• Korean Journal of Animal Reproduction
• /
• v.26 no.3
• /
• pp.215-221
• /
• 2002

Antioxidants(N-acetyl-1-cysteine, ebselen and glutathione) and growth factors(EGF, PDGF) were studied as a mean of increasing the development of porcine embryos produced by in vitro maturation (IVM) and in vitro fertilization(IVF). Porcine embryos developed to the 2~8 cell stage after IVF were cultured fer 6 to 7 days at 38.5$^{\circ}C$ in NCSU 23 medium containing antioxidants plus growth factors. Cell numbers of blastocysts were counted by fluorescence staining method. The developmental rate beyond morula stages in NCSU 23 containing NAC(1nm) or NAC(1nm) plus EGF(100ng/$m\ell$) or PDGF(5ng/$m\ell$) were 28.1, 32.3 and 35.3%, respectively. NAC plus PDGF group was slightly higher than control group(P>0.05). The developmental capacity in NCSU 23 containing ebselen(10 $\mu$M) or ebselen(10 $\mu$M) plus EGF(100ng/$m\ell$) or PDGF(5ng/$m\ell$) were 17.8, 36.9 and 40.3%, respectively Ebselen plus growth factor groups were significantly higher than control group(P<0.05). The developmental capacity in NCSU 23 containing glutathione(100 $\mu$M) or glutathione(100 $\mu$M) plus EGF (100ng/$m\ell$) or PDGF(5ng/$m\ell$) were 24.1, 30.5 and 27.7%, respectively. There were not difference in all experimental groups(P>0.05). In all experimental groups, there was no significantly differences on the cell number of blastocysts, but ebselen plus growth factor groups were significantly higher than control group. These studies indicate that antioxidants plus growth factors can increase the proportion of embryos that developed beyond morulae stage.

• Proceedings of the KSAR Conference
• /
• 2000.10a
• /
• pp.10-12
• /
• 2000

Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$-subunits. The subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$ - and $\beta$ -subunits in the placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was. efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to consist of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t63I or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632-653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agonist-occupied receptors ~2- and ~17-fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.

• Korean journal of applied entomology
• /
• v.41 no.3
• /
• pp.211-216
• /
• 2002

The present study was aimed to examine the effect of different host plants on the biological property of cotton caterpillar, outbreak pest of cucurbit vegetables. The feeding and oviposition preferences toward cucumber and pumpkin were greater than those toward watermelon, oriental melon, and melon. The periods of egg and larva fed on oriental melon, melon, and pumpkin were longer than those on watermelon and cucumber. The longest pupal period was 10.5 days from cucumber. The hatching rate, pupation rate, emergence rate, and survival rate were observed from different host plants. The hatching rate was the highest on cucumber (87.2%) and the lowest on melon (72.8%). The pupation rate on watermelon and oriental melon (90.0% and 89.1%, respectively) was higher than that on cucumber (62.0%). The emergence rate on cucumber and pumpkin (93.5% and 92.0%, respectively) was higher than that on oriental melon (78.7%). The survival rate from hatching to emergence on watermelon (76.0%) was the highest, and the lowest on cucumber (50.0%). The adult period on melon (21.0 days) was the longest, and the shortest on cucumber (15.5 days) among 5 host plants. Average number of eggs per female on cucumber (281.8 eggs) was the highest, and the least on oriental melon (96.6 eggs). The survivorship on cucumber was the longest (30 days), and the shortest on pumpkin (17 days). The preoviposition period on oriental melon and melon (3.4 days) was longer than that on watermelon (2.1 days) and mean generation time in day (T) on melon was the longest (47.2 days) though they were not significant. The net reproductive rate per generation (R$_{o}$) on cucumber, 191.3, was the highest and the highest intrinsic rate of natural increase (r$_{m}$) was observed from cucumber as 0.127. All these results suggest that the growth ana reproduction of cotton caterpillar varies depending upon the kind of host plants. The analysis of the life table revealed that cucumber and pumpkin were the favorable host plants of cotton caterpillar.

