• Title, Summary, Keyword: Follicular Fluid

Search Result 189, Processing Time 0.042 seconds

N-glycoproteomic analysis of human follicular fluid during natural and stimulated cycles in patients undergoing in vitro fertilization

  • Lim, Hee-Joung;Seok, Ae Eun;Han, Jiyou;Lee, Jiyeong;Lee, Sungeun;Kang, Hee-Gyoo;Cha, Byung Heun;Yang, Yunseok
    • Clinical and Experimental Reproductive Medicine
    • /
    • v.44 no.2
    • /
    • pp.63-72
    • /
    • 2017
  • Objective: Hyperstimulation methods are broadly used for in vitro fertilization (IVF) in patients with infertility; however, the side effects associated with these therapies, such as ovarian hyperstimulation syndrome (OHSS), have not been well studied. N-glycoproteomes are subproteomes used for the remote sensing of ovarian stimulation in follicular growth. Glycoproteomic variation in human follicular fluid (hFF) has not been evaluated. In this study, we aimed to identify and quantify the glycoproteomes and N-glycoproteins (N-GPs) in natural and stimulated hFF using label-free nano-liquid chromatography/electrospray ionization-quad time-of-flight mass spectrometry. Methods: For profiling of the total proteome and glycoproteome, pooled protein samples from natural and stimulated hFF samples were selectively isolated using hydrazide chemistry to obtain the total proteomes and glycoproteomes. N-GPs were validated by the consensus sequence N-X-S/T (92.2% specificity for the N-glycomotif at p<0.05). All data were compared between natural versus hyperstimulated hFF samples. Results: We detected 41 and 44 N-GPs in the natural and stimulated hFF samples, respectively. Importantly, we identified 11 N-GPs with greater than two-fold upregulation in stimulated hFF samples compared to natural hFF samples. We also validated the novel N-GPs thyroxine-binding globulin, vitamin D-binding protein, and complement proteins C3 and C9. Conclusion: We identified and classified N-GPs in hFF to improve our understanding of follicular physiology in patients requiring assisted reproduction. Our results provided important insights into the prevention of hyperstimulation side effects, such as OHSS.

Studies on the Effects of Estrous Cow Serum, Follicular Fluids and Matured Cumulus Cells on In Vitro Maturation and Fertilization of Bovine Follicular Oocytes (발정우 혈청, 난포액 및 난구세포의 첨가가 우난포란의 체외성숙 및 수정에 미치는 영향에 관한 연구)

  • 김상근;이만휘;김무강;박항균;한방근
    • Korean Journal of Animal Reproduction
    • /
    • v.14 no.3
    • /
    • pp.183-190
    • /
    • 1990
  • These studies were carried out ot investigate the effects of estrous cow serum(ECS), fetal calf serum(FCS), bovine follicular fluid(BFF) and matured cumulus cell(MCC) on in vitro maturation and fertilization of bovine follicular oocytes. The ovaries were obtained from slaughtered Korean native cows. The follicular oocytes surrounded with cumulus cells were recovered by aspirating follicular fluid from the visible follicles of diameter 3-5mm. The follicular oocytes were cultured in TCM-199 medium containing hormones, FCS, ECS, BFF and MCC for 24~48 hrs. in a incubator with 5% CO2 in air at 38.5$^{\circ}C$ and then matured oocytes were again cultured for 18$^{\circ}C$20 hrs. with motile capacitated sperm in the TCF(Tyrode calcium-free) solution containing 200$\mu\textrm{g}$/ml of heparin. The results obtained in these experiments were summarized as follows : 1. The oocytes classified as "A, B, C, D and Degenerative" depending morphological integrity and those were 61.4%, 12.1%, 19.2%, 4.2% and 3.0% of the total oocytes harvested, respectively. The maturation and fertilization rate of the A, B, C class follicular oocytes, cultured in the TCM-199 medium supplemented with 10% FCS were 89.1%, 78.0%, 52.6% and 78.1%, 33.3%, respectively. 2. The maturation and fertilization rate of the follicular oocytes cultured in TCM-199 medium supplemented with 5%~20% ECS and FCS were 74.0%~80.6, 26.2%~30.0% and 71.7%~76.9%, 51.9%~58.0%, and those values were higher the supplement of ECS than FCS. 3. The maturation rate(68.0%~64.6%) and fertilization rate(59.6%~60.4%) of follicular oocytes cultured in TCM-199 medium supplemented with 10% FCS and 20~30% BFF were higher than those of follicular oocytes cultured TCM-199 medium supplemented with 10% FCS and 10% and 50% BFF. 4. The maturation rate(76.5%) and fertilization rate(61.7%) of follicular oocytes cultured in TCM-199 medium supplemented with 10% FCS and 1$\times$106/ml cumulus cells were higher than those of follicular oocytes cultured in TCM-199 medium supplemented with 10% FCS and 1$\times$104~5/ml and 1$\times$108/ml cumulus cells.lus cells.

