The effects of bisphenol A(BPA) on gonadal sex differentiation and maturation in Oryzias latipes ... more The effects of bisphenol A(BPA) on gonadal sex differentiation and maturation in Oryzias latipes were investigated. Fish were exposed to aqueous solutions of BPA at nominal concentrations of 50, 100 and 200 µg/l from 2 days to 70 days of age. In process of sex differentiation, advanced oocyte development was observed when compared to the testicular growth. Ovaries were composed of the oocyte of the chromatin-nucleolus stage and peri-nucleolus stage in 20 days after hatching. Otherwise testes contained a number of the spermatogonia and spermatocytes in 30 days after hatching. In the process of sex differentiation, gonadal development was not different in the controls and BPA treatment groups until 30 days after hatching. In contrast, 70 days after hatching advanced development of oocytes in the ovary was observed from BPA treatment groups when compared to the controls, and inhibition of development of spermatogenesis in the testis was observed from BPA treatment groups when compared ...
The micropyles on the prefertilized and artificial fertilized eggs of Epinephelus fasciatus were ... more The micropyles on the prefertilized and artificial fertilized eggs of Epinephelus fasciatus were investigated using scanning electron microscope (SEM). The micropyles () of E. fasciatus eggs were found in the animal pole and theirs shape were observed as a flat crateriform of cylindrical shape. The micropylar vestibule arranged by 6~7 thickened spiral annuli on the ridge and contributed to differentiate and form fertilization cone for blocking to polyspermy by presenting swollen vestibule structure. As E. fasciatus eggs was pelagic, so chorionic surface was an uneven structures such as circular and fillar form nodules. Especially, various pores (0.15~0.55 , 230~270 pores) distributed at the only around micropyles, those pores radiately exhibited regular projection structures showing gill filament-shape. These ultrastructural characters of E. fasciatus eggs can be utilized in a taxonomical cue of grouper species.
To effects of sex maturation in olive flounder by regulating long photoperiod, gonadal developmen... more To effects of sex maturation in olive flounder by regulating long photoperiod, gonadal development and GTH mRNA expression in the pituitary were investigated. Photoperiod was treated natural photoperiod and long photoperiod (15L:9D) conditions from September 2011 to March 2012. The results showed that natural photoperiodic group showed a higher gonadosomatic index (GSI) than long photoperiodic group during the spawning season (March 2012). The histological analysis of ovarian tissue showed that natural photoperiod group of ovaries contained vitellogenic oocytes, but long photoperiod group of ovaries mainly contained perinucleolus staged oocyte and oil-drop staged oocytes. The FSH mRNA of olive flounder, under natural photoperiod group, showed a significantly higher expression but no significant difference under long photoperiod group. The LHβ mRNA showed a significantly higher expression only under natural photoperiod group. These results may suggest that long photoperiodic informat...
This study examines the effects on fertilization rate (FR), hatching rate (HR), and normal indivi... more This study examines the effects on fertilization rate (FR), hatching rate (HR), and normal individual rate after artificial fertilization using frozen thawed sperm according to the cryoprotectant (DMSO) concentration and the period of cryopreserved sperm of longtooth grouper, Epinephelus bruneus. Performing artificial fertilization using frozen-thawed sperm, after freezing the sperm at different DMSO concentration of 5.0%, 7.5%, 10.0% respectively, FR were (DMSO 5.0%: 99.5±0.8%, DMSO 7.5%: 99.5±0.7%, and DMSO 10.0%: 99.6±0.6%). The results are not significantly different from the control fresh sperm (100%). HR also (DMSO 5.0%: 96.2±2.3%, DMSO 7.5%: 95.3±3.6%, 10.0%: 96.6±1.8%) were not significantly different in each group. The normal individual rate after hatching using with control fresh sperm (98.4%±0.5) and DMSO concentration level of 5.0% (97.8±0.1%) were not significantly different. However, with 7.5% (97.2±0.6%) and 10.0% DMSO concentrations (95.9±0.2%) are lower than the nor...
The fertilized eggs of E. septemfasciatus are spherical and transparent with buoyancy at 790 to 8... more The fertilized eggs of E. septemfasciatus are spherical and transparent with buoyancy at 790 to 890 μm (average 821.8±2.0 μm) in diameter with 170 to 230 μm oil globules (average 192.9±0.93 μm). Hatching began approximately 46 and 35 hours after fertilization at 22.0°C and 25.0°C water temperature, respectively. The average total length of newly hatched larvae was 1.75±0.03 mm. Most of the yolk and oil globules were absorbed within 3 to 4 days after hatching. The larvae reached 2.48 to 2.72 mm in total length, and their mouths and anuses opened at 3 to 4 days after hatching. In this time, the mouth diameters of the larvae were 0.209 to 0.238 mm. The larvae reached 3.24 to 4.15 mm in total length at 11 to 17 days after hatching, and began to metamorphose at the time the second dorsal and pelvic spines appeared and elongated. The abdominal cavity was densely lined with melanophores. The larvae reached 5.12 mm in total length at 24 days after hatching.
