74.19%, p<0.05). Most of the oocytes survived from vitrification using the MD methods. Cleavage and blastocyst development of parthenogenetically activated oocytes were higher in MD (45.05% and 10.81%, respectively; p<0.05)) than those in Straw method (27.17% and 6.52%, respectively; p<0.05). Male and female pronuclear formation of vitrified-thawed oocytes with or without cumulus cells (CC) after IVF were examined, respectively. The survival rate of vitrified oocytes by MD without CC was no difference between MD and Straw (80.368.14% vs. 67.31%). Normal fertilization (2PN) rates were not different among groups (Fresh; 54.55% vs. MD; 42.22% vs. Straw; 37.14%, p>0.05). While no fertilization (<1PN) rates were significantly different between fresh and vitrified-thawed groups (Fresh; 32.47% vs. MD; 57.78% and Straw 62.86%, p<0.05). The polyspermy (3PN) was appeared in the fresh (12.99%), but no appeared in the vitrified-thawed groups. In the without CC, normal fertilization (2PN) rates were significantly different between fresh and vitrified-thawed oocytes (Fresh; 59.38% vs. MD; 17.31% and Straw; 30.43%, p<0.05). Moreover, no fertilization (<1PN) rates were significantly different between fresh and vitrified-thawed groups (Fresh; 23.44% vs. MD; 73.08% and Straw 58.70%, p<0.05). The polyspermy (3PN, >4PN) was appeared not only fresh but vitrified-thawed groups. After IVF, two-cell developmental rates of vitrified oocytes with CC by MD and Straw were significantly low compared to fresh oocytes (Fresh; 81.76% vs. MD; 22.22% and Straw; 11.36%, p<0.05). Blastocyst developmental rates of vitrified oocytes also were significantly low compared to fresh oocytes (Fresh; 28.38 vs. MD; 1.71% and Straw 0%, p<0.05). In the without CC, two-cell developmental rates were no difference between Fresh and MD (27.59% vs. 19.25%, p<0.05), while blastocyst rates were difference between Fresh and MD or Straw (4.31% vs. 0.62% and 0%, respectively; p<0.05). In conclusion, the results indicate that the vitrified bovine oocytes have the ability to develop to the blastocyst stage after IVF.

Outcome of Preimplantation Genetic Diagnosis in Patients with Klinefelter Syndrome (클라인펠터 증후군 환자에서 착상전 유전진단의 결과)

  • Kim, Jin-Yeong;Lim, Chun-Kyu;Jun, Jin-Hyun;Park, So-Yeon;Seo, Ju-Tae;Cha, Sun-Hwa;Koong, Mi-Kyoung;Kang, Inn-Soo
    • Clinical and Experimental Reproductive Medicine
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    • v.31 no.4
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    • pp.253-260
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    • 2004
  • Objectives: Klinefelter syndrome is the most common genetic cause of male infertility and presents with 47, XXY mainly or 46, XX/47, XXY mosaicism. It is characterized by hypogonadism and azoospermia due to testicular failure, however, sporadic cases of natural pregnancies have been reported. With the development of testicular sperm extraction (TESE) and intracytoplasmic sperm injection (ICSI), sperm can be retrieved successfully and ART is applied in these patients for pregnancy. It has been suggested that the risk of chromosome aneuploidy for both sex chromosome and autosome is increased in the sperms from 47, XXY germ cells. Considering the risk for chromosomal aneuploidy in the offspring, preimplantation genetic diagnosis (PGD) could be applied as a safe and more effective treatment option in Klinefelter syndrome. The aim of this study is to assess the outcome of PGD cycles by using FISH for sex chromosome and autosome in patients with Klinefelter syndrome. Materials and Methods: From Jan. 2001 to Dec. 2003, PGD was attempted in 8 cases of Klinefelter syndrome but TESE was failed to retrieve sperm in the 3 cases, therefore PGD was performed in 8 cycles of 5 cases (four 47, XXY and one 46, XY/47, XXY mosaicism). In one case, ejaculated sperm was used and in 4 cases, TESE sperm was used for ICSI. After fertilization, blastomere biopsy was performed in $6{\sim}7$ cell stage embryo and the chromosome aneuploidy was diagnosed by using FISH with CEP probes for chromosome X, Y and 17 or 18. Results: A total of 127 oocytes were retrieved and ICSI was performed in 113 mature oocytes. The fertilization rate was $65.3{\pm}6.0%$ (mean$\pm$SEM) and 76 embryos were obtained. Blastomere biopsy was performed in 61 developing embryos and FISH analysis was successful in 95.1% of the biopsied blastomeres (58/61). The rate of balanced embryos for chromosome X, Y and 17 or 18 was $39.7{\pm}6.9%$. The rate of aneuploidy for sex chromosome (X and Y) was $45.9{\pm}5.3%$ and $43.2{\pm}5.8%$ for chromosome 17 or 18, respectively. Embryo transfer was performed in all 8 cycles and mean number of transferred embryos was $2.5{\pm}0.5$. In 2 cases, clinical pregnancies were obtained and normal 46, XX and 46, XY karyotypes were confirmed by amniocentesis, respectively. Healthy male and female babies were delivered uneventfully at term. Conclusion: The patients with Klinefelter syndrome can benefit from ART with TESE and ICSI. Considering the risk of aneuploidy for both sex chromosome and autosome in the sperms and embryos of Klinefelter syndrome, PGD could be offered as safe and more effective treatment option.

