본 연구는 밀어 (Rhinogobius brunneus)의 산란행동 및 초기생활사를 밝히기 위하여 4가지 수온조건 하에서 실험을 수행되었다. 본 실험을 위해 밀어는 금강 지류중의 하나인 갑천에서 성숙된 수컷과 암컷을 채집하였으며, 일정기간의 순치과정을 거친 후에 실험을 실시하였다. 본 실험 결과에 따르면, 성숙된 성어들의 구애행동 및 산란행동의 유형은 다음과 같이 요약된다. 밀어의 성숙된 수컷은 산란에 앞서 세력권을 형성한 후 자신의 세력권 안에 산란을 위한 둥지를 형성한 후, 성숙된 암컷을 유인하여 둥지에 하부에 타원형 난을 단층으로 붙이는 부착난 종으로 파악되었다. 수정된 알들은 수컷에 의해 보호받는 종(Parental care species)임 이 관찰되었으며, 수정난의 평균 장경은 1.5${\pm}$0.1 mm, 단경은 0.7${\pm}$0.1 mm였다. 실험실내 4개의 수온처리군에 따르면, 고온처리군 보다는 저온 처리군에서 부화까지 더 긴 시간이 소요되었으며, 높은 수온일수록 부화까지 소요되는 시간이 단축되는 것으로 나타났다. 또한, $18^{\circ}C$의 낮은 실험수온에서 부화된 자어는 약 3.6 mm의 전장을, $25^{\circ}C$ 이상의 높은 실험수온에서 부화된 자어는 3.1${\sim}$3.2 mm의 전장을 보였다. 또한, 처리군에서 수온이 높을수록 빠른 성장을 보여 부화까지 소요되는 시간이 단축되는 결과를 보였으나, 부화 시 자어의 크기는 높은 수온 상태에서 보다는 낮은 수온상태에 서 부화된 개체가 큰 것으로 나타났다. 본 밀어에 대한 발생생태학적 기초연구는 수질오염 및 서식지 파괴에 의해 어종의 다양성이 빠르게 감소하는 상황에서 어류 개체군 보호 및 생태계보호에 기초자료를 제공할 것으로 사료된다.
The genetic defects in human gametes and embryos can cause adverse effects on overall reproductive events. Biopsy of embryos for preimplantation genetic diagnosis (PGD) offers a new possibility of having children free of the genetic disease. In addition, advanced embryo culture method may enhance the effectiveness of embryo biopsy for the practical application of PGD. This experimental study was undertaken to evaluate the effects of coculture on the development in vitro of biopsied mouse embryos as a preclinical model for PGD of human embryos. Embryos were obtained after in vitro fertilization (IVF) from F1 hybrid mice (C57BLfemale/CBAmale). Using micromanipulation, 1, 2, 3 or 4 blastomeres of 8-cell stage embryos were aspirated through a hole made in the zona pellucida by zona drilling (ZD) with acidic Tyrode's solution (ATS). After biopsy of blastomeres, embryos were cultured in vitro for 110 hours in Ham's F-10 supplemented with 0.4% BSA or cocultured on the monolayer of Vero cells in the same medium. The frequence of blastocyst formation were recorded, and the embryos beyond blastocyst stage were stained with 10% Giemsa to count the total number of nuclei in each embryo. There was no significant difference in the blastocyst formation between the zona intact control group and the zona drilling (ZD) only, or biopsied groups. The hatching rate of all the treatment groups except 4/8 group was significantly higher than that of control group. In all the treatment groups, there was a significant reduction in the mean cell number of embryos beyond blastocyst stage ($50.2{\pm}14.0$ in control group vs. $41.2{\pm}7.9$ in ZD, $39.3{\pm}8.8$ in 7/8, $29.7{\pm}6.4$ in 6/8, $25.1{\pm}5.7$ in 5/8, and $22.1{\pm}4.3$ in 4/8 groups, p<0.05). When the same treatments were followed by coculture with Vero cells, a similar pattern was seen in the blastocyst formation and the hatching rate. However, in all the treatment groups, there was a significant increase in the mean cell number of embryos beyond blastocyst stage with coculture, compared with the parallel groups without coculture. In the cleavage rate of biopsied blastomeres cultured for 110 hours after IVF, there was no significant difference between coculture and non-coculture groups (87.2% vs. 78.7%). However, the mean cell number of embryos developed from the biopsied blastomeres was significantly higher in coculture group ($11.5{\pm}4.7\;vs.\;5.9{\pm}1.9$, p<0.05). In conclusion, biopsy of mouse embryos after ZD with ATS is a safe and highly efficient method for PGD, and coculture with Vero cells showed a positive effect on the development in vitro of biopsied mouse embryos and blastomeres as a preclinical model for PGD of human embryos.