To understand the sex reversal characteristics in the longtooth grouper (Epinephelus bruneus), th... more To understand the sex reversal characteristics in the longtooth grouper (Epinephelus bruneus), this study examined the sex reversal and artificial masculinization of wild caught E. bruneus reared in indoor rearing tank after a 17α-methyltestosterone injection. To domesticate a broodstock, 64 wild caught E. bruneus, between 47.0 to 110.0 cm in total length and from 1.5 to 21.4 kg in body weight, were reared in indoor rearing tank (4.0 to 5.0 m wide, and 2.5 to 3.0 m depth) for four years. Seven specimens showed sex reversal from female to male during indoor rearing condition, whose total length and body weights were from 63.0 to 99.0 cm and from 4.4 to 13.2 kg, respectively. After inducing artificial masculinization in 14 female E. bruneus with a 17α-methyltestosterone (2.0 mg/kg BW) implants for 3 years, spermiation occurred in 9 specimens (total length: 54.0 to 68.0 cm, body weight: 2.3 to 4.3 kg). Among the female to male sex reversals, two specimens returned back to being female,...
We investigated the androgenic effects of 17α- methyltestosterone (MT) on gonadal sex reversal in... more We investigated the androgenic effects of 17α- methyltestosterone (MT) on gonadal sex reversal in juvenile red spotted grouper Epinephelus akaara. The fish were immersed in 17α-MT at 1 and 5 mg/L. Treatment method of 17α-MT was once weekly for 4 and 8 weeks. Fish were sampled at 12 months after end of the treatment period in order to histological analysis. At the initiation of an experiment (70 day after hatching), juvenile red spotted grouper have the paired primordial gonads with somatic cells bellow kidney in the posterior portion of the body cavity. Formation of ovarian cavity indicates that the ovarian differentiation beginning at 70 DAH in red spotted grouper. At 12 months after end of the treatment period, control group, 17α-MT 1 mg/L treatment group for 4 and 8 weeks, and 17α-MT 5 mg/L treatment group for 4 weeks were all female. However, sex-changed males without ovarian cavity were observed in the 17α-MT 5 mg/L treatment group for 8 weeks. In grouper, we firstly reported that the red spotted grouper be able to induce the primary males by hormone treatment prior to gonadal sex differentiation.
The Cre/LoxP system is a well-established approach to spatially and temporally control genetic in... more The Cre/LoxP system is a well-established approach to spatially and temporally control genetic inactivation. The calcium/calmodulin-dependent protein kinase II alpha subunit (CaMKIIα) promoter limits expression to specific regions of the forebrain and thus has been utilized for the brain-specific inactivation of the genes. Here, we show that CaMKIIα-Cre can be utilized for simultaneous inactivation of genes in the adult brain and in male germ cells. Double transgenic Rosa26(+/stop-lacZ)::CaMKIIα-Cre(+/Cre) mice generated by crossing CaMKIIα-Cre(+/Cre) mice with floxed ROSA26 lacZ reporter (Rosa26(+/stop-lacZ)) mice exhibited lacZ expression in the brain and testis. When these mice were mated to wild-type females, about 27% of the offspring were whole body blue by X-gal staining without inheriting the Cre transgene. These results indicate that recombination can occur in the germ cells of male Rosa26(+/stop-lacZ)::CaMKIIα-Cre(+/Cre) mice. Similarly, when double transgenic Gnao(+/f)::C...
This study was investigated spawning behavior, structure of egg masses and egg development in Apl... more This study was investigated spawning behavior, structure of egg masses and egg development in Aplysia kurodai inhabiting the coastal waters of Jeju Island, Korea. The mating and courtship behavior of A. kurodai occurred in the form of unilateral copulating with chain formation. In chain copulation, only the first animal acted as a female; the second and succeeding animals acted as males (sperm donors) to the animals in front and as females to the animals behind. The fertilized eggs were packaged in capsules that are embedded in jelly to form a cylindrical string called an egg masses. The number of capsule per cm of the egg masses was 55 to 60 capsules and each capsule within the egg masses held 15 to 25 eggs. After spawning, the egg masses were bright yellow or orange in color. This egg masses color not changed until embryos developed into trochophore stage. Thereafter, as embryo developed from trochophore stage to veliger stage the egg masses color became brownish. The fertilized eggs were spherical, with a diameter of approximately 80±1 μm at spawning. At 5 to 6 days after spawning, the embryo developed into trochophore stage and began to rotate within the egg capsule. In the trochophore stage, the precursor of the velum, called the prototroch or prevelum, developed. At 10 days after spawning, the prevelum is transformed into the velum, and the trochophore developed into veliger stage. Between 10 to 15 days after spawning, the veligers broke out of the egg capsule, and hatched as free-swimming larvae.
