Dynamics of plant DNA replication based on PCNA visualization

Yokoyama, Ryohei; Hirakawa, Takeshi; Hayashi, Seri; Sakamoto, Takuya; Matsunaga, Sachihiro

doi:10.1038/srep29657

Cited by 39 publications

(51 citation statements)

References 29 publications

(43 reference statements)

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“…In the cell cycle, the synthesis of PCNA is important for the entry of cells into the S phase from the G0 and G1 phases. PCNA expression has been demonstrated to be increased in the late stages of cell entry into the G1 phase, reaches its peak in the S phase, and decreases in the G2 and M phases (23). The results of the present study demonstrated that expression of PCNA was significantly increased in the surgery group compared with the normal and the medical adhesive spray groups, indicating that the medical adhesive inhibited cell proliferation following surgery.…”

Section: Discussionsupporting

confidence: 48%

Application of medical adhesive inhibits intimal hyperplasia involving the downregulation of ERK1/2 and eNOS levels

Chai

et al. 2018

Mol Med Report

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Vein graft remains the most broadly applied vascular material in coronary artery bypass surgery. However, the restenosis rate of the vein bridge following angioplasty is high. The present study investigated the effect of medical adhesive on vascular intimal hyperplasia, in addition to the signal transduction mechanism. A total of 36 New Zealand white rabbits were divided into three groups at random, including the normal group, the surgery group and the medical adhesive spray group. Following surgery for transplantation of the left external jugular vein to the ipsilateral common carotid artery for 4 weeks, the thickness and area of the intima and media of the vessel were measured on formalin‑fixed, paraffin wax‑embedded pathological sections using hematoxylin‑eosin staining, and alterations in the expression of proliferating cell nuclear antigen (PCNA), platelet endothelial cell adhesion molecule 1 (PECAM‑1), vascular cell adhesion protein 1 (VCAM‑1), extracellular signal‑regulated kinase (ERK)1/2, and endothelial nitric oxide synthase (eNOS) were detected by immunohistochemical staining, reverse transcription‑quantitative polymerase chain reaction analysis and western blotting. The levels of intimal hyperplasia in the medical adhesive spray group were markedly decreased compared with the surgery group. Consistently, PCNA, PECAM‑1 and VCAM‑1 were underexpressed in the medical adhesive spray group compared with the surgery group. ERK1/2 and eNOS were underexpressed in the medical adhesive spray group compared with the surgery group. Therefore, the application of medical adhesive may inhibit intimal hyperplasia, which may be associated with the restriction of the over‑distension of the vein graft by downregulating the ERK1/2 and eNOS levels, reducing injury to the vascular intima and inhibiting the signaling pathway involved in intimal hyperplasia.

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Section: Discussionsupporting

confidence: 48%

Application of medical adhesive inhibits intimal hyperplasia involving the downregulation of ERK1/2 and eNOS levels

Chai

et al. 2018

Mol Med Report

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“…To monitor the progression of the cell cycle in living cells, we used transgenic lines expressing RAD54‐EYFP and PCNA1‐tdTomato (Figure c). Arabidopsis PCNA1 shows distinct subcellular localization patterns, which are whole, dotted or speckled indicating G 1 or G 2 phase, early S phase and late S phase during the cell cycle, respectively (Yokoyama et al ., ). The whole pattern is defined by the uniform distribution of PCNA1 in the nucleoplasm.…”

Section: Resultsmentioning

confidence: 97%

RAD54 forms DNA repair foci in response to DNA damage in living plant cells

et al. 2017

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Plants have various defense mechanisms against environmental stresses that induce DNA damage. Genetic and biochemical analyses have revealed the sensing and signaling of DNA damage, but little is known about subnuclear dynamics in response to DNA damage in living plant cells. Here, we observed that the chromatin remodeling factor RAD54, which is involved in DNA repair via the homologous recombination pathway, formed subnuclear foci (termed RAD54 foci) in Arabidopsis thaliana after induction of DNA double-strand breaks. The appearance of RAD54 foci was dependent on the ATAXIA-TELANGIECTASIA MUTATED-SUPPRESSOR OF GAMMA RESPONSE 1 pathway, and RAD54 foci were co-localized with γH2AX signals. Laser irradiation of a subnuclear area demonstrated that in living cells RAD54 was specifically accumulated at the damaged site. In addition, the formation of RAD54 foci showed specificity for cell type and region. We conclude that RAD54 foci correspond to DNA repair foci in A. thaliana.

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“…Cells in early S-phase, characterized by small PCNA dots, could not be clearly distinguished under our imaging conditions. Nevertheless, the duration of the speckle-stage is comparable to mitotic cells 43 , indicating that the pre-meiotic S-phase is not substantially longer than the S-phase of mitotic cells. This is in contrast to observations in other organisms, where pre-meiotic S-phase was reported to be at least twice as long as mitotic S-phase 44,45 .…”

Section: Use Of Live Imaging In Gene Function Studiesmentioning

confidence: 87%

“…S-phase in Arabidopsis can be monitored by proliferating cell nuclear antigen (PCNA). PCNA exhibits a disperse nuclear localization throughout the cell cycle but forms nuclear foci ranging from small dots to large nuclear speckles during S-phase 43 . Live imaging of pre-meiotic cells expressing ASY1:eYFP and PCNA:tagRFP markers showed a reorganization of PCNA from a diffused signal to nuclear speckles, which were detectable for approximately 90 min ( Fig.…”

Section: Use Of Live Imaging In Gene Function Studiesmentioning

confidence: 99%

Imaging plant germline differentiation within Arabidopsis flower by light sheet microscopy

Valuchova

Mikulková

Pecinkova

et al. 2019

Preprint

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In higher plants, germline differentiation occurs during a relatively short period within developing flowers. Understanding of the mechanisms that govern germline differentiation lags behind other plant developmental processes. This is largely because the germline is restricted to relatively few cells buried deep within floral tissues, which makes them difficult to study. To overcome this limitation, we have developed a methodology for live imaging of the germ cell lineage within floral organs of Arabidopsis using light sheet fluorescence microscopy. We have established reporter lines, cultivation conditions, and imaging protocols for high-resolution microscopy of developing flowers continuously for up to several days. We used multiview imagining to reconstruct a three-dimensional model of a flower at subcellular resolution. We demonstrate the power of this approach by capturing male and female meiosis, asymmetric pollen division, movement of meiotic chromosomes, and unusual restitution mitosis in tapetum cells. This method will enable new avenues of research into plant sexual reproduction.

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Dynamics of plant DNA replication based on PCNA visualization

Cited by 39 publications

References 29 publications

Application of medical adhesive inhibits intimal hyperplasia involving the downregulation of ERK1/2 and eNOS levels

Application of medical adhesive inhibits intimal hyperplasia involving the downregulation of ERK1/2 and eNOS levels

RAD54 forms DNA repair foci in response to DNA damage in living plant cells

Imaging plant germline differentiation within Arabidopsis flower by light sheet microscopy

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