• Journal of Animal Science and Technology
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• 제60권6호
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• pp.11.1-11.7
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• 2018

After pubertal, cohort of small antral follicles enters to gonadotrophin-sensitive development, called recruited follicles. This study was aimed to identify candidate genes in follicular cyclic recruitment via analysis of protein-protein interaction (PPI) network. Differentially expressed genes (DEGs) in ovine granulosa cells of small antral follicles between follicular and luteal phases were accumulated among gene/protein symbols of the Ensembl annotation. Following directed graphs, PTPN6 and FYN have the highest indegree and outdegree, respectively. Since, these hubs being up-regulated in ovine granulosa cells of small antral follicles during the follicular phase, it represents an accumulation of blood immune cells in follicular phase in comparison with luteal phase. By contrast, the up-regulated hubs in the luteal phase including CDK1, INSRR and TOP2A which stimulated DNA replication and proliferation of granulosa cells, they known as candidate genes of the cyclic recruitment.

• 한국수정란이식학회지
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• 제30권4호
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• pp.327-333
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• 2015

The objectives of the present study were to select an effective basic medium including its hormone and protein supplementation for IVM of oocytes of indigenous zebu cows. The ovaries of cows were collected from slaughter house and the follicular fluid was aspirated from 2 to 8 mm diameter follicles. The COCs with more than 3 cumulus cell layers and homogenous cytoplasm were selected for maturation. The oocytes were matured in media for 24 hrs at $39^{\circ}C$ with 5% $CO_2$ in humidified air. The maturation of oocytes was evaluated by examining the presence of first polar body under microscope. An efficient basic medium was determined after culturing COCs in either TCM 199 or SOF medium in Experiment 1. An efficient hormone supplementation was determined after culturing COCs in either FSH or gonadotrophin supplemented TCM 199 in Experiment 2. An efficient protein supplementation was determined after culturing COCs in either FBS or Oestrous cow serum (OCS) supplemented TCM 199 in Experiment 3. The oocyte recovery rate per ovary was 3.35. The overall rate of IVM was 74.6%. The maturation rate was $75.5{\pm}3.9$ and $62.2{\pm}20.2%$ in TCM and SOF medium, respectively (P>0.05). The maturation rate of oocytes was significantly higher ($76.6{\pm}13.2%$) in FSH supplemented medium than gonadotrphin supplemented counterpart ($69.7{\pm}10.8%$) (P<0.05). The maturation rates of oocytes were $81.7{\pm}12.9$ and $85.7{\pm}12.7%$ in medium supplemented with FBS and OCS, respectively (P>0.05). In conclusions, both TCM 199 and SOF supplemented with either FBS or OCS, and FSH may be used as medium for IVM of indigenous zebu oocytes in Bangladesh.

• Asian-Australasian Journal of Animal Sciences
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• 제24권9호
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• pp.1211-1216
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• 2011

Sexual maturity in poultry is controlled by a complex neural circuit located in the basal forebrain, which integrates the central and peripheral signals to activate hypothalamic gonadotrophin-releasing hormone (GnRH) secretion. This study demonstrated the changes of GnRH-I, POMC and NPY mRNA transcription in hypothalamus and IGF-I and leptin levels in serum of Shaoxing ducks during puberty. Body weight increased progressively from d30 to d120 and at d120 the flock reached 5% of laying rate. A significant upregulation of hypothalamic GnRH-I mRNA expression was observed from d60, reaching the peak at d120. POMC and NPY mRNA expression in hypothalamus showed a similar pattern, which increased from d30 to d60, followed by a significant decrease towards sexual maturity. Serum IGF-I levels exhibited two peaks at d30 and d120, respectively. Serum leptin displayed a single peak at d90. The results indicate that the down-regulation of POMC and NPY genes in hypothalamus coincides with the up-regulation of GnRH-I gene to initiate sexual maturation in ducks. In addition, peripheral IGF-I and leptin may relay the peripheral metabolic status to the central system and contribute to the initiation of the reproductive function in ducks.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.33-33
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• 2003

