• 한국발생생물학회지:발생과생식
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• 제6권1호
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• pp.55-66
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• 2002

배아줄기세포(embryonic stem cell, ESC)는 미분화상태로 지속적인 계대가 가능하며, 정상 핵형과 전 분화능(pluripotency)을 가져 생체내-외에서 분화 유도시 삼배엽성의 모든 세포로 분화 가능하다. ESC를 feeder 세포 없이 부유배양하면 배아체(embryoid body, EB)를 형성하고, 초기 배아 발생과 유사한 분화 양상을 갖는다. ESC의 분화 유도가 초기배아 발생처럼 생식호르몬(GTH: FSH, LH; steroids)의 영향을 받는지는 불명하다. 본 연구는 ESC가 분화과정중 생식호르몬처리에 의해 그들 수용체가 발현되는가를 알아보고자 하였다. 순계혈통 생쥐인 C57BL/6J에서 과배란 유도후 포배를 수획하고, 유사분열적으로 불활성화된 feeder 세포와 공배양하여, 계대배양 하는 중 배아줄기세포주(JHYl)를 확립하였다. JHY1의 alkaline phosphatase 활성과 SSEA-1, 3, 4 발현을 통해 ESC임을 확인하였다. Feeder 세포 없이 ESC를 계대배양 후 호르몬처리(FSH LH E$_2$, P$_4$, T)하에서 5일 동안 부유배양하여 배아체를 형성시키고, 이후 7일 동안 부착배양하여 분화를 유도하였다. GTH와 스테로이드의 수용체 발현 실험에서 ESC에 E$_2$ 처리에 의한 LHR의 발현 증가를 제외한 나머지 호르몬 처리군에서 ESC보다 낮은 생식호르몬의 수용체 발현이 관찰되었다. 생식호르몬을 농도별 수용체 발현 정도는 증감되지 않았다. 미분화 ESC 표지유전자인 Oct-4는 호르몬 처리군에서도 발현되었다. 각 배엽의 표지유전자들(영양세포, handl; 외배엽성, keratin와fgf-5; 중배엽성, enolase와 $\alpha$ -globin; 내배엽성, gata-4와 $\alpha$ -fetoprotein) 등의 발현 양상을 조사한 결과 호르몬 처리후 내배엽성 표지유전자외에는 발현 증가가 관찰되지 않았다. 즉 생식호르몬에 의해 gata-4, $\alpha$-fetoprotein의 발현이 증가되는 것으로 보아 내배엽성 계열로의 분화 유도가 이루어진 것으로 사료된다.

• 한국수정란이식학회지
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• 제31권1호
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• pp.47-52
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• 2016

본 연구는 희소한우의 증식을 위해 성 감별 처리된 정자를 체내수정란 생산 및 이식 기술에 적용하고자 정자의 농도별 수정란 회수 결과를 조사하고, 생산된 수정란은 이식을 통해 후대 생산 기술을 정립하고자 실시하였다. 가축유전자원센터에서 보유 중인 칡소 암소를 선발하여 공란우로 공시하였으며, 과배란처리는 발정주기에 관계없이 CIDR-Plus(Intravaginal Progesterone Releasing Device)를 질 내에 삽입하고, 4일 후부터 FSH(Antorin)를 12시간 간격으로 4일간 총 28AU(Amour unit) 절감 주사하고, FSH 주사 3일째 CIDR-Plus을 제거함과 동시에 $PGF_{2{\alpha}}$(Lutalyse) 3.0 ml를 주사해 과배란을 유기하였다. 인공수정은 발정이 확인된 후 12시간 간격으로 3회 실시하였다. 대조구의 경우, 보편적으로 사용하는 인공수정용 스트로우인 2,000만 농도의 칡소 동결정액 스트로우를 이용하였으며, 성감별 처리구의 경우, 스트로우 당 400만과 1,000만의 두 실험군으로 인공수정을 실시하였다. 수정란의 회수는 인공수정 후 7일째 되는 날 실시하였으며, 이식가능 수정란은 IETS의 기준에 따라 판정하였다. 대조구에서 체내수정란 회수 결과는 이식가능 수정란이 두 당 $6.20{\pm}2.28$개로 전체 회수 수정란의 67.39%로 나타났으나, 성감별 정자처리구에서는 $4{\times}10^6$ 농도에서 $4.33{\pm}5.39$개(26.53%), $10{\times}10^6$ 농도에서 $1.57{\pm}1.72$개(24.44%)로, 대조구에 비해 유의적으로 낮은 이식가능 수정란 결과를 나타냈다. 미수정란의 비율은 성감별 처리구가 각각 52.04%와 57.78%로 대조구의 8.70%에 비해 유의적으로 매우 높았다(p<0.05). 이식가능 수정란의 발달 단계에 대한 조사 결과, 대조구의 경우 초기배반포배 단계의 수정란이 30.43%로 가장 높은 비율을 차지했으며, 그 다음이 확장배반포배(19.57%), 상실배(17.39%) 단계였다. 반면에 처리구의 경우, 두 처리구 모두 상실배 단계의 수정란이 15.31%와 24.44%로 가장 높은 비율을 나타냈다. 생산된 체내수정란을 한우 수란우에 이식한 결과, 대조구의 경우 35.00%, 처리구의 경우 12.50%의 수태율을 나타내었다. 성감별 정자 유래 암송아지의 염색체 핵형분석 결과, 2n=60으로 일반 한우와 다르지 않은 염색체 분포 양상을 나타내었으며, 29개의 상동염색체와 XX 성 염색체를 가진 정상 암컷 개체임을 확인하였다.

