The virulent Wolbachia strain wMelPop increases the frequency of apoptosis in the female germline cells of Drosophila melanogaster
© Zhukova and Kiseleva; licensee BioMed Central Ltd. 2012
Published: 18 January 2012
Wolbachia are bacterial endosymbionts of many arthropod species in which they manipulate reproductive functions. The distribution of these bacteria in the Drosophila ovarian cells at different stages of oogenesis has been amply described. The pathways along which Wolbachia influences Drosophila oogenesis have been, so far, little studied. It is known that Wolbachia are abundant in the somatic stem cell niche of the Drosophila germarium. A checkpoint, where programmed cell death, or apoptosis, can occur, is located in region 2a/2b of the germarium, which comprises niche cells. Here we address the question whether or not the presence of Wolbachia in germarium cells can affect the frequency of cyst apoptosis in the checkpoint.
Our current fluorescent microscopic observations showed that the wMel and wMelPop strains had different effects on female germline cells of D. melanogaster. The Wolbachia strain wMel did not affect the frequency of apoptosis in cells of the germarium. The presence of the Wolbachia strain wMelPop in the D. melanogaster w1118 ovaries increased the number of germaria where cells underwent apoptosis in the checkpoint. Based on the appearance in the electron microscope, there was no difference in morphological features of apoptotic cystocytes between Wolbachia-infected and uninfected flies. Bacteria with normal ultrastructure and large numbers of degenerating bacteria were found in the dying cyst cells.
Our current study demonstrated that the Wolbachia strain wMelPop affects the egg chamber formation in the D. melanogaster ovaries. This led to an increase in the number of germaria containing apoptotic cells. It is suggested that Wolbachia can adversely interfere either with the cystocyte differentiation into the oocyte or with the division of somatic stem cells giving rise to follicle cells and, as a consequence, to improper ratio of germline cells to follicle cells and, ultimately, to apoptosis of cysts. There was no similar adverse effect in D. melanogaster Canton S infected with the Wolbachia strain wMel. This was taken to mean that the observed increase in frequency of apoptosis was not the general effect of Wolbachia on germline cells of D. melanogaster, it was rather induced by the virulent Wolbachia strain wMelPop.
Apoptosis, a form of programmed cell death, is a process needed for normal development and maintenance of tissue homeostasis in multicellular organisms [1, 2]. Cells undergoing apoptosis show characteristic changes, such as chromatin and cytoplasm condensation, chromosomal DNA fragmentation, breaking up of nuclei and then of cells into fragments called apoptotic bodies [3, 4]. Individual cells apoptose, while the neighboring cells remain undamaged [3, 4]. Apoptosis is a complex process whereby a proteolytic cascade of caspases is activated in cells .
The occurrence of apoptosis is a feature of female germline development common to vertebrate and invertebrate species [6, 7]. In the Drosophila melanogaster ovaries, there are two checkpoints where programmed cell death occurs. One is in the germarium (region 2a/2b), where apoptosis probably regulates the proper ratio of germline cells to follicle cells . The other checkpoint is located in the vitellarium (stages 7-8 of oogenesis) . The number of egg chambers undergoing apoptosis increased in D. melanogaster fed a diet lacking protein , under the effect of 900-MHz and 1800-MHz radiation , and after exposure to chemical agents . The normal development of mature egg is consistently associated with apoptosis of 15 nurse cells in the egg chamber . It is noteworthy that apoptosis and autophagy coexist at all the above mentioned stages of oogenesis in D. melanogaster [13, 14].
It has been also hypothesized that the apoptotic process had a symbiotic origin . In terms of the endosymbiotic theory, mitochondria, which play a major role at the early stages of apoptosis, evolved from the free-living prokaryotes . One of the symbionts may be involved in the regulation of apoptosis in partner cells. To illustrate, extracellular parasites, particularly such worms as filarial nematodes, schistosomes and the cestode Taenia crassiceps, are able to induce apoptosis in host immune cells . Bacterial pathogens (Chlamydia, Neisseria, Legionella pneumophila) can either block or induce apoptosis in host cells, depending on the stage of infection [17, 18]. At the early stage of infection, bacteria replicate in the host cell, using different mechanisms to prevent apoptosis. At the late stages of infection, the bacteria induce apoptosis in the host cell, thereby facilitating egress and ensuring infection of neighboring cells.