This study was performed to investigate the effect of desalinated underground seawater (named as ... more This study was performed to investigate the effect of desalinated underground seawater (named as 'magma seawater', MSW) of Jeju Island in Korea on lipid metabolism and antioxidant activity. MSW was collected from underground of Han-Dong in Jeju Island, and freely given to high fat diet (HFD)-fed C57BL/6 mice for 10 weeks. Although there were no significant differences in the body weight changes and plasma lipid levels, hepatic triglyceride levels were significantly lower in the MSW group than in the normal tap water (TW)-drunken control group. Furthermore, the activity of fatty acid synthase (FAS) was significantly decreased and carnitine palmitoyltransferase (CPT) activity was increased in MSW group compared to TW group. Similarly, real-time PCR analysis revealed that mRNA expressions of lipogenic genes were lowered in MSW groups compared to the control group. In a morphometric observation on the liver tissue, accumulation of fats was remarkably reduced in MSW group. Meanwh...
Salmosin is a snake venom-derived novel disintegrin that antagonizes platelet aggregation. In thi... more Salmosin is a snake venom-derived novel disintegrin that antagonizes platelet aggregation. In this study, we investigated its functional specificity in tumor angiogenesis. Salmosin significantly inhibited bovine capillary endothelial cell proliferation induced by basic fibroblast growth factor but had no effect on normal growth of the cell. The basic fibroblast growth factor-induced in vivo angiogenesis in the chorioallantoic membrane was disrupted by salmosin treatment without affecting normal embryonic angiogenesis. Adhesion of the bovine capillary endothelial cells to vitronectin was also inhibited by the binding of salmosin to the alpha(v)beta3 integrin. Both the metastatic-tumor growth and the solid-tumor growth that developed in mice were effectively suppressed by salmosin treatment. Several lines of experimental evidence strongly suggest that the tumor-specific antiangiogenic activity of salmosin disrupts tumor growth by blocking the alpha(v)beta3 integrin that is expressed o...
In teleosts, gonadotropin-releasing hormone (GnRH) and gonadotropin hormone (GTH) play important ... more In teleosts, gonadotropin-releasing hormone (GnRH) and gonadotropin hormone (GTH) play important roles in regulating gonadal development and maturation. In Southeast Asia, the longtooth grouper, Epinephelus bruneus, is a commercially important aquaculture fish. In this study, we cloned and characterized the longtooth grouper GnRH1 gene and cDNAs of three gonadotropin subunits (GTHα, FSHβ, LHβ). The GnRH1 gene of longtooth grouper was 4,032 bp long, and contained four exons, 61, 151, 99, and 423 bp long. GTHα, FSHβ, and LHβ cDNAs were 509, 576, and 579 bp, respectively. Phylogenetic and Southern hybridization analyses revealed that the longtooth grouper GTH subunits were evolutionarily close to those of groupers and are one-copy genes. RT-PCR analyses showed that GTH subunit mRNAs were expressed at a higher level in the pituitary glands of immature fish than in those of mature fish, suggesting a role in gonadal maturation.
Stroke is one of the common causes of death and disability. Despite extensive efforts in stroke r... more Stroke is one of the common causes of death and disability. Despite extensive efforts in stroke research, therapeutic options for improving the functional recovery remain limited in clinical practice. Experimental stroke models using genetically modified mice could aid in unraveling the complex pathophysiology triggered by ischemic brain injury. Here, we optimized the procedure for generating mouse stroke model using an intraluminal suture in the middle cerebral artery and verified the blockage of blood flow using indocyanine green coupled with near infra-red radiation. The first week after the ischemic injury was critical for survivability. The survival rate of 11% in mice without any treatment but increased to 60% on administering prophylactic antibiotics. During this period, mice showed severe functional impairment but recovered spontaneously starting from the second week onward. Among the various behavioral tests, the pole tests and neurological severity score tests remained rel...
