This study explored the changes in senescence patterns and vase life of cut roses grown in summer and autumn, aiming to identify the relationship between harvest seasons and flower longevity. We analyzed gene expression profiles associated with lignin, pectin, ethylene, auxin, and sucrose transport to understand the molecular mechanisms underlying senescence symptoms, such as the bent neck, petal abscission, and petal wilting in cut rose flowers. Our results revealed season-dependent occurrences of bent neck and petal abscission, with higher incidence rates in autumn-harvested rose flowers. These increases in bent neck and petal abscission contributed to a shortened vase life for the cut flowers. Gene expression analysis indicated that elevated levels of ethylene biosynthesis genes and reduced expression of lignin, pectin biosynthesis, auxin response factor, and sucrose transport genes accelerated the increased senescence symptoms. Notably, the incidence rates of the bent neck were highly negatively correlated with the transcript levels of key genes involved in lignin and pectin biosynthesis, RhPRXPX and RhGAUT1, in pedicels. These findings contribute to our understanding of the molecular factors influencing the mechanical strength of flower pedicels and provide insights for postharvest strategies to enhance the ornamental value of cut flowers across seasons.
Mesenchymal stem cells (MSCs) have been widely used as donor cells for somatic cell nuclear transfer (SCNT) to increase the efficiency of embryo cloning. Since replicative senescence reduces the efficiency of embryo cloning in MSCs during in vitro expansion, transfection of telomerase reverse transcriptase (TERT) into MSCs has been used to suppress the replicative senescence. Here, TERT-transfected MSCs in comparison with early passage MSCs (eMSCs) and sham-transfected MSCs (sMSCs) were used to evaluate the effects of embryo cloning with SCNT in a porcine model. Cloned embryos from tMSC, eMSC, and sMSC groups were indistinguishable in their fusion rate, cleavage rate, total cell number, and gene expression levels of OCT4, SOX2 and NANOG during the blastocyst stage. The blastocyst formation rates of tMSC and sMSC groups were comparable but significantly lower than that of the eMSC group (p < 0.05). In contrast, tMSC and eMSC groups demonstrated significantly reduced apoptotic incidence (p < 0.05), and decreased BAX but increased BCL2 expression in the blastocyst stage compared to the sMSC group (p < 0.05). Therefore, MSCs transfected with telomerase reverse transcriptase do not affect the overall development of the cloned embryos in porcine SCNT, but enables to maintain embryo quality, similar to apoptotic events in SCNT embryos typically achieved by an early passage MSC. This finding offers a bioengineering strategy in improving the porcine cloned embryo quality.
신체활동은 노화로 의한 인지기능 저하에 긍정적 영향을 미친다. 하지만 신체활동 참여가 정서인지기능에 미치는 영향을 과학적으로 밝혀 낸 연구는 드물다. 따라서 본 연구의 목적은 청년기와 노년기 성인의 정서인지 기능에 신체활동 참여가 어떠한 영향을 미치는지를 뇌 과학적으로 연구하는 것이다. 연구의 피험자는 D 광역시 성인 남성 60명으로 선정하였으며, 청년 운동 집단(25∼35살), 청년 비 운동 집단(26∼35살), 노년 운동 집단(60∼70살), 노년 비 운동 집단(60∼70살) 4개의 집단으로 구성하였다. 실험도구는 EEG 측정 장비와 국제정서사진체계(International Affective Picture System: IAPS)를 사용하였다. 본 연구의 실험과제는 사진으로 제시하는 세 가지 정서자극(긍정적, 중립, 부정적)에 적합한 버튼을 눌러 정서적인지 검사를 실시하였다. EEG 측정은 8개(Fp1, Fp2, Fz, C3, C4, Cz, T3, T4)의 영역을 측정하였다. 통계처리는 4(집단)×3(자극)×8(영역)에 대한 삼원분산분석을 실시하였다. 결과: 본 연구 결과, 반응시간과 반응정확률 그리고 P3 잠재기는 집단간 주효과가 나타났다. 그리고 P3 진폭은 집단과 자극간 상호작용이 나타났다. 결론: 신체활동은 청년층과 노년층의 정서적 인지기능에 긍정적인 영향을 미치는 것으로 나타났다.