This study was conduct to compare the efficacy to produce male and female somatic cloned piglets. Maturation of porcine COCs was accomplished by incubation in NCSU-23 medium supplemented with 0.6 mM cysteine, 10% porcine follicular fluid, 1mM dibutyryl cyclic adenosine monophosphate (dbc-AMP, Sigma, USA), and 0.1 IU/ml human menopausal gonadotrophin (hMG, Teikokuzoki, Japan) for 20h and then cultured without dbcAMP and hMG for another 18 to 24 h. Female and male fetal cells were isolated from each fetus, cultured in ES-DMEM medium containing 10% FCS. Enucleated oocytes were fused with fetal fibroblasts (passage 4 to 15). Reconstructed embryos were cultured in NCSU-23 with 4 mg/ml BSA under mineral oil at 39$^{\circ}C$ in 5% $CO_2$ in air. A total of 12,328 nuclear-transferred embryos (1- to 4-cell stage) were surgically transferred into 69 surrogate gilts. Three recipients aborted during the period of conception. Three gilts delivered eleven female piglets, and five recipients gave rise to birth 22 male piglets. The average birth weigh of the cloned piglets was 1.52 kg (1.38~1.83 kg) in female piglets and 0.84 kg (0.45~1.25 kg) in male piglets. Alive cloned pigs was seven in female piglets (63.6%) and four in male piglets (18.2%). The other two recipients is ongoing. This study suggests that female-derived fetal cell as a nuclear donor has more capability on production of cloned piglets than male.

• Clinical and Experimental Reproductive Medicine
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• 제18권2호
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• pp.181-187
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• 1991

Epidermal growth facter(EGF)는 내열성이 강하고 분자량이 6045 dalton인 단쇄상의 polypeptide로써, Cohen에 의해 생쥐의 악하선에서 처음 발견된 이래, 여러학자들에 의해 많은 연구가 되어왔다. 인체의 EGF는 urogastrone이라고도 불리우며, 인체의 소변에서 처음 검출되었고 분자구조 및 생리작용이 생쥐의 EGF와 매우 유사한 것으로 판명되었다. EGF의 자세한 작용기전은 확실히 규명되어있지는 않지만 세포의 증식과 분화를 촉진시키며 위산의 분비를 억제시킨다고 알려져 있다. 또한 EGF receptor는 분자량이 170,000${\sim}$180,000dalton인 세포표면의 polypeptide로써 인체, 쥐, 닭, 소 등의 세포막조직에 특이하게 결합되어 있다. 최근 수년동안 몇몇 학자들에 의해 EGF가 배아와 태아 및 태반의 성장을 촉진시키고 chorionic gonadotrophin과 placental lactogen의 분비를 증진하는데 기여할 것이라고 가정되어 왔다. 그러나 아직까지 배아착상에 대한 EGF의 작용여부에 관해서는 발표된 문헌이 없어 저자는 radioreceptor assay를 이용하여 EGF receptor binding과 토끼의 배아착상과의 관계를 규명하고자 임신경과에 따른 착상부위와 비착상부위의 자궁 및 태아측 태반과 모체측 태반을 분리취득하고 receptor binding assay를 시행하여 다음과 같은 결론을 얻었다. 1. 전임신군과 비임신군의 자궁조직의 membrane fraction으로부터 specific한 EGF receptor binding이 관찰되었다. 2. 착상전 임신 3일에 자궁조직의 EGF receptor수는 4.72 +0.16($10\;mol/{\mu}g$)로 비임신시보다 의의있게 증가되어 있었고(p<0.01), 착상시기인 임신 7일에는 착상된 부위에서 20.33+6.58로 훨씬 더 높은 측정치를 나타내었다(p<0.05). 3. 착상이후 가장 먼저 취득된 임신 14일의 태아측 태반은 모체측 태반의 1.39+0.49에 비해 훨씬 높은 11.94+1.97의 EGF receptor 측정치를 보였다 (p<0.01). 4. 이상의 소견들로 보아 EGF가 토끼의 배아착상에 밀접한 관련이 있을 것으로 추측되며, 이러한 착상전후의 EGF의 작용은 태아측으로부터 일 것으로 예상된다.