In Pleuronectiformes, blind-side malpigmentation (hypermelanosis) is common in cultured flatfishes, and is economically important. To understand the mechanism of blind-side hypermelanosis in flatfishes, we examined when the malpigmentation initially occurred, and studied how the symptoms proceeded during early development of the starry flounder, Platichthys stellatus. To assess quantitative pattern changes of blind-side skin, we observed morphological development of the whole body from 22 (total length [TL] $10.0{\pm}0.2$ mm and body weight [BW] $8.8{\pm}0.57$ mg) to 110 days (TL $23.4{\pm}0.7$ mm, BW $193.6{\pm}23.3$ mg) after hatching (DAH), and also examined the malpigmented area rate of blind-side skin and the malpigmented fish ratios. The experimental animals were reared in fiberglass-reinforced plastic (FRP) tanks in water at a temperature of $18.9{\pm}1.9^{\circ}C$ and salinity of $32.6{\pm}0.6$ psu and were fed with rotifer and Artemia nauplii from 22 to 48 DAH, and with A. nauplii and commercial feed from 49 to 110 DAH. As results, the first staining patch seen by the naked eye was observed around the area between the anus and pelvic fin or caudal edge of the trunk at 80 DAH (TL $20.6{\pm}0.5$ mm, BW $112.5{\pm}8.8$ mg). The pigmented area and the pigmented fish ratios were significantly increased from 80 to 110 DAH. These results indicated that malpigmentation on the blind side of starry flounder was initially observed at about 2 cm in length and 100 mg in weight, and the pigmented domain on the blind-side skin was continually broadened by the differentiation of pigmented cells (melanophores and xanthophores) with growth.
The pattern of economic integration in the broiler industry can be grouped into three categories; 1) non-integration, 2) quasi-integration and 3) complete-integration. It is general to see that the non-integration is quite common under the market conditions of perfect competition, whereas the complete integration is more preferable in the imperfect competition. The quasi-integration, however, exists at all phases where the complete integration is not fully formed and implemented, but the non-integration has begun to alter its nature into integrated structure. The broiler industry in Korea has been characterized with the typically non-integrated independent operation, resulting in considerable price fluctuation and unstable industry as a whole. As a means of solving out the problem stemed from the non-integrated, growers and agribusinessmen involved in broiler industry have tended to develope the regular customer relationship prevailed between two parties. In fact, it has been practiced for years that most growers have been dealt with factor suppliers or processors on a regular basis for advantages of better price and quality, useful information, management help and so forth. Under the customary transaction, no formal contract has been made due to simple buyers and sellers relations, not like the one used to be performed in the form of contractual agreement. The broiler industry realizes the direction to go ahead toward the formal arrangement of integrated system from current regular transactions. As more Vowers, suppliers and processors recognize the necessity of it, the non-integrated industry appears to become the partially integrated by developing the existing customer relationship in such a way that functions of integrators are. further expanded and better organized. As a result, a type of quasi-integration started to show up by an integrator dominated in the field of hatching, feedmilling, dressing and by a grower's coop, It is concluded, therefore, that the evolution of quasi-integration in Korea's broiler industry is continuously taking place, implying the close approach to the completely integrated broiler production and marketing system.
