The apoptosis cascade -- morphological and immunohistochemical methods for its visualization

Huppertz, Berthold; Frank, Hans-Georg; Kaufmann, Peter

doi:10.1007/s004290050254

Cited by 275 publications

(187 citation statements)

References 102 publications

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“…We performed additional assays using a substrate (Phi Phi Lux) for mitochondrial caspases associated with early events in apoptosis (25). As demonstrated in Fig.…”

Section: Evaluation Of Thymocyte Apoptosis As a Results Of Siv Infectionmentioning

confidence: 99%

Mechanisms Associated with Thymocyte Apoptosis Induced by Simian Immunodeficiency Virus

Rosenzweig¹,

Connole²,

Forand-Barabasz³

et al. 2000

The Journal of Immunology

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Despite considerable research, the mechanisms by which HIV disrupts thymic function remain controversial. We have described the phenotypic changes that occur in the thymus of SIV-infected macaques during acute SIV infection. In this study, we analyzed the effects of SIV infection on apoptotic pathways in thymic tissue from newborn macaques infected with SIV. Thymocyte apoptosis was accompanied by a modest increase in surface Fas expression, a profound decrease in the frequency of bcl-2-positive cells, as well as the amount of bcl-2 per cell. With control of viral replication, levels of bcl-2 and Fas returned to baseline together with a return to basal levels of apoptosis. In the thymus, SIV infection resulted in depletion of CD4+CD8+ thymocytes, an increase in apoptosis of thymocytes, and a down-regulation of MHC class I molecules. These changes peaked 14–21 days after infection at or just after peak viremia. This data further suggests disruption of the antiapoptotic pathway regulated by bcl-2 plays a critical role in SIV-induced apoptosis of thymocytes.

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“…We performed additional assays using a substrate (Phi Phi Lux) for mitochondrial caspases associated with early events in apoptosis (25). As demonstrated in Fig.…”

Section: Evaluation Of Thymocyte Apoptosis As a Results Of Siv Infectionmentioning

confidence: 99%

Mechanisms Associated with Thymocyte Apoptosis Induced by Simian Immunodeficiency Virus

Rosenzweig¹,

Connole²,

Forand-Barabasz³

et al. 2000

The Journal of Immunology

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“…During the analysis of cells in mitotic processes, apoptotic cells were also observed in the RM. Thus, several sections were carefully observed, and the morphological characteristics of cells in mitosis (Alberts et al, 2002) were distinguished from apoptotic cells (Wyllie et al, 1980;Huppertz et al, 1999;Cerri, 2005;Cerri and Katchburian, 2005;Gonçalves et al, 2008).…”

Section: Discussionmentioning

confidence: 99%

Area of Rests of Malassez in Young and Adult Rat Molars: Evidences in the Formation of Large Rests

Cerri

Gonçalves

Sasso-Cerri

2008

The Anatomical Record

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Recently, we have demonstrated that the reduction in the number of rests of Malassez (RM) according to age may be due to epithelial cell death by apoptosis. It is known that, under experimental or pathological conditions, the epithelial cells of RM can proliferate, resulting in the formation of odontogenic cysts and tumors. In this study, the area of RM profiles and the number of profiles of epithelial cell nuclei/RM were investigated in young and adult rats. Fragments containing upper molars of rats aged 29, 45, and 120 days were fixed, decalcified, and embedded for light microscopy. In the sections stained by hematoxylin and eosin, the numerical density of RM profiles/mm 2 of periodontal ligament, the area of each RM profile and the number of epithelial cell nuclei profile/RM were obtained. Moreover, detection of cytokeratin by immunohistochemistry and ultrastructural analysis of RM were carried out. Smaller RM were found in all groups, but the very large RM were only observed in the 120-day-old rats. The presence of cells in close juxtaposition exhibiting cytokeratin filaments in their cytoplasm, including in the large RM, characterized their epithelial nature. The number of epithelial cells/RM varied proportionally to the area. Moreover, epithelial cells in mitosis were found in small and large RM, indicating that cellular proliferation can be responsible for the enlargement of RM, under physiological conditions. The results suggest that a possible disequilibrium between the proliferative and apoptotic processes can be related to the enlargement of some RM in the 120-day-old rats.

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“…Annexin V is a Ca ++ dependant phospholipid-binding protein with a high affinity for phosphatidylserine (PS) [21,22]. In an early apoptotic state, PS is known to translocate from the inner to the outer leaflet of the plasma membrane.…”

Section: Evaluation Of Apoptosismentioning

confidence: 99%

Impact of integrin-matrix matching and inhibition of apoptosis on the survival of purified human beta-cells in vitro

et al. 2002

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Aims/hypothesis. Human islet cells survive poorly in culture and are overgrown by non-endocrine cells. The aims of this study were to sort human beta cells and to develop approaches for their improved survival in culture. Methods. Human islets were infected with recombinant adenovirus expressing green fluorescent protein (GFP) under the control of the rat insulin promoter such that only beta cells expressed GFP. GFP-positive beta cells were sorted by flow cytometry, and expression of select integrins evaluated by RT-PCR. Beta cells were cultured on different extracellular matrices for up to 15 days. Apoptosis was measured by annexin V binding and ELISA. Insulin secretion was measured by ELISA. Results. Sorted beta cells survived less well in culture than unsorted islet cells. This did not appear to be due to adenoviral infection and/or GFP expression. Purified beta cells expressed the integrins α3, α5, α6, αV, β1, but not β4. Of the various matrices tested, sorted beta cells attached and spread best on a lawn of lysed human bladder carcinoma cells (5637 cells). However, survival remained poor. Cell death was decreased but not prevented by continued presence of 10 mmol/l nicotinamide and apoptosis decreased by 24 h incubation with 20 µmol/l Z-VAD. Insulin secretion was maintained over 6 days following treatment with both agents. Conclusions/interpretation. Purification of human beta cells induces marked apoptosis limiting their function and survival in vitro. This was improved by matching the extracellular matrix to the specific expression of integrins and by addition of nicotinamide and Z-VAD. [Diabetologia (2002) 45:841-850]

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The apoptosis cascade -- morphological and immunohistochemical methods for its visualization

Cited by 275 publications

References 102 publications

Mechanisms Associated with Thymocyte Apoptosis Induced by Simian Immunodeficiency Virus

Mechanisms Associated with Thymocyte Apoptosis Induced by Simian Immunodeficiency Virus

Area of Rests of Malassez in Young and Adult Rat Molars: Evidences in the Formation of Large Rests

Impact of integrin-matrix matching and inhibition of apoptosis on the survival of purified human beta-cells in vitro

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