Mesenchymal stem cells are able to trans-di¡erentiate into nonmesodermal lineage cells. Here, we ... more Mesenchymal stem cells are able to trans-di¡erentiate into nonmesodermal lineage cells. Here, we identi¢ed downstream signaling molecules required for acquisition of neuron-like traits by mesenchymal stem cells following the elevation of intracellular cAMP levels. We found that forskolin induced neuron-like morphology and expression of neuron-speci¢c enolase and neuro¢lament-200 in mesenchymal stem cells. Forskolin sequentially activated protein kinase A and B-regulation of alpha-fetoprotein (Raf), which led to phosphorylation of extracellular signal-regulated kinase. Importantly, blockade of extracellular signal-regulated kinase phosphorylation with a mitogen-activated protein kinase (MAPK) kinase inhibitor abrogated the forskolin-induced morphological changes and induction of neuronal proteins. These results indicate that extracellular signal-regulated kinase/MAPK mediates both cAMP-induced early cytoskeletal rearrangement and the later induction of neuronal markers in mesenchymal stem cells. NeuroReport 16:1357^1361 c 2005 Lippincott Williams & Wilkins.
Reelin, an extracellular glycoprotein has an important role in the proper migration and positioni... more Reelin, an extracellular glycoprotein has an important role in the proper migration and positioning of neurons during brain development. Lack of reelin causes not only disorganized lamination of the cerebral and cerebellar cortex but also malpositioning of mesencephalic dopaminergic (mDA) neurons. However, the accurate role of reelin in the migration and positioning of mDA neurons is not fully elucidated. In this study, reelin-deficient reeler mice exhibited a significant loss of mDA neurons in the substantia nigra pars compacta (SNc) and a severe alteration of cell distribution in the retrorubal field (RRF). This abnormality was also found in Dab1-deficinet, yotari mice. Stereological analysis revealed that total number of mDA neurons was not changed compared to wild type, suggesting that the loss of mDA neurons in reeler may not be due to the neurogenesis of mDA neurons. We also found that formation of PSA-NCAM-positive tangential nerve fibers rather than radial glial fibers was greatly reduced in the early developmental stage (E14.5) of reeler. These findings provide direct evidence that the alteration in distribution pattern of mDA neurons in the reeler mesencephalon mainly results from the defect of the lateral migration using tangential fibers as a scaffold.
Ultrastructural characteristics of late spermatids and spermatozoa during spermiogenesis in longt... more Ultrastructural characteristics of late spermatids and spermatozoa during spermiogenesis in longtooth grouper Epinephelus bruneus from Jeju, Korea were investigated using transmission and scanning electron microscopy. The spermiogenesis type of E. bruneus belongs to type I because of nuclear rotation occurrence. The spherical head of late spermatids and spermatozoa ultrastructurally consist of apple-shaped nucleus, nuclear notch (Y or branch-shaped) and nuclear fossa. The midpiece contains the proximal centriole (9+0 microtubule structure), two centrioles connection structure (three osmophilic filaments), the distal centriole (9+0 microtubule structure), an osmophilic basal foot, two necklaces (six or seven osmophilic particles) and osmophilic alar sheets. It contains six to nine spherical mitochondria showing two rows arrangement type. Also a flagellum (9+2 microtubule structure) presents six swelling shaped segments type lateral fins with regular interval. Consequently, ultrastructural features of E. bruneus spermatids and spermatozoa contribute to provide useful systematic characters for taxonomic purpose. It may contribute to study that whether ultrastructural features of cellular organelles (cytoplasmic canal, necklace and lateral fins) correlate to physiological function or not on the duration of motility and survival in fish spermatozoa.
The characteristics and distributions of cholecystokinin (CCK)-producing cells and mucus-secretin... more The characteristics and distributions of cholecystokinin (CCK)-producing cells and mucus-secreting goblet cells were investigated in the digestive tract of the blacktip grouper (Epinephelus fasciatus). CCKproducing cells were scattered throughout the digestive tract. The highest frequency of CCK-producing cells was observed in the anterior intestine portion and pyloric ceca, with a very small number of cells distributed as far as the rectum. Mucus-secreting goblet cells were found to differ remarkably in their regional distributions and relative frequencies. High frequencies of mucus-secreting goblet cells were found in the digestive tract, mainly in the anterior intestine portion and pyloric ceca, but not the esophagus; the frequency decreased slightly toward the rectum. Our result suggests that food digested by gastric acid in the stomach moves on the anterior (including the pyloric ceca) and mid intestine portion, thereby ensuring effective stimulation of the CCK-producing cells. In addition, the distribution pattern of the CCK-producing cells closely resembled that of mucus-secreting goblet cells. In E. fasciatus, CCKproducing cells and mucus-secreting goblet cells seem to be well adapted to promoting optimal control of the digestive process.