• Korean journal of applied entomology
• /
• v.24 no.1 s.62
• /
• pp.15-18
• /
• 1985

Studies were made on the morphology and life cycle of the mite, Pyemotes tritici $Lagr{\acute{e}}z$-Fossot & $Montagn{\acute{e}}$ (Trombidiformes; Pyemotidea) ,parasitic on larvae of cigarette beetle, Lasioderma serricorne F., which is one of the most serious pests of stored tobacco, cigar, and cigarette in Korea. One generation time was $20.9{\pm}0.7$ days out of which $9.5{\pm}0.3$ days were spent for feeding and $10.3{\pm}0.8$ days for reproduction of progenies. A female of this insect-parasitic mite produced $56.7{\pm}6.9$ progenies during her reproduction period. The body size of a newly-laid male or female was $280{\mu}m$ long and $85{\mu}m$ wide. As female of this mite sucked on, their abdomen grew larger and larger to reach $825{\mu}m$ in width and $0.346mm^3$ in volume after $9{\sim}10$ days. By sucking the humor of a host, the abdomen of a female mite became almost a global shape in two days. The increase rate of abdominal width was the biggest on the second or the third day of feeding while the volume of abdomen reached to the largest on $6{\sim}8$ days after feeding. The largest number of the daily young produced on 4-th day after a female began to reproduce.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.33 no.6
• /
• pp.533-540
• /
• 2000

Gonadal development, gametogenesis, reproductive cycle, gonad index, flesh weight rate, and first sexual maturity of the turban shell, Lunella coronata coreensis were investigated by histological observation. The materials used were collected monthly from the rocky intertidal zone of Daehang-ri, Buan-gun, Jeollabuk-do, on the west coast of Korea, from July 1998 to June 1999. Sex of L coronata coreensis was separate. The gonad was widely located in the spirals of the visceral mass buried in the digestive gland. The ovary and testis were composed of a number of oogenic follicles and speymatogenic follicles, respectively. Monthly variations in the gonad index increased from March ($23.86{\pm}3.73$) when the water temperature increased and reached the maximun in July ($49.76{\pm}6.47$). And then, the gonad index sharply decreased in September ($15.58{\pm}2.33$). The flesh weight rate ranged from $25.2{\%}$ to $32.3{\%}$, and its variation showed a similar pattern to the gonad index. Individuals $<5.9 mm$ in shell height could not take part in reproduction in both sexes. Percentages of first sexual maturity of female and male specimens ranging from $7.0{\~}7.9 mm$ in shell heights were $84.6{\%}\;and\;91.7{\%}$, respectively, and $100{\%}$ in those over 8.0 mm in shell height in both sexes took part in reproduction. By studying the monthly changes of the morphological features and sizes of germ cells during gametogenesis in the gonad, the reproductive cycle of this species could be devided into five successive stages: early active (December to April), late active (January to July), ripe (May to August), spawning (July to September), and recovery (September to March). The spawning period of this species was once a year between July and September, and the main spawning occurred in July when the seawater temperature reached above $24.8^{\circ}C$. The fully ripe eggs were $150{\~}160\;{\mu}m$ in diameter.

• Korean Journal of Animal Reproduction
• /
• v.26 no.1
• /
• pp.79-85
• /
• 2002

We investigated the optimal concentration and exposure time of cycloheximide(CHX) on development of activated porcine oocytes following electrical pulse(EP). After 42~44 h maturation, oocytes were treated with 0.1% hyaluronidase, and denuded cumulus cells by pipetting. Oocytes were stimulated by electric pulse (1.2 kV/cm, 30 $\mu$sec, 1 pulse) or incubated for 3, 5 and 7 h in cycloheximide (1, 5 and 10 $\mu\textrm{g}$/$m\ell$, respectively) following electric pulse, and cultured for 8 days. Cleavage rate of oocytes activated with 10 $\mu\textrm{g}$/$m\ell$ CHX following EP was significantly (P＜0.05) higher than those of 1 $\mu\textrm{g}$/$m\ell$ (86.8% vs. 74.4%). The developmental competence of oocytes incubated to 5 $\mu\textrm{g}$/$m\ell$ of CHX was significantly (P＜0.05) higher development to blastocysts (13.3%), compared with 10 $\mu\textrm{g}$/$m\ell$ of CHX (5.6%). When the oocytes were activated with 5$\mu\textrm{g}$/$m\ell$ CHX for 3, 5, and 7 h following EP, the cleavage rate of oocytes in 5 h group(86.6%) was significantly (P＜0.05) higher than that in 3 h group(73.2%). The developmental rate of oocytes to morula in 5 and 7 h groups(26.7% and 16.4%) were significantly (P＜0.05) high than that in 3 h group(14.5%). Matured oocytes were activated with electric pulse (EP) or electric pulse combined with cycloheximide (EP + CHX) and cultured for 8 days. The rate of cleavage and development to blastocyst (80.1% and 11.6%) of activated with EP group were similar to EP combined with CHX group. When activated with EP or EP combined with CHX, the mean cell number of blastocysts were less in the activated with EP (18.67$\pm$5.53) than in the activated EP combined CHX (20.71$\pm$6.16), but not significantly different. This results suggest that, when the porcine oocytes were activated with CHX following EP, the developmental rate of activated oocytes can be improved by treated with a concentration of 5 $\mu\textrm{g}$/$m\ell$ CHX for 5 h exposure time.

