Differential capacity of the parthenogenetic embryonic stem cells (PESCs) is still under controversy and the mechanisms of its neural induction are yet poorly understood. Here we demonstrated neural lineage induction of PESCs by addition of insulin-like growth factor-2 (Igf2), which is an important factor for embryo organ development and a paternally expressed imprinting gene. Murine PESCs were aggregated to embryoid bodies (EBs) by suspension culture under the leukemia inhibitory factor-free condition for 4 days. To test the effect of exogenous Igf2, 30 ng/ml of Igf2 was supplemented to EBs induction medium. Then neural induction was carried out with serum-free medium containing insulin, transferrin, selenium, and fibronectin complex (ITSFn) for 12 days. Normal murine embryonic stem cells derived from fertilized embryos (ESCs) were used as the control group. Neural potential of differentiated PESCs and ESCs were analyzed by immunofluorescent labeling and real-time PCR assay (Nestin, neural progenitor marker; Tuj1, neuronal cell marker; GFAP, glial cell marker). The differentiated cells from both ESC and PESC showed heterogeneous population of Nestin, Tuj1, and GFAP positive cells. In terms of the level of gene expression, PESC showed 4 times higher level of GFAP expression than ESCs. After exposure to Igf2, the expression level of GFAP decreased both in derivatives of PESCs and ESCs. Interestingly, the expression level of $Tuj1$ increased only in ESCs, not in PESCs. The results show that IGF2 is a positive effector for suppressing over-expressed glial differentiation during neural induction of PESCs and for promoting neuronal differentiation of ESCs, while exogenous Igf2 could not accelerate the neuronal differentiation of PESCs. Although exogenous Igf2 promotes neuronal differentiation of normal ESCs, expression of endogenous $Igf2$ may be critical for initiating neuronal differentiation of pluripotent stem cells. The findings may contribute to understanding of the relationship between imprinting mechanism and neural differentiation and its application to neural tissue repair in the future.
Litter production, nutrient concentration of each component of litterfall and amount of nutrients input to forest floor via litterfall were investigated for four years from May 2005 through April 2009 in Quercus variabilis forest at Mt. Worak National Park. Amount of litterfall in 2005, 2006, 2007, 2008 were 5.181, 5.372, 5.314, 5.361 ton $ha^{-1}yr^{-1}$, respectively. Average amount of litterfall for four years was 5.307${\pm}$0.09 ton $ha^{-1}yr^{-1}$. Average percentage of leaf litter, branch and bark, reproductive organ and the miscellaneous for four years were 67.5${\pm}$1.9, 13.3${\pm}$0.8, 10.1${\pm}$0.5 and 9.1${\pm}$1.2%, respectively. Average amount of N, P, K, Ca and Mg returned to forest floor via litterfall for four years in this Q. variabilis forest were 44.47${\pm}$6.70, 2.50${\pm}$0.25, 12.26${\pm}$0.30, 17.23${\pm}$2.32 and 9.56${\pm}$0.54 kg $ha^{-1}yr^{-1}$, respectively.
In recent years the number of patients waiting for organ transplantation has greatly outpaced the supply of human organs available, which leads to a renewed interest in pig-to-human xenotransplantation as an alternative. However, one of the biggest barriers in the xenotransplantation is presence of porcine endogenous retroviruses(PERV) that can infect human cells. In this study, to present a possible solution for this problem we tried to inhibit expression of PERVs using shRNAs(short hairpin RNA) at the level of RNA synthesis and virus release. The shRNA targeting the sequence of PERV A, B type was cloned into pSIREN-RetroQ vector under the control of polymerase-III U6-RNA gene promoter. Quantitative real-time PCR was performed to detect my alterations in mRNA production of PERV A, B targeted by the shRNA in each done. Depending on the target sequence of the shRNA, the transcription of PERV was decreased to as much as 4% and the number of progeny viruses was reduced to less than 1/200,000. Transgenic pigs producing such shRNAs may result in a highly reduced PERV expression in cells and organs, which is a prerequisite for safe xenotransplantations.