The effects of bisphenol A(BPA) on gonadal sex differentiation and maturation in Oryzias latipes ... more The effects of bisphenol A(BPA) on gonadal sex differentiation and maturation in Oryzias latipes were investigated. Fish were exposed to aqueous solutions of BPA at nominal concentrations of 50, 100 and 200 µg/l from 2 days to 70 days of age. In process of sex differentiation, advanced oocyte development was observed when compared to the testicular growth. Ovaries were composed of the oocyte of the chromatin-nucleolus stage and peri-nucleolus stage in 20 days after hatching. Otherwise testes contained a number of the spermatogonia and spermatocytes in 30 days after hatching. In the process of sex differentiation, gonadal development was not different in the controls and BPA treatment groups until 30 days after hatching. In contrast, 70 days after hatching advanced development of oocytes in the ovary was observed from BPA treatment groups when compared to the controls, and inhibition of development of spermatogenesis in the testis was observed from BPA treatment groups when compared ...
The micropyles on the prefertilized and artificial fertilized eggs of Epinephelus fasciatus were ... more The micropyles on the prefertilized and artificial fertilized eggs of Epinephelus fasciatus were investigated using scanning electron microscope (SEM). The micropyles () of E. fasciatus eggs were found in the animal pole and theirs shape were observed as a flat crateriform of cylindrical shape. The micropylar vestibule arranged by 6~7 thickened spiral annuli on the ridge and contributed to differentiate and form fertilization cone for blocking to polyspermy by presenting swollen vestibule structure. As E. fasciatus eggs was pelagic, so chorionic surface was an uneven structures such as circular and fillar form nodules. Especially, various pores (0.15~0.55 , 230~270 pores) distributed at the only around micropyles, those pores radiately exhibited regular projection structures showing gill filament-shape. These ultrastructural characters of E. fasciatus eggs can be utilized in a taxonomical cue of grouper species.
To effects of sex maturation in olive flounder by regulating long photoperiod, gonadal developmen... more To effects of sex maturation in olive flounder by regulating long photoperiod, gonadal development and GTH mRNA expression in the pituitary were investigated. Photoperiod was treated natural photoperiod and long photoperiod (15L:9D) conditions from September 2011 to March 2012. The results showed that natural photoperiodic group showed a higher gonadosomatic index (GSI) than long photoperiodic group during the spawning season (March 2012). The histological analysis of ovarian tissue showed that natural photoperiod group of ovaries contained vitellogenic oocytes, but long photoperiod group of ovaries mainly contained perinucleolus staged oocyte and oil-drop staged oocytes. The FSH mRNA of olive flounder, under natural photoperiod group, showed a significantly higher expression but no significant difference under long photoperiod group. The LHβ mRNA showed a significantly higher expression only under natural photoperiod group. These results may suggest that long photoperiodic informat...
This study examines the effects on fertilization rate (FR), hatching rate (HR), and normal indivi... more This study examines the effects on fertilization rate (FR), hatching rate (HR), and normal individual rate after artificial fertilization using frozen thawed sperm according to the cryoprotectant (DMSO) concentration and the period of cryopreserved sperm of longtooth grouper, Epinephelus bruneus. Performing artificial fertilization using frozen-thawed sperm, after freezing the sperm at different DMSO concentration of 5.0%, 7.5%, 10.0% respectively, FR were (DMSO 5.0%: 99.5±0.8%, DMSO 7.5%: 99.5±0.7%, and DMSO 10.0%: 99.6±0.6%). The results are not significantly different from the control fresh sperm (100%). HR also (DMSO 5.0%: 96.2±2.3%, DMSO 7.5%: 95.3±3.6%, 10.0%: 96.6±1.8%) were not significantly different in each group. The normal individual rate after hatching using with control fresh sperm (98.4%±0.5) and DMSO concentration level of 5.0% (97.8±0.1%) were not significantly different. However, with 7.5% (97.2±0.6%) and 10.0% DMSO concentrations (95.9±0.2%) are lower than the nor...
The fertilized eggs of E. septemfasciatus are spherical and transparent with buoyancy at 790 to 8... more The fertilized eggs of E. septemfasciatus are spherical and transparent with buoyancy at 790 to 890 μm (average 821.8±2.0 μm) in diameter with 170 to 230 μm oil globules (average 192.9±0.93 μm). Hatching began approximately 46 and 35 hours after fertilization at 22.0°C and 25.0°C water temperature, respectively. The average total length of newly hatched larvae was 1.75±0.03 mm. Most of the yolk and oil globules were absorbed within 3 to 4 days after hatching. The larvae reached 2.48 to 2.72 mm in total length, and their mouths and anuses opened at 3 to 4 days after hatching. In this time, the mouth diameters of the larvae were 0.209 to 0.238 mm. The larvae reached 3.24 to 4.15 mm in total length at 11 to 17 days after hatching, and began to metamorphose at the time the second dorsal and pelvic spines appeared and elongated. The abdominal cavity was densely lined with melanophores. The larvae reached 5.12 mm in total length at 24 days after hatching.
