scholarly journals In vivo time-lapse imaging of cell proliferation and differentiation in the optic tectum of Xenopus laevis tadpoles

2011 ◽  
Vol 520 (2) ◽  
pp. 401-433 ◽  
Author(s):  
Jennifer E. Bestman ◽  
Jane Lee-Osbourne ◽  
Hollis T. Cline
Keyword(s):  
Cell Proliferation ◽  
Xenopus Laevis ◽  
Optic Tectum ◽  
Time Lapse ◽  
Cell Proliferation And Differentiation ◽  
Proliferation And Differentiation ◽  
Time Lapse Imaging

The Cdx-1 and Cdx-2 homeobox genes in the intestine

1998 ◽  
Vol 76 (6) ◽  
pp. 957-969 ◽  
Author(s):  
Jean-Noël Freund ◽  
Claire Domon-Dell ◽  
Michèle Kedinger ◽  
Isabelle Duluc
Keyword(s):  
Cell Proliferation ◽  
Ex Vivo ◽  
Homeobox Genes ◽  
Intestinal Cell ◽  
Primary Role ◽  
Colon Tumorigenesis ◽  
Cell Proliferation And Differentiation ◽  
Proliferation And Differentiation

The past years have witnessed an increasing number of reports relative to homeobox genes in endoderm-derived tissues. In this review, we focus on the caudal-related Cdx-1 and Cdx-2 homeobox genes to give an overview of the in vivo, in vitro, and ex vivo approaches that emphasize their primary role in intestinal development and in the control of intestinal cell proliferation, differentiation, and identity. The participation of these genes in colon tumorigenesis and their identification as important actors of the oncogenic process are also discussed.Key words: caudal, epithelial cell proliferation and differentiation, cancer.

Depletion in nuclear spermine during human spermatogenesis, a natural process of cell differentiation

1992 ◽  
Vol 263 (2) ◽  
pp. C343-C347 ◽  
Author(s):  
V. Quemener ◽  
Y. Blanchard ◽  
D. Lescoat ◽  
R. Havouis ◽  
J. P. Moulinoux
Keyword(s):  
Cell Proliferation ◽  
Cell Differentiation ◽  
Mammalian Cells ◽  
Gradient Centrifugation ◽  
Cell Nuclei ◽  
Cell Proliferation And Differentiation ◽  
Proliferation And Differentiation ◽  
Human Spermatogenesis

Polyamines (PA), polycations present in all mammalian cells, are essential for cell proliferation and differentiation. In vitro, PA are known to bind to DNA with a high affinity. In vivo, the intimate association of endogenous PA with highly condensed chromatin has been reported. During spermatogenesis, when processes of cell proliferation and differentiation take place, the potential role of polyamines has not been studied in depth. We report here the PA levels measured in human spermatogenic cell nuclei at different stages of differentiation. Cell populations (spermatocytes and round, elongating, or elongated spermatids) were obtained after submitting human testes to a trypsin-deoxyribonuclease digestion, then to a centrifugal elutriation and Percoll gradient centrifugation. A significant and progressive nuclear spermine level decrease was observed from primary spermatocytes to elongated spermatids. This release of spermine from nuclei was concomitant with three major events in mammalian spermiogenesis: the reduction of DNA transcription activity, the replacement of histone proteins by protamines, and the compaction of chromatin. This is the first report arguing a release of nuclear spermine during an in vivo physiological cell differentiation process.

scholarly journals Quantifying endothelial cell proliferation in the zebrafish embryo

F1000Research ◽  
2021 ◽  
Vol 10 ◽  
pp. 1032
Author(s):  
George Bowley ◽  
Timothy JA Chico ◽  
Jovana Serbanovic-Canic ◽  
Paul C Evans
Keyword(s):  
Cell Proliferation ◽  
Endothelial Cell ◽  
Zebrafish Embryo ◽  
Time Lapse ◽  
Endothelial Cell Proliferation ◽  
Small Scale ◽  
Zebrafish Embryos ◽  
Time Lapse Imaging