  • PDF

Fatty Acid constituents and Relative Compositions of Reproductive Tract Fluids in Korean Native Cows (한우에 있어서 생식기관액 중 지방산 조성과 조성율)

  • 신원집;정진우;최광수;신수길
    • Journal of Embryo Transfer
    • /
    • v.15 no.2
    • /
    • pp.147-156
    • /
    • 2000
  • To investigate fatty acid constituents and relative compositions in the fluid of the follicles, oviducts, uterine body and uterine body in Korean native cow, the fluids of the reproductive tract were analyzed using gas chromatography. The samples were taken from various reproductive tract of 23 Korean native cows. q. Caprylic acid (C8:0), myristic acid(C14:0), palmitic acid(C16:0), palmitoleic acid(C16:1), stearic acid(C18:0), oleic acid(C18:1), linoleic acid(C18:2), arachidonic acid(C20:4) were found in the reproductive tracts of the cows, which made 8 kinds of fatty acid in total. 2. Palmitic acid, oleic acid, stearic acid were predominant with 35.67%, 24.98% and 17.52%, respectively. while low levels of fatty acids(<5%) were myristic acid, palmitoleic acid and caprylic acid with 1.75%, 1.28% and 2.69%, respectively. 3. Two kinds of polyunsaturated fatty acids, linoleic acid arachidonic acid were found in the reproductive tracts of cows. 4. Palmitic acid among saturated fatty acids and oleic acid among unsaturated fatty acids were the highest level in all of the reproductive tracts. 5. The Highest level of arachidonic acid was found in the uterine horn. 6. The sum of the palmitic acid and oleic acid were 61.72%, 63.72%, 57.66% and 57.65% for the fluid of follicle, oviduct, uterine horn and uterine body of the cows, respectively. 7. The relative compositions of palmitic acid, palmitoleic acid and caprylic acid were higher during the luteal phase than during the follicular phase. 8. The relative compositions of arachidonic acid was higher during the follicular phase in the fluid of uterine horn and uterine body of the cows. 9. The long chain fatty acid, the palmitic acid, stearic acid, oleic acid and linoleic acidshowed higher relative compositions during the follicular phase(86.49%∼95.51%) than during the luteal phase(85.64%∼88.93%).

  • PDF

Fatty Acid Constituents and Relative Compositions of Reproductive Tract Fluids in Sow (돼지에 있어서 생식기관액 중 지방산 조성과 조성율)