To understand the sex reversal characteristics in the longtooth grouper (Epinephelus bruneus), th... more To understand the sex reversal characteristics in the longtooth grouper (Epinephelus bruneus), this study examined the sex reversal and artificial masculinization of wild caught E. bruneus reared in indoor rearing tank after a 17α-methyltestosterone injection. To domesticate a broodstock, 64 wild caught E. bruneus, between 47.0 to 110.0 cm in total length and from 1.5 to 21.4 kg in body weight, were reared in indoor rearing tank (4.0 to 5.0 m wide, and 2.5 to 3.0 m depth) for four years. Seven specimens showed sex reversal from female to male during indoor rearing condition, whose total length and body weights were from 63.0 to 99.0 cm and from 4.4 to 13.2 kg, respectively. After inducing artificial masculinization in 14 female E. bruneus with a 17α-methyltestosterone (2.0 mg/kg BW) implants for 3 years, spermiation occurred in 9 specimens (total length: 54.0 to 68.0 cm, body weight: 2.3 to 4.3 kg). Among the female to male sex reversals, two specimens returned back to being female,...
We investigated the androgenic effects of 17α- methyltestosterone (MT) on gonadal sex reversal in... more We investigated the androgenic effects of 17α- methyltestosterone (MT) on gonadal sex reversal in juvenile red spotted grouper Epinephelus akaara. The fish were immersed in 17α-MT at 1 and 5 mg/L. Treatment method of 17α-MT was once weekly for 4 and 8 weeks. Fish were sampled at 12 months after end of the treatment period in order to histological analysis. At the initiation of an experiment (70 day after hatching), juvenile red spotted grouper have the paired primordial gonads with somatic cells bellow kidney in the posterior portion of the body cavity. Formation of ovarian cavity indicates that the ovarian differentiation beginning at 70 DAH in red spotted grouper. At 12 months after end of the treatment period, control group, 17α-MT 1 mg/L treatment group for 4 and 8 weeks, and 17α-MT 5 mg/L treatment group for 4 weeks were all female. However, sex-changed males without ovarian cavity were observed in the 17α-MT 5 mg/L treatment group for 8 weeks. In grouper, we firstly reported that the red spotted grouper be able to induce the primary males by hormone treatment prior to gonadal sex differentiation.
The Cre/LoxP system is a well-established approach to spatially and temporally control genetic in... more The Cre/LoxP system is a well-established approach to spatially and temporally control genetic inactivation. The calcium/calmodulin-dependent protein kinase II alpha subunit (CaMKIIα) promoter limits expression to specific regions of the forebrain and thus has been utilized for the brain-specific inactivation of the genes. Here, we show that CaMKIIα-Cre can be utilized for simultaneous inactivation of genes in the adult brain and in male germ cells. Double transgenic Rosa26(+/stop-lacZ)::CaMKIIα-Cre(+/Cre) mice generated by crossing CaMKIIα-Cre(+/Cre) mice with floxed ROSA26 lacZ reporter (Rosa26(+/stop-lacZ)) mice exhibited lacZ expression in the brain and testis. When these mice were mated to wild-type females, about 27% of the offspring were whole body blue by X-gal staining without inheriting the Cre transgene. These results indicate that recombination can occur in the germ cells of male Rosa26(+/stop-lacZ)::CaMKIIα-Cre(+/Cre) mice. Similarly, when double transgenic Gnao(+/f)::C...
This study was investigated spawning behavior, structure of egg masses and egg development in Apl... more This study was investigated spawning behavior, structure of egg masses and egg development in Aplysia kurodai inhabiting the coastal waters of Jeju Island, Korea. The mating and courtship behavior of A. kurodai occurred in the form of unilateral copulating with chain formation. In chain copulation, only the first animal acted as a female; the second and succeeding animals acted as males (sperm donors) to the animals in front and as females to the animals behind. The fertilized eggs were packaged in capsules that are embedded in jelly to form a cylindrical string called an egg masses. The number of capsule per cm of the egg masses was 55 to 60 capsules and each capsule within the egg masses held 15 to 25 eggs. After spawning, the egg masses were bright yellow or orange in color. This egg masses color not changed until embryos developed into trochophore stage. Thereafter, as embryo developed from trochophore stage to veliger stage the egg masses color became brownish. The fertilized eggs were spherical, with a diameter of approximately 80±1 μm at spawning. At 5 to 6 days after spawning, the embryo developed into trochophore stage and began to rotate within the egg capsule. In the trochophore stage, the precursor of the velum, called the prototroch or prevelum, developed. At 10 days after spawning, the prevelum is transformed into the velum, and the trochophore developed into veliger stage. Between 10 to 15 days after spawning, the veligers broke out of the egg capsule, and hatched as free-swimming larvae.
