Recent findings indicate that Type 2 taste receptors (T2Rs) are expressed outside the gustatory system, including in the gastrointestinal tracts and the exocrine glands, such as the submandibular (SM), parotid (P), lacrimal (L) glands and pancreas (PC). Specifically, T2Rs are found in some of the gastrointestinal endocrine cells, and these cells secreted peptide hormones in response to stimulation by bitter-tasting compounds. The results show that T2Rs may have significant physiological roles besides bitter taste reception. The functions of the T2Rs in the exocrine glands remain poorly understood. An expression levels analysis of T2Rs will help to determine those functions in the exocrine glands. The expression levels of the T2Rs in the exocrine glands were discovered via the qPCR. C57BL/6J mice of 42~60-day-old were used. Messenger RNAs were extracted from S, P, L and PC. Cloned DNAs were synthesized by reverse transcription. Quantitative PCRs were performed using the SYBR Green method. The expression levels of the T2Rs were calculated as relative expression levels to that of the GAPDH. The statistical significance among the observed exocrine glands was tested using the variance analysis (ANOVA test). Tas2r108, out of murine 35 T2Rs, was the most highly expressed in every observed exocrine gland. This finding was similar to previous results from tongue papillae, but the expression levels were lower than those of the tongue papillae. Tas2r137 of SM, P, L and PC were expressed a little lower than that of tongue papillae. The T2Rs in the exocrine glands may play slightly different roles from those in the tongue. We suggest that physiological studies such as a patch clamp and functional Ca2+ imaging of acinar cells are necessary for understanding the Tas2r108 functions.

Cryopreservation affects osmotic tolerance and intracellular ion concentration through changes in expression levels of water and ion channels. Control of these changes is important for cell survival after cryopreservation. Relatively little is known about changes in K+ channel expression compared to water channel expression. This study was performed to investigate changes in TASK-2 channel (KCNK5: potassium channel, subfamily K, member 5), a member of two-pore domain K+ channel family, in cryopreserved mouse ovaries. Cryopreservation increased TASK-2 mRNA expression in mouse ovaries. In addition, TASK-2 protein expression was upregulated in vitrified and slowly frozen ovaries. TASK-2 protein was expressed in all area of granulosa cells that surround the oocyte within the follicle, except nucleus. Viability of cells overexpressed with TASK-2 was higher than that of vector-transfected cells. Our results found that TASK-2 expression was increased by cryopreservation and overexpression of TASK-2 decreased cryopreservation-induced cell death. These results suggest that TASK-2 upregulation might reduce cryodamage.

Muscular satellite cell (SC), which is stem cell of postnatal pig, is an important for study of differentiation into adipogenesis, myogenesis, and osteoblastogenesis. In this study, we isolated and examined from pig muscle tissue to determine capacity in proliferate, differentiate, and expression of various genes. Porcine satellite cells (PSC) were isolated from semimembranosus (SM) muscles of 90∼100 days old pigs according to standard conditions. The cell proliferation increased in multi-potent cell by Masson’s, oil red O, and Alizarin red staining respectively. We per-formed the expression levels of differentiation related genes using real-time PCR. We found that the differentiation into adipocyte increased expression levels of both fatty acid binding protein 4 (FABP4) and peroxisome proliferator- acti-vated receptor gamma (PPARγ) genes (p<0.01). Myocyte increased the expression levels of the myosin heavy chain (MHC), myogenic factor 5 (Myf5), myogenic regulatory factor (MyoD), and Myogenic factor 4 (myogenin) (p<0.01). Osteo-blast increased the expression levels of alkaline phosphatase (ALP) (p<0.01). Finally, porcine satellite cells were indu-ced to differentiate towards adipogenic, myogenic, and osteoblastogenic lineages. Our results suggest that muscle satellite cell in porcine may influence cell fate. Understanding the progression of PSC may lead to improved strat-egies for augmenting meat quality.

Immune-inducible antimicrobial peptides were produced using transgenic silkworms that expressed Rel family transcription factor, truncated BmRelish1 (BmRelish1t) genes under the control of the BmA3 promoter using the piggyBac vector. BmRelish1t gene contains all domains of Bmrelish: a Rel homolog domain (RHD), nuclear localization signal (NLS), acidic and hydrophobic amino acids (AHAA) rich region except the Ankyrin repeat domain (ANK) and the death domain (DD). (1:1) Mixtures of the donor vector (pG-3xP3EGFP-BmA3BmRelish1t) and helper vector were micro-injected into 1,800 eggs of bivoltin silkworms, Baegokjam and EGFP-induced fluorescence was observed for 25 broods of transgenic silkworms under a florescence stereomicroscope. Analysis by real-time PCR indicated that transgenic silkworms expressing BmRelish1t recombinant proteins displayed higher mRNA expression levels of the Bombyx mori antimicrobial peptides such as lebocin, moricin, and nuecin than the normal silkworms. Moreover, transgenic silkworms expressing BmRelish1t showed antibacterial activity against Escherichia coli. We suggest that transgenic expression of BmRelish1t may find useful applications for the production of various antimicrobial peptides at the same time in transgenic silkworms.

