Cell death and the development of limb form and skeletal pattern in normal and wingless (ws) chick embryos

The wingless condition resulting from the action of the sex-linked wingless (ws) gene arises from the precocious appearance of cell death in the anterior necrotic zone (ANZ) of the forelimb-bud at stage 19 (3 days) and its progressive extension beyond its normal area during stages 20–23. A similar though less pronounced effect occurs in the hindlimb-bud. Although some wingless hindlimb-buds are normal, others are affected by the precocious appearance of cell death in the ANZ. The ws wingless mutant resembles the different wingless mutant investigated by Zwilling (1956) in that the apical ectodermal ridge (AER) is absent in most ws wing-buds. AER absence could be due to ws mesenchymal cell death interfering with the production of apical ectodermal maintenance factor (AEMF), which Zwilling claims is necessary to maintain the AER which plays an essential role in inducing limb outgrowth. Wingless mutant phenotypes range from birds with rudimentary wings and normal legs through a modal type with forelimbs absent and hindlimbs normal to wingless and legless forms showing a high degree of expressivity. Individual wingless embryos vary in the degree to which the precocious ANZ appearing at 3 days is extended into the limb-bud and the wide range of wingless phenotypic expression is attributed to this variation. Electron microscopic and histochemical analysis of the cell death process in wingless wing-buds revealed the presence of both isolated dead cells and macrophages, which contained intense acid phosphatase activity. These findings are interpreted as showing that isolated dead cells are ingested by neighbouring mesenchymal cells which thus become transformed into macrophages, first ingesting and then digesting further dead cells. A study was made of the origin of the anomalous hindlimb condition, including absence or reduction of the tibia and digits, characteristic of severely affected wingless embryos. Autoradiographic analysis of the pattern of 35SO4 uptake revealed that at stage 24/5 (4½ days) wingless hindlimb-buds which were smaller than normal had a normal prospective fibula region, but that the prospective tibia region was small or absent. Thus the effect of a precocious hindlimb ANZ at stages 19–22 is to reduce or delete the pre-axial prospective tibia at stage 24/5.

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Vascular regression during the formation of the free digits in the avian limb bud: a comparative study in chick and duck embryos

Development ◽

10.1242/dev.85.1.239 ◽

1985 ◽

Vol 85 (1) ◽

pp. 239-250

Author(s):

J. M. Hurle ◽

E. Colvee ◽

M. A. Fernandez-Teran

Keyword(s):

Cell Death ◽

Blood Vessels ◽

Mesenchymal Cell ◽

Limb Bud ◽

Death Process ◽

Electron Microscopic ◽

Transmission Electron ◽

Transmission Electron Microscopic ◽

Microscopic Studies ◽

Cell Death Process

The pattern and structure of the blood vessels of the interdigital spaces of the leg bud have been studied by means of Indian ink injections and transmission electron microscopy in the chick and duck embryos. The results show that in the chick the interdigital necrotic process responsible for the freeing of the digits is followed by regression of the blood vessels. In the webbed foot of the duck, the interdigital necrotic processes are not followed by vascular regression. Transmission electron microscopic studies show that both in the chick and in the duck, interdigital blood vessels are immature structures lacking basal lamina. Dead cells of presumably endothelial origin were detected in the lumen of the regressing blood vessels of the chick but not in the duck. However, the intensity of this cell death process does not appear to be high enough to account by itself for the disappearance of the interdigital blood vessels. The possible relationships between interdigital mesenchymal cell death and vascular regression are discussed.

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Genetic inhibition of mesenchymal cell death and the development of form and skeletal pattern in the limbs of talpid3 (ta3) mutant chick embryos

Development ◽

10.1242/dev.31.3.747 ◽

1974 ◽

Vol 31 (3) ◽

pp. 747-760

Author(s):

J. R. Hinchliffe ◽

P. V. Thorogood

Keyword(s):

