Gametogenesis, the reproductive cycle, and the condition index of the soft clam, Mya arenaria were investigated monthly based on histological observation at Sachon Bay, south coast of Korea from May 1998 to September 1999 . M. arenaria is dioceious. The ovary and testis were composed of a number of ovarian sacs and testicular tubules, respectively. Ripe oocytes were characterized by germinal vesicles with nucleoli, and their sizes about $60{\mu}m$ in diameter. Monthly changes in condition index and water temperature were closely related to the annual reproductive cycle. The reproductive cycle can be classified into 5 stages: early active stage (february to March), late active stage (April to August), ripe stage (September), partially spawned and spent stage (September to October), inactive stage (November and January).
Park, Sang-Mi;Kwon, Ki-Sang;Goo, Tae-Won;Yun, Eun-Young;Kang, Seok-Woo;Kim, Sung-Wan;Yu, Kweon;Kwon, O-Yu
Biomedical Science Letters
/
v.15
no.1
/
pp.37-45
/
2009
Insect cell cultures have become important tools in the production of biological substances for use in a variety of research, human and veterinary medicine, and pest control applications. These applications often require the introduction of foreign DNA into the cells and have generally used methods originally developed for use with human and other mammalian cell cultures. While these methods can be successfully employed, they are often less efficient with insect cells and frequently involve complex procedures or require specialized equipment. Even when they do work, they may require substantial modification because of differences in the culture medium or growth patterns of insect cells. In this study, We have optimized transfection conditions of Sf9 cell line using insect expression vector pIZT/V5-His which expresses green fluorescent protein effectively. Human stem cell factor (hSCF) is a glycoprotein that plays a key role in hematopoiesis acting both as a positive and negative regulator, often in synergy with other cytokines. It also plays a key role in mast cell development, gametogenesis, and melanogenesis. It can exist in membrane-bound form and in proteolytically released soluble form. As determined by an enzyme-linked immunosorbent assay performed, hSCF level in supernatant averaged 995ng/ml. The human hSCF was partially purified by immunoaffinity chromatography and analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. The results show that the hSCF has N-linked carbohydrate and corresponds to the soluble form, at or about 223 amino acids in length. The findings suggest functional importance for soluble hSCF in cells.
Closely correlated expression patterns between ubiquitin specific peptidase 9X-linked (USP9X) and adherens junction formation factor (Afadin) in mouse testis development suggests that Usp9x regulates the deubiquitination of Af-6 (also known as Afadin, AFDN), and subsequently, the cell adhesion dynamics during gametogenesis. However, this relationship has not yet been tested in other domestic animals. The study was examined the temporal and spatial expression patterns of porcine USP9X and AFDN from the pre-pubertal to adult stages using real time-PCR and immunohistochemistry. Furthermore, we detected the transcripts of USP9X and AFDN in the testis of 1-, 6- and 12-months old boar, respectively. USP9X and AFDN were found to have similar expressions patterns, with basal expression after 1 month followed by a significant up-regulation from 6 months (puberty) onwards. In addition, neither the AFDN or USP9X proteins were detected in spermatogenic cells but they were expressed in the leydig cells and sertoli cells. USP9X was detected around the basal lamina during pre-puberty, and predominantly expressed in the leydig cells at puberty. Finally, in adult testis, USP9X was increased at the sertoli cell-cell interface and the sertoli cell-spermatid interface. In summary, closely correlated expression patterns between USP9X and AFDN in boar testis supports the previous findings in mice. Furthermore, the junction connections between the sertoli cells may be regulated by the ubiquitination process mediated via USP9X.
Augyte, Simona;Yarish, Charles;Neefus, Christopher D.