• Korean Journal of Animal Reproduction
• /
• v.22 no.1
• /
• pp.11-17
• /
• 1998

This study was carried out to investigate the in vivo developmental potential of mouse zona-hatched blastocysts (HBs). The HBs were cultured in vitro until day 5 and day 6 from zygotes produced in vivo and classified to small (S-HBs), medium (M-HBs) and large (L-HBs) on the basis of embryo diameters. The results obtained in these experiments were summarized as follows ; 1) when the blastocysts at day 4 were further cultured for $24\sim48hr$, HBs obtained at day 5 and day 6 culture in vitro were 29.1% and 22.8%, respectively. 2) Also, when the total cell number of HBs were counted, cell numbers of classified HBs on day 5 and day 6 to small ($77.3\pm5.3$, $59.6\pm4.4$), medium ($83.7\pm4.0$, $66.8\pm3.5$) and large ($100.7\pm2.6$, $88.9\pm3.8$) were increased as their size increases. Especially, there were significantly different between S-HBs and L-HBs (p<0.01). 3) In addition, when the classified HBs were transferred into when the classified HBs were transferred into day 3 pseudopregnant recipients, the pregnancy and implantation rates of S-HBs (28.6%, 15.7%), M-HBs (44.4%, 30.9%) and L-HBs (62.5%, 49.1%) at day 5 were increased as their size increases. However, this pattern was not showed in embryo transfer of day 6 HBs. But, when the live fetuses formation against total implantation rates were observed, the result (87.5%) of S-HBs of day 5 was significantly higher than that of the others (p<0.01). Therefore, this study demonstrates that in vitro cultured healthy HBs can not only be developed normally with good pregnancy rates, implantation rates and live fetuses formation, but also served as a fundamental data for utility of supernumerary HBs in human blastocyst transfer.

• Korean Journal of Animal Reproduction
• /
• v.21 no.3
• /
• pp.239-246
• /
• 1997

The objective of this study was to evaluate the variation of fertilizing ability following the morphologically normal sperm ratio in porcine IVF using epididymal sperm The results obtained in this experiment were summarized as follows: 1. When the penetration rate (PR), polysper my rate (PSR), pronuclei formation (2PNF) and mean number of sperm (MNS) per oocyte were evaluated according to the percentage of morphologically normal epididyrnal sperm at insemination($\leq$lO%, 10~30% and $\geq$50%). the PR and PSR of $\leq$50% group (82.4, 87.4%) were significantly higher than those of other two groups ($\leq$lO%; 29.7%, 22.6% and 10~30%; 20.3, 37.0%) (p<0.01). Also, the 2PNF per examined oocytes was significantly high in $\geq$ 50% group (p<0.01). 2. When the $\geq$50% group in epididymal sperm was adjusted to 100% (5x1$^5$ cells/ml) , the PSR and 2PNF were not different between epididymal sperm (86.7, 35.1%) and frozen-thawed ejaculated sperm (86.0. 39.4%) although the PR in epididymal sperm (79.7%) was significantly lower than that in frozen-thawed ejaculated sperm (95.5%)(p<0.01). 3. Also. when the PR, PSR, 2PNF and MNS of epididymal sperm were evaluated according to the oocyte: sperm ratio (1:6000, 1:6650. 1:7700 and 1: 10000) at insemination. the PR, PSR and MNS were increased as the oocyte:sperm ratio increases. However, this result indicated that the 2PNF was high in the oocyte:sperm ratio (1:6000 and 1:6650). Therefore. these results suggested that when the percentage of morphologically normal epididymal sperm was more than 50. the fertilizing a ability was very similar to that of frozen-thawed ejaculated sperm and that the detailed evalu¬a ation of morphological normality in porcine IVF using epididymal sperm should be prerequisite to obtain the more effective fertilizing ability.