Cell Death ◽

Soft Tissue ◽

Limb Development ◽

Chondroitin Sulphate ◽

Vital Staining ◽

Mesenchymal Cell ◽

Equivalent Position ◽

Skeletal Pattern ◽

Mesenchyme Cells ◽

Interdigital Cell Death

Vital staining reveals that in homozygous (ta3/ta3) talpid3 embryos, the areas of mesenchymal cell death which occur regularly in normal limb development are absent or reduced. The necrotic locus in the central mesenchyme (the ‘opaque patch’) which in the normal chick limb reaches maximum development at stages 24 and 25 (4½–5 days) is absent or much reduced in talpid3 fore- and hindlimb-buds. Autoradiographic studies, following application of a 2 h pulse of 40 μCi of 35SO4 to the vitelline circulation, show that normal tibia and fibula incorporate 35SO4 into chondroitin sulphate at stage 24 and more strongly at stage 26 during the process of chondrogenesis. The mesenchyme in the opaque patch region of normal limbs ceases to incorporate 35SO4 into chondroitin sulphate at stage 24. Talpid3 mesenchyme cells in the equivalent position at stages 24 and 26 continue to incorporate 35SO4, remain viable and become chondrogenic. It is suggested that absence or reduction of this central necrotic locus in talpid3 is causally related to the fusion of radius/ulna and (in some cases) of tibia/fibula characteristic of the later stages (28–35) of talpid3 limb development. This evidence supports the hypothesis that cell death in the opaque patch plays a morphogenetic role in separation of radius/ulna and tibia/fibula. The digital plate of stage 32 (7½ days) normal limbs is characterized by massive necrosis of the interdigital tissue. In talpid3 forelimbs of stages 30–35 interdigital necrosis is absent, and there is no regression of the tissue between the digits (‘soft tissue syndactyly’). In talpid3 hindlimbs of stage 30–35 interdigital necrosis is either absent or much reduced, and there is little or no erosion of the soft tissue between the digits. This evidence supports the hypothesis that the morphogenetic role of interdigital cell death is in causing separation of the digits through shaping and remodelling the contours of the digital plate.

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BMP-2/-4 mediate programmed cell death in chicken limb buds

Development ◽

10.1242/dev.122.12.3725 ◽

1996 ◽

Vol 122 (12) ◽

pp. 3725-3734 ◽

Cited By ~ 17

Author(s):

Y. Yokouchi ◽

J. Sakiyama ◽

T. Kameda ◽

H. Iba ◽

A. Suzuki ◽

...

Keyword(s):

Cell Death ◽

Programmed Cell Death ◽

Mesenchymal Cells ◽

Dominant Negative ◽

Limb Bud ◽

Death Process ◽

Signal Molecules ◽

Opaque Zone ◽

Limb Buds ◽

Bmp Receptor

During limb development, the mesenchymal cells in restricted areas of limb bud, anterior necrotic zone, posterior necrotic zone, opaque zone and interdigital necrotic zones, are eliminated by programmed cell death. The transcripts of bone morphogenetic protein (Bmp)-2 and −4 were first detected in the areas where cell death was observed, then showed overlapping expression with the programmed cell death zones except the opaque zone. To investigate the function of BMP-2 and BMP-4 during limb pattern formation, the dominant negative form of BMP receptor was overexpressed in chick leg bud via a replication-competent retrovirus to block the endogenous BMP-2/-4 signaling pathway. This resulted in excess web formation at the anterior and posterior regions of limb buds in addition to marked suppression of the regression of webbing at the interdigital regions. Significant reductions in the number of apoptotic cells in these three necrotic zones were found in the limb buds which received the virus carrying dominant negative BMP receptor. This indicates that extra tissue formation is due to suppression of programmed cell death in the three necrotic zones. Moreover, BMP-2/-4 protein induced apoptosis of mesenchymal cells isolated from the interdigital region in vitro. Other TGFbeta family proteins as TGFbeta1 and Activin did not show this effect. These results suggest that BMP-2 and BMP-4 are the apoptotic signal molecules of the programmed cell death process in the chick limb buds.

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Faculty Opinions recommendation of A nonapoptotic cell death process, entosis, that occurs by cell-in-cell invasion.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1097183.572616 ◽

2008 ◽

Author(s):

Gary Bokoch

Keyword(s):

Cell Death ◽

Cell Invasion ◽

Death Process ◽

Cell Death Process ◽

Nonapoptotic Cell Death

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Reverse Screening Bioinformatics Approach to Identify Potential Anti Breast Cancer Targets Using Thymoquinone from Neutraceuticals Black Cumin Oil

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520619666190124155359 ◽

2019 ◽

Vol 19 (5) ◽

pp. 599-609 ◽

Cited By ~ 1

Author(s):

Sumathi Sundaravadivelu ◽

Sonia K. Raj ◽

Banupriya S. Kumar ◽

Poornima Arumugamand ◽

Padma P. Ragunathan

Keyword(s):