The spawning behavior and developmental pattern of the Korean eleotrid fish (Odontobutis obscurus) was investigated in one of the tributary of Han River. The spawining of this species begins on April and continues up to August, whereas, the spawning. of same species were between the end of June and the middle of July in Kyushu, Japan. August The spawning places were the margin of river, its the bottom composed of sand and pebble (Table 5). The velocity of water in the spawning places was 0-40cm/sec, and the depth of waters was 10-40cm. The egg was spawned on underside of stone in the spawning places with one or two folds and ellipsoid in shape, 3.4mm in long diameter and 2.0mm in short diameter. The yolk is 1.2mm in diameter and yellow in color. For hatching of the egg, it takes one month in, $16^{circ}C~19^{\circ}C$ water, in Japan, whereas it takes only 14 days in $20^{circ}C~30^{\circ}C$ wate rin the study area. The growth rate of larva and juvenile fish were as follows: new hatched larva is 45mm in length; after 5 days it became 5.0 to 5.5mm in length and 1.2 to 1.5mm in height after 20 days. it became 11.0mm in length and 2.3mm in length. The mean size of the mature female is 110mm in total length and larger 10mm than that of the same fish found in Japan.
The work was conducted with tole purpose of investigation on the development pattern of epididymis in accordance with the growth of meat-type cockerels. 1. Histological features of various ductules in epididymis of the cockerel on the age of weeks were as follow: within 10 weeks after hatching rete testis and connecting ductules were well developed but efferent ductules were observed in immature form. During 10th to 20th week, the lining epithelium of various ductules in epididymis was in the developing stage near to the mature form. From 21th week, various ductules were abruptly matured. Lumen of rete testis was lined by simple squamous or simple columnar epithelial cells and that of efferent ductules, having many folds and being larger than any others, were lined by ciliated pseudostratified columnar epithelium with ciliated columnar cells, clear cells and basal cells were noted. Luminal epithelium of connecting ductules was composed of ciliated low pseudostratified columnar epithelial cells, ciliated columnar cells, clear cells and basal cells. The luminal surface of epididymal ducts was pseudostratified columnar epithelium and which was composed of high columnar cells and basal cells. 2. In the India ink absorption test, India ink granules were noted above the nucleus of some cells in the efferent ductules and the connecting ductules at 7 hours after administration of India ink to the mature epididymis, but not absorbed in the other ductules. The granules reactive to acid phosphatase were most abundant in some epithelial cells of efferent ductules and connecting ductules, especially above the nucleus of cells. The granules reactive to alkaline phosphatase were noted on the luminal border of efferent ductules. The granules reactive to PAS were scattered in the epithelial cells of efferent ductules and connecting ductules.
Mun, Seong Hee;You, Jin Ho;Oh, Hyeon Ji;Lee, Chi Hoon;Baek, Hea Ja;Lee, Young-Don;Kwon, Joon Yeong
한국발생생물학회지:발생과생식
/
제23권1호
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pp.35-42
/
2019
Fish shows great difference in growth rate between individuals during larval development and early growth. This difference seriously reduces the production efficiency in fish culture. Growth hormone (GH)/Insulin-like growth factor 1 (IGF1) system is said to play some pivotal roles in fish growth. In this study, we investigated differences of GH, IGF1 and GHR gene expressions in juvenile red spotted grouper (Epinephelus akaara) with different growth performance. Red spotted groupers were reared under the same environmental condition (water temperature $24{\pm}1^{\circ}C$, natural light) for 96 days after hatching. They were divided into 3 groups by size (fast growing, middle growing and slow growing groups: FGG, MGG, and SGG, respectively). RNA was extracted from the brain, liver and muscle tissues from each group, and target gene expression was examined by real-time PCR. In the brain with pituitary gland, expression of GH gene in FGG was significantly higher than the expression in SGG, but the expression of IGF1 and GHR genes in the muscle was highest in SGG. Difference of GHR and IGF1 mRNA in the liver between groups with different growth performance was less clear than that in other tissues, although level of IGF1 mRNA was higher in SGG than in MGG. These results suggest that hormonal governing of growth is not the same in fast growing and slow growing fish, and size grading could cause a shift of hormonal state and growth pattern in this species.
