• 한국발생생물학회지:발생과생식
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• 제7권2호
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• pp.119-126
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• 2003

본 연구실에서는 이전의 실험에서 suppression subtractive hybridization(SSH)을 통하여 생쥐의 생후 1일자 난소와 5일자 난소에서 차이 나게 발현하는 유전자들의 목록을 얻었고 그 중에서 MT transposon-like element, clone MTi7(MTi7)이 성장하는 난포에서 더 높게 발현한다는 것을 알아냈다(Park et al., 2002). In situ hybridization과 RNA interference를 이용한 연구결과, MTi7은 난자에서 특이 적으로 발현하는 유전자로 특히 난자성숙에 관여하는 것으로 관찰되었다(Park et at., 2003). 그러나 현재까지 MTi7의 염기서열은 생쥐에서만 알려져 있다. 따라서 본 연구는 두 부분으로 나누어서 첫째, MTi7이 다른 포유류에도 존재하는지 알아보기 위해 소,돼지, 흰쥐 그리고 사람 등 각기 다른 네 종의 난소 cDNA를 사용하여 새로운 MTi7을 분리하고자 하였으며, 둘째, 생쥐의 MTi7이 transposon의 특징을 갖고 있어 다른 유전자에 삽입되어 있는지를 알아보기 위해 inverse PCR을 시행하여 MTi7주변의 유전자가 있는지를 조사하였다. 네 종의 난소 cDNA를 사용하여 생쥐의 MTi7과 매우 유사한 염기서열을 갖고 있음을 알았다(87%∼98%). Inverse PCR 결과, 생쥐의 MTi7은 beta-carotene 15, 15'-monooxygenase(Bcdo) 유전자 혹은 serine protease inhibitor, Kunitz type I(Spint 1) 유전자에 삽입되어 있음을 알 수 있었다. 본 연구의 결과로 여러 포유류의 MTi7 sequence를 알아내고, 또한 생쥐의 MTi7이 삽입되어 있는 유전자를 알아냄으로써 머지 않은 장래에 난자형성 및 난포형성과정에 있어서 MTi7의 역할을 밝혀 낼 수 있을 것으로 기대된다.

• Archives of Pharmacal Research
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• 제29권8호
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• pp.666-676
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• 2006

Gangliosides are widely distributed in mammalian cells and play important roles in various functions such as cell differentiation and growth control. In addition, diabetes and obesity cause abnormal development of reproductive processes in a variety of species. However, the mechanisms underlying these effects, and how they are related, are not fully understood. This study examined whether the differential expression of gangliosides is implicated in the abnormal follicular development and uterine architecture of streptozotocin (STZ)-induced and db/db diabetic mice. Based upon the mobility on high-performance thin-layer chromatography, mouse ovary consisted of at least five different ganglioside components, mainly gangliosides GM3, GM1, GD1a and GT1b, and diabetic ovary exhibited a significant reduction in ganglioside expression with apparent changes in the major gangliosides. A prominent immunofluorescence microscopy showed a dramatic loss of ganglioside GD1a expression in the primary, secondary and Graafian follicles of STZ-induced and db/db diabetic mice. A significant decrease in ganglioside GD3 expression was also observed in the ovary of db/db mice. In the uterus of STZ-induced diabetic mice, expression of gangliosides GD1a and GT1b was obviously reduced, but gangliosides GM1, GM2 and GD3 expression was increased. In contrast, the uterus of db/db mice showed a significant increase in gangliosides GM1, GD1a and GD3 expression. Taken together, a complex pattern of ganglioside expression was seen in the ovary and uterus of normoglycemic ICR and $db/^+$ mice, and the correspoding tissues in diabetic mice are characterized by appreciable changes of the major ganglioside expression. These results suggest that alterations in ganglioside expression caused by diabetes mellitus may be implicated in abnormal ovarian development and uterine structure.

• Asian-Australasian Journal of Animal Sciences
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• 제23권8호
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• pp.1000-1006
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• 2010

Phosducin (PDC) is a photoreceptor cell-specific protein that is phosphorylated by cyclic nucleotide-dependent protein kinase. PDC and PDC-like proteins (PDCL, PDCL2, and PDCL3) are members of a conserved family of small thioredoxin-like proteins that modulate the ${\beta}$- and ${\gamma}$-subunits of G-proteins. In mammals, Pdc, Pdcl, and Pdcl3 genes show ubiquitous expression; however, Pdcl2 gene expression is limited to the testis and ovary. The aim of the present study was to examine the expression patterns of chicken Pdcl2 (cPdcl2) during testicular and ovarian development. Protein sequence comparisons performed using the CLUSTAL X program revealed that the amino acid sequences and potential phosphorylation sites of cPDCL2 and mammalian PDCL2 proteins were highly conserved. Quantitative real-time PCR analysis revealed that cPdcl2 was differentially expressed in the testis and ovary. Specifically, cPdcl2 expression was detected at low levels in the ovary at all time points. In the testis, cPdcl2 expression was detected at low levels until 5 weeks of age. At 8 weeks of age, however, cPdcl2 showed increased expression levels in the testis. Using in situ hybridization, we detected high levels of cPdcl2 expression in the testis, particularly in the spermatocytes and round spermatids. In summary, our data describe expression patterns of germ cell-specific Pdcl2 during testicular and ovarian development in chickens.