ALGAE
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v.34
no.2
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pp.153-162
/
2019
Anthropogenic disturbances, including coastal habitat modification and climate change are threatening the stability of kelp beds, one of the most diverse and productive marine ecosystems. To test the effect of temperature and irradiance on the microscopic gametophyte and juvenile sporophyte stages of the rare kelp, Saccharina angustissima, from Casco Bay, Maine, USA, we carried out two sets of experiments using a temperature gradient table. The first set of experiments combined temperatures between $7-18^{\circ}C$ with irradiance at 20, 40, and $80{\mu}mol\;photons\;m^{-2}\;s^{-1}$. The second set combined temperatures of $3-13^{\circ}C$ with irradiance of 10, 100, and $200{\mu}mol\;photons\;m^{-2}\;s^{-1}$. Over two separate 4-week trials, in 2014 and again in 2015, we monitored gametogenesis, the early growth stages of the gametophytes, and early sporophyte development of this kelp. Gametophytes grew best at temperatures of $8-13^{\circ}C$ at the lowest irradiance of $10-{\mu}mol\;photons\;m^{-2}\;s^{-1}$. Light had a significant effect on both male and female gametophyte growth only at the higher temperatures. Temperatures of $8-15^{\circ}C$ and irradiance levels of $10-100{\mu}mol\;photons\;m^{-2}\;s^{-1}$ were conditions for the highest sporophyte growth. Sporophyte and male gametophyte growth was reduced at both temperature extremes-the hottest and coldest temperatures tested. S. angustissima is a unique kelp species known only from a very narrow geographic region along the coast of Maine, USA. The coupling of global warming with high light intensity effects might pose stress on the early life-history stages of this kelp, although, as an intertidal species, it could also be better adapted to temperature and light extremes than its subtidal counterpart, Saccharina latissima.
Kim, Chung Yoo;Hur, Young Baek;Han, Jong Chul;Park, Young chul
The Korean Journal of Malacology
/
v.32
no.3
/
pp.231-240
/
2016
Techniques were developed for holding and conditioning of Pacific oysters, Crassostrea gigas, in a closed recirculating system. Experimental adults were used 500 oysters ( x two system, total 1,000 oysters) which were collected in $20^{th}$ March 2016 from long-line aquaculture farm at the south coast of Korea. During conditioning periods concentrated live microalgae as Isochrysis sp. $15{\times}10^7cells/mL$, Tetraselmis sp. $2{\times}10^7cells/mL$ and Pheaodactylum sp. $18{\times}10^7cells/mL$ were added 5 L every day, respectively which micro algae were functioned as diets and biological filter. Over all experimental periods total water exchange rate was 21.3% (daily 0.5%). Over 42 days conditioning, female and male oysters were maturated 90.9% and 94.4%, respectively. Survival rate was 98.7%. Mean shell hight (8.3 mm), total wet weight (19.2 g), meat wet weight (5.0 g) and shell wet weight (13.6 g) were significantly increased (P < 0.05). Water quality parameters including the water temperature ($22.1{\pm}0.4^{\circ}C$), salinity ($24.9{\pm}04$), dissolved oxygen (5.1-7.9 mg/L) and pH ($7.93{\pm}0.15$) were kept stable. Concentration of dissolved inorganic nutrient as ammonia (1.96-0.35 mg/L), nitrite (0.03-0.16 mg/L), nitrate (1.34-0.47 mg/L), DIP (0.42-0.03 mg/L) and silicate (3.83-0.00 mg/L) were significantly decreased throughout experiment except nitrite which was increased (P < 0.05), but nitrogenous components stayed below toxic levels (ammonia 0.0-5.5 mg/L, nitrite 0.0-460.0 mg/L) which indicated that closed recirculation system with microalgae based bio-filter could supply sufficiently environment condition to holding and conditioning of oyster.
