Cardiac cell proliferation assessed by EdU, a novel analysis of cardiac regeneration

Zeng, Bin; Tong, Shuping; Ren, Xiaofeng; Xia, Hao

doi:10.1007/s10616-014-9827-8

Cited by 11 publications

(7 citation statements)

References 32 publications

(35 reference statements)

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“…Hence, this method can be used to quantify proliferation in cultured cells as well as organs of whole animals [ 1 ]. For these reasons, the use of EdU in animals is growing and has been reported in experimental models of neoplastic diseases [ 4 – 11 ], cardiovascular and metabolic diseases [ 12 – 15 ], and neuroimmunologic development [ 13 , 15 , 16 ].…”

Section: Introductionmentioning

confidence: 99%

A Simple Method to AssessIn VivoProliferation in Lung Vasculature with EdU: The Case of MMC-Induced PVOD in Rat

Antigny

Ranchoux

Nadeau

et al. 2015

Analytical Cellular Pathology

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5-Ethynyl-2′-deoxyuridine (EdU) incorporation is becoming the gold standard method for in vitro and in vivo visualization of proliferating cells. The small size of the fluorescent azides used for detection results in a high degree of specimen penetration. It can be used to easily detect DNA replication in large tissue samples or organ explants with low proliferation and turnover of cells formerly believed to be in a “terminal” state of differentiation. Here we describe a protocol for the localization and identification of proliferating cells in quiescent or injured pulmonary vasculature, in a model of pulmonary veno-occlusive disease (PVOD). PVOD is an uncommon form of pulmonary hypertension characterized by progressive obstruction of small pulmonary veins. We previously reported that mitomycin-C (MMC) therapy is associated with PVOD in human. We demonstrated that MMC can induce PVOD in rats, which currently represents the sole animal model that recapitulates human PVOD lesions. Using the EdU assay, we demonstrated that MMC-exposed lungs displayed areas of exuberant microvascular endothelial cell proliferation which mimics pulmonary capillary hemangiomatosis, one of the pathologic hallmarks of human PVOD. In vivo pulmonary cell proliferation measurement represents an interesting methodology to investigate the potential efficacy of therapies aimed at normalizing pathologic angioproliferation.

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Section: Introductionmentioning

confidence: 99%

A Simple Method to AssessIn VivoProliferation in Lung Vasculature with EdU: The Case of MMC-Induced PVOD in Rat

Antigny

Ranchoux

Nadeau

et al. 2015

Analytical Cellular Pathology

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“…After P1 surgery, a subset of neonatal rats in the sham (n = 4) and MI groups (n = 5) received EdU on postoperative days 1, 7, and 14, followed by heart explant at 3 weeks post-surgery. EdU, a thymidine analog and marker of DNA replication during cell proliferation [34], was found to be incorporated into the DAPI-stained nuclei of troponin + cardiomyocytes at a significantly greater density in P1 MI hearts ( Figure 5B,C) compared to P1 sham hearts ( Figure 5A) in both the peri-infarct region (4.6 ± 1.3 cells per ROI vs. 14.1 ± 4.9 cells per ROI, p = 0.0072) and remote uninjured region of the LV (2.8 ± 1.5 cells per ROI vs. 13.9 ± 4.4 cells per ROI, p = 0.0021, Figure 5D). In order to more directly assess cell cycle activation, a subset of neonatal rats in the P1 sham (n = 3) and P1 MI groups (n = 3) were sacrificed at 1 week after surgery for analysis of Ki-67 staining.…”

