• Title/Summary/Keyword: Histochemical analysis

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Antagonistic Regulation of Arabidopsis Growth by Brassinosteroids and Abiotic Stresses

  • Chung, Yuhee;Kwon, Soon Il;Choe, Sunghwa
    • Molecules and Cells
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    • v.37 no.11
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    • pp.795-803
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    • 2014
  • To withstand ever-changing environmental stresses, plants are equipped with phytohormone-mediated stress resistance mechanisms. Salt stress triggers abscisic acid (ABA) signaling, which enhances stress tolerance at the expense of growth. ABA is thought to inhibit the action of growth-promoting hormones, including brassinosteroids (BRs). However, the regulatory mechanisms that coordinate ABA and BR activity remain to be discovered. We noticed that ABA-treated seedlings exhibited small, round leaves and short roots, a phenotype that is characteristic of the BR signaling mutant, brassinosteroid insensitive1-9 (bri1-9). To identify genes that are antagonistically regulated by ABA and BRs, we examined published Arabidopsis microarray data sets. Of the list of genes identified, those upregulated by ABA but downregulated by BRs were enriched with a BRRE motif in their promoter sequences. After validating the microarray data using quantitative RT-PCR, we focused on RD26, which is induced by salt stress. Histochemical analysis of transgenic Arabidopsis plants expressing RD26pro:GUS revealed that the induction of GUS expression after NaCl treatment was suppressed by co-treatment with BRs, but enhanced by co-treatment with propiconazole, a BR biosynthetic inhibitor. Similarly, treatment with bikinin, an inhibitor of BIN2 kinase, not only inhibited RD26 expression, but also reduced the survival rate of the plant following exposure to salt stress. Our results suggest that ABA and BRs act antagonistically on their target genes at or after the BIN2 step in BR signaling pathways, and suggest a mechanism by which plants fine-tune their growth, particularly when stress responses and growth compete for resources.

Effects of Isokinetic Exercise on the Composition of Muscle Fibers in Rats Lower Limb (등속성 운동이 흰쥐 하퇴의 근섬유조성에 미치는 영향)

  • Lee, Yong-Deok;Cho, Hyun-Gug;Kim, Han-Soo;Park, Youn-Ki;Suh, Tae-Soo
    • The Journal of Korean Physical Therapy
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    • v.3 no.1
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    • pp.75-90
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    • 1991
  • This study is to characterize the effects of swimming exercise on the composition of muscle fibers in rats lower limb tissue. By comparing the exercised group (4, 8 weeks) with the control group, we analyzed the difference histologically, histochemically and morphometrically. The obtained results are as follows ; 1. In the histological view, the distance between the individual muscle fibers and the muscle bundles in exercised group was shorter than that of control group. The irregular shape of sarcolemma in exercised group was observed. 2, In the histochemical view(NADH-TR. and Myosin ATPase reaction analysis), the increased populations of red and intermediate fibers were observed, whereas the white fiber was decreased. 3. In the morphometrical view, the red and intermediate fibers of tibialis anterior and gastrocnemius in exercised group were quantitatively increased, compared with control group. However, the white fiber and connective tissue in exercised group were decreased.

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Genetic Transformation and Plant Regeneration of Codonopsis lanceolata Using Agrobacterium (Agrobacterium에 의한 더덕의 형질전환과 식물체 재분화)

  • 최필선;김윤성;유장렬;소웅영
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.5
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    • pp.315-318
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    • 1994
  • To obtain transformed plants, we cocultured cotyledonary explants of Codonopsis lanceolata with Agrobacterium tumefaciens LBA4404, a disamed strain harboring a binary vector pBI121 carrying the CaMV35S promoter-$\beta$-glucuronidase (GUS) gene fusion used as a reporter gene and NOS promoter-neomycin phosphotransferase gene as a positive selection marker in MS liquid medium with 1mg/L BA. After 48 h of culture, explants were transferred onto MS solid medium with Img/L BA, 250mg/L carbenicillin, and 100mg/L kanamycin sulfate and cultured in the dark. Numerous adventitious buds formed on the cut edges of the explants after 2 weeks of culture. When subjected to GUS histochemical assay buds showed a positive response at a frequency of 15%. Explants formed adventitious shoot at a frequency of 56.7%, after 6 weeks of culture. Upon transfer onto the basal medium, most of the shoots were rooted and subsequently the regenerants were transplanted to potting soil. Southern blot analysis confirmed that the GUS gene was incorporated into the genomic DNA of the GUS-positive regenerants.