This study was performed to investigate the effect of desalinated underground seawater (named as ... more This study was performed to investigate the effect of desalinated underground seawater (named as 'magma seawater', MSW) of Jeju Island in Korea on lipid metabolism and antioxidant activity. MSW was collected from underground of Han-Dong in Jeju Island, and freely given to high fat diet (HFD)-fed C57BL/6 mice for 10 weeks. Although there were no significant differences in the body weight changes and plasma lipid levels, hepatic triglyceride levels were significantly lower in the MSW group than in the normal tap water (TW)-drunken control group. Furthermore, the activity of fatty acid synthase (FAS) was significantly decreased and carnitine palmitoyltransferase (CPT) activity was increased in MSW group compared to TW group. Similarly, real-time PCR analysis revealed that mRNA expressions of lipogenic genes were lowered in MSW groups compared to the control group. In a morphometric observation on the liver tissue, accumulation of fats was remarkably reduced in MSW group. Meanwh...
Salmosin is a snake venom-derived novel disintegrin that antagonizes platelet aggregation. In thi... more Salmosin is a snake venom-derived novel disintegrin that antagonizes platelet aggregation. In this study, we investigated its functional specificity in tumor angiogenesis. Salmosin significantly inhibited bovine capillary endothelial cell proliferation induced by basic fibroblast growth factor but had no effect on normal growth of the cell. The basic fibroblast growth factor-induced in vivo angiogenesis in the chorioallantoic membrane was disrupted by salmosin treatment without affecting normal embryonic angiogenesis. Adhesion of the bovine capillary endothelial cells to vitronectin was also inhibited by the binding of salmosin to the alpha(v)beta3 integrin. Both the metastatic-tumor growth and the solid-tumor growth that developed in mice were effectively suppressed by salmosin treatment. Several lines of experimental evidence strongly suggest that the tumor-specific antiangiogenic activity of salmosin disrupts tumor growth by blocking the alpha(v)beta3 integrin that is expressed o...
In teleosts, gonadotropin-releasing hormone (GnRH) and gonadotropin hormone (GTH) play important ... more In teleosts, gonadotropin-releasing hormone (GnRH) and gonadotropin hormone (GTH) play important roles in regulating gonadal development and maturation. In Southeast Asia, the longtooth grouper, Epinephelus bruneus, is a commercially important aquaculture fish. In this study, we cloned and characterized the longtooth grouper GnRH1 gene and cDNAs of three gonadotropin subunits (GTHα, FSHβ, LHβ). The GnRH1 gene of longtooth grouper was 4,032 bp long, and contained four exons, 61, 151, 99, and 423 bp long. GTHα, FSHβ, and LHβ cDNAs were 509, 576, and 579 bp, respectively. Phylogenetic and Southern hybridization analyses revealed that the longtooth grouper GTH subunits were evolutionarily close to those of groupers and are one-copy genes. RT-PCR analyses showed that GTH subunit mRNAs were expressed at a higher level in the pituitary glands of immature fish than in those of mature fish, suggesting a role in gonadal maturation.
Stroke is one of the common causes of death and disability. Despite extensive efforts in stroke r... more Stroke is one of the common causes of death and disability. Despite extensive efforts in stroke research, therapeutic options for improving the functional recovery remain limited in clinical practice. Experimental stroke models using genetically modified mice could aid in unraveling the complex pathophysiology triggered by ischemic brain injury. Here, we optimized the procedure for generating mouse stroke model using an intraluminal suture in the middle cerebral artery and verified the blockage of blood flow using indocyanine green coupled with near infra-red radiation. The first week after the ischemic injury was critical for survivability. The survival rate of 11% in mice without any treatment but increased to 60% on administering prophylactic antibiotics. During this period, mice showed severe functional impairment but recovered spontaneously starting from the second week onward. Among the various behavioral tests, the pole tests and neurological severity score tests remained rel...