• Korean Journal of Animal Reproduction
• /
• v.24 no.1
• /
• pp.19-33
• /
• 2000

Eighty dairy farms (38 farms responded) in Palmerston North area of New Zealand were questioned by mail questionnaire on: 1) general characteristics, 2) milk yield and feed supplementary, 3) reproductive efficiencies (12 questions) and 4) reproductive disorders (12 questions) from February to July, 1998. Among those 4 items, the reproductive efficiencies (3) and disorders (4), various diseases and culling rates were surveyed and analyzed for Korean dairy farmers (especially in Cheju island) and compared with New Zealand. The results are as follows: 1. Fifteen farms in 38 dairy farms relied entirely on artificial insemination, the rest of 23 dairy farms (60.5%) raised 5∼6 bulls to increase conception rate. The dairy farmers in Palmerston North used artificial insemination from Oct 4th to Dec 10th for 42.8 days, and then used bulls from that point to coming Jan 10th for 41.4 days. The submission rate within 3, 6 and 10 weeks following the initiation of AI season was 84.7, 93.9 and 97.9% respectively. 2. The average age of heifers at the first estrus, pregnancy and calving was 11.0, 18.0 and 24.7 months respectively, and an average 1.4 estrus cycles were required for conception. The intervals of estrus recurrence and the following conception after calving were 38 and 68 days respectively. 3. Among inseminated cows, calving, abortion and empty cow was 90.9, 1.6 and 7.4% respectively. Calving rate decreased according to increasing farm size, while the number of empty cows decreased. 4. Stillbirth, retained placenta and delivery abnormalities were 5.3, 3.7 and 5.5% respectively, not different depend on herd size. 5. The incidence of milk fever, grass tetany, and ketosis was 3.6, 3.0 and 1.0%, respectively. The delivery abnormality and mastitis treated with medicine were 3.1 and 6.7%, but decreased according to farm size. Lameness was 8.6% on average, but over 10% in farms which has more than 400 milking cows. 6. Among the culled cows (15.5% of the total), those culled due to an old age, lameness and other diseases were 2.9, 1.8 and 4.3% respectively and those culled due to low milk production, reproductive abnormality reduced with farm size. 7. Compared with the data collected in Korea, the reproductive efficiency was better, and lameness, metabolic problem and culling rate were higher in New Zealand

• Korean Journal of Animal Reproduction
• /
• v.24 no.1
• /
• pp.97-108
• /
• 2000

The objectives of this study were performed to increase the efficiency of the culture conditions of embryos produced in vitro, and to assess the developmental potential after transfer of those embryos into recipients. The mean number of folliclular oocytes recovered from an ovary was 10.7. The rates of maturation and fertilization in Grade I oocytes were significantly (P＜0.05) higher than Graden and III. Developmental rate into blastocyst in the culture group of TCM-199 with BOEC were significantly higher (P＜0.05) than the groups of TCM-199 and conditioned medium (24.7% vs. 12.4% and 18.2%). The survivability of post-thawed blastocysts equilibrated for 3 min in EFS solution was significantly (P＜0.05) lower than l0 for 1 and 2 min (32.1% vs. 82.9% and 73.3%). Significantly higher (P＜0.05) survival rate in blastocysts was seen after freezing than in morulae stage embryos. Out of all 105 recipients, 49 (46.7%) were confirmed in pregnant. On pregnancy of cattle, 48 calves were born from 40 recipients. The ratio of twin and single calves was 30.5% (32/40 and 7.6% (8/40), respectively. However, the others composed of abnormal, as judging as 6 (12.2%) for abortion and 3 (6.1%) for stillbirth during the pregnant period.