Wolbachia associated with various hosts in which it manipulates viability and reproduction causing parthenogenesis, feminization, male killing and cytoplasmic incompatibility, provides a unique model for studying mechanisms of symbiont interactions [19, 20]. The Wolbachia strain wMel is widely spread in natural populations of D. melanogaster [21, 22]; in contrast, wMelPop has been detected in a laboratory stock of D. melanogaster . It is possibly not encountered in nature. In D. melanogaster, the wMelPop strain reduces lifespan, proliferating widely in the brain, muscle and retina cells . In certain insect species, the presence of Wolbachia is required for oogenesis . Removal of the Wolbachia strain wAtab3 from parasitic wasps Asobara tabida resulted in copious apoptosis of the egg chambers in the ovarioles and led to sterility . The mechanisms whereby the endosymbiont Wolbachia impacts apoptosis in host cells have been poorly studied. Preferential infection and high accumulation of Wolbachia in region 2a of the germarium  where the checkpoint is located in Drosophila was thought-provoking. We raised the question: Can bacteria Wolbachia in region 2a of the germarium affect the frequency of apoptosis there? Using fluorescence and transmission electron microscopy we compared germaria from ovaries of two D. melanogaster stocks infected with either the wMel or wMelPop strains with germaria from two uninfected counterparts. It was established that the presence of wMel did not alter apoptosis frequency in germaria from D. melanogaster Canton S. In contrast, the number of germaria containing apoptotic cells in the checkpoint was considerably increased in the wMelPop-infected flies as compared with their uninfected counterparts. Thus, evidence was obtained indicating that the virulent Wolbachia strain wMelPop has an effect on the fate of germline cells during D. melanogaster oogenesis.
Frequency of apoptosis in germaria from ovaries of the uninfected and Wolbachia-infected D. melanogaster
Details of statistical analysis (two-way ANOVA)
Source of variation
Canton S/Canton ST
% of total variation
% of total variation
Type of food
Quantification of TUNEL staining in region 2a/2b of the germaria
Total % of TUNEL-positive
Cluster of spots
w 1118 T
Ultrastructure of germaria from ovaries of the uninfected and the Wolbachia-infected D. melanogaster
This is, to our knowledge, the first study that demonstrated by using AO- and TUNEL staining an increase in the frequency of apoptosis in the germarium checkpoint in wMelPop-infected D. melanogaster w1118 . This increase is possibly caused by the specific effect of the Wolbachia strain wMelPop, since it was not observed in wMel-infected D. melanogaster Canton S. Our current electron microscopic observations allowed us to identify changes in Wolbachia morphology in apoptotic germline cells.
Morphological evidence of apoptosis in germarium cells
The ultrastructural features of apoptosis in the cyst cells of higher eukaryotes have gained wide recognition. They include cytoplasmic and nuclear condensation (pyknosis); nuclear fragmentation (karyorrhexis); normal morphological appearance of cytoplasmic organelles; an intact plasma membrane [3, 4]. The ultrastructural changes we identified here in D. melanogaster cyst cells are consistent with the above hallmarks. Furthermore, we revealed mitochondria of two types: intact morphology in one type and markedly swollen with a few cristae in the other. A similar heterogeneity of mitochondrial ultrastructure has been observed during apoptosis in granulose cells of Japanese quail (Coturnix coturnix japonica) , lymphocytes from leukemia patients , and megakaryocytes from patients with idiopathic thrombocytopenic purpura . It has been suggested that the swollen mitochondria release cytochrome c, which activates a cascade of proteolytic reactions, while the normal ones retain their capacity for ATP synthesis, a process apoptosis requires [30, 31, 33]. According to our qualitative analysis using EM, morphological evidence of apoptosis was revealed in germline cells from uninfected flies and those infected with wMel and wMelPop. Thus, there are reasons for inferring that the endosymbiont Wolbachia in D. melanogaster cystocytes has no effect on sequential passage of intracellular organelles through apoptosis. To reveal the possible differences between the effect of the wMel and wMelPop strains on apoptosis in the germaria, additional morphometric analysis of the number of apoptotic structures and of Wolbachia density in the cystocytes is required.
