본 연구는 간척지에서 재배가 가능한 내염성 보리 품종육성을 위한 기초정보를 얻고자 겉보리 두 품종을 대상으로 생육초기 염 스트레스에 따른 생리적 반응과 잎 프로테옴의 발현양상 변화를 분석한 결과는 아래와 같다. 1. 토양의 염 농도가 증가함에 따라 보리의 건물중은 무처리구에 비해 유의적으로 감소하는 경향이었으며, 상록보리는무처리구에 비해 건물중 감소가 작았으며, 선우보리는 컸다. 2. 염처리에 따른 잎의 엽록소 함량을 나타내는 SPAD 값은상록보리가 57.6으로 47.6인 선우보리보다 높았으며, Na+의 함량은 선우보리에서 유의적으로 높았고, K+/Na+의 비율은 상록보리에서 높은 경향을 보였다. 3. 이차원전기영동에 의하여 염 스트레스에 의한 잎 프로테옴의 발현양상을 분석한 결과 47개 단백질 spot이 발현양의차이를 나타냈다. 품종별로 발현양이 증가한 단백질 spot은 상록보리와 선우보리에서 각각 17개와 14개로 나타났고, 발현양이 감소한 단백질 spot은 상록보리와 선우보리에서 각각 28개및 27개로 확인되었다. 4. 염처리에 따른 발현양의 차이를 보이는 18개 단백질을 동정한 결과 ribosomal protein 등 기능과 스트레스와의 관련성이 보고된 10개의 단백질과 ankyrin repeat domain protein등 스트레스 조건에서의 역할이 명확하지 않은 4개의 단백질및 Os02g0753300 등 기능 및 스트레스와의 관련성을 알 수없는 2개의 단백질이 동정되었다.

In the present study, the effect of cysteine and NT or bisphenol A (BP) on in vitro aturation (IVM) of porcine oocytes were examined. COCs was cultured in NCSU-23 medium supplement with 10% FCS which had previously been covered with mineral oil and equilibrated in a humidified atmosphere of 5% CO2 and 95% air at 38℃. The IVM rate of oocytes cultured for 48 hrs in NCSU-23 medium supplement with 0.5～10.0 mM cysteine were 34.0±3.2%, 36.0±3.5%, 48.0±3.8%, 22.0±3.2%, respectively. The IVM rate of oocytes cultured in NCSU-23 medium supplement with 0.5～5.0 mM NT for 48 hrs were 24.0±4.2%, 18.0±4.9%, 8.0±2.2%, respectively. NT affects oocyte in vitro maturation rate in a dose-dependent. This result were significantly lower than the control group. The IVM rate of oocytes cultured for 48 hrs in NCSU-23 medium supplement with 1.0 mM NT+5.0 mM cysteine (38.0±4.3%) were significantly higher than that of NT treatment. The IVM rate of oocytes cultured in NCSU-23 medium supplement with 0.05～5.0 mM BP for 48 hrs were 20.0±4.7%, 10.0±5.3%, 6.0±3.2%, respectively. The IVM rate of oocytes cultured in NCSU-23 medium supplement with BP was significantly lower cultured non supplement of BP (44.0±3.5%). BP affects porcine oocyte maturation rate in a dose-dependent manner. The IVM rate of oocytes cultured for 48 hrs in NCSU-23 medium supplement with 1.0 mM BP+5.0 mM cysteine (32.0±3.2%) were increased than that of BP treatment.