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Induction of Transgenic Shoots in Tomato (토마토의 형질전환 신초 유기)

  • Ryu, Soo-Kyung;Park, Young-Doo;Sul, Ill-Whan;Choi, Geun-won
    • Horticultural Science & Technology
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    • v.16 no.2
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    • pp.213-214
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    • 1998
  • This study was conducted to provide useful information for improvement on the efficency of transformation mediated by Agrobacterium tumefaciens. The result from the sensitivity test of cotyledon explants of tomato to kanamycin suggested that 50mg/L could be a proper concentration for selection media. Two hundred mg/L of cefotaxime was selected as a proper concentration to remove Agrobacteria from media without any negative effect on explants. Both callus formation and shoot regeneration from cotyledon explants of tomato were significantly suppressed by the cocultivation with Agrobacterium. Three days of cocultivation was effective on callus formation and shoot regeneration in all of tomato cultivars tested. Confirmation of transformation for regenerated shoots was carried out by histochemical GUS assay and PCR analysis using NPTII primer, and transgenic shoots were obtained from all of 3 tomato cultivars tested.

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Differential Expression of Adipocyte-Related Molecules in the Distal Epididymal Fat of Mouse during Postnatal Period

  • Lee, Ki-Ho;Kim, Nan Hee
    • Development and Reproduction
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    • v.23 no.3
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    • pp.213-221
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    • 2019
  • The epididymal fat of mouse is a part of visceral fat deposit and is divided into the distal or proximal part based on its histochemical characteristics. Even though the formation of the epididymal fat pad begins before the birth, a detailed adipogenic procedure of the epididymal fat has not been revealed. The epididymal fat pad becomes enlarged and expended with age, and expressional changes of numerous genes are associated with the maturation of fat tissues. In the present research, expressional patterns of adipose tissue-related genes in the distal epididymal fat of mouse at 2, 5, 8, and 12 months of postnatal age were determined by a quantitative real-time polymerase chain reaction (PCR) analysis. The lowest transcript levels of fatty acid binding protein 4 (Fabp4), lipoprotein lipase (Lpl), delta like non-canonical Notch ligand 1 (Dlk1), peroxisome proliferator-activated receptor gamma (Pparg), leptin (Lep), adiponectin (Adipoq), and resistin (Retn) were detected at 2 months of age, except fatty acid synthase (Fasn) showing the lowest level at 5 months of age. Even though expression of Lep and Fabp4 were gradually increased until 12 months of age, significant increases of Pparg and Adipoq transcript levels were continued until 8 months of age. The transcript levels of Lpl, Rent, Dlk1, and Fasn were significantly increased at 8 months of age, compared with those at 2 months of age. The current findings suggest that the expansion of the distal epididymal fat of mouse during postnatal period would be companied with differential expression of various adipocyte-associated molecules.

Hydrogen Peroxide Prompted Lignification Affects Pathogenicity of Hemi-biotrophic Pathogen Bipolaris sorokiniana to Wheat