The dovelopment of the gonads, gametogenesis and the reproductive cycle of the topshell, Turbo cornutus Solander, which is one of valuable food animals fom Korean waters were studied by photomicroscophy. The materials were monthly collected from Bangeojin, Jeongjari and Dangweol, all these places being located in the south-eastern part of Korea, for one year from March 1979 to February 1980. Topshell is dioecious and oviparous. Gonad is situated on the surface of liver, which lies posteriorly. The surface of ovary and testis is covered with a fibrous membrane, membrane of connective and muscular fibers and then an outermost layer of simple-columnar epithelial cells which are composed of cuboidal and columnar mucous gland cells. Primordial germ cells develop on the germinal epithelium of ovarian and testicular lobuli which are originated from the fibrous membrane and extend toward hepatic gland. Undifferentiated mesenchymal tissue and pigment granular cells are abundantly distributed between the growing oocytes and spermatocytes in the early development stages. With the further development of the ovary and testis these tissue and cells gradually disappear. Then the undifferentiated mesenchymal tissue and pigment granular cells are considered to be related to the growing of the oocytes and spermatocytes. Early multiplicating oogonium is ca. $10\mu$ in diameter and nucleushaving a central nucleolus is ra. $8\mu$. As the oocytea grow to ca. $50-60\mu$ by the increase of cytoplasm, the oocytes become look like bunches of grapes which are attached to ovarian lobuli. Mature eggs are ca. $180-210\mu$ in diameter and it is surrounded by a gelatinous membrane of ca. $10\mu$ in thickness. After spawning, undischarged ripe eggs and spermatozoa remain in the ovary and testis respectively for some time. Then they finally degenerate, and proliferation of new oogonia and spermatogonia occur along the germinal epithelia of newly developed ovarian and testicular lobuli. Reprocuctive cycle of Turbo cornutus could be classified into five successive stages: multiplicative, growing, maturer spent and recovery stages. Spawning occurs from August to November with Peak spawning from early September to late October.
The structure of gonads, gametogenesis and reproductive cycle of the cockle, Fulvia mutice, were studied mainly by histological observation. The materials were monthly sampled in the southern area of Yeosu from October 1980 to September 1981. F. mutica was monoecious. The gonads were situated between the liver tissues and the outer fibronuscular layers compacted by the connective tissue fibers and muscle fibers beneath the outermost layer of simple cuboidal epithelium. The gonad was composed of a number of the ovarian sacs and the testicular tubules which form the tubular structure. Testicular tubules in the mature stage sometimes contained 'testis-ova' The undifferentiated mesenchymal tissues and the eosinophilic cells were abundantly distributed on the germinal epithelium in the early development stage. With the further development of the ovary and testis, these tissues and cells gradually disapprared. The undifferentiated mesenchymal tissues and the eosinophilic cells are related to the growing of the oocytes and spermatocytes . Early multiplicating oogonium was about $10{\mu}m$ in diameter. As the oocytes grow to $27-34\times50-58{\mu}m$ by increasing cytoplasm, the oocytes connected to the basement membrane by their egg-stalks. The ripe eggs were about $60{\mu}m$ in diameter and they were surrounded by gelatinous membrane. Most male germ cells in mature stage were transformed into the spermatozoa and they formed the sperm bundles. After spawning, undischarged ripe eggs and spermatozoa remained in the ovarian sac and the testicular tubule respectively for some time, then they finally degenerated. Especially the early spent ovarian sacs in May did not contract significantly and then they took part in the secondary maturation within two or three months during the summer season. The monthly changes of the fatness well agreed with the reproductive cycle. The reproductive cycle of F. mutica could be classified into six successive stages : multiplicative, growing, mature, spent, degenerative and recovery stage. It seems that the spawning season is closely rotated to the water temperature, and the spawning occurs from May to October at about $20^{\circ}C$ in water temperature. The peak spawning seasons appeared twice a year between June and July and in September. Acknowledgement The authors wish to express their gratitude to Dr. Kim, In Bae, Dr. Chun, Seh Kyu and Dr. Yoo, Sung Kyoo of National Fisheries University of Busan and Mr. Min, Byoung Seo of National fisheries Research and Development Agency for their critical reading of the manu script.
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.