Introduction: Endothelial cell (EC) proliferation is a fundamental determinant of vascular development and homeostasis, and contributes to cardiovascular disease by increasing vascular permeability to blood-borne lipoproteins. Rodents have been traditionally used to analyse EC proliferation mechanisms in vascular health and disease; however, alternative models such as the zebrafish embryo allow researchers to conduct small scale screening studies in a physiologically relevant vasculature whilst reducing the use of mammals in biomedical research. In vitro models of EC proliferation are valuable but do not fully recapitulate the complexity of the in vivo situation. Several groups have used zebrafish embryos for vascular biology research because they offer the advantages of an in vivo model in terms of complexity but are also genetically manipulable and optically transparent. Methods: Here we investigated whether zebrafish embryos can provide a suitable model for the study of EC proliferation. We explored the use of antibody, DNA labelling, and time-lapse imaging approaches. Results: Antibody and DNA labelling approaches were of limited use in zebrafish due to the low rate of EC proliferation combined with the relatively narrow window of time in which they can label proliferating nuclei. By contrast, time-lapse imaging of fluorescent proteins localised to endothelial nuclei was a sensitive method to quantify EC proliferation in zebrafish embryos. Discussion: We conclude that time-lapse imaging is suitable for analysis of endothelial cell proliferation in zebrafish, and that this method is capable of capturing more instances of EC proliferation than immunostaining or cell labelling alternatives. This approach is relevant to anyone studying endothelial cell proliferation for screening genes or small molecules involved in EC proliferation. It offers greater biological relevance than existing in vitro models such as HUVECs culture, whilst reducing the overall number of animals used for this type of research.

004.Control of skeletal muscle cell proliferation and differentiation

2005 ◽  
Vol 17 (9) ◽  
pp. 63
Author(s):  
M. Grounds
Keyword(s):  
Skeletal Muscle ◽  
Cell Proliferation ◽  
Multinucleated Cells ◽  
Cell Proliferation And Differentiation ◽  
Proliferation And Differentiation ◽  
The Matrix ◽  
Key Issues

Skeletal muscle is formed by mononucleated precursor cells (myoblasts) that cease cell proliferation to start differentiation; this results in fusion between the myoblasts to form multinucleated cells (myotubes) that continue to differentiate (and fuse with more muscle cells) and mature into myofibres. Myogenesis has been widely used as a model to study in vitro factors controlling cell proliferation and differentiation. Condition in vitro may not reflect what happens in the more complex in vivo environment. Some of the key issues are what activates quiescent myoblasts in mature skeletal muscle in vivo, and what controls the switch between proliferation and differentiation? The role of the matrix, and molecules such as MyoD, p53, NFAT and IGF-1 will be considered.

scholarly journals Differential regulation of the retinoblastoma family of proteins during cell proliferation and differentiation

1998 ◽  
Vol 333 (3) ◽  
pp. 645-654 ◽  
Author(s):  
Judit GARRIGA ◽  
Ana LIMÓN ◽  
Xavier MAYOL ◽  
Sushil G. RANE ◽  
Jeffrey H. ALBRECHT ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Protein Expression ◽  
Cultured Cells ◽  
Protein Levels ◽  
Pocket Protein ◽  
Cell Proliferation And Differentiation ◽  
Mouse Tissues ◽  
Proliferation And Differentiation