• 한국수정란이식학회지
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• 제21권3호
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• pp.191-198
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• 2006

본 연구는 $2001{\sim}2005$년(5년간) 제주도에서 사육중인 제주 한우와 흑우를 체내 수정란의 생산과 수정란 이식 기술을 통하여 조기 증식하고자 실시하였다. 한우 고등 등록우 286두와 흑우 경산우 69두(총 355두)에 대하여 발정 주기에 관계없이 CIDR를 질내에 삽입 후 7일째부터 성선 자극 호르몬($Folltropin^{(R)}-V$) 400 mg을 50 mg씩 균등하게 나누어 4일간 12시간 간격으로 근육주사하였다. 투여 6회째에 CIDR를 제거하였으며, 동시에 $PGF_{2{\alpha}}$ 25mg을 근육 주사하여 과배란을 유기하였다. 공란우의 인공수정은 $PGF_{2{\alpha}}$투여 후 발정을 확인하고 12 시간 간격으로 3회 인공 수정하였으며, 2회 인공 수정시 GnRH $250{\mu}g$을 근육 주사하였다. 수정란 회수는 1차 인공 수정 후 $7{\sim}8$일째에 비외과적 방법으로 채란하였다. 수란우는 CIDR를 7일간 삽입하였다가 제거하는 동시에 $PGF_{2{\alpha}}$ 25 mg을 근육 주사하여 공란우와 발정을 동기화 시켰다. 수정란 이식은 발정이 동기화된 수란우 1,219두에 대하여 6명의 수정란 이식 시술자가 비외과적 방법으로 황체가 존재하는 자궁각에 이식하였다. 한우 및 흑우 공란우의 평균 회수 총 난자수 및 이식 가능 수정란 수는 각각 7.4개 및 4.7개 였으며, 회수 총 난자수는 2 품종간에 차이가 없었으나, 이식 가능 수정란 수는 한우(5.0개)가 흑우(3.5개)에 비해 많았다(p<0.05). 공란우의 반복 과배란 처리에 따라 흑우에서는 과배란처리 두수 대비 수정란 채란우 비율이 감소되었으며(p<0.05), 한우에서는 회수 총 난자수 및 이식 가능 수정란 수의 감소가 나타났다(p<0.05). 과배란 처리 계절에 따른 수정란 회수 성적은 여름(5.6개)이 겨울(2.9개)에 비해 이식 가능 수정란의 수가 많았다(p<0.01). 수정란 이식 후 수태율은 평균 40%를 나타내었으며, 공란우의 품종간 차이가 없었으며, 이식년도가 2001년에서 2004년으로 경과될 때 수태율이 증가하였으며(p<0.05), 수정란 이식 시술자에 따른 수태율의 차이가($33.0{\sim}45.9%$) 인정되었다(p<0.01). 본 연구의 결과는 제주 한우 및 흑우에서 CIDR를 이용한 과배란 처리 및 수란우의 발정동기화 처리는 한우 체내 수정란의 안정적인 생산과 수란우의 준비가 가능함을 보여 주었다.