  • Poudel, Ajit;Navathe, Sudhir;Chand, Ramesh;Mishra, Vinod K.;Singh, Pawan K.;Joshi, Arun K.
    • The Plant Pathology Journal
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    • v.35 no.4
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    • pp.287-300
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    • 2019
  • Spot blotch caused by Bipolaris sorokiniana has spread to more than 9 million ha of wheat in the warm, humid areas of the Eastern Gangetic Plains (EGP) of South Asia and is a disease of major concern in other similar wheat growing regions worldwide. Differential lignin content in resistant and susceptible genotypes and its association with free radicals such as hydrogen peroxide ($H_2O_2$), superoxide ($O_2{^-}$) and hydroxyl radical ($OH^-$) were studied after inoculation under field conditions for two consecutive years. $H_2O_2$ significantly influenced lignin content in flag leaves, whereas there was a negative correlation among lignin and $H_2O_2$ to the Area Under Disease Progress Curve (AUDPC). The production of $H_2O_2$ was higher in the resistant genotypes than susceptible ones. The $O_2{^-}$ and $OH^-$ positively correlated with AUDPC but negatively with lignin content. This study illustrates that $H_2O_2$ has a vital role in prompting lignification and thereby resistance to spot blotch in wheat. We used cluster analysis to separate the resistant and susceptible genotypes by phenotypic and biochemical traits. $H_2O_2$ associated lignin production significantly reduced the number of appressoria and penetration pegs. We visualized the effect of lignin in disease resistance using differential histochemical staining of tissue from resistant and susceptible genotypes, which shows the variable accumulation of hydrogen peroxide and lignin around penetration sites.

The relationship between myofiber characteristics and meat quality of Chinese Qinchuan and Luxi cattle

  • Lu, Xiao;Yang, Yuying;Zhang, Yimin;Mao, Yanwei;Liang, Rongrong;Zhu, Lixian;Luo, Xin
    • Animal Bioscience
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    • v.34 no.4
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    • pp.743-750
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    • 2021
  • Objective: The objectives of this study were to explore the expression patterns of myosin heavy chain (MyHC) genes of different skeletal muscles from Chinese cattle, and to investigate the relationship between myofiber characteristics and meat quality of M. longissimus lumborum (LL), M. psoas major (PM), and M. semimembranosus (SM) from Chinese Luxi and Qinchuan cattle. Methods: Three major muscles including LL, PM, and SM from Chinese Luxi cattle and Chinese Qinchuan cattle were used in this study. The myofiber characteristics were measured by histochemical analysis. The MyHC isoforms expression was evaluated by real-time quantitative polymerase chain reaction. Quality traits including pH value, meat color, cooking loss, Warner-Bratzler shear force (WBSF) and sarcomere length were determined at day 5 postmortem. Results: PM muscle had higher pH value, a* value, sarcomere length and lower WBSF value compared to LL and SM muscles (p<0.05). Numbers of type I myofiber and the relative expression of MyHC I mRNA in PM muscle were higher than those of LL and SM muscles (p<0.05). Myofiber diameter of PM muscle was lower than that of LL and SM muscles, regardless of myofiber types (p<0.05). Conclusion: According to the stepwise linear regression analyses, tenderness was influenced by myofiber characteristics in all three examined muscles. Tenderness of beef muscles from Qinchuan and Luxi cattle could be improved by increasing numbers of type I myofiber.

Influence of freeze-thawed cycles on pork quality

  • Tippala, Tiprawee;Koomkrong, Nunyarat;Kayan, Autchara
    • Animal Bioscience
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    • v.34 no.8
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    • pp.1375-1381
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    • 2021
  • Objective: This study was conducted to determine the effect of freeze-thawed cycles (Fresh meat, F-T 1 cycle and F-T 2 cycles) on the quality characteristics of porcine longissimus dorsi muscle. Methods: A total of 20 three-crossbred pigs (Duroc×[Large White×Landrace]) were randomly obtained from a commercial slaughterhouse in Thailand. Muscle samples were immediately taken from 10 to 11th of the longissimus dorsi for histochemical analysis. The muscles were cut into 2.54 cm-thick chops. A minimum of 20 chops were used for each treatment (fresh meat, freeze-thawed 1 and 2 cycles). Individually chops were packaged in polyethylene bags and frozen at -20℃ for 6 months followed by thawing in refrigerator at 4℃ for 24 h (the 1st freeze-thawed cycle). The freeze-thawed procedure was repeated for two cycles (the 2nd freeze-thawed cycle). Thawing loss, shear force value, citrate synthase activity and muscle fiber characteristics were determined on the muscles. Results: Results showed that increasing of freeze-thawed cycle increased the thawing loss (p<0.01) and citrate synthase activity (p<0.001). Shear force value of fresh meat was higher than freeze-thawed 1 and 2 cycles (F-T 1 cycle and F-T 2 cycles). Freeze-thawed cycles affected muscle characteristics. Muscle fiber area and muscle fiber diameter decreased with an increasing number of freeze-thawed cycles (p<0.001), while the thickness of endomysium and perimysium were increased (p<0.001). Conclusion: Repeated freeze-thawed cycles degraded muscle fiber structure and deteriorated pork quality.