• Journal of Microbiology and Biotechnology
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• 제23권8호
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• pp.1116-1122
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• 2013

Cell line development is the most critical and also the most time-consuming step in the production of recombinant therapeutic proteins. In this regard, a variety of vector and cell engineering strategies have been developed for generating high-producing mammalian cells; however, the cell line engineering approach seems to show various results on different recombinant protein producer cells. In order to improve the secretory capacity of a recombinant tissue plasminogen activator (t-PA)-producing Chinese hamster ovary (CHO) cell line, we developed cell line engineering approaches based on the ceramide transfer protein (CERT) and X-box binding protein 1 (XBP1) genes. For this purpose, CERT S132A, a mutant form of CERT that is resistant to phosphorylation, and XBP1s were overexpressed in a recombinant t-PA-producing CHO cell line. Overexpression of CERT S132A increased the specific productivity of t-PA-producing CHO cells up to 35%. In contrast, the heterologous expression of XBP1s did not affect the t-PA expression rate. Our results suggest that CERT-S132A-based secretion engineering could be an effective strategy for enhancing recombinant t-PA production in CHO cells.

• Asian-Australasian Journal of Animal Sciences
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• 제17권11호
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• pp.1496-1500
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• 2004

In the mammalian ovary the follicular fluid contains proteins and peptides which play an important role in growth, development and maturation of oocytes. The gonadotropins and some other factors work synergistically and regulate ovarian functions. In the present study the effect of follicular fluid proteins (FFP) and gonadotropins on progesterone secretion by granulosa cells (GC) from buffalo ovary, was investigated during culture. The follicular fluid was collected from small (<5 mm), and medium (5-8 mm) follicles obtained from buffalo ovaries. The follicular fluid from medium follicles was fractionated with ammonium sulphate at 80% saturation. The precipitated protein fraction was further resolved in to minor (peaks I, III) and major (peak II) proteins using gel filtration (Sephadex G-200). The FFP from small follicles and major FFP (peak II) at a dose of 200 $\mu$g/well, significantly stimulated progesterone secretion by pooled GC (3${\times}10^{5}$ cells/2 ml medium/well). The minor FFP did not show any stimulatory effect. There was a significant increase in progesterone secretion by pooled GC in presence of FFP and LH (10 ng/well), however, FSH (20 ng/well) with FFP exhibited an inhibitory effect. The major FFP and gonadotropins were also studied for their effect on progesterone production by GC isolated from medium and large size follicles. The GC from medium follicles were more responsive to FSH and FFP whereas GC from large follicles exhibited enhanced progesterone secretion with LH and FFP. These results indicated that FFP have their own stimulatory effect and also act synergistically with gonadotropins. The significantly different response shown by GC, for steroid hormone secretion, is based on their stage of growth and differentiation. The purification and characterization of such steroidogenic proteins may help in elucidating their role in growth and differentiation of granulosa cells.

• 한국동물학회지
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• 제39권4호
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• pp.410-418
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• 1996

Gangiloside는 포유동물세포에 편재하는 막성분으로서, 이들은 세포상호간의 접착, 분화 및 정보전달과정에 과여하는 것으로 알려지고 있다. Rat난소는 주요한 Gangiloside로서 GM3를 함유하고 있으며, 본 연구에서는 폐쇄난포에서 이들의 분포여부와 난포의 발달 과정에서의 변화여부를 조사하기 위하여, Rat 난소의 동결절편을 이용해 GM3를 포함 11종류의 Gangilo-series Gangiloside에 대해 특이한 단일항체로서 염색시켰다. 폐쇄난포들에서 GM3는 난포발달이 진행되는 동안 시간적, 공간적으로 서로 다른 양식으로 발현하였다. 그러나 GM1을 포함한 다른 종류의 Gangiloside들은 면역조직화학적으로 검출되지 않았다. 일차난포에서 관찰되는 폐쇄난포들에서 GM3는 모든 교막세포와 난자에 인접한 과립막세포의 일부에서 발현하였다. 이차난포의 시기에서 이들 폐쇄난포의 GM3는 모든 교막세포와 과립막세포들에서 발현하였다. 이어서 발달하고 있는 그라프난포의 시기에서 관찰되는 폐쇄난포의 GM3발현은 이차난포에서의 분포패턴과 유사함을 보여 주었다.