• Journal of Nutrition and Health
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• 제36권2호
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• pp.154-166
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• 2003

To investigate the effects of isoflavone supplementation on both bone mineral density and hormone variation in premenopausal women who had decreased bone mass, the 24 subjects were divided into two groups: one was the underweight group, consisting of 13 subjects, and the other was the normal weight group consisting of 11 subjects. For each group, we investigated the effects of isoflavone supplementation of 90 mg/day on both bone mineral density and hormone variation during 3 menstrual cycles. Anthropometric measurements, dietary recall, and analyses of blood and urine were assessed from baseline to post-treatment. The results were as follows: The average age of the underweight group was 21.8 years old and that of the normal weight group was 23.2 years old. The comparative results for the two groups at baseline were as follows: Onset of menarche, menstrual cycle, and menstrual length were not significantly different between the groups. Serum protein, total, HDL-, LDL-cholesterol, triglyceride, Ca, P, Mg, Cu, and Zn level were not significantly different between the groups. Serum estradiol, SHBG, LH, and FSH level were also not significantly different between the groups. Lumbar spine BMD by T scores of the underweight group was significantly lower than that of the normal weight group. Serum osteocalcin, urinary DPD, and urinary pH were not significantly different between the groups. The comparative results for the two groups at post-treatment were as follows: From baseline to post-treatment, the intake of energy, nutrients and isoflavone in food did not significantly change in either group. Serum protein, total cholesterol, HDL-, LDL-cholesterol, and triglyceride levels did not significantly change in either group. Serum Ca, Cu, and Zn levels were significantly lower in both groups and serum Mg level significantly decreased only in the underweight group. Serum estradiol levels were significantly lower in both groups, but serum SHBG, LH, and FSH levels did not significantly change in either group. Lumbar spine BMD by T score of the underweight group significantly increased to 15%, but that of the normal weight group did not significantly change. Serum osteocalcin of the underweight group significantly increased to 28%, while that of the normal weight group significantly increased to 40%. Urinary DPD of the normal weight group significantly increased to 12%. The results show that the BMD of the underweight group was lower than that of the normal weight group. Therefore, the underweight group had a disadvantage in obtaining maximum bone mineral density. The results also show that isoflavone supplementation during 3 menstrual cycles was effective in increasing the bone mineral density of the lumbar spine and affected bone metabolism markers in premenopausal underweight women. Therefore, it can be concluded that sufficient intake of isoflavone could be helpful in preventing decreases in bone mass among premenopausal women, especially underweight women.

• Clinical and Experimental Reproductive Medicine
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• 제31권4호
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• pp.235-244
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• 2004

Objective: The effects on spermatogenesis by expression of vascular endothelial growth factor (VEGF) and endothelin-1 (ET-1) were investigated. Materials and Methods: Testicular specimens were obtained from 40 infertile males due to primary testicular failure and from 10 fertile males with other urologic problems. The specimens of infertile males were devided into 4 groups according to histologic findings; Sertoli cell only syndrome (A), maturation arrest (B), hypospermatogenesis (C) and sloughing and disorganization (D). VEGF and ET-1 expression were detected with immunohistochemical stain. Results: VEGF expression on Leydig cell was detected in all cases. But, VEGF expression rates on germ cell were significantly higher in infertile group B, C, D compared to that of the control group (p<0.05). ET-1 expression rates on Leydig cell was significantly lower in all infertile group compared to that of the control group (p<0.05). But, ET-1 expression rates on Sertoli cell was significantly higher in all infertile group compared to that of the control group (p>0.05). In germ cell of infertile group, LH, FSH and prolactin were significantly decreased, and estradiol is increased in positive stain group on ET-1 immunohistochemical stain (p<0.05). VEGF and ET-1 expression were not correlated mean seminiferous tubule diameter (p>0.05). Conclusions: Abnormal spermatogenesis would be reflected in VEGF expression in germ cell.