Breast Cancer ◽

Gene Expression ◽

Cell Cycle ◽

Cell Death ◽

In Silico ◽

Chemotherapeutic Agents ◽

Normal Cells ◽

Black Cumin ◽

In Silico Docking ◽

Wide Range

Background: Functional foods, neutraceuticals and natural antioxidants have established their potential roles in the protection of human health and diseases. Thymoquinone (TQ), the main bioactive component of Nigella sativa seeds (black cumin seeds), a plant derived neutraceutical was used by ancient Egyptians because of their ability to cure a variety of health conditions and used as a dietary food supplement. Owing to its multi targeting nature, TQ interferes with a wide range of tumorigenic processes and counteracts carcinogenesis, malignant growth, invasion, migration, and angiogenesis. Additionally, TQ can specifically sensitize tumor cells towards conventional cancer treatments (e.g., radiotherapy, chemotherapy, and immunotherapy) and simultaneously minimize therapy-associated toxic effects in normal cells besides being cost effective and safe. TQ was found to play a protective role when given along with chemotherapeutic agents to normal cells. Methods: In the present study, reverse in silico docking approach was used to search for potential molecular targets for cancer therapy. Various metastatic and apoptotic targets were docked with the target ligand. TQ was also tested for its anticancer activities for its ability to cause cell death, arrest cell cycle and ability to inhibit PARP gene expression. Results: In silico docking studies showed that TQ effectively docked metastatic targets MMPs and other apoptotic and cell proliferation targets EGFR. They were able to bring about cell death mediated by apoptosis, cell cycle arrest in the late apoptotic stage and induce DNA damage too. TQ effectively down regulated PARP gene expression which can lead to enhanced cancer cell death. Conclusion: Thymoquinone a neutraceutical can be employed as a new therapeutic agent to target triple negative breast cancer which is otherwise difficult to treat as there are no receptors on them. Can be employed along with standard chemotherapeutic drugs to treat breast cancer as a combinatorial therapy.

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Control by the Posterior Border of Cell Death Patterns in Limb Bud Development of Amniotes: Evidence from Experimental Amputations and from Mutants

Teratology of the limbs ◽

10.1515/9783110861082-006 ◽

1980 ◽

pp. 27-34 ◽

Cited By ~ 1

Author(s):

J.R. Hinchliffe

Keyword(s):

Cell Death ◽

Limb Bud ◽

Posterior Border ◽

Bud Development

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Individual Expression of Hepatitis A Virus 3C Protease Induces Ferroptosis in Human Cells In Vitro

International Journal of Molecular Sciences ◽

10.3390/ijms22157906 ◽

2021 ◽

Vol 22 (15) ◽

pp. 7906

Author(s):

Alexey A. Komissarov ◽

Maria A. Karaseva ◽

Marina P. Roschina ◽

Andrey V. Shubin ◽

Nataliya A. Lunina ◽

...

Keyword(s):

Cell Death ◽

Hepatitis A ◽

Hepatitis A Virus ◽

Proteolytic Enzymes ◽

Morphological Changes ◽

Human Cells ◽

Mitochondrial Potential ◽

3C Protease ◽

Wide Range

Regulated cell death (RCD) is a fundamental process common to nearly all living beings and essential for the development and tissue homeostasis in animals and humans. A wide range of molecules can induce RCD, including a number of viral proteolytic enzymes. To date, numerous data indicate that picornaviral 3C proteases can induce RCD. In most reported cases, these proteases induce classical caspase-dependent apoptosis. In contrast, the human hepatitis A virus 3C protease (3Cpro) has recently been shown to cause caspase-independent cell death accompanied by previously undescribed features. Here, we expressed 3Cpro in HEK293, HeLa, and A549 human cell lines to characterize 3Cpro-induced cell death morphologically and biochemically using flow cytometry and fluorescence microscopy. We found that dead cells demonstrated necrosis-like morphological changes including permeabilization of the plasma membrane, loss of mitochondrial potential, as well as mitochondria and nuclei swelling. Additionally, we showed that 3Cpro-induced cell death was efficiently blocked by ferroptosis inhibitors and was accompanied by intense lipid peroxidation. Taken together, these results indicate that 3Cpro induces ferroptosis upon its individual expression in human cells. This is the first demonstration that a proteolytic enzyme can induce ferroptosis, the recently discovered and actively studied type of RCD.

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