This study was conducted to compare the expression pattern of the specific factors associated with pregnancy and angiogenesis during early pregnancy in Hanwoo. Synchronized female Hanwoo (4～6 year‐old) were inseminated artificially. After 10 weeks after artificial insemination (AI), the pregnancy was tested by rectal palpation method. Three pregnant and non‐pregnant Hanwoo were used in this experiment, respectively. The plasma progesterone level was measured by ELISA. Western blot analysis was performed to detect the expression of pregnancy associated glycoprotein (PAG) or angiogenic factors (VEGF, B‐FGF, ANP‐1, and TIE‐2). The plasma P4 level was increase gradually in pregnant group and maintained high level. The concentration of PAG was significantly higher from 5th weeks in pregnant group compared to that of non‐pregnant group (p<0.05). The concentrations of the VEGF (p<0.05), B‐FGF (p<0.05), and ANP‐1 (p<0.05) were significantly increased from 6th or 7th week after AI in pregnant group, respectively. And the intensity of TIE‐2, ANP‐1 receptor, was well matched with ANP‐1 (p<0.05). Taken together, it can be postulated that the blood vessels connected with fetus and dam were formed dramatically around 40 days after AI, because the expression levels of the angiogenic factors were increased significantly from this time in pregnant Hanwoo.

Our previous research has identified granulin (grn) and p130 genes as sex steroidinducible genes in the rat hypothalamus, which might be involved in sexual differentiation of the brain. Phthalate esters that are used as plasticizers and also found at low levels in foods such as dairy products are often mentioned as suspected endocrine disrupters. The purpose of the present study is to elucidate whether perinatal exposure to dinbutyl phthalate (DBP), diisononyl phthalate (DINP) and di2ethylhexyl adipate (DEHA) affects hypothalamic sex steroidinducible genes. The present study assessed the effects of perinatal exposure to DBP, DINP and DEHA on sex steroid hormones levels and hypothalamic grn and p130 mRNA expressions at postnatal day (PND) 3 and 7. Pregnant rats were fed a soyfree diet containing 20, 200, 2,000 and 10,000 ppm of DBP, 40, 400, 4,000 and 20,000 ppm of DINP, or 480, 2,400 and 12,000 ppm of DEHA from gestational day (GD) 15 to GD 3 or 7. At PND 3 and 7, perinatal exposure to these chemicals did not substantially affect serum concentrations of testosterone and estradiol. At PND 3, the expression of grn mRNA levels in males was decreased by DEHA, and that of p130 was decreased by DBP, DINP and DEHA, though the effects were not dosedependent. At PND 7, the expression of grn gene in female pups was increased by higher doses of DBP and all the doses, except for 4,000 ppm, of DINP, while that in male pups decreased by 480 and 12,000 ppm of DEHA. Hypothalamic expression of p130 mRNA in males was increased by lower doses of DBP and all the doses of DINP, whereas that of females was decreased by 480 and 2,400 ppm of DEHA. These results suggest that these chemicals may affect the expression of grn and p130 genes by directly acting on the hypothalamus, thus leading to inappropriate expression of these genes.

To evaluate the correlations between the expression of cyclin B1 mRNA and protein after stimulation and oocyte activation and development of nuclear transferred mouse embryos, this study was performed. The oocyte activation was induced by 7% ethanol or 10/ Ca-ionophore without (single) or with (combined) 10/ cycloheximide (CH). Cyclin B1 mRNA and protein in mouse oocytes was evaluated by PCR and western blot. The activation and blastocyst development in both single (P<0.05) and combined (P<0.01) stimulation was higher than in non-activated group. The cyclin B1 mRNA and protein levels were significantly reduced in both single and combined stimulation groups (P<0.05), respectively. Cyclin B1 mRNA expression showed a negative correlation between activation and blastocyst development in both single and combined stimulation groups. And also the expression of cyclin B1 protein showed a negative correlation with between oocyte activation and blastocysts development in both single and combined stimulation groups. In conclusion, it may suggest that single and combined stimulation increases the oocyte activation and blastocyst development of nuclear transferred embryos, because it induces the degradation of cyclin B1 mRNA and protein after activation in enucleated mouse oocytes.