These study was carried out to investigate the effects of the supplementation with sodium nitroprusside (SN) and nitric oxide (NO) of canine oocytes on IVM rates. Oocytes were incubated in TCM-199 supplement with at 0.03～0.10 mM SN and 0.3～1.0 mM NO for 48 hrs. Oocytes were transferred to 50 ul drops of maturation medium covered mineral oil and cultured in a CO2 incubator (5% CO2, 95% air, 38℃). The in vitro maturation rate of oocytes cultured for 48 hrs in TCM-199 medium supplement with 0.03, 0.05, 0.07, 0.10 mM SN were 25.9±3.5%, 36.4±3.2%, 33.3±3.5%, 28.8±3.2%, respectively. The in vitro maturation rate of oocytes cultured for 48 hrs in TCM-199 medium supplement with 0.03～0.07 mM SN were significantly increased compare to the control (26.0±2.2%). The in vitro maturation rates of oocytes cultured for 48 hrs in TCM-199 medium supplement with 0.3, 0.5, 0.7, 1.0 mM NO were 28.0±4.2%, 36.5± 3.6%, 30.0±3.8%, 19.2±3.5%, respectively. The in vitro maturation rate of oocytes in TCM-199 medium supplemented with 0.3 and 0.5 mM NO were significantly increased compare to the control (26.0±2.2%). The in vitro maturation rates of oocytes cultured for 12～48 hrs in TCM-199 medium supplement with 0.05 mM SN were 26.0±3.2%, 28.0±3.4%, 38.0±3.2%, respectively. The in vitro maturation rate of oocytes cultured for 12～48 hrs in TCM-199 medium supplement with 0.5 mM NO were 22.0±3.0%, 30.0±3.8%, 36.0±4.2%, respectively. These result was significantly increased compare to the control.

In this study, we analyzed expression patterns of apoptotic and autophagic gene products in culture follicular cells of normal and miniature pigs to assess the effect of hormones on the choice for programmed cell death. Autophagic activity progressively increased from control cultures to luteinizing hormone (LH)-treated cultures of follicular cells of normal pigs, but decreased from the LH to follicle stimulating hormone (FHS) +LH-treated cultures. Expression of Casp-3 protein in follicular cells was highest in LHtreated cultures, but the activity of Casp-3 decreased in the control, FSH-treated, and FSH+LH-treated cultures. The activity of the apoptosis protein was highly expressed in the control, LH-treated, and FSH+LH-treated follicular cells of miniature pigs, but autophagy- associated proteins were expressed at low levels in all treatments groups of the miniature pig. The expression of autophagy and apoptosis proteins appeared similar in control and rapamycin-treated cells. In addition, stimulation with FSH triggered the activation of autophagy in the follicular cells of normal pigs, but induced apoptosis in the follicular cells of miniature pigs. A similar effect was obtained when LH was applied. These results suggest that the autophagy process and FSH stimulation is more effective for stable and innovative follicular cell development.

These study was to investigate the in vitro fertilization and viability of fresh and vitrified oocytes. Also, the developmental capacity of IVF and intracytoplasmic sperm injection (ICSI) oocytes were investigated. Then vitrification was performed with the use of 20% ethylene glycol + 20% DMSO + 0.5 M sucrose + 10% FCS + TCM-199 medium. Vitrification immature oocytes are cultured in vitrification solution for 10 min afterwards transferred to expose at room temperature for 5 min. and transferred to the ice water for 5 min. The oocytes were sealed in a 1.0 mm straw and placed in a LN2 container. Frozen oocytes were rapidly thawed in a water bath at 30～35℃, and then placed in TCM-199 medium containing 0.5 M sucrose for 5 min each, respectively, at 38℃. After being washed for 2～3 times, using fresh medium the oocytes were cultured in TCM-199 medium supplemented with 5% FCS at 38℃ in 5% CO2 and air. The normal morphology of fresh and vitrified-thawed oocytes were 87.1±2.1% and 54.8±2.5%, respectively. The viability rates of fresh and vitrified-thawed oocytes were 70.0±2.2% and 41.9±2.6%, respectively. Viability rates of vitrified-thawed oocytes were lower than that of fresh follicular oocytes (p<0.05). The in vitro maturation rates of fresh and vitrified oocytes were 45.1±3.6% and 28.9±4.4%, respectively. The IVF rates of fresh follicular and vitrified-thawed oocytes were 34.0±2.2% and 20.2±2.6%, respectively. The in vitro maturation and fertilization rates of vitrified-thawed oocytes were lower than those of the fresh follicular oocytes (p<0.05). A total of 350 oocytes were fixed and stained after co-incubation with spermatozoa, of which 88 had identifiable nuclear material. After IVF for 20 hrs, 25.1±3.4% of the oocytes found to have been penetrated by spermatozoas. Oocytes were fixed and stained after ICSI, and 105 oocytes contained identifiable nuclear material. After IVF and ICSI for 20 hrs, 34.3±3.4% and 59.0±2.0% of the oocytes were found to have been penetrated by spermatozoas. The developmental rates upon ICSI were significantly higher than those of the IVF method (p<0.05).