• Asian-Australasian Journal of Animal Sciences
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• 제23권8호
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• pp.1049-1056
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• 2010

This study was conducted to investigate the effects of dietary glutamine (Gln) on the productive performance and egg quality of laying hens. A total of four hundred Lingnan Yellow laying hens aged 34 weeks were randomly assigned into four groups (100 laying hens/group), and fed, respectively, with diets supplemented with 0% (control group), 0.2%, 0.4%, and 0.8% Gln during the 6-week feeding period. The results were as follows. First, the productivity of laying hens fed with 0.8% Gln in diet was significantly increased (p<0.05); however, the egg quality (egg weight, yolk weight, shell weight, egg shape index, shell thickness, shell density, shell breaking strength, yolk color, yolk index, and Haugh unit) was not affected compared with that of the control group (p>0.05). Second, luteinizing hormone (LH) (p<0.01), follicle stimulating hormone (FSH) (p<0.01), triiodothyronine ($T_3$), and tetraiodothyronine ($T_4$) contents (p<0.05) in blood of laying hens fed with 0.8% Gln in diets were also significantly improved, and greater improvement in the duodenum and oviduct structure was observed in that treatment group. This study indicated for the first time that diets with 0.8% Gln were able to increase the productive performance of laying hens through stimulating hormone secretion and better development of both the duodenum and oviduct structure in laying hens.

• Clinical and Experimental Reproductive Medicine
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• 제28권2호
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• pp.131-140
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• 2001

Objective: To know the effects of xenoestrogen on spermatogenesis, we investigated the expression of cytochrome P450s enzymes (CYPscc, $CYP_{17{\alpha}}$, CYP19) and $3{\beta}$-HSD genes involved in steroidogenesis. Methods: Mouse testicular cells were prepared from 15-day-old ICR mice which had only pre-meiotic germ cells by enzyme digestion using collagenase and trypsin. Testicular cells were cultured in DMEM supplemented with FSH (0.1 IU/ml) and 10% FBS or medium with estrogen ($E_2$), bisphenol-A (BPA), octylphenol (OP; $10^{-9},\;10^{-7},\;10^{-6},\;10^{-5},\;10^{-4}M$, respectively) and aroclor 1254 (A1254) known as PCBs for 48 hours. The gene expression of cytochrome P450 enzymes were examined by semi-quantitive RT-PCR. The production of estrogen and testosterone was examined by RIA. Results: As results, expression of CYPscc mRNA was not significantly decreased, but $3{\beta}$-HSD and $CYP_{17{\alpha}}$. mRNA were significantly dose-dependent decreased. And production of testosterone and estrogen were not different except BPA and OP group ($10^{-5}M$). Conclusion: BPA, OP and A1254 might inhibit steroidogenesis by decreasing CYPscc, $3{\beta}$-HSD and $CYP_{17{\alpha}}$. mRNA expression in the mouse testis. These results suggest that BPA, OP and PCBs like as an endocrine disruptors inhibit the productions of steroidogenic enzymes and decrease the production of T and E by negative feedback mechanism. Therefore, these might disrupt steroidogenesis in Leydig cells of testis and would disturb testicular function and subsequently impair spermatogenesis.

• Clinical and Experimental Reproductive Medicine
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• 제38권4호
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• pp.234-237
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• 2011