Mesenchymal stem cells are able to trans-di¡erentiate into nonmesodermal lineage cells. Here, we ... more Mesenchymal stem cells are able to trans-di¡erentiate into nonmesodermal lineage cells. Here, we identi¢ed downstream signaling molecules required for acquisition of neuron-like traits by mesenchymal stem cells following the elevation of intracellular cAMP levels. We found that forskolin induced neuron-like morphology and expression of neuron-speci¢c enolase and neuro¢lament-200 in mesenchymal stem cells. Forskolin sequentially activated protein kinase A and B-regulation of alpha-fetoprotein (Raf), which led to phosphorylation of extracellular signal-regulated kinase. Importantly, blockade of extracellular signal-regulated kinase phosphorylation with a mitogen-activated protein kinase (MAPK) kinase inhibitor abrogated the forskolin-induced morphological changes and induction of neuronal proteins. These results indicate that extracellular signal-regulated kinase/MAPK mediates both cAMP-induced early cytoskeletal rearrangement and the later induction of neuronal markers in mesenchymal stem cells. NeuroReport 16:1357^1361 c 2005 Lippincott Williams & Wilkins.
Reelin, an extracellular glycoprotein has an important role in the proper migration and positioni... more Reelin, an extracellular glycoprotein has an important role in the proper migration and positioning of neurons during brain development. Lack of reelin causes not only disorganized lamination of the cerebral and cerebellar cortex but also malpositioning of mesencephalic dopaminergic (mDA) neurons. However, the accurate role of reelin in the migration and positioning of mDA neurons is not fully elucidated. In this study, reelin-deficient reeler mice exhibited a significant loss of mDA neurons in the substantia nigra pars compacta (SNc) and a severe alteration of cell distribution in the retrorubal field (RRF). This abnormality was also found in Dab1-deficinet, yotari mice. Stereological analysis revealed that total number of mDA neurons was not changed compared to wild type, suggesting that the loss of mDA neurons in reeler may not be due to the neurogenesis of mDA neurons. We also found that formation of PSA-NCAM-positive tangential nerve fibers rather than radial glial fibers was greatly reduced in the early developmental stage (E14.5) of reeler. These findings provide direct evidence that the alteration in distribution pattern of mDA neurons in the reeler mesencephalon mainly results from the defect of the lateral migration using tangential fibers as a scaffold.
Ultrastructural characteristics of late spermatids and spermatozoa during spermiogenesis in longt... more Ultrastructural characteristics of late spermatids and spermatozoa during spermiogenesis in longtooth grouper Epinephelus bruneus from Jeju, Korea were investigated using transmission and scanning electron microscopy. The spermiogenesis type of E. bruneus belongs to type I because of nuclear rotation occurrence. The spherical head of late spermatids and spermatozoa ultrastructurally consist of apple-shaped nucleus, nuclear notch (Y or branch-shaped) and nuclear fossa. The midpiece contains the proximal centriole (9+0 microtubule structure), two centrioles connection structure (three osmophilic filaments), the distal centriole (9+0 microtubule structure), an osmophilic basal foot, two necklaces (six or seven osmophilic particles) and osmophilic alar sheets. It contains six to nine spherical mitochondria showing two rows arrangement type. Also a flagellum (9+2 microtubule structure) presents six swelling shaped segments type lateral fins with regular interval. Consequently, ultrastructural features of E. bruneus spermatids and spermatozoa contribute to provide useful systematic characters for taxonomic purpose. It may contribute to study that whether ultrastructural features of cellular organelles (cytoplasmic canal, necklace and lateral fins) correlate to physiological function or not on the duration of motility and survival in fish spermatozoa.
The characteristics and distributions of cholecystokinin (CCK)-producing cells and mucus-secretin... more The characteristics and distributions of cholecystokinin (CCK)-producing cells and mucus-secreting goblet cells were investigated in the digestive tract of the blacktip grouper (Epinephelus fasciatus). CCKproducing cells were scattered throughout the digestive tract. The highest frequency of CCK-producing cells was observed in the anterior intestine portion and pyloric ceca, with a very small number of cells distributed as far as the rectum. Mucus-secreting goblet cells were found to differ remarkably in their regional distributions and relative frequencies. High frequencies of mucus-secreting goblet cells were found in the digestive tract, mainly in the anterior intestine portion and pyloric ceca, but not the esophagus; the frequency decreased slightly toward the rectum. Our result suggests that food digested by gastric acid in the stomach moves on the anterior (including the pyloric ceca) and mid intestine portion, thereby ensuring effective stimulation of the CCK-producing cells. In addition, the distribution pattern of the CCK-producing cells closely resembled that of mucus-secreting goblet cells. In E. fasciatus, CCKproducing cells and mucus-secreting goblet cells seem to be well adapted to promoting optimal control of the digestive process.
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