Early development Linuparus trigonus(von Siebold) has been studied based on the samples collected monthly in Je-ju Island, Korea from February, 1975 to January, 1977. Gametogenesis, reproductive cycle, embryonic development were investigated by histological mettled, and morphological description was made on the first phyllosoma larva which reared in the laboratory. Testis is composed of two tubular duct which are symmetrical with H-shaped appearance. Outer layer of testis is of fibrous connective tissue capsule. In the lumen there is a convoluted seminiferous tubule with interstitial tissue. Ovary is a pair of symmetrical blind tubular lobes, and the midportions are connected each other. The ovary consists of a couple of ovarian sacs partitioned by two-layered connective tissue fibers. Proliferation of spermatogonia are observed all the year around on the germinal epithelium of seminiferous tubule. Partial spermatogenesis is always in progress, and the spermatozoa appear all the year around in the tubules. Nutrition of early oogonia is supplied by fibrous mesenchyme which is abundantly distributed in ovarian sacs. Oocytes grow and couplete maturation divisions in the follicle layers. They finally develop into mature ova before spawning. Reproductive cycle is classified into four successive stages; multiplication stage from September to December, growing stage from January to March, maturation division stage from April to May and mature stage from June to August. Spawning takes place from May to August with peak spawning from Into July to early August. Cleavage type is superficial. Blastopore is formed in blasto-disc region which is proliferation of blastoderm cells. Germinal layers are also derived from tile region. Mesoderm formation is originated from endodermal cells which are formed front the blasto-disc region. The endodermal cells are separated by the process of delamination from yolk sac and take part in the formation of the mid-gut. Morphological characteristics of first phyllosoma larva are different from the larvae of other Palinurid and Scyllarid species.
Fifty-five local collections of buck wheat, Fagopyrum esculentum, were investigated their ratios of long-styled (LS) and short-styled (SS) flowers, fertility, meiosis of megaspore and microspore mother cell, female and male gametogenesis, and egg apparatus in accordance with the sowing seasons (spring, summer), altitudes (20m, 50-100m, 300m), and parent style types (L, S). Also they were embryologically investigated the fertility, fertilizing phenomenon and proembryogenesis by the legitimate and illegitimate pollination. There were no differences in the ratios of long-styled and short-5tyled flowers along with altitudes, but more irregularness was observed in plain area than that in the mountaineous or coastal area. LS versus SS ratios by sowing seasons were significantly separated into 1 : 1 in the summer sowing (P 0.1), but they were irregularly separated in the spring sowing. The segregating ratios by parent style types showed more number of short-styled flower in the spring sowing, and were statistically seperated into 1 : 1 in the summer sowing (P 0.25), regardless to parent style types. In the artificial legitimate union, the seed setting rates of the summer sowing (59-61%) were much higher than those of the spring sowing (about 30%), but in the artificial illegitimate union the seed setting rates were only fructified about 0.8-1.8% in the spring sowing. The seed setting rates in accordance with flowering stages were larger in turn early, middle, late, in the summer sowing. The grain number and grain weight per plant of short-styled flower were more than those of long-styled one regardless to style types. The 1,000 grain weight of long-styled flower was heavier than that of short-styled one in large grain, but it was lighter than that of short-styled flower in small or medium grain. The percentage of normal female and male gametogenesis in the summer sowing were higher than those in the spring sowing. The ovule was atropous and two polar nuclei were a synkarion before flowering. The pollens germinated at 30 minuts after pollination and the pollen tube grew continually and penetrated into micropyle at 1.5-2 hours and the two male nuclei fertilized with egg nucleus at 3 -5 hours after pollination. Flertilizing times in summer were shorter than in autumn. The fertilized egg was divided in a small apical cell toward the interior of the embryo sac and a large basal cell toward the micropyle cell at 15-24 hours after pollination, and division times in summer were shorter than in autumn. The proembryo began the embryogenesis at 7-8 days and formed itself into the perfect embryo at 15 days after pollination.
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