Although anti-aging activities of melatonin, a hormone secreted by the pineal gland, have been reported in senescence-accelerated mouse models and several types of cells, its impact and mechanism on the senescence of human dental pulp cells (HDPCs) remains unknown. In this study, we examined the impact of melatonin on cellular premature senescence of HDPCs. Here, we found that melatonin markedly inhibited senescent characteristics of HDPCs after exposure to hydrogen peroxide (H2O2), including the increase in senescence-associated β-galactosidase (SA-β-gal)-positive HDPCs and the upregulation of p21 protein, an indicator for senescence. In addition, as melatonin attenuated H2O2-stimulated phosphorylation of c-Jun N-terminal kinase (JNK), while selective inhibition of JNK activity with SP600125 significantly attenuated H2O2-induced increase in SA-beta-gal activity. Results reveal that melatonin antagonizes premature senescence of HDPCs via JNK pathway. Thus, melatonin may have therapeutic potential to prevent stress-induced premature senescence, possibly correlated with development of dental pulp diseases, and to maintain oral health across the life span.
노화란 시간경과에 따라 생체기능의 불가역적인 저하로 치매, 뇌혈관질환, 고혈압, 암 등과 같은 노화관련 질병이 생겨나며, 세포들이 생명력을 잃어가는 일련의 현상이다. 노화로 인한 대표적인 질병은 퇴행성 뇌 질환에 의한 알츠하이머, 치매와 파킨슨병이 있다. 이 병들은 초기의 변화가 두드러지지 않아 조기 진단이 어렵고, 질병의 진행을 막는데 어려움이 있다. 이를 해결할 방법으로 유전자 지표를 개발하는 것이 중요한 과제라 할 수 있는데, 본 연구는 노화 동물모델인 SAMP1 마우스를 대조군인 SAMR1 마우스를 비교하여 노화의 초기단계에 차등 발현하는 유전자를 확인하여 향후 노화와 연관 지표를 발굴하기 위하여 실시되었다. 먼저, 성장단계인 8주령과 노화초기진행단계인 26주령의 뇌 조직에서의 차등발현 유전자를 발굴하고자 cDNA chip 분석을 수행하였고, 신뢰성을 가지고 생물학적 변이를 검출할 수 있는 유전자는 13,939개의 유전자를 발굴하였다. 이들 중 발현이 2-fold 이상 증감하는 유전자들을 조사한 결과, 2,029개의 유전자를 선별할 수 있었다. 또한, 기능별로 분류한 결과 노화가 진행되는 과정에서 총 증가 혹은 감소한 유전자의 개수는 660개였으며, 이 중 노화와 연관성이 높은 유전자는 74개였다. 26주령에 증가양상을 보인 것이 46개, 감소양상을 보인 유전자가 28개로 밝혀졌다. 이를 검증하기 위해 노화와 관련된 유용한 유전자인 Wnt7a, Prkcb, Dgke가 선별되어 real-time qPCR을 수행하였다. 그 결과 Wnt7a 유전자는 노화가 진행됨에 따라 발현양이 급격히 감소하여 노화 초기 단계의 지표로 활용이 가능할 것으로 사료된다. 반면에 Prkcb 유전자는 성장단계의 발현량에는 관계없이 노화초기단계에서 억제됨을 확인하였고, Dgke는 성장단계부터 유전자 발현이 억제되어 노화에 영향을 미칠 것으로 판단된다.