  • 신원집;정진우;최광수;신수길
    • Journal of Embryo Transfer
    • /
    • v.15 no.2
    • /
    • pp.137-145
    • /
    • 2000
  • To investigate fatty acid constituents and relative compositions in the fluid of the follicles, oviducts, uterine ho군 and uterine body in sows, the fluids of the reproductive tract were analyzed using Gas chromatography. The samples were taken from various reproductive tract of 21 sows slaughtered. 1. Caprylic acid(C8: 0), capric acid(C10:0), lauric acid(C12:0), myristic acid(C14:0), palmitic acid(C16:0), plamitolele acid(C16:1), stearic acid(C18:0), oleic acid(C18:1), linoleic acid(C18:2) and arachidonic acid(C20:4) were found in the reproductive tracts of the sows, which made 10 kinds of fatty acid intotal. 2. Two kinds of polyunsaturated fatty acids, linoleic acid and archidonic acid were found inthe reproductive tracts. 3. Palmitic acid among saturated fatty acids and oleic acid among unsaturated fatty acids were the hihgest level in all of the reproductive tracts. 4. Palmitic acid, oleic acid and stearic acid showed higher rate with 44.89%, 23.69% and 14.36%, respectively, and lauric acid, capric acid, palmitoleic acid, arachidonic acid ad myristic acid showed lower rate with 0.62%, 1.13%, 1.65%, 1.97% and 2.24%, respectively in the reproductive fluid. 5. The highest level of arachidonic acid was found in the uterine horn. 6. The sum of the palmitic acid and oleic acid were 66.91%, 70.41%, 66.14% and 73.36% in the fluid of follicle, oviduct, uterine horn and uterine body, respectively. 7. The relative composition of arachidonic acid was higher during the follicular stage than during the luteal phase in the fluid of oviduct and uterine. 8. The long chain fatty acids such as the palmitic acid, stearic acid, oleic acid and linoleic acid showed higher relative compositions during the follicular phase(93.18%∼96.83%) than during the luteal phase(82.56%∼88.37%). 9. Caprylic acid, luric acid and palmitoleic acid were undetected in the fluid of all of the reproductive tracts during the follicular phase. Low relative compositions of capric acid, myristic acid andarachidonic acid were found during the follicular phase, while the low relative compositions (<5%)of capric acid, lauric acid, myristic acid, plamitoleic acid and arachidonic acid were found during the luteal phase.

  • PDF

Iozyme Patterns of Lactate Dehydrogenase in Follicular Components (돼지체조직 및 난포구성분에 있어 Lactate Dehydrogenase Isozyme 양식)

  • 이중한;변태호;유형진;이상호
    • Korean Journal of Animal Reproduction
    • /
    • v.17 no.3
    • /
    • pp.257-262
    • /
    • 1993
  • Various tissue and follicular components were analyzed for the determination of lactate dehydrogenase(LDH) isozyme patterns by electrophoretic technique with chromogen reaction in the pig. Optimum conditions for the tissue homogenate and the storage were finally established. Small quantities of follicular components were analysed for typing of LDH isozymes by microelectrophoresis. Microelectrophoretic analysis showed that only LDH-1 was visible in the oocytes, all isozymes in cumulus masses, and LDH-1, 2 and 3 in follicular fluid. The results provide critical information on the LDH activity of various tissues and follicular components. Furthermore, t he developed methods should be useful the analysis of LDH in the small quantity of samples, especially in the oocyte, and easily applicable to the oocyte and early embryos of other domestic species.

  • PDF

Effects of Characteristics of Ovarian follicular Fluid and Ant-Inhibin Serum on Steroid Hormone Secretion by Hanwoo Granulosa Cells In Vitro (한우 난소의 Follicular Fluid의 특징과 과립막 세포의 스테로이드호르몬 분비에 대한 Anti-Inhibin Serum의 첨가효과)