Background : Agastache rugosa (A. rugosa), belonging to the Lamiaceae family, is a medicinal plant mainly distributed in Korea and contains various phenolic compounds revealing anti-fungal and anti-HIV properties. This study is aim to investigate change in phenylpropanoid content of flowers at different developmental stages using high performance liquid chromatography (HPLC) and quantitative real time polymerase chain reaction (qRT-PCR). Methods and Results : The variation in the transcriptional level of phenylpropanoid biosynthetic genes and phenylpropanoid contents in the flowers of A. rugosa at different developmental stages was analyzed. The transcript levels of phenylpropanoid biosynthesis genes, including ArPAL (phenylalanine ammonia-lyase), ArC4H (cinnamate 4-hydroxylase), and ArCHS (Chalcone synthase), were high in flowers at 1st stage compared with flowers at 2nd and 3rd stages. On the other hand, the expression levels of flavonoid biosynthesis genes, including ArTAT (tyrosine amino transferase), ArHPPR (hydroxyl phenylpyruvate reductase), and ArRAS (rosmarinic acid synthase), were higher in flowers at 3rd stage than those of flowers at 1st and 2nd. These results were consistent with HPLC analysis revealing that most phenolic compounds were higher in flowers at 1st and 2nd stage but the level of rosmarinic acid was the highest in 3rd stage. Conclusion : Our findings provide the information on change in phenylpropanoid biosynthesis in A. rugosa flowers at different developmental stages.

Background : Lycoris radiata belongs to the Amaryllidaceae family and is a bulbous plant native to South Korea, China, and Japan. Galantamine, a representative alkaloid of Amaryllidaceae plants, including L. radiata, exhibits selective and dominant acetylcholinesterase inhibition. In this study, transcriptome analysis of L. radiata was performed. Methods and Results : Genes for galantamine biosynthesis were used to design primers for qRT-PCR. Quantitative RT-PCR was performed with LrActin as a reference gene for normalization. The RT-PCR results reveal the expression of LrPAL A and LrC4H at an early stage in the pathway. Interestingly, the expression of these genes was significantly higher in roots. However, the expression levels of LrNNR and LrN4OMT, which are closely involved in galantamine biosynthesis, were significantly higher in bulbs than leaves and roots. The expression levels of LrPAL B, LrTYDC, LrCYP98A3 and LrCYP76T were not significantly different among the different parts of the plants tested. HPLC analysis confirmed the presence of galantamine in all the organs, including the root (0.53 ± 0.07 ㎎/g dry weight), bulb (0.27 ± 0.04 ㎎/g dry weight), and leaf (0.75 ± 0.09 ㎎/g dry weight). The galantamine level in the bulb was 1.42 and 2.78 times higher than that in the root and leaf, respectively. The results of qRT-PCR for the eight galantamine genes revealed relatively high levels of genes expressed early, including LrPAL A, LrPAL B, LrC4H, and LrTYDC in the roots. However, in the bulbs, the levels of LrNNR and LrN4OMT were higher, which are crucial for galantamine biosynthesis. It also explains why bulbs contain high amounts of galantamine, which is likely due to the increased expression of LrNNR and LrN4OMT and the high levels of LrCYP96T, although the genes expressed early were expressed at high levels in the root. Conclusion : Transcript data of plants grown in a growth chamber revealed high expression levels of LrNNR and LrN4OMT genes that are closely involved in galantamine biosynthesis, and, as expected, we observed higher amounts of galantamine in the bulbs than in the root and leaves.

The process of spontaneous abortion involves a complex mechanism with various cytokines, growth factors, and hormones during the pregnancy. However, the mechanism underlying spontaneous abortion by pro- and antiinflammatory cytokines in the serum during the pregnancy is not fully understood. Therefore, the purpose of this study was to examine the relationship between the serum levels of pro- and anti-inflammatory cytokines and spontaneous abortion using the CBA/j × DBA/2 mouse model. Serum levels of pro-inflammatory cytokines, such as IFN-γ, IL-1α and TNF-α were not increased in abortion model mice, but anti-inflammatory cytokines, such as IL-4, IL-13 and IL-1ra were decreased compared to normal pregnant mice. In addition, serum levels of chemokine, such as SDF-1, G-CSF, M-CSF, IL-16, KC and MCP-1 were decreased in abortion model mice compared to normal pregnant mice. However, the expression levels of nesfatin-1/NUCB2 mRNA and protein in the uteri of implantation sites were significantly higher in abortion model mice than normal pregnant mice. These results suggest that uterine nesfatin-1/NUCB2 expression may be down-regulated by inflammatory cytokines and chemokines in the serum of pregnant mice. Moreover, this study suggests the possibility that nesfatin-1/NUCB2 expressed in the implantation sites may be associated with the maintenance of pregnancy.

Simazine은 triazine계 제초제로서, 잡초와 일년생 풀들을 통제하는데 우리나라를 포함하여 세계적으로 널리 사용되고 있으며, 미국, 유럽, 오스트레일리아의 경우, 물에서 두 번째로 많이 검출되는 살충제로 알려져 있다. Simazine에오염된 토양과 물을 통하여 사람에게 노출되어 이후 생체 내에 수년간 잔존하며 특히, 생물체의 지방 및 조직에 농축되는특징을 갖는 내분비계 장애물질로 분류되어졌다. 하지만 simazine이 정소세포의 사멸 또는 생존