• Asian-Australasian Journal of Animal Sciences
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• 제13권5호
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• pp.595-599
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• 2000

Fine wool sheep (n=18) maintained in a tropical environment were allocated to three treatment groups. Estrus was induced with two injections of $PGF_{2{\alpha}}$ (10 mg. im) at 10 days interval. Superovulation treatment started 2 days prior to the second injection of $PGF_{2{\alpha}}$. Each ewe was treated with a total dose of 25 units FSH (Super-OV) i.m. every 12 hover 3 days; Group 2 were also injected i.m. with 200 IU PMSG at the first injection of FSH; Group 3 was treated as in Group 2 and also with GnRH ($4{\mu}g$ Buserelin) at the onset of estrus. The ewes in estrus were mated with a fertile ram. Ovarian examination and recovery of embryo and ova were performed at laparoscopy and laparotomy on day 3 or 4 after mating. Data for onset of estrus, duration of estrus, number of corpora lutea (CL), number of unnovulated large follicle (LF), embryo recovery rate, embryo quality and fertilization recorded for the 3 groups. Ewes in the Group 1 set in estrus later (p<0.05; $50.0{\pm}7.29h$) than the ewes in Group 2 ($24.5{\pm}3.58$) and 3 ($32.5{\pm}3.58h$). The duration of estrus, ovarian size and ovarian response (number of CL and LF) did not differ significantly (p>0.05) among the 3 groups. The proportion of ewes with a superovulatory response (${\geq}2$ CL) was the lowest (50%) in Group 1 treated with FSH alone but ova/embryo recovery (100%) and fertilization (100%) was significantly (p<0.05) higher than Group 2 (58.3 and 85.7%, respectively) and Group 3 (48.6 and 50%, respectively). It is concluded that in tropical fine wool sheep, there is no difference in the 3 treatments for yield of good quality embryos but ovarian response and ovulation rate increased on additional use of PMSG and GnRH respectively to FSH alone.

• Asian-Australasian Journal of Animal Sciences
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• 제17권12호
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• pp.1652-1656
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• 2004

The aim of this study was to obtain mature ova or embryos at a single cell stage, which can be used in avian transgenesis and nuclear transfer through multiple ovulations, in vitro fertilization and culture. Chicken anterior pituitary extract (CAPE) or acetone-dried chicken anterior pituitary extract (ACAPE) was used to induce multiple ovulations in hens pretreated with pregnant mare' serum gonadotrophin (PMSG). In vitro fertilization of the multiple ovulated ova was performed by inseminating sperm onto the germinal disks in m-Ringer' solution and incubating the ova at 41$^{\circ}C$, 5% $CO_2$ for 10 h in DME-F12 medium containing 20% liquid albumen. The in vitro fertilization process was observed using an environmental scanning electron microscope. When normal laying hens (white Leghorn) were administered daily with PMSG (100 IU), egg laying ceased in most hens within 3 to 8 days. Ovulation began to occur about 7.5 h after injection of CAPE and ACAPE. The number of ovulated ova was 1.00${\pm}$0.00, 2.33${\pm}$0.52 and 2.20${\pm}$0.45, respectively, after receiving 100, 200 and 300 mg CAPE. The number of ovulated ova was 2.00${\pm}$0.00, 2.86${\pm}$0.69 and 3.00${\pm}$1.22, respectively, after receiving 10, 15 and 20 mg ACAPE. The fertilized and cultured ova were able to develop into embryos up to the 32 cell stage. The present experiments demonstrated that multiple ovulations can be induced by CAPE and ACAPE successfully, and the ova resulted from the treatment retained the capability for further fertilization and embryonic development. These data provide new information to support the establishment of an in vitro culture system for future avian transgenesis studies.

• Asian-Australasian Journal of Animal Sciences
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• 제13권7호
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• pp.897-900
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• 2000