In the present study we have analysed the regulation of pocket protein expression and post-transcriptional modifications on cell proliferation and differentiation, both in vivo and in vitro. There are marked changes in pocket protein levels during these transitions, the most striking differences being observed between p130 and p107. The mechanisms responsible for regulating pocket protein levels seem to be dependent on both cell type and pocket protein, in addition to their dependence on the cell growth status. Changes in retinoblastoma protein and p107 levels are independent of their state of phosphorylation. However, whereas p130 phosphorylation to forms characteristic of quiescent/differentiated cells results in the accumulation of p130 protein, phosphorylation of p130 to one or more forms characteristic of cycling cells is accompanied by down-regulation of its protein levels. We also show here that the phosphorylation status and protein levels of p130 and p107 are regulated in vivo as in cultured cells. In vivo, changes in p130 forms are correlated with changes in E2F complexes. Moreover, the modulation of p130 and p107 status during cell differentiation in vitro is consistent with the patterns of protein expression and phosphorylation status found in mouse tissues. Thus in addition to the direct disruption of pocket protein/E2F complexes induced by cyclin/cyclin-dependent kinase, the results we report here indicate that the differential modulation of pocket protein levels constitutes a major mechanism that regulates the pool of each pocket protein that is accessible to E2F and/or other transcription factors.

scholarly journals In Vivo Time-Lapse Imaging and Analysis of Dendritic Structural Plasticity in Xenopus laevis Tadpoles

2021 ◽  
Vol 2022 (1) ◽  
pp. pdb.prot106781
Author(s):  
Hai-yan He ◽  
Chih-Yang Lin ◽  
Hollis T. Cline
Keyword(s):  
Data Analysis ◽  
Xenopus Laevis ◽  
Dynamic Analysis ◽  
Structural Changes ◽  
Structural Plasticity ◽  
Time Lapse ◽  
High Temporal Resolution ◽  
Dendritic Arbors ◽  
Time Lapse Imaging

In vivo time-lapse imaging of complete dendritic arbor structures in tectal neurons of Xenopus laevis tadpoles has served as a powerful in vivo model to study activity-dependent structural plasticity in the central nervous system during early development. In addition to quantitative analysis of gross arbor structure, dynamic analysis of the four-dimensional data offers particularly valuable insights into the structural changes occurring in subcellular domains over experience/development-driven structural plasticity events. Such analysis allows not only quantifiable characterization of branch additions and retractions with high temporal resolution but also identification of the loci of action. This allows for a better understanding of the spatiotemporal association of structural changes to functional relevance. Here we describe a protocol for in vivo time-lapse imaging of complete dendritic arbors from individual neurons in the brains of anesthetized tadpoles with two-photon microscopy and data analysis of the time series of 3D dendritic arbors. For data analysis, we focus on dynamic analysis of reconstructed neuronal filaments using a customized open source computer program we developed (4D SPA), which allows aligning and matching of 3D neuronal structures across different time points with greatly improved speed and reliability. File converters are provided to convert reconstructed filament files from commercial reconstruction software to be used in 4D SPA. The program and user manual are publicly accessible and operate through a graphical user interface on both Windows and Mac OSX.

Bulk Dye Loading for In Vivo Calcium Imaging of Visual Responses in Populations of Xenopus Tectal Neurons

2021 ◽  
Vol 2022 (1) ◽  
pp. pdb.prot106831
Author(s):  
Peter W. Hogg ◽  
Kurt Haas
Keyword(s):  
Optic Tectum ◽  
Image Plane ◽  
Time Lapse ◽  
Visual Responses ◽  
Easy Method ◽  
Tectal Neurons ◽  
Evoked Activity ◽  
Sensory Evoked ◽  
Time Lapse Imaging

Bulk loading of neurons with fluorescent calcium indicators in transparent albino Xenopus tadpoles offers a rapid and easy method for tracking sensory-evoked activity in large numbers of neurons within an awake developing brain circuit. In vivo two-photon time-lapse imaging of an image plane through the optic tectum allows defining receptive field properties from visual-evoked responses for studies of single-neuron and network-level encoding and plasticity. Here, we describe loading the Xenopus tadpole optic tectum with the membrane-permeable AM ester of Oregon Green 488 BAPTA-1 (OGB-1 AM) for in vivo imaging experiments.

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