Effect of Human Follicular Fluid and Bovine Oviductal Tissue Extract on the Mouse Oocyte-Cumulus Complex (사람 난포액과 소의 수란관 조직추출액이 생쥐 난구세포에 미치는 영향)

  • 홍민정;김지수;심명선;김해권
    • Development and Reproduction
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    • v.6 no.2
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    • pp.97-104
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    • 2002
  • In most mammals, mature oocyte-cumulus complexer(OCCs) ovulate into the oviduct where fertilization by sperm takes place. However, the complex that fail to fertilize eventually undergoes degeneration while they reside in the oviduct. Yet there is no blown mechanism how both oocyte and cumulus cells degenerate. Using human follicular fluid (hFF), bovine oviductal tissue extract (BOX) and mouse OCC, the present study aimed to find how the oviduct influence the viability of the oocyte and cumulus cells in vitro. There was no difference of oocyte maturation rate between the control and BOX-treated groups. However, there was a significant difference in the survival of cumulus cells between two groups. Cumulus cells cultured in the presence of hFF alone underwent initially expansion and then they formed monolayer in the culture dish. Even after 72 hr, they proliferated well and showed fibroblast-like morphology. Cumulus cells cultured in the presence of both hFF and BOX also expanded after 24 hr, however, after 72 hr culture, they eventually detached and degenerated. Cumulus cells cultured in the BOX alone gave a similar drastic result. When the cumulus cells cultured in the presence of BOX were stained with DAPI, their nuclei showed partial condensation and fragmentation. After detailed analysis of these cells by TUNEL assay, many nuclei of them exhibited well stained spots indicating the signs of apoptosis. Based upon these observations, it is suggested that BOX might possess a factor that leads mouse cumulus cells to undergo apoptosis in vitro.

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Effects of Estrogen on the Transcriptional Activities of Catecholamine Biosynthesizing Enzymes in the Brain and Adrenal Gland of Ovariectomized Rats (난소 절제 흰쥐의 뇌와 부신에서의 Catecholamine Biosynthesizing Enzyme들의 전사에 미치는 Estrogen의 효과)