Lentinula edodes (shiitake mushroom) is a very popular edible, cultivated mushroom in Japan. There are post-storage problems with shiitake mushrooms, such as gill browning and cell wall lysis of the fruiting body, which can result in loss of fresh food quality and consequent loss of value. Lentinan is a cell wall component of beta-1, 3-linked-D-glucan with beta-1, 6 branches, which was isolated as an anti-tumor active-substance from L. edodes. Lentinan content decreases following harvest as a result of increased glucanase activity. We isolated one exo-glucanase encoding genes, exg21) and two endo-glucanase encoding gene tlg12) and glu1 from L. edodes. Transcription level of the exg2, tlg1 and glu1 gene increased after harvesting. Enzymes encoded by the genes have lentinan degrading activity, therefore, these genes are involved in lentinan degradation after harvesting. We also identified several cell wall degradation- related enzyme-encoding genes3), such as mixed-linked glucanase (mlg1), chitinases (chi1, chi29), chitin deacetylase (chd1), and chitosanase (cho1). It is revealed that transcriptional levels of these genes increased after harvesting, by real-time PCR. Glucanase and chitinase activity increased following harvest as results of increased transcription of these cell wall degradation-related enzyme-encoding genes. Increase of these cell wall degradation- related enzyme activities would cause cell wall lysis and lentinan degradation during post-harvest preservation. We identified laccase and tyrosinase encoding genes (lcc4 and tyr, respectively) by PCR-subtraction. The lcc4 was a novel laccase-encoding gene in L. edodes. Transcription levels of lcc4 and tyr increased after harvesting, and these genes would be involved in browning of the fruiting body. 1) Sakamoto et al. (2005) Current Genetics, 48: 195-203 2) Sakamoto et al. (2006) Plant Physiology 141: 793-801 3) Sakamoto et al. (2009) Current Genetics 55: 409-423
Tooth loss in elderly is mainly caused by alveolar bone loss via severe periodontitis. Although the severity of periodontitis is known to be affected by age, the aging process or the genetic changes during the aging of periodontal tissue cells are not well characterized. In this study, we investigated the effect of in vitro aging on the change of gene expression pattern in periodontal fibroblasts. Gingival fibroblasts (GF) and periodontal ligament fibroblasts (PDL) were obtained from two young patients and replicative senescence was induced by sequential subcultivation. When more than 90% cells were positively stained with senescence-associated β-galactosidase, those cells were regarded as aged cells. In aged GF and PDL, the level of phosphorylated retinoblastoma (RB) and p16INK4A protein was significantly decreased and increased, respectively. However, the protein level of p53 and p21, well known senescence-inducing genes, did not increase in aged GF and PDL. Although P27Kip1 and p15INK4B, another cyclin-dependent kinase inhibitors, were reported to be involved in replicative senescence of human cells, they were decreased in aged GF and PDL. Because senescent cells showed flattened and enlarged cell shape and are known to have increased focal adhesion, we examined the protein level of several integrins. Aged GF and PDL showed increased protein level of integrin α2, αu, and β1. When the gene expression profiles of actively proliferating young cells and aged cells were compared by cDNA microarray of 3,063 genes and were confirmed by reverse transcription-polymerase chain reaction, 7 genes and 15 genes were significantly and commonly increased and decreased, respectively, in aged GF and PDL. Among them, included are the genes that were known to be involved in the regulation of cell cycle, gene transcription, or integrin signaling. The change of gene expression pattern in GF and PDL was minimally similar to that of oral keratinocyte. These results suggest that p16INK4A/RB might be involved in replicative senescence of periodontal fibroblasts and the change of gene expression profile during aging process is cell type specific.
토양 미생물 A. tumefaciens로부터 cytokinin 합성효소 (ipt) 유전자를 분리하여 형질전환 식물을 작성하였다. 도입된 ipt 유전자는 모든 조직에서 발현되었으며, 잎에서의 발현량이 서로 다른 3개의 형질전환 식물체를 선발하였다. 선발한 식물체는 도입유전자의 고정을 위하여 2회 자가수정을 거쳐 homozygous 식물체를 작성한 후 해석하였다. 형질전환 식물체는 노화가 지연되어 녹엽을 유지하였으며, 측아로부터 측지가 분얼하여 그 수가 증가하였을 뿐만아니라, 각 마디에서 복수의 본엽이 생성되었다. 식물체의 노화가 가장 많이 진행된 잎에서의 chlorophyll 함량은 형질전환 식물체에서 1.5~4배 정도로 wild-type에 비하여 월등히 높았다. 이러한 결과는 축적된 cytokinin이 식물의 노화를 지연시킬 뿐만 아니라, 분얼 및 잎수를 증가시켜 식물의 수량을 증가시켰음을 나타낸다.