Structural features of Wolbachiain apoptotic cysts
Wolbachia with matrix of moderate and low electron density in apoptotic cells in region 2a/2b of the germarium have been previously encountered in other types of D. melanogaster ovaries  and they presumably reflect different functional states of bacteria. Wolbachia with disrupted envelopes and light matrix are possibly dying bacteria in apoptotic cells. Such appearance has not been observed in Wolbachia injured or killed by heat stress  and tetracycline . The electron-dense bacteria-like structures at the periphery of region 1 of the germarium may be evidence of changes in dying Wolbachia. Large masses of structures of this kind resembling the bacteria endospores have been found in the brain cells of the wMelPop-infected D. melanogaster w1118 . In our view, the electron-dense structures, which we revealed at the periphery of region 1 of the germarium, are presumably autophagosome encapsulated dying Wolbachia. A supporting line of evidence came from Wright and Barr , who on the basis of their observations on degenerating germaria cysts from mosquitoes Aedes scutellaris suggested that these structures represented degenerating Wolbachia.
Cell fragments containing dying bacteria and autophagosomes and appearing as numerous smaller puncta in regions 2a/2b and 1 of the germarium may represent autophagy, not apoptosis. This appears plausible when recalling that AO stains not only apoptotic cells, also lysosomes . TUNEL did not reveal such puncta in these regions.
The possible role of the Wolbachiastrain wMelPop in programmed cell death in region 2a/2b of the germarium
Our current estimates of apoptosis in region 2a/2b of the germarium from the ovaries of the uninfected D. melanogaster w1118T raised on standard food are consistent with those reported elsewhere . It is of interest that apoptosis level in the germaria decreased in D. melanogaster w1118T , but not in D. melanogaster Canton ST after transfer to rich food. This may be indicative of differences in sensitivity to changes in food composition between different fly stocks. AO- and TUNEL staining demonstrated that the virulent Wolbachia strain wMelPop increased the percentage of germaria containing apoptotic cells in D. melanogaster w1118 ovaries, while wMel strain was without such an effect. The effect of wMelPop on cystocytes in ovaries was observed in flies maintained on standard and rich food. Evidence was provided that the effect of Wolbachia on D. melanogaster is not general, being rather specific to the pathogenic strain wMelPop.
On the other hand, the increase in the number of germaria containing apoptotic cysts may result from the action of the bacteria on the SSCs, which gives rise to follicle cells in region 2b of the germarium (Figure 7A, C). Drummond-Barbosa and Spradling  have suggested that apoptosis in region 2a/2b of the germarium serves to maintain the proper ratio of germline cells to somatic follicle cells. In poorly fed flies, follicle cells slow down their proliferation, the germline cells to somatic follicle cell ratio becomes skewed, resulting in cyst apoptosis in region 2a/2b which corrects this ratio . It has been established that stem cells are maintained in specialized microenvironment called the niche . The abundance of Wolbachia in the SSCN  is of interest in this context. Thus reasoning, it may be assumed that the presence of Wolbachia in the SSCN decreases the SSC proliferation rate, the ratio of germline cells to follicle cells becomes imbalanced and, as a consequence, cysts undergo apoptotic death. Judging from our current data, the ultrastructural appearance of follicle cells in region 2b of the germarium from ovaries of wMelPop-infected D. melanogaster w1118 was normal, thereby indicating that Wolbachia presumably did not negatively affect follicle cells. It should be noted that the fecundity of the wMelPop infected D. melanogaster w1118 was not decreased as compared with their uninfected counterparts [43, 44]. This was evidence of insect plasticity, rendering them capable to adapt to diverse factors.