본 연구는 고품질 비단잉어 생산을 위해 비단잉어와 홍잉어 교배종의 성장과 적색색소의 빈도분석을 통해 비단잉어 색도를 분석하였다. 색소 및 성장도 분석을 위해 부화 후 170일령 비단잉어를 각 교배 집단별로[비단잉어${\times}$비단잉어(KK), 비단잉어${\times}$홍잉어(KR)와 홍잉어${\times}$비단잉어(RK)] 100개체씩 분석하였다. 성장도 분석 결과 비단잉어${\times}$홍잉어(KR) 교배집단에서 전장과 전중의 성장이 가장 높았다. 교배 그룹 모두 흰색, 적색-흰색, 적색의 3가지 색소 패턴을 보였다. 적색색소 분포는 KK 집단이 64%, KR 집단이 56%, RK 집단이 36%였다. 적색 색소 범위의 빈도분석 결과 적색색소 30~50%를 차지하는 비율이 KK 집단이 10%, RK 집단이 12% 그리고 KR 집단이 14%로 가장 높았다. 비단잉어 적색색소 빈도 분석에 관한 연구는 비단잉어 선별을 위한 분석 도구로서 활용이 가능할 것으로 판단된다.
Nutritional regulation of gene expression associated with growth and feeding behavior in avian species can become an important technique to improve poultry production according to the supply of nutrients in the diet. Insulin-like growth factor-I (IGF-I) found in chickens has been characterized to be a 70 amino acid polypeptide and plays an important role in growth and metabolism. Although it is been well known that IGF-I is highly associated with embryonic development and post-hatching growth, changes in the distribution of IGF-I gene expression throughout early- to late-embryogenesis have not been studied so far. We revealed that the developmental pattern of IGF-I gene expression during embryogenesis differed among various tissues. No bands of IGF-I mRNA were detected in embryonic liver at 7 days of incubation, and thereafter the amount of hepatic IGF-I mRNA was increased from 14 to 20 days of incubation. In eyes, a peak in IGF-I mRNA levels occurred at mid-embryogenesis, but by contrast, IGF-I mRNA was barely detectable in the heart throughout all incubation periods. In the muscle, no significant difference in IGF-I gene expression was observed during different stages of embryogenesis. After hatching, hepatic IGF-I gene expression as well as plasma IGF-I concentration increases rapidly with age, reaches a peak before sexual maturity, and then declines. The IGF-I gene expression is very sensitive to changes in nutritional conditions. Food-restriction and fasting decreased hepatic IGF-I gene expression and refeeding restored IGF-I gene expression to the level of fed chickens. Dietary protein is also a very strong factor in changing hepatic IGF-I gene expression. Refeeding with dietary protein alone successfully restored hepatic IGF-I gene expression of fasted chickens to the level of fed controls. In most circumstances, IGF-I makes a complex with specific high-affinity IGF-binding proteins (IGFBPs). So far, four different IGFBPs have been identified in avian species and the major IGFBP in chicken plasma has been reported to be IGFBP-2. We studied the relationship between nutritional status and IGFBP-2 gene expression in various tissues of young chickens. In the liver of fed chickens, almost no IGFBP-2 mRNA was detected. However, fasting markedly increased hepatic IGFBP-2 gene expression, and the level was reduced after refeeding. In the gizzard of well-fed young chickens, IGFBP-2 gene expression was detected and fasting significantly elevated gizzard IGFBP-2 mRNA levels to about double that of fed controls. After refeeding, gizzard IGFBP-2 gene expression decreased similar to hepatic IGFBP-2 gene expression. In the brain, IGFBP-2 mRNA was observed in fed chickens and had significantly decreased by fasting. In the kidney, IGFBP-2 gene expression was observed but not influenced by fasting and refeeding. Recently, we have demonstrated in vivo that gizzard and hepatic IGFBP-2 gene expression in fasted chickens was rapidly reduced by intravenous administration of insulin, as indicated that in young chickens the reduction in gizzard and hepatic IGFBP-2 gene expression in vivo stimulated by malnutrition may be, in part, regulated by means of the increase in plasma insulin concentration via an insulin-response element. The influence of dietary protein source (isolated soybean protein vs. casein) and the supplementation of essential amino acids on gizzard IGFBP-2 gene expression was examined. In both soybean protein and casein diet groups, the deficiency of essential amino acids stimulated chickens to increase gizzard IGFBP-2 gene expression. Although amino acid supplementation of a soybean protein diet significantly decreased gizzard IGFBP-2 mRNA levels, a similar reduction was not observed in chickens fed a casein diet supplemented with amino acids. This overview of nutritional regulation of IGF-I and IGFBP-2 gene expression in young chickens would serve for the establishment of the supply of nutrients to diets to improve poultry production.