The experiment was conducted to study the effect of GnRH administration at induced estrus on pituitary and ovarian response in buffalo heifers. Eight Murrah river buffaloes of 12 to 13 months of age were treated with $PGF_{{2}{\alpha}}$ and eCG combination. GnRH (Fertagyl) 200 ug was injected (iv) at estrus in four heifers (treated group) while saline (2 ml, iv) was injected in remaining four heifers (control group). Blood was collected through jugular catheter to estimate plasma FSH, LH and estradiol level. The pretreatment plasma FSH, LH and estradiol values ranged from $8.46{\pm}1.97ng/ml$ to $12.31{\pm}1.30ng/ml$, $0.87{\pm}0.21ng/ml$ to $1.19{\pm}0.29ng/ml$ and $19.09{\pm}2.38pg/ml$ to $20.24{\pm}1.00pg/ml$ respectively. The plasma estradiol concentration elevated significantly (p<0.05) within 24 hr after eCG administration and reached its peak levels of $154.09{\pm}17.28pg/ml$ and $181.95{\pm}31.82pg/ml$ at estrus in respectively treatment and control groups. The plasma FSH and LH concentrations did not increase during follicular development after eCG administration while initial significant (p<0.05) increases in both plasma FSH and LH concentrations occured within 5 and 10 min, reaching peak levels of respectively $110.06{\pm}23.56ng/ml$ and $13.15{\pm}3.13ng/ml$ within 90 min after GnRH injection was detected. A sharp and significant decline in plasma estradiol concentration ($59.27{\pm}8.78pg/ml$) associated with synchronized ovulation within 24 hours after GnRH injection was recorded. The observation suggest that the hypophysis of prepubertal buffaloes treated with eCG have gonadotrophins awaiting the releasing factor to evoke release of gonadotrophin during the follicular phase to induce synchronized ovulation.

• Asian-Australasian Journal of Animal Sciences
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• 제10권5호
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• pp.514-518
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• 1997

This study was designed to record the ovarian response towards a combined administration of heterologous buffalo FSH (buFSH) and LH (buLH) in goats. The impact of such a treatment on ovarian structures and on the plasma profile of the ovarian sex steroids (estradiol $17-{\beta}$ and progesterone) was studied. The buFSH and buLH were isolated from the buffalo pituitaries involving a procedure of ethanolic extraction, acetone precipitation followed by metaphosphoric acid - ammonium sulphate fractionation. Both gonadotrophin samples prepared were found biologically active and potent. There was an increase in the total number of follicles in the treated group ($12.66{\pm}1.24$) vis-a-vis the control group ($8.50{\pm}2.06$). However, the percentage ($51.48{\pm}6.37$) of large follicles were found reduced ($23.74{\pm}5.93$) following the treatment. Again the number of corpora lutea were observed significantly higher ($2.33{\pm}0.47C.L.$) in the treated group than (1 C. L.) in the control group. The peak plasma estradiol- $17{\beta}$ levels achieved, were much higher ($17.16{\pm}9.52pg/ml$) in the treated group, than the peak ($7.22{\pm}1.67pg/ml$) achieved in the control group. Similar trend was observed with respect to the progesterone levels (higher in the treated group). This study thus indicated that, a combined administration of heterologous buffalo FSH and LH to goats speeded up development of larger follicles nearing the ovulation stage. This population of the follicles subsequently got reduced and lead to the formation of the increased number of the corpora lutea observed in this study.

• 한국동물학회지
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• 제31권2호
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• pp.81-86
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• 1988

생쥐혹은 돼지의 난자-난구 복합체를 인공배양하면서 뇌하수체호르몬 혹은 세포내 cAMP의 농도를 높이는 시약을 사용하여 난자의 성숙과 난구세포의 분산을 조절하고 이 때 두 세토들 사이에 상호협력(metabolic cooperativity)관계가 어떻게 변하는지를 조사하여 보았다. 생귀와 돼지의 난구세포들은 뇌하수체호르몬이나 cAMP의 증가에 의해 분산이 유도됨과 동시에 배양액 내에서 있는 uridine의 흡수가 크게 촉진되었다. (대조군의 약 4배). 그러나 난구세포에 흡수된 uridine이 난자로 전달되는 물질이동율(transfer ratio)은 대조군과 같이 시간이 지남에 따라 감소하였으며 cAMP의 영향도 거의 받지 않았다. 또한 물질이동율의 감소는 난구세포의 부산여부나 난자의 성숙(핵붕괴) 여부에 크게 영향을 받지 않았다.단지 생쥐의 경우 호르몬에 의해 물질이동율의 감소가 더욱 두드러지게 나타나는 경우를 볼수 있었다. 따라서 물질이동율의 변화가 난구세포의 분산이나 난자의 성숙과 직접 관련이 없는 것으로 보여지며 두 세포들 사이의 metabolic cooperativity가 난자의 성숙조절에 중요한 요인이 되지 않는다는 것을 알았다.