Kim, Joo-Heon;Shim, Cheol-Soo;Won, Jin-Young;Park, Young-Ji;Park, Soo-Kyoung;Kang, Jae-Seon;Hong, Yong-Geun
Reproductive and Developmental Biology
/
v.33
no.3
/
pp.163-169
/
2009
Many biological systems are regulated by an intricate set of feedback loops that oscillate with a circadian rhythm of roughly 24 h. This circadian clock mediates an increase in body temperature, heart rate, blood pressure, and cortisol secretion early in the day. Recent studies have shown changes in the amplitude of the circadian clock in the hearts and livers of streptozotocin (STZ)-treated rats. It is therefore important to examine the relationships between circadian clock genes and growth factors and their effects on diabetic phenomena in animal models as well as in human patients. In this study, we sought to determine whether diurnal variation in organ development and the regulation of metabolism, including growth and development during the juvenile period in rats, exists as a mechanism for anticipating and responding to the environment. Also, we examined the relationship between changes in growth factor expression in the liver and clock-controlled protein synthesis and turnover, which are important in cellular growth. Specifically, we assessed the expression patterns of several clock genes, including Per1, Per2, Clock, Bmal1, Cry1 and Cry2 and growth factors such as insulin-like growth factor (IGF)-1 and -2 and transforming growth factor (TGF)-${\beta}1$ in rats with STZ-induced diabetes. Growth factor and clock gene expression in the liver at 1 week post-induction was clearly increased compared to the level in control rats. In contrast, the expression patterns of the genes were similar to those observed after 5 weeks in the STZ-treated rats. The increase in gene expression is likely a compensatory change in response to the obstruction of insulin function during the initial phase of induction. However, as the period of induction was extended, the expression of the compensatory genes decreased to the control level. This is likely the result of decreased insulin secretion due to the destruction of beta cells in the pancreas by STZ.
This study was carried out to investigate the effects of changes of ionic strength according to growth stage on growth and flowering of Dendranthema grandiflorum (Ramat.) Kitamura 'Seiun' grown hydroponically in perlite. The stage I, II, and III covered early vegetative growth (27-40 days after planting), latter vegetative growth (41-54 days), and reproductive growth (55-80 days), respectively. The 2 strength (1S and 2S) of nutrient solution were treated in stage I, whereas 3 strengths (1S, 1-2S, and 2S) were treated in stage II. Then, total 9 treatments in stage III were designated by 3 treatments (tap water, 1S, and 2S) for each 3 strengths in stage II. Each nutrient solution was applied 8 times per day. At vegetative growth stage (54 days after planting), stem length was highest when irrigated 8 times a day with 1S nutrient solution. Both photosynthesis and transpiration rate were higher in 1S than those in other treatments (1-2S, 2S), whereas leaf chlorophyll content was highest in 2S treatment. Ion content of plant treated with 2S was higher than other treatments. Growth (plant height, leaf area, stem length), fresh weight, and dry weight of each plant organ after flower bud formation were better in tap water treatment (1-1-0) than other 1S treatments (1-1-1, 1-1-2). Regarding the number of days to flowering, tap water treatment was the most effective. Thus, after flower bud formation supplying tap water or lower concentration of nutrient solution than those used during the vegetative growth stage was economical in saving chemical fertilizers, shortening the number of days to flowering, reducing salt accumulation in media, saving efforts of leaching, and reducing ground water contamination.