• Clinical and Experimental Reproductive Medicine
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• 제49권1호
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• pp.16-25
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• 2022

In mammalian species, females are born with a number of oocytes exceeding what they release via ovulation. In humans, an average girl is born with over a thousand times more oocytes than she will ovulate in her lifetime. The reason for having such an excessive number of oocytes in a neonatal female ovary is currently unknown. However, it is well established that the oocyte number decreases throughout the entire lifetime until the ovary loses them all. In this review, data published in the past 80 years were used to assess the current knowledge regarding the changing number of oocytes in humans and mice, as well as the reported factors that contribute to the decline of oocyte numbers. Briefly, a collective estimation indicates that an average girl is born with approximately 600,000 oocytes, which is 2,000 times more than the number of oocytes that she will ovulate in her lifetime. The oocyte number begins to decrease immediately after birth and is reduced to half of the initial number by puberty and almost zero by age 50 years. Multiple factors that are either intrinsic or extrinsic to the ovary contribute to the decline of the oocyte number. The inflammation caused by the ovulatory luteinizing hormone surge is discussed as a potential contributing factor to the decline of the oocyte pool during the reproductive lifespan.

• Applied Microscopy
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• 제40권4호
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• pp.211-218
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• 2010

포유동물의 난소는 호르몬과 감각신경 및 교감신경의 지배를 받아 난소 내 혈류의 조절, 스테로이드 호르몬 생성 및 난포의 발달과 관계된 난소의 고유기능이 조절되고 있다. 본 실험에서는 통상적인 추적자인 CT-HRP와 Pseudorabies바이러스(PRV)의 Bartha strain을 신경추적자로 이용하여 뇌하수체 후엽 및 난소와 연결된 중추신경부위를 밝히고자 하였다. 또 oxytocin을 난소 지배 신경축속에서 동정함으로써 신경축내 oxytocinergic neuron들의 존재를 확인하고, 이들이 배란을 중심으로 한 난소생식주기에 따라 보이는 중추 내 oxytocin신경세포의 변화를 조사하고자 하였다. Sprague-Dawley 흰쥐를 대상으로 난소 내에 PRV를 주사하고 48시간 후 실험동물들은 4% paraformaldehyde-lysine periodae로 고정하였으며, 뇌를 적출하여 $30{\mu}m$ 두께의 관상연속 절편을 만들어 CTHRP, PRV 및 oxytocin에 대한 삼중염색을 시행하였다. 본 실험 결과 후뇌에서부터 전뇌에 이르기까지 PRV에 양성반응을 보인 신경핵들이 관찰되어 난소를 지배하는 신경축을 구성할 수 있었다. 또 시상하부의 뇌실옆핵에서 oxytocin, PRV와 CT-HRP에 삼중으로 염색된 세포가 관찰됨으로써 신경내분비축과 자율신경축이 공동으로 기원하고 있다는 것을 형태학적으로 보여주었다. 따라서 oxytocin은 이 두 계통 내에서 호르몬의 역할과 신경전달물질의 역할을 겸할 것이라는 것을 추측할 수 있었다.

• 한국잠사학회:학술대회논문집
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• 한국잠사학회 2003년도 International Symposium of Silkworm/Insect Biotechnology and Annual Meeting of Korea Society of Sericultural Science
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• pp.128-129
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• 2003

Cricket (Gyilus bimacutus) is mass-bred in 6 time cycles per one year in insect farms. They are used as dry or live foods for animals, tropical fish, reptile and amphibians. Therefore, it is necessary to study the genotoxicity of whole bodies of G. bimaculatus. The aim of this present study was to evaluate the genotoxicity of the G bimaculatus extract with three methods, Ames test, chromosome aberration test in Chinese hamster ovary cells in vitro and micronucleus (MN) test in vivo which involve the different test systems (bacteria, mammalian cells and mice nuclei). (omitted)

• 한국환경성돌연변이발암원학회지
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• 제11권2호
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• pp.107-117
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• 1991

The effect of 3-aminobenzamide (3AB), an inhibitor of poly (ADP-ribose) polymerase, on ethyl methanesulfonate (EMS)-or bleomycin (BLM)-induced DNA repair synthesis and chromosome aberrations was examined during the cell cycle of Chinese hamster ovary (CHO)-K$_1$ cells. The synchronized cells were obtained by using thymidine double block method and mitotic selection method. Three assays were employed in this study: unscheduled DNA synthesis, alkaline elution and chromosome aberrations. 3AB alone did not induce DNA repair and chromosome aberrations in all phases. The post-treatment with 3AB inhibited DNA repair synthesis induced by EMS or BLM in G$_2$ phase, whereas 3AB did not affect chromosome aberrations induced by EMS or BLM in all phases. These results suggest that 3AB aggravates the cell cycle disturbance which occur after DNA damage, and leads to an accumulation of cells at G$_2$ phase, and inhibits DNA repair synthesis, while the effect 3AB on chromosome aberrations may need reevaluated.