Objective: During stimulated IVF cycles, up to 15% of oocytes are recovered as immature. The purpose of this study was to investigate the trend of oocyte maturity in repeated ovarian stimulation for IVF. Methods: One hundred forty-eight patients were selected who underwent two consecutive IVF cycles using same stimulation protocol during 2008 to 2010. Ovarian stimulation was performed with FSH and human menopausal gonadotropin and flexible GnRH antagonist protocol in both cycles. Oocyte maturity was assessed according to presence of germinal vesicle (GV) and the first polar body. Immature oocyte was defined as GV stage or metaphase I oocyte (GV breakdown with no visible polar body) and cultured up to 48 hours. If matured, they were fertilized with ICSI. Results: Percentages of immature oocytes were 30.8% and 32.9% ($p$=0.466) and IVM rates of immature oocytes were 36.2% and 25.7% ($p$=0.077), respectively. A significant correlation was noted between percentage of immature oocytes in the two cycles (R=0.178, $p$=0.03). Women with >40% immaturity in both cycles (n=21) showed lower fertilization rate of $in$ $vivo$ matured oocytes (56.4% vs. 72.0%, $p$=0.005) and lower pregnancy rate (19.0% vs. 27.1%, $p$=0.454) after the second cycle when compared with women with <40% immaturity (n=70). In both groups, female age, number of total retrieved oocyte and embryos transferred were similar. Conclusion: In repeated ovarian stimulation cycles for IVF, the immature oocyte tended to be retrieved repetitively in consecutive IVF cycles.

• Asian-Australasian Journal of Animal Sciences
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• 제23권2호
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• pp.169-174
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• 2010

A meta-analysis was conducted to summarize the results of studies which have described the profiles of hormones during the oestrous cycle in buffalo using a fixed effect model and a random effect model. Plasma progesterone concentrations were lowest (0.30${\pm}$0.06 ng/ml) during the peri-oestrous phase and increased (p = 0.067) through the early luteal phase to a maximum concentration (1.94${\pm}$0.03 ng/ml) during the mid-luteal phase. Circulating plasma inhibin and estradiol concentrations were lowest (0.31${\pm}$0.01 and 11.04${\pm}$0.13 ng/ml) during the mid-luteal phase, increased through the late luteal phase to maximum concentrations (0.44${\pm}$0.02 and 22.48${\pm}$0.32 ng/ml) during the peri-oestrous phase. Plasma FSH concentrations were lowest during the early luteal phase and increased through the mid-luteal phase to a maximum concentration during the peri-oestrous phase. Peripheral prolactin concentrations were lowest during the late luteal phase and increased to a maximum concentration during the peri-oestrous phase which then declined (p = 0.716) during the early luteal phase. Peripheral plasma cortisol concentrations decreased from 2.68${\pm}$0.14 ng/ml during the early luteal phase to 1.43${\pm}$0.27 ng/ml during the mid-luteal phase (p<0.001) which then increased to 2.06${\pm}$0.17 ng/ml during the late luteal phase. Plasma $T_{5}$ concentrations decreased from the late luteal phase to the peri-oestrous phase (p<0.001) which then increased during the early luteal phase. $T_{4}$ concentrations increased from the late luteal phase to the peri-oestrous phase which then decreased during the early luteal phase.

• 한국수정란이식학회지
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• 제15권1호
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• pp.57-65
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• 2000

The objective of this study was to apply the multiple ovulation and embryo transfer (MOET) program practically in dairy herds. Forty five superior Holstein cows ranked in 5% according to Type-Production Index(TPI) in Korea were selected as donors. The donors were superovulated with pFSH and the embryos collected from donors were frozen and preserved. The preserved embryos and frozen Holstein embryo imported from foreign country were thawed and transferred to recipients. The results obtained were as follows; 1. The total number of ova and freezable embryos collected per donor was 6.5 and 2.8, respectively. 2. The freezable embryos were obtained more(p<0.05) when the body condition score (BCS) of donors was in range of 2.50∼3.25(4.1) than in range of 3.50∼4.00(1.9), while the total number of ova was not changed. 3. The season affected on the collected number of freezable embryos(6.1 in winter, 4.5 in fall, 1.1∼1.5 in spring and summer, P<0.05), and the total number of obtained ova were more in winter than in other seasons(P<0.05). 4. Embryos were transferred to 343 recipients and 152 cows were confirmed pregnant(44.3%). 5. The higher pregnancy was obtained (P<0.05) when embryos were transferred in summer(53.3%) than in fall(36.0%), while the pregnancy rate was not affected by the origin and developmental stage of embryos, and the parity, BCS and estrus induction of recipients. From these results, the pregnancy rate was considered to be acceptable for the embryo transfer with domestic or imported Holstein embryos, however embryo production from superior Holstein donors was unsatisfactory for application of MOET scheme.