Section: Increased Cardiomyocyte Proliferation Is Observed After Myocmentioning

confidence: 99%

Natural Heart Regeneration in a Neonatal Rat Myocardial Infarction Model

Wang

Paulsen

Hironaka

et al. 2020

Cells

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Newborn mice and piglets exhibit natural heart regeneration after myocardial infarction (MI). Discovering other mammals with this ability would provide evidence that neonatal cardiac regeneration after MI may be a conserved phenotype, which if activated in adults could open new options for treating ischemic cardiomyopathy in humans. Here, we hypothesized that newborn rats undergo natural heart regeneration after MI. Using a neonatal rat MI model, we performed left anterior descending coronary artery ligation or sham surgery in one-day-old rats under hypothermic circulatory arrest (n = 74). Operative survival was 97.3%. At 1 day post-surgery, rats in the MI group exhibited significantly reduced ejection fraction (EF) compared to shams (87.1% vs. 53.0%, p < 0.0001). At 3 weeks post-surgery, rats in the sham and MI groups demonstrated no difference in EF (71.1% vs. 69.2%, respectively, p = 0.2511), left ventricular wall thickness (p = 0.9458), or chamber diameter (p = 0.7801). Masson’s trichome and picrosirius red staining revealed minimal collagen scar after MI. Increased numbers of cardiomyocytes positive for 5-ethynyl-2′-deoxyuridine (p = 0.0072), Ki-67 (p = 0.0340), and aurora B kinase (p = 0.0430) were observed within the peri-infarct region after MI, indicating ischemia-induced cardiomyocyte proliferation. Overall, we present a neonatal rat MI model and demonstrate that newborn rats are capable of endogenous neocardiomyogenesis after MI.

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“…EdU is a nucleoside analogue of thymidine and is incorporated into DNA during only DNA synthesis, thus enabling the visualization of newly synthesized DNA [ 25 ]. This technique is a less toxic alternative to a BrdU incorporation assay.…”

Section: Methodsmentioning

confidence: 99%

A Loss of Nuclear—Cytoskeletal Interactions in Vascular Smooth Muscle Cell Differentiation Induced by a Micro-Grooved Collagen Substrate Enabling the Modeling of an In Vivo Cell Arrangement

Nagayama

2021

Bioengineering

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Vascular smooth muscle cells (VSMCs) remodel vascular walls actively owing to mechanical cues and dedifferentiate to the synthetic phenotype from contractile phenotype in pathological conditions. It is crucial to clarify the mechanisms behind the VSMC phenotypic transition for elucidating their role in the vascular adaptation and repair and for designing engineered tissues. We recently developed novel micro-grooved collagen substrates with “wavy wrinkle” grooves to induce cell–substrate adhesion, morphological polarization, and a tissue-like cell arrangement with cytoskeletal rearrangements similar to those in vascular tissue in vivo. We found that cultivation with this micro-grooved collagen significantly induced VSMC contractile differentiation. Nonetheless, the detailed mechanism underlying the promotion of such VSMC differentiation by micro-grooved collagen has not been clarified yet. Here, we investigated the detailed mechanism of the cell arrangement into a tissue and contractile-differentiation improvement by our micro-grooved collagen substrates in terms of nuclear–cytoskeletal interactions that possibly affect the nuclear mechanotransduction involved in the activation of transcription factors. We found that VSMCs on micro-grooved collagen manifested significant cell arrangement into a tissue and nucleus slimming with a volume reduction in response to the remodeling of the actin cytoskeleton, with consequent inhibition of nuclear shuttling of a transcriptional coactivator, Yes-associated protein (YAP), and improved contractile differentiation. Furthermore, VSMC nuclei rarely deformed during macroscopic cell stretching and featured a loss of nesprin-1–mediated nuclear–cytoskeletal interactions. These results indicate that our micro-grooved collagen induces a cell alignment mimicking in vivo VSMC tissue and promotes contractile differentiation. In such processes of contractile differentiation, mechanical interaction between the nucleus and actin cytoskeleton may diminish to prevent a nuclear disturbance from the excess mechanical stress that might be essential for maintaining vascular functions.

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Cardiac cell proliferation assessed by EdU, a novel analysis of cardiac regeneration

Cited by 11 publications

References 32 publications

A Simple Method to AssessIn VivoProliferation in Lung Vasculature with EdU: The Case of MMC-Induced PVOD in Rat

A Simple Method to AssessIn VivoProliferation in Lung Vasculature with EdU: The Case of MMC-Induced PVOD in Rat

Natural Heart Regeneration in a Neonatal Rat Myocardial Infarction Model

A Loss of Nuclear—Cytoskeletal Interactions in Vascular Smooth Muscle Cell Differentiation Induced by a Micro-Grooved Collagen Substrate Enabling the Modeling of an In Vivo Cell Arrangement

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