  • 유경신;이종화;최돈찬;이성호
    • Development and Reproduction
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    • v.6 no.2
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    • pp.117-122
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    • 2002
  • Dopamine(DA), norepinephrine(NE), and epinephrine(E) belong to a class of neurotransmitters known as catecholamine (CA) which are synthesized and secreted by mammalian brain and adrenal medulla. CA regulate several behavior patterns connected with breeding, and regulate GnRH-gonadotropin hormone axis' vitality between hypothalamus-pituitary gland linking with reproduction freeze. The present study examined effects of sex steroid hormone on the transcriptional activities of CA biosynthesis enzymes, tyrosine hydroxylase(TH), dopamine $\beta$ -hydroxylase(DBH), and phenylethaolamine-N-methyl transferase(PNMT). Mature female rats were ovariectomized(OVX) and implanted with 17 $\beta$-estradiol(E$_2$: 500 $\mu\textrm{g}$/ml) or sesame oil. Forty-eight hours after implantation all the animals were sacrificed. Total RNAs were extracted immediately and were applied to semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR). The expression level of TH was appeared by hypothalamus > SNc> adrenal medulla orders in OVX+Oil group, and by SNc > hypothalamus) adrenal medulla orders in OVX+E$_2$ group. Treatment with E$_2$ significantly increased TH expression in SNc and adrenal medulla but in hypothalamus, the reduced TH expression was observed. The expression level of DBH was appeared by adrenal medulla > SNc > hypothalamus orders in OVX+Oil group and in OVX+E$_2$ group. Administration of E$_2$ significantly reduced DBH expression in SNc, and increased in adrenal medulla. Two cDNA products, large(PNMT1) and small(PMNTs) species of 110bp difference, were amplified in SNc and hypothalamus, but only PNMTs was observed in adenal medulla. The PNMTs expression level was in the order of adrenal medulla > hypothalamus > SNc in both OVX+Oil and OVX+E$_2$ group. The PNMTs expression in SNc and adrenal medulla was significantly increased byE$_2$. The present report demonstrated that estrogen effects on transcriptional activities for CA biosynthethic enzymes were tissue specific in adrenal medulla as well as different region of brain. These results suggest that it might be crucial relationship between the type of estrogen receptor and CA enzyme gene expression.

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Effect of cytokines in breast milk on infant during the first two-years of life (모유 내 사이토카인 함량과 2 년간 추적관찰 후 알레르기 발생)

  • Kim, Woo Kyung;Yoon, Hae-Sun
    • Clinical and Experimental Pediatrics
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    • v.50 no.9
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    • pp.912-918
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    • 2007
  • Purpose : Breast milk contains several components that provide specific immunity and affect the maturation of the infant's immune system. Allergic disease (AD), including atopic eczema, asthma, allergic rhinitis, and food allergy is characterized by an imbalance between cytokines produced by distinct T-helper cell subtypes. The aim of the study was to investigate the concentrations of cytokines and chemokines that were involved in allergic reactions in breast milk from allergic and nonallergic mothers and then analyse the effect of breastfeeding duration on the prevalence of allergic disease in the age of two. Methods : The breast milk samples were collected from mothers with AD (n=88) and without AD (n=47). Breast milk was collected at the second day (colostrum) and four weeks later (mature milk).The level of Interlukine (IL)-4, IL-5, IL-6, IL-8, IL-10, IL-13, $TGF-{\beta}1$, $TGF-{\beta}2$, RANTES in breast milk were determined by commercially available enzyme-linked immunosorbent assay (ELISA) kits according to the manufacturer's instructions. Results : Mothers with AD had a higher concentration of IL-8 in colostrum compared with those without AD (P=0.021). But, $TGF-{\beta}1$ and $TGF-{\beta}2$ were higher concentrated in colostrum of mother without AD (P=0.013, P=0.001). Whereas concentrations of other cytokines were not significantly different between the two groups. There was no association between levels of cytokines and chemokines in the breast milk and allergic development during the first 2 years of life in the infants. Conclusion : The higher concentration of $TGF-{\beta}1$ and $TGF-{\beta}2$ in colostrum from non-allergic mothers may explain the protective effect. But, the higher concentrations of IL-8 in colostrum from allergic mothers may in part explain the controversial results on the protective effect of breastfeeding against allergic diseases. We conclude that there is no convincing evidence form a relation between cytokines in breast milk and allergic diseases in infants. Longer follow-up are necessary to evaluate the effects of breast milk components on AD.