Background: Excessive stress causes varied physiological and psychological disorders including male reproductive problems. Here, we attempted to investigate the protective effects of Korean Red Ginseng (Panax ginseng Meyer; KRG) against sub-acute immobilization stress-induced testicular damage in experimental rats. Methods: Male rats (age, 4 wk; weight, 60-70 g) were divided into four groups (n = 8 in each group): normal control group, immobilization control group, immobilization group treated with 100 mg/kg of KRG daily, and immobilization group treated with 200 mg/kg of KRG daily. Normal control and immobilization control groups received vehicle only. KRG (100 mg/kg and 200 mg/kg) was mixed in the standard diet powder and fed daily for 6 mo. Parameters such as organ weight, blood chemistry, sperm kinematic values, and expression levels of testicular-related molecules were measured using commercially available kits, Western blotting, and reverse transcription polymerase chain reaction. Results: Data revealed that KRG restored the altered testis and epididymis weight in immobilization stress-induced rats significantly (p < 0.05). Further, KRG ameliorated the altered blood chemistry and sperm kinematic values when compared with the immobilization control group and attenuated the altered expression levels of spermatogenesis-related proteins (nectin-2, cAMP responsive element binding protein 1, and inhibin-${\alpha}$), sex hormone receptors (androgen receptor, luteinizing hormone receptor, and follicle-stimulating hormone receptor), and antioxidant-related enzymes (glutathione S-transferase m5, peroxiredoxin-4, and glutathione peroxidase 4) significantly in the testes of immobilization stress-induced rats. Conclusion: KRG protected immobilization stress-induced testicular damage and fertility factors in rats, thereby indicating its potential in the treatment of stress-related male sterility.
The morphological study on the parenchymal cells in the adult planaria performed to observe their cytochemical and ultrastructural characteristics. The results are as follows. Nine types of cells are found in parenchyma. 1. Free parenchymal cell: These cells contain several chromatoid bodies around the nucleus. Heterochromatins are evenly dispersed in large nucleus. These cells are abundant in free ribosomes. 2. Fixed parenchymal cells: These cells have well-developed granular endoplasmic reticulum, mitochondria and Golgi complex but they contain the cytosols exhibiting electron-lucencies. 3. Rhabdite-forming cells: These cells contain the electron-dense rhabdite granules of up to about 0.3 x 0.9 $\mu$m in size. Rhabdite-forming cells have well-developed cell organelles, granular endolplasmic reticulum, mitochondria and Golgy complex. 4. A-type of basophilic granule cells: These cells contain irregularly-shaped granules exhibiting alcianophilia. These granules surrounded by a limited membrane, approximately 1.4 x 0.7 $\mu$m in size, are accumulated in the cytoplasm. 5. C-type of basophilic granule cells: These cells contain electron-dense granules of less than 0.2 $\mu$m in size, which exhibit PAS- positive reaction. This type of granule is also found in the muscle layer of parenchyma. 6. D-type of basophilic granule cells: This type of granule cell occurs only in the parenchyma around reproductive organ. The granules have cytochemical characteristics that they exhibit strongly positive reaction with PAS and weakly eosinophilic property. These electron-dense granules, which are 0.2 to 0.6 $\mu$m in length, have oval shapes. 7. E-type of basophilic granule cells: These cells are found only in the parenchyma around re productive organ. The granules contained in a small number in the cell, exhibit PAS-positive reaction and have an average size of 0. 2pm. 8. Eosinophilic granule cells: These cells contain a large number of eosinophilic granules which have relatively diverse sizes from 0.3 x 0.2 to 0.8 x 0.4 $\mu$m. Most of granules are round or irregularly-shaped and highly electrondense. These cells have an array of well-developed granular endoplasmic reticulum of which cisternae are distened. 9. Transparent granule cells contain electron-lucent granules which exhibit negative reactions with three kinds of cytochemical methods used in this experiment.
This study was conducted to determine the effect of the adrenal function on the reproductive organs in immature rats treated with PMS. Two hundred and ten female rats (Wistar-Imamichi albino rats) of 21 days old (body weight : $58.7{\pm}3.53g$) were disposed in the intact rat group (Int.-) and adrenalectomized rat group (Adx.-) and then each group was devided into 3 subgroups, such as control (-Cont.), PMS treated (-PMS) was administered subcutaneously with 25 IU PMS, and and PMS cortisol treated groups (-PMS+Corti.) with 25 IU PMS and $30.0{\mu}g$ cortisol on 5 th day (aged 26 days old) after adrenalectomy, while the control groups with physiological salt solution by the same way. The reprodutive organs were observed at 48, 54, 60, 66, 72, 78 and 84 hours after hormone treatments. The results obtained were as follows ; 1. The measurments of time average ovary weight in all treated groups were increased with the elapse of time after treatment, and the difference among the treatments was significant (p<0.01) in the all observation time. But the difference of those was not recognized in Int.-Cont. and Adx.-Cont. groups. In the multiple range test. ovary weight of adrenalectomized rat groups (Adx.-PMS and Adx.-PMS+Corti. groups) was significantly (p<0.05) lighter than those of intact rat groups (Int.-PMS and Int.-PMS+Corti. groups), and the effect of cortisol administration was not reconized. 2. The difference of uterus weight was significantly reconized (p<0.01) in all observation time. The weight in Int.-PMS and Int.-PMS+Corti. groups was heavier until 66 hours after treatment, but the values in the adrenalectomized Adx.-PMS and Adx.-PMS+Corti. groups were heavier after 72 hours. The multiple range test showed that the significant difference was not found between Int.-PMS and Int.-PMS+Corti. groups, and Adx.-PMS and Adx.-PMS+Corti. groups. 3. The adrenal weight was not significantly different among the compared groups.