Taken together, our findings clearly demonstrated that the Wolbachia strain wMelPop has an effect on the egg chamber formation in the D. melanogaster germarium. However, the underlying mechanism is still unclear. We intend to perform a comparative morphometric analysis of apoptotic structures and bacteria in cystocytes of wMel- and wMelPop-infected flies. The results would be helpful in deciding whether the increase in apoptosis frequency is due to high bacterial density or to particular pathogenic effect of the Wolbachia strain wMelPop on female germline cells.
The results of this study showed that the presence of the Wolbachia strain wMelPop in D. melanogaster ovaries led to an increase in the frequency of apoptosis in the germarium checkpoint. Two possible pathways along which Wolbachia affect egg chamber formation in region 2a/2b of the germarium have been suggested. Future research should be conducted to clarify the mechanism underlying this phenomenon.
Drosophilastocks and maintenance
The Drosophila melanogaster Canton S infected with the Wolbachia strain wMel (IC&G, Russia) and D. melanogaster w1118 infected with wMelPop (a kind gift from prof. S. O’Neill, The University of Queensland, Australia) were used in these experiments. Flies were maintained at 25 °C either on a standard yeast-agar medium or on daily replaced rich food (standard medium covered with wet yeast paste). To obtain uninfected D. melanogaster w1118T , flies were raised on food supplemented with tetracycline at 0.03% for two generations, then on standard food for more than three generations . Confirmation of the infection status of each stock was provided by PCR. For this purpose, total DNA extracted from fly ovaries and wsp 81F/wsp 691R primers for amplifying a Wolbachia surface protein gene fragment were used .
Acridine orande staining
Acridine orange (AO), a vital stain highly specific to apoptotic nuclei, was used . Ovaries were dissected from 5-day old flies in EBR buffer (130 mM NaCl, 4.7 mM KCl, 1.9 mM CaCl2, 10 мM Hepes pH 6.9), stained with AO (Merck), 5 μg/ml, in 0.1 M sodium phosphate buffer, pH 7.2, for 3 min at room temperature [12, 47]. Samples were placed onto glass slides and covered with halocarbon oil (KMZ Chemicals Ltd.). They were viewed under an Axioscop 2 plus fluorescence microscope (Zeiss) using an appropriate filter (Zeiss filter set 02). Time elapsed from dissection to the end of viewing was restricted, 20 min. Staining of nuclei varied from bright yellow to brilliant orange, depending on the stage of degeneration . The percentage of AO-staining germaria was expressed as the ratio of the number of AO-stained germaria containing apoptotic cells to the total number of analysed germaria. Three experiments were performed for each of the 4 D. melanogaster groups (w1118, w1118T stocks, standard food; w1118, w1118T, rich food). In each replicate, ovaries were dissected from 6 flies, 7-12 germaria per fly were analysed. In all, about 1350 AO-stained germaria were analysed. Bartlett’s test was used to check homogeneity of variances. Two-way ANOVA was used to determine the significance of the difference between the frequency of apoptosis of the uninfected and Wolbachia-infected flies maintained on different food.
TUNEL was the independent assay of detection of apoptotic cells. TUNEL is advantageous because preferentially labeling apoptotic cells relatively late in the apoptotic process . Ovaries were dissected from 5-day old flies in phosphate-buffered saline (PBS), fixed in PBS containing 4% formaldehyde plus 0.1% Triton X-100 for 25 min. Then, they were separated into individual ovarioles, rinsed briefly in PBS twice and washed in PBS three times for 5 min each. Ovarioles were made permeable with 20 μg/ml proteinase K in PBS for 20 min at room temperature, this was followed by 3 washes in PBS for 5 min each. The TUNEL reaction and all the subsequent steps were performed using the FragEL DNA Fragmentation Detection Kit (Calbiochem) according to the manufacturer’s protocol. Samples were viewed with an Axioscop 2 plus fluorescent microscope (Zeiss), images were captured with a high resolution microscopy camera AxioCam HRc and AxioVision software. Germaria from ovaries of 10 flies were counted in each of the 4 groups. The total number of germaria analysed was about 850. The data were compared using a Chi-square test (χ2).