Effects of Calcium, Vitamin D and Egg Yolk Peptide Treatment on the Retardation of Longitudinal Bone Growth Induced by Low-Calcium Diets (저칼슘 식이로 유발한 성장 저하 흰쥐에 대한 칼슘, 비타민 D 및 난황 펩타이드의 투여가 장골 길이 성장에 미치는 영향)

  • Kim, Myung-Gyou;Kim, Hye Kyung;Leem, Kang-Hyun
    • The Korea Journal of Herbology
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    • v.29 no.5
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    • pp.31-38
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    • 2014
  • Objectives : Egg yolk is composed of various important chemical substances for human health. A calcium shortage causes the growth retardation on the body growth. In this study, we examined the therapeutic effects of calcium, vitamin D and egg yolk peptide (EYP) treatment on the retardation of the longitudinal bone growth induced by low-calcium diet in adolescent rats. Methods : Low calcium diets were administrated for 15 days. During the last five days, calcium and/or vitamin D and/or EYP were administrated. The body weights, longitudinal bone growth rates, the heights of growth plates, and bone morphogenetic protein (BMP)-2 and insulin-like growth factor (IGF)-1 expressions were measured using histochemical analysis. Results : Low calcium diets caused the significant reduction in body weight gains and the longitudinal bone growth. The heights of growth plates and the expressions of BMP-2 and IGF-1 showed the impairment of body growth as well. Calcium and/or vitamin D administration could not significantly increase the longitudinal bone growth. However, calcium, vitamin D, and EYP administration significantly increased the bone growth, the growth plate height, and BMP-2 and IGF-1 expressions. Conclusions : These results suggest that EYP enhances the longitudinal bone growth in the calcium and/or vitamin D deficiency and it could be a promising agent for the treatment of children suffering from malnutrition.

Testicular fat deposition attenuates reproductive performance via decreased follicle-stimulating hormone level and sperm meiosis and testosterone synthesis in mouse

  • Miao Du;Shikun Chen;Yang Chen;Xinxu Yuan;Huansheng Dong
    • Animal Bioscience
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    • v.37 no.1
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    • pp.50-60
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    • 2024
  • Objective: Testicular fat deposition has been reported to affect animal reproduction. However, the underlying mechanism remains poorly understood. The present study explored whether sperm meiosis and testosterone synthesis contribute to mouse testicular fat deposition-induced reproductive performance. Methods: High fat diet (HFD)-induced obesity CD1 mice (DIO) were used as a testicular fat deposition model. The serum hormone test was performed by agent kit. The quality of sperm was assessed using a Sperm Class Analyzer. Testicular tissue morphology was analyzed by histochemical methods. The expression of spermatocyte marker molecules was monitored by an immuno-fluorescence microscope during meiosis. Analysis of the synthesis of testosterone was performed by real-time polymerase chain reaction and reagent kit. Results: It was found that there was a significant increase in body weight among DIO mice, however, the food intake showed no difference compared to control mice fed a normal diet (CTR). The number of offspring in DIO mice decreased, but there was no significant difference from the CTR group. The levels of follicle-stimulating hormone were lower in DIO mice and their luteinizing hormone levels were similar. The results showed a remarkable decrease in sperm density and motility among DIO mice. We also found that fat accumulation affected the meiosis process, mainly reflected in the cross-exchange of homologous chromosomes. In addition, overweight increased fat deposition in the testis and reduced the expression of testosterone synthesis-related enzymes, thereby affecting the synthesis and secretion of testosterone by testicular Leydig cells. Conclusion: Fat accumulation in the testes causes testicular cell dysfunction, which affects testosterone hormone synthesis and ultimately affects sperm formation.