The egg bags of Korean salamander(Hynobius leechii) were collected from farmlands in Gyeongsangnam-do area. The assumed breeding time, numerical variation of embryos in each egg bag, mortality and the rates of abnormalities were investigated. The toxicity of benomyl, the metabolite carbendazim and BIC which were frequently spread in agricultural area and caused spontaneous embryonic malformation was investigated. The assumed breeding time between the end of February and the end of March has the difference about a month because of a habitat and it takes about 2 or 3 weeks from laying eggs to hatching. The length of each egg bag and the number of embryos were very varied in each area. It is due to geographical variation. Among egg bags in total study area, only 406 of egg bags(17.70% of total egg bags) developed all of embryos to normal larvae, and 78.49% of total embryos were normally developed. The patterns of spontaneous embryonic malformation were 26 species from A to Z and the abnormal patterns in individual were 8 species and above. the geographical differences about the abnormal pattern were identified and 11 habitats categorized 4 groups. The most frequent abnormality in Gyeongsangnam-do area is the dysplasia of external gill. The caudal dysplasia, abdominal blister and dysplasia of fin were also frequently observed. Individuals showing severe external defect were histologically studied and they showed retinal hypo-pigmentation, thyroid carcinoma, somatic muscular dysplasia, degeneration of cephalic neuron and various organ dysplasia. Benomyl and carbendazim were treated by 10pM$^{\sim}$10uM and BIC was treated by 1$^{\sim}$40ppm to know the effect of toxicity about toxic substance of salamander. After benomyl was treated, a survival rate was sharply dropped from 2 to 8 days. $LC_{100}$ identified in $1{\mu}M$, $LC_{50}$ identified between 100nM and $1{\mu}M$. $EC_{50}$ was assumed between 10nM and 100nM. The prevalent external malformation was abdomen swelled abnormally and histo-pathological effects were abdomen, neural tube and lens hernia. This suggests that benomyl is the toxicitic substance which inhibits the development of digestive system and nervous system. The result of treated carbendazim was similar to that of the treated benomyl. The survival rate is sharply dropped between 2 and 6 days. $LC_{100}$ was identified $1{\mu}M$ and $LC_{50}$ was identified between 10nM and 100nM. This shows that cabendazim has stronger lethal toxicity than benomyl. Ventral blister, eye dysplasia and cephalic dysplasia in the individual of external malformation mean that cabendazim affected nervous system much more than benomyl. Because the toxicity of BIC affected less in the beginning but affected more in the near hatching period, the period causing toxicity is somewhat different. $LC_{100}$ identified near 40ppm and $LC_{50}$ identified near 25ppm. The external defect shows mainly ventral blister and histo-pathological results show intestinal deformities. This result suggests the BIC inhibited strongly the development of digestive system. These abnormal developments may be caused by antimitotic action, inhibition of tubulin complex, destruction of microtubule, inhibitions of neurulation and closing of neural fold, and by the inhibition of movement of neural crest cells of benomyl. These abnormal developments may be caused by the rupture of epithelium, the loss of microtubule, the reduction of spindle size, the inhibition of spindle assembly formation, the destruction of spindle poles of carbendazim. These abnormal developments may be caused cytotoxicity by inhibition of the synthesis of a number of macromolecules and similar reaction the inhibition of benomyl.
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