Fixation of the D. melanogaster ovaries was carried out using the method described previously [49, 35]. Briefly, 5 day-old females were dissected in 0.1 M phosphate buffer, pH 7.4, fixed in 2.5% glutaraldehyde (Sigma) in 0.1 M sodium cacodylate buffer, pH 7.4, for 2.5 h. This was followed by washings in the same buffer and postfixation in 1% OsO4 and 0.8% potassium ferrocyanide for 1 h. After washings, samples were placed in 1% aqueous solution of uranyl acetate (Serva) for 12 h at 4 °C. Then they were dehydrated in ethanol series and acetone, finally samples were embedded in Agar 100 Resin (Agar Scientific Ltd.). Ultra-thin sections were stained with uranyl acetate and Reynolds lead citrate. They were examined with a transmission electron microscope (JEM 100 SX, JEOL). The number of flies analysed in each of the 4 groups was 8-12.
We thank Prof. S. O’Neill (The University of Queensland, Australia) for kindly supplying us with D. melanogaster stock. We are also grateful to the staff of the IC&G SB RAS, particularly to Dr. A.A. Ogienko for sharing her experience with AO-staining of the D. melanogaster ovaries, Prof. I.K. Zakharov for providing conditions for fly maintenance, A.N. Fadeeva for translating the manuscript from Russian into English. This work was supported by the Program of Basic Research of the RAS Presidium “Biodiversity” (26.30), “Molecular and Cellular biology” (6.12) and a grant from the Russian Foundation for Basic Research.
This article has been published as part of BMC Microbiology Volume 11 Supplement 1, 2012: Arthropod symbioses: from fundamental studies to pest and disease mangement. The full contents of the supplement are available online at http://0-www.biomedcentral.com.brum.beds.ac.uk/1471-2180/12?issue=S1.
- Jacobson MD, Weil M, Raff MC: Programmed cell death in animal development. Cell. 1997, 88: 347-354. 10.1016/S0092-8674(00)81873-5.View ArticlePubMedGoogle Scholar
- Shen J, Tower J: Programmed cell death and apoptosis in aging and life span regulation. Discov Med. 2009, 8 (43): 223-226.PubMedGoogle Scholar
- Kerr JF, Wyllie AH, Currie AR: Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer. 1972, 26: 239-257. 10.1038/bjc.1972.33.PubMed CentralView ArticlePubMedGoogle Scholar
- Taatjes DJ, Sobel BE, Budd RC: Morphological and cytochemical determination of cell death by apoptosis. Histochem Cell Biol. 2008, 129: 33-43. 10.1007/s00418-007-0356-9.PubMed CentralView ArticlePubMedGoogle Scholar
- Green DR, Reed JC: Mitochondria and apoptosis. Science. 1998, 281 (5381): 1309-1312.View ArticlePubMedGoogle Scholar
- McCall K: Eggs over easy: cell death in the Drosophila ovary. Dev Biol. 2004, 274: 3-14. 10.1016/j.ydbio.2004.07.017.View ArticlePubMedGoogle Scholar
- Aitken RJ, Findlay JK, Hutt KJ, Kerr JB: Apoptosis in the germ line. Reproduction. 2011, 141: 139-150. 10.1530/REP-10-0232.View ArticlePubMedGoogle Scholar
- Drummond-Barbosa D, Spradling AC: Stem cells and their progeny respond to nutritional changes during Drosophila oogenesis. Dev.Biol. 2001, 231: 265-278. 10.1006/dbio.2000.0135.View ArticlePubMedGoogle Scholar
- Giorgi F, Deri P: Cell death in ovarian chambers of Drosophila melanogaster. J Embryol Exp Morphol. 1976, 35: 521-533.PubMedGoogle Scholar
