Real-time observation of autophagic programmed cell death of Drosophila salivary glands in vitro

Myohara, Maroko

doi:10.1007/s00427-003-0374-0

Cited by 9 publications

(8 citation statements)

References 16 publications

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“…The induction of autophagy that anticipates salivary gland histolysis may act as part of a novel killing mechanism in these cells (Lee and Baehrecke, 2001;Myohara, 2004;Thummel, 2001) , 2004). However, in other circumstances, 'self-digestion' clearly promotes survival when apoptosis in prevented (Lum et al, 2005), and, consequently, it is important to understand how links between autophagy and cell death may instruct cell fates (Levine and Klionsky, 2004).…”

Section: Discussionmentioning

confidence: 99%

“…These observations are consistent with effects produced by p35, a broad-spectrum caspase inhibitor (Jiang et al, 1997;Lee and Baehrecke, 2001;Martin and Baehrecke, 2004), and with animals mutated for the apical caspase dronc (Daish et al, 2004). Because apoptotic and histolytic forms of cell death are similarly impacted by the same mutation, we conclude that common effector pathways, regulated by the apoptosome, underlie morphologically distinct forms of PCD.The induction of autophagy that anticipates salivary gland histolysis may act as part of a novel killing mechanism in these cells (Lee and Baehrecke, 2001;Myohara, 2004;Thummel, 2001) , 2004). However, in other circumstances, 'self-digestion' clearly promotes survival when apoptosis in prevented (Lum et al, 2005), and, consequently, it is important to understand how links between autophagy and cell death may instruct cell fates (Levine and Klionsky, 2004).…”

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confidence: 90%

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Autophagy occurs upstream or parallel to the apoptosome during histolytic cell death

et al. 2006

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Histolysis refers to a widespread disintegration of tissues that is morphologically distinct from apoptosis and often associated with the stimulation of autophagy. Here, we establish that a component of the apoptosome, and pivotal regulator of apoptosis, is also required for histolytic cell death. Using in vivo and ex vivo assays, we demonstrate a global apoptogenic requirement for dark, the fly ortholog of Apaf1, and show that a required focus of dark -organismal lethality maps to the central nervous system. We further demonstrate that the Dark protein itself is a caspase substrate and find that alterations of this cleavage site produced the first hypermorphic point mutation within the Apaf1/Ced-4 gene family. In a model of 'autophagic cell death', dark was essential for histolysis but dispensable for characteristic features of the autophagic program, indicating that the induction of autophagy occurs upstream or parallel to histolytic cell death. These results demonstrate that stimulation of autophagy per se is not a 'killing event' and, at the same time, establish that common effector pathways, regulated by the apoptosome, can underlie morphologically distinct forms of programmed cell death.KEY WORDS: Autophagy, Apoptosis, Drosophila, Histolysis, dark Development 133, 1457Development 133, -1465Development 133, (2006 DEVELOPMENT 1458 per se is not the mechanism of cell killing but lies upstream, or parallel to dark. These data establish that common effector pathways, regulated by the apoptosome, specify apoptotic and histolytic forms of PCD. MATERIALS AND METHODS MutagenesisTo isolate deletions that eliminate dark without compromising the function of adjacent neighboring genes, a P insertion associated with dark CD4 (Rodriguez et al., 1999) was remobilized and candidates were tested in trans to existing alleles and against lethal mutations in flanking genes. Promising 'hits' were screened by PCR. dark 82 failed to complement dark CD4 , but complements adjacent lethal alleles in the neighboring genes, RhoGEF and a new lethal P mutation in CG8963 that we fortuitously obtained in our first round of mutagenesis. Genomic PCR across the deletion junction and RT-PCR were used to validate the mutation and define the dark 82 lesion. yw was the parental wild-type strain for molecular analysis and for ex vivo hemocyte studies. RNA extraction and QRT-PCR were conducted as described by Gorski et al. (Gorski et al., 2003). Ages at 25°C were normalized from 18°C (Park et al., 1996). Genomic PCR and RT-PCR were performed as described by Chew et al. (Chew et al., 2004), with relevant gene-specific primers. Transgenic 'rescue' and genetic manipulationFull-length dark with 8ϫHis-tags at the N terminus and 3ϫMyc-tags at the C terminus was cloned into the BamHI/XhoI sites of the pFastBac1 vector (Invitrogen). The BamHI/XhoI insert was then subcloned into the pUAST vector to produce pUAST-dark WT . pUAST-dark V was generated by changing Aspartate 1292 to Alanine using a QuikChange Site-Directed Mutagenesis Kit (Stratagene). The ...