  • 성환후;민관식;양병철;노환국;최선호;임기순;장유민;박성재;장원경
    • Korean Journal of Animal Reproduction
    • /
    • v.25 no.2
    • /
    • pp.119-124
    • /
    • 2001
  • This study was performed to investigate the effects of the peptide to carrier ratio on the immune and biological functions to inhibin immunization in Hanwoo. A peptide sequence kom the alpha -subunit (19~32 peptide) of porcine inhibin was synthesized for antigen and conjugated to human serum albumin(HSA) for carrier protein. Anti-inhibin sera(AI) were produced 52 day later from rabbit after injection of inhibin-$\alpha$ -subunit peptide conjugator for antigen with the interval of 2 weeks. Immune-blotting analysis using antibody specific fur inhibin-$\alpha$ subunits revealed that the inhibin was detected at 1.0 cm bovine follicular fluid(bFF). However, each stage of corpus lutea and 0.1 cm of follicular fluid were not detected. The maximal contents of estradiol-17 $\beta$ in Hanwoo ovarian follicular fluid were detected at 2.0 cm of follicular size(diameter), but the mean total contents of these hormone decreased significantly with decreasing diameter of follicles. However, progesterone contents of follicular fluid were high at 1.0 cm of follicle. Progesterone secretion by Hanwoo granulosa cell cultured for 48 hr in vitro was significantly (p<0.05) inhibited in 5% bFF and 5% bFF + 5% AI addition group compared with control group. Estradiol-17 $\beta$ secretion by Hanwoo granulosa cell cultured for 48 hr in vitro was significantly (p<0.05) increased in 5% AI and 5% AI + 5% bFF addtion group compared with control group. However, the groups added 5% AI were not changed compared to control groups in progesterone and estradiol-17 $\beta$. Taken together, we suggested that inhibin in the mature FF plays a pivotal role on the biosynthesis of steroid hormone of follicular cells during follicular development.

  • PDF

Studies on Investigation and Change of Protein Factors in Porcine Follicular Fluids (돼지의 난포액 내 단백질인자의 탐색과 변화에 관한 연구)

  • Ji, Mi-Ran;Cheong, Hee-Tae;Yang, Boo-Keun;Lee, Chae-Sik;Park, Choon-Keun
    • Reproductive and Developmental Biology
    • /
    • v.34 no.3
    • /
    • pp.217-221
    • /
    • 2010
  • When fully grown oocytes are removed from their follicles, they can resume meiosis and mature spontaneously under in vitro conditions. However, nuclear maturation under in vitro condition is not accompanied by complete cytoplasmic maturation, which is essential for successful fertilization and the initiation of zygotic development. This study analyzed change of proteins in follicular fluids during the porcine follicular development. Follicular fluids were collected from follicles of diameter 1~2 mm, 2~6 mm and 6~10 mm in ovary of slaughtered pigs. Total proteins were extracted from follicular fluids by M-PER Mammalian Protein Extraction Reagent. We confirmed totally 27 same spots, 1 spot from follicle fluid of 2~6 mm follicle and 5 spots from follicle fluid of 6~10 mm in diameter were analyzed by MALDI mass spectrometry and searched on NCBInr. In results, spot No. 28 from 2~6 mm follicle was Ig lambda chain C region, and spot No.32 and 33 from 6~10 mm was Apolipoprotein A-(APOA4). Spot No.29 and 31 were failed to analyze. These results indicate that the porcine oocyte during in vitro maturation depend on specific different expressed proteins may play an important roles in the sequence of molecular events in porcine oocyte maturation and follicular development.

Separation of Follicular Fluid Components Stimulating Sperm Migration with Chromatographic Paper, $=mu$RPC and Superose Columns (Chromatography용 Paper, $\mu$RPC Column 및 Superose Column을 이용한 정자의 이동을 자극하는 난포액 성분의 분리)

