Microenvironments surrounded with various extracellular matrix (ECM) components can decide specifically the fate of spermatogonial stem cells (SSCs) and integrin heterodimers recognizing directly ECM proteins play an important role in transporting ECM-derived signals into cytoplasm, resulting in inducing a variety of biological functions such as cell attachment, self-renewal and differentiation. However, to date, studies on type of integrin heterodimers expressed functionally on the undifferentiated SSCs derived from mouse with hybrid strain remain unclear. Therefore, we tried to investigate systematically what kind of integrin heterodimers are expressed transcriptionally, translationally and functionally in the SSCs derived from testis of hybrid (B6CBAF1) mouse. For these, magnetic activated cell sorting (MACS) using Thy1 antibody was used for isolating SSCs from testis, and real-time PCR or fluorescence immunoassay was conducted for measuring transcriptional or translational level of integrin α and β subunits in the isolated SSCs. Subsequently, antibody inhibition assay was conducted for confirming functionality of presumed integrin heterodimers. As the results, transcriptional levels of genes encoding total 25 integrin subunits were quantified, 7 integrin α (α4, α6, α7, α9, αV, αL and αE) and 2 integrin β (β1 and β5) subunit genes showed significantly increased transcriptional up-regulation, compared to the other integrin subunit genes. In contrast, integrin α3, α5, α10 and α11, and integrin β2, β3, β4 and β7 were weakly transcribed. When translational levels of the integrin α subunits showing high transcription level (α4, α6, α7, α9, αV, αL and αE) were measured, significantly strong translational up-regulation of integrin α6, α7, α9, αV and αL subunit genes were detected, whereas integrin α4 and αE subunit genes were weakly. In case of integrin β subunit, β1 evaluated more expression than β5. Based on these results, we speculated that the undifferentiated SSCs derived from B6CBAF1 mouse might express integrin α4β 1, α6β1, α7β1, α9β1, αVβ1 or αVβ5 on plasma membrane. Subsequently, the hybrid strain SSCs showed significantly increased adhesion to fibronectin, laminin, tenascine-C and vitronectin and functional blocking of integrin α4β1, α6β1, α9β1, and αVβ1 or αVβ5 in SSCs significantly inhibited attachment to fibronectin, laminin, tenascin-C and vitronectin, respectively. Accordingly, we could identify that the hybrid (B6CBAF1) mouse-derived SSCs had integrin α4β1, α6β1, α9β1, αVβ1 or αVβ5 on plasma membrane. Moreover, this information will greatly contribute to constructing non-cellular niche supporting self-renewal of SSCs in the future.
Glutathione S-transferase (GST) is a key gene involved in multiple stress tolerance in all living organisms, though it is still to be disclosed the gene function in teff grass [Eragrostis tef (Zucc.)Trotter].The objectives of this study were to clone and molecular characterization of GST gene in teff grass. We characterized GST1 from teff grass (EtGST1), it composed of a 645-bp open reading frame (ORF) that encoded 195 amino acid residue. Further, we transformed EtGST1 in E.coli BL21 (DE3) cells. This recombinant EtGST1 in E.coli BL21(DE3) induced at 37°C temperature. In addition, Growth of cells overexpressing EtGST1 rapidly increased in the presence of polyethylene glycol (5%), heat (46°C), NaCl (0.6%), and arsenic (1 mM) than that of cells harboring an empty vector. These results suggest that EtGST1 would be suitable candidate for improving tolerance in forages and/or grasses species against multiple abiotic stresses.
1. 국내에서 수집한 수수의 성숙기 간장이 250 cm 이상되는자원은 5 종으로 전체 62 수집종의 8.1 %를 차지하였다.
2. 수형 특성 조사에서는 3개 분얼수를 가지는 자원은 전체수집종의 56.6%를 나타낸 것으로 보아, 평균 분얼수는 3개 인것으로 나타났다.
3. 잎의 특성조사에서 엽장이 90 cm 이상되는 자원은 14.5%로 분류되었다.
4. 분얼형에 있어서는 반밀수타원형이 전체 수수 수집종의32.3%를 차지하였다.
5. 전체 수집종 수장의 평균값은 29.2 cm로 나타났다.
Spermatogonial stem cells (SSCs) developed into sperms through spermatogenesis have been utilized as a useful tool in the field of regenerative medicine and infertility. However, a small number of highly qualified SSCs are resided in the seminiferous tubule of testis, resulted in developing effective in-vitro culture system of SSCs for solving simultaneously quantitative and qualitative problems. Presently, SSCs can be enriched on testicular stromal cells (TSCs), but there are no systematic researches about TSC culture. Therefore, we tried to optimize culture condition of TSCs derived from mouse with different strains. For these, proliferation and viability were measured and compared by culturing ICR outbred or DBA/2 inbred mouse-derived TSCs at 35 or 37℃. In case of ICR strain, primary TSCs cultured at 37℃ showed significantly higher proliferation and viability than those at 35℃ and significant increase of proliferation and viability in sub-passaged TSCs was detected in the 35℃ culture condition. Moreover, sub-passage of primary TSCs at 35℃ induced no significant effects on proliferation and viability. In contrast, in case of DBA/2 strain, significantly improved proliferation were detected in the primary TSCs cultured at 35℃, which showed no significant difference in the viability, compared to those at 37℃. Furthermore, sub-passaged TSCs cultured at 37℃ showed no significant differences in proliferation and viability, compared to those at 35℃. However, with significant decrease of proliferation induced by sub-passage of primary TSCs at 35℃, no significant effects on proliferation and viability were resulted from sub-passage of primary TSCs at 37℃. From these results, culture temperature of primary TSCs derived from outbred and inbred strain of mouse could be separately optimized in primary culture and subculture.
Background : This study was performed to investigate by antioxidant activity, total phenolic contents, total flavonoid contents, and effective component of Astragalus membranaceus treated with different artificial light Sources (fluorescent lamp, red, blue, green, white, LEP). Methods and Results : We investigated the effects of various artificial light sources on the DPPH radical activity, total phenol and flavonoid contents, tyrosinase activity and main flavonoid compounds contents (formononetin and calycosin) and other biological activities in A. membranaceus. Antioxidant activities were 53.6% as the highest level of activity under LEP light. Growth under LEP light also produced the highest total phenolic and flavonoid contents of 36.05 and 5.94 mg/ml, respectively. Extracts from plants grown under LEP light caused the highest inhibition of tyrosinase activity with inhibition of 35.37, 61.87, and 65.49%, respectively, for extract concentrations of 100 μg/ml, 500 μg/ml, and 1000 μg/ml compared with other artificial light treatments. Conclusion : Little information is available on the influence of LED and LEP light sources on antioxidant production or other biological activities in A. membranaceus. Our goal in this study was to determine the effects of LED and LEP artificial light sources on the production of new functional compounds in A. membranaceus.
This study was investigated to evaluate germination rate of Astragalus membranaceus B. in Korea as affectedby storage temperature, germination temperature and storage period of seed. The highest germination rate was obtainedfrom condition of 25℃ in germination temperature. Seeds were stored at −20℃ and 5℃ for 8 weeks has showed higher ger-mination rate than one at room temperatures. The germination rates showed significantly difference by harvested year of2010, 2011 and 2012. The seed of A. membranaceus in harvested year of 2011 and 2012 had germinated well. On the otherhand, seeds in harvested year of 2010 were not nearly germinated. Consequently, the longer storage period after seed har-vest lower germination rate and seed vigor as well.