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Section: Discussionmentioning

confidence: 99%

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confidence: 90%

Autophagy occurs upstream or parallel to the apoptosome during histolytic cell death

et al. 2006

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“…Thus, it must be acknowledged that DNA fragmentation is neither universal nor necessarily predominant in the cell death process (Oberhammer et al, 1993;Lockshin and Zakeri, 1992;Zakeri et al, 1993). In addition, the action of endonucleases in the cell death of insects remains controversial, with its participation being confirmed by some authors (Gregorc and Bowen, 1996;Jochová et al, 1997;Foley and Cooley, 1998;Sheng-Hao and Nagoshi, 1999;Myohara, 2004) and denied by others (Lockshin, 1985;Bowen et al, 1993). In the review of Robertson et al (2000) regarding the nuclear events of apoptosis, the authors showed that the chromatin condensation and degradation of the DNA are controlled separately.…”

Section: Introductionmentioning

confidence: 93%

Nuclear alterations associated to programmed cell death in larval salivary glands of Apis mellifera (Hymenoptera: Apidae)

Silva-Zacarín

Taboga

Moraes

2008

Micron

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“…Several aspects of programmed cell death in larval insect salivary glands have been studied, mainly in the Lepidoptera and Diptera (Aidells et al 1971; Schin and Laufer 1973;Jurand and Pavan 1975;Beaulaton and Lockshin 1982;Levy and Bautz 1985;Armbruster et al 1986;Sehnal and Akai 1990;Jones and Bowen 1993;Bowen et al 1993Bowen et al , 1996Jochová et al 1997a, b;Dorstyn et al 1999;Lee and Baehrecke 2001;Myohara 2004). In the blow-fl y Calliphora, programmed cell death of the salivary glands does not emulate classical apoptosis because the cells are seen to vacuolate and swell rather than condense and shrink (Bowen et al 1996); additionally, there is basal fragmentation (Levy and Bautz 1985) and the nucleus does not show a distinct chromatin margination and blebbing (Bowen et al 1996).…”

Section: Introductionmentioning

confidence: 99%

“…Histolysis of the silk gland during metamorphosis of Bombyx mori is carried out by autophagy, with lysosomal proteinases playing the key role in degrading cytoplasmic organelles in the autophagosomes (Lockshin et al 1998;Shiba et al 2001). However, real-time observation of autophagic programmed cell death of Drosophila salivary glands showed that the gland shrank due to cytoplasmic loss caused by blebbing (Myohara 2004).…”

Section: Introductionmentioning

confidence: 99%

Programmed cell death in the larval salivary glands of Apis mellifera (Hymenoptera, Apidae)

et al. 2007

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The morphological and histochemical features of degeneration in honeybee (Apis mellifera) salivary glands were investigated in 5th instar larvae and in the pre-pupal period. The distribution and activity patterns of acid phosphatase enzyme were also analysed. As a routine,the larval salivary glands were fixed and processed for light microscopy and transmission electron microscopy.Tissue sections were subsequently stained with haematoxylin -eosin,bromophenol blue,silver,or a variant of the critical electrolyte concentration (CEC) method.Ultrathin sections were contrasted with uranyl acetate and lead citrate.Glands were processed for the histochemical and cytochemical localization of acid phosphatase,as well as biochemical assay to detect its activity pattern. Acid phosphatase activity was histochemically detected in all the salivary glands analysed.The cytochemical results showed acid phosphatase in vesicles, Golgi apparatus and lysosomes during the secretory phase and,additionally, in autophagic structures and luminal secretion during the degenerative phase. These findings were in agreement with the biochemical assay. At the end of the 5th instar, the glandular cells had a vacuolated cytoplasm and pyknotic nuclei, and epithelial cells were shed into the glandular lumen.The transition phase from the 5th instar to the pre-pupal period was characterized by intense vacuolation of the basal cytoplasm and release of parts of the cytoplasm into the lumen by apical blebbing; these blebs contained cytoplasmic RNA, rough endoplasmic reticule and, occasionally, nuclear material. In the pre-pupal phase, the glandular epithelium showed progressive degeneration so that at the end of this phase only nuclei and remnants of the cytoplasm were observed.The nuclei were pyknotic,with peripheral chromatin and blebs. The gland remained in the haemolymph and was recycled during metamorphosis. The programmed cell death in this gland represented a morphological form intermediate between apoptosis and autophagy.

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Real-time observation of autophagic programmed cell death of Drosophila salivary glands in vitro

Cited by 9 publications

References 16 publications

Autophagy occurs upstream or parallel to the apoptosome during histolytic cell death

Autophagy occurs upstream or parallel to the apoptosome during histolytic cell death

Nuclear alterations associated to programmed cell death in larval salivary glands of Apis mellifera (Hymenoptera: Apidae)

Programmed cell death in the larval salivary glands of Apis mellifera (Hymenoptera, Apidae)

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