Reproductive cycle with the gonadal development of Ruditapes philippinarum can be classified into five successive stages by histological observations: early active stage (January to March), late active stage (February to May), ripe stage (April to August), partially spawned stage (May to October), and spent/inactive stage (August to February). Changes in total protein contents in the adductor muscle tissues reached the maximum in the early and late active stages (February) and appeared the minimum in the ripe and partially spawned stages (May), while changes in their contents in the visceral mass tissues reached the maximum in the ripe and partially spawned stages (June) and gradually decreased in the partially spawned stage (June to October). On the whole, changes in total protein contents showed a negative correlationship between the adductor muscle and visceral mass tissues (r = -0.292, p < 0.05). Changes in total lipid contents in the adductor muscle tissues reached the maximum in the inactive and early active stages (January) and sharply decreased in the early and late active stages (February), while their contents in the visceral mass tissues reached the maximum in the ripe and partially spawned stages (April) and gradually decreased in the partially spawned stage (to October). On the whole, changes in total lipid contents showed a negative correlationship between the adductor muscle and visceral mass tissues (r = -0.699, p<0.05). Changes in glycogen contents in the adductor muscle tissues reached the maximum in the late active and ripe stages (April) and rapidly decreased in the partially spawned stage (May to October), while their contents in the visceral mass tissues reached the maximum in the early and late active stages (February) and rapidly decreased in the late active stage (March). Thereafter, their levels gradually increased in the ripe and partially spawned stages (April to July). On the whole, changes in glycogen contents appeared no correlationship between the adductor muscle and visceral mass tissues (r = 0.062, p > 0.05). These results indicate that the adductor muscle and visceral mass tissues are an important energy storage and nutrient supply organ in the Manila clams, and the nutrient contents of the adductor muscle and visceral muscle tissues change in response to gonadal energy needs.
There are numerous reports on the relative risk of pelvic inflammatory disease among the users versus the nonusers of intrauterine device. Reported relative risk varied from no difference between the two groups to 3-9 fold increase in the users. In an attempt to define this relative risk of pelvic inflammatory disease and related microorganisms ,pelvic organ observation and bacteriological study were done through laparoscopy. Specimens for microbiologic culture were obtained simultaneously from the fallopian tubes via laparoscopy and from the endocervix via regular pelvic examination method. The study population was consisted of 30 I.U.D.users and 35 J.U.D.nonusers who visited the Yonsei University Severance Hospital and the Sung-Ga Hospital for laparoscopic sterilization. The results obtained were as follows: 1. There was no difference in age distribution, economic status and numbers of parity and abortion between I.U.D. users and I.U.D. nonusers. 2. The pelvic inflammatory findings were noted on laparoscopy in 2 cases of I.U.D. users, with an incidence of 6.6%. And no pelvic inflammatory finding was noted in any of the nonusers,but this difference was not statistically significant (p>0.005). 3. All the bacteriologic culture of the specimens from the fallopian tubes of both groups yielded negative results. 4. The bacteriologic culture of the spec imens f rom the endocervix revealed more frequent isolation of possible pathogen such as Hem ophilus ,alpha-Streptococcus ,Corynebacteria, Bacteroides in the I.U.D.users than in the nonusers.But,this difference was also not statistically significant (p>0.005).
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