  • 박영식
    • Journal of Embryo Transfer
    • /
    • v.13 no.3
    • /
    • pp.301-312
    • /
    • 1998
  • To efficiently separate a protein stimulating sperm swim-up migration and movement from follicular proteins, the effect of paper chromatography and liquid chromatography with reverse phase column and superose column on protein separation was examined. And the results obtained were as follows; 1. The band component that was separated with paper chromatography stimulated sperm migration and movement depending on its additional levels. Especially, band I component significantly increased sperm migration. But, all components of bands 1, 2 and 3 showed lower sperm migration and movement, compared to follicular fluid at the same additional level. 2. Among the components separated from follicular protein of 2~5mm follicles with reverse phase column ($\mu$RPC), components at retention time (RT) of 3.33, 7.00, 13.87, and 16.6A minutes stimulated sperm migration within a limited range. 3. All components separated from follicular protein of 10mm follicles with $\mu$RPC column didn't stimulate sperm migration and movement. 4. Among the components separated from follicular protein of 2~5m follicles with superose column, components at retention volume (RV) of 1.35 and 0.82 ml significantly stimulated sperm migration and movement. In conclusion, protein components stimulating sperm migration and movement were efficiently separated with superose column in Smart system. Especially, components of RV 1.35 and RV0.82 stimulated sperm swim-up separation.

  • PDF

Effect of Follicular Fluid on Attraction and Motility of Human Spermatozoa (사람 정자에 대한 유인능과 운동성에 미치는 난포액의 영향)

  • 곽대오;전병균;문진수;김광철
    • Development and Reproduction
    • /
    • v.5 no.2
    • /
    • pp.159-165
    • /
    • 2001
  • Follicular fluid has pivotal effects on attraction and motility of spermatozoa for successful fertilization. The effect of samples of human follicu1ar fluid(hFF) on attraction and motility of spermatozoa was investigated. Capillary tubes loaded with one of these samples, hFF sample A collected from patients with tubal factor, hFF sample B collected from patients with male factor, m-HTF and heated hFF sample were used for assessment of attraction and motility of spermatozoa following culture of 1, 2, and 4 hrs. Number and motile rate of spermatozoa in the tubes loaded with hFF sample A were significantly(P<0.05) higher than those of m-HTF, hFF sample B and heated hFF. Although the fresh hFF tended to increase the attraction of spermatozoa as compared to inactivated hFF, there was no significant difference between treatments.

  • PDF

Effect of Mature Human Follicular Fluid on the Development of Mouse Embryos in vitro (성숙난포액을 이용한 생쥐배아의 발달에 관한 연구)

  • Park, S.Y.;Lee, J.J.;Kim, S.H.;Ku, P.S.
    • Clinical and Experimental Reproductive Medicine
    • /
    • v.19 no.2
    • /
    • pp.125-131
    • /
    • 1992
  • The possible effect of human follicular fluid(hFF) on the growth and development of fertilized oocytes and embryos is important because the fallopian tubes are exposed to FF after follicular rupture and the processes of fertilization and embryo cleavage occur inside the fallopian tubes. Previously, it was suggested that human FF might adversely affect on the development of early mouse embryos. In order to investigate the effect of hFF on the development of embryos, early mouse embryos were cultured in media containing various protein sources as bovine serum albumin(BSA), fetal cord serum(FCS) and FF. And we evaluated the development of early mouse embryos in terms of the morphology, cleavage rate, and cell count of blastcysts. There were no significant differences in the morula and blstocyst formation rates of 2-cell mouse embryos cultured in the media containg three different protein sources and three different concentrations of FF. The blastocyst formation rate of 1-cell mouse embryo cultured in FF group was significantly higher than that cultured in BSA group(P<0.05). The morula and blastocyst formation rates of 2-cell mouse embryos of the group cultured in the media containing FF were comparable with those of other two groups, in addition, the cell count of blastocysts of FF group in the 2-cell embryo culture was higher than those of BSA group and HCS group(P<0.01), and this finding was also noted in 1-cell embryo culture. There was no difference in the morula and blastocyst formation rates of the 2-cell mouse embryos cultured in the media containing different concentrations of FF. These results suggest that mature human follicular fluid has no inhibitory activity on the development of early mouse embryos even in high concentration and may be a good protein source which is positively associated with the development of mouse embryos in vitro especially in 1 cell embryo culture.

  • PDF