- Panagopoulos DJ, Chavdoula ED, Nezis IP, Margaritis LH: Cell death induced by GSM 900-MHz and DCS 1800-MHz mobile telephony radiation. Mutat Res. 2007, 626: 69-78.View ArticlePubMedGoogle Scholar
- Nezis IP, Stravopodis DJ, Papassideri I, Robert-Nicoud M, Margaritis LH: Stage-specific apoptotic patterns during Drosophila oogenesis. Eur J Cell Biol. 2000, 79 (9): 610-620. 10.1078/0171-9335-00088.View ArticlePubMedGoogle Scholar
- Foley K, Cooley L: Apoptosis in late stage Drosophila nurse cells does not require genes within the H99 deficiency. Development. 1998, 125: 1075-1082.PubMedGoogle Scholar
- Velentzas AD, Nezis IP, Stravopodis DJ, Papassideri IS, Margaritis LH: Apoptosis and autophagy function cooperatively for the efficacious execution of programmed nurse cell death during Drosophila virilis oogenesis. Autophagy. 2007, 3 (2): 130-132.View ArticlePubMedGoogle Scholar
- Nezis IP, Lamark T, Velentzas AD, Rusten TE, Bjørkøy G, Johansen T, Papassideri IS, Stravopodis DJ, Margaritis LH, Stenmark H, Brech A: Cell death during Drosophila melanogaster early oogenesis is mediated through autophagy. Autophagy. 2009, 5 (3): 298-302. 10.4161/auto.5.3.7454.View ArticlePubMedGoogle Scholar
- Kroemer G: Mitochondrial implication in apoptosis. Towards an endosymbiont hypothesis of apoptosis evolution. Cell Death Differ. 1997, 4: 443-456. 10.1038/sj.cdd.4400266.View ArticlePubMedGoogle Scholar
- James ER, Green DR: Manipulation of apoptosis in the host-parasite interaction. Trends Parasitol. 2004, 20 (6): 280-287. 10.1016/j.pt.2004.04.004.View ArticlePubMedGoogle Scholar
- Faherty CS, Maurelli AT: Staying alive: bacterial inhibition of apoptosis during infection. Trends Microbiol. 2008, 16 (4): 173-180. 10.1016/j.tim.2008.02.001.PubMed CentralView ArticlePubMedGoogle Scholar
- Lancellotti M, Pereira RF, Cury GG, Hollanda LM: Pathogenic and opportunistic respiratory bacteria-induced apoptosis. Braz J Infect Dis. 2009, 13 (3): 226-231. 10.1590/S1413-86702009000300014.View ArticlePubMedGoogle Scholar
- Yen JH, Barr AR: New hypothesis of the cause of cytoplasmic incompatibility in Culex pipiens. Nature. 1971, 232 (5313): 657-658. 10.1038/232657a0.View ArticlePubMedGoogle Scholar
- Werren JH, Baldo L, Clark ME: Wolbachia: master manipulators of invertebrate biology. Nat Rev Microbiol. 2008, 6 (10): 741-751. 10.1038/nrmicro1969.View ArticlePubMedGoogle Scholar
- Riegler M, Sidhu M, Miller WJ, O'Neill SL: Evidence for a global Wolbachia replacement in Drosophila melanogaster. Curr Biol. 2005, 15 (15): 1428-1433. 10.1016/j.cub.2005.06.069.View ArticlePubMedGoogle Scholar
- Ilinsky YuYu, Zakharov IK: The endosymbiont Wolbachia in Eurasian populations of Drosophila melanogaster. Russ J Genet. 2007, 43 (7): 748-756. 10.1134/S102279540707006X.View ArticleGoogle Scholar
- Min KT, Benzer S: Wolbachia, normally a symbiont of Drosophila, can be virulent, causing degeneration and early death. Proc Natl Acad Sci U S A. 1997, 94: 10792-10796. 10.1073/pnas.94.20.10792.PubMed CentralView ArticlePubMedGoogle Scholar
- Dedeine F, Vavre F, Fleury F, Loppin B, Hochberg ME, Boulétreau M: Removing symbiotic Wolbachia bacteria specifically inhibits oogenesis in a parasitic wasp. Proc Natl Acad Sci U S A. 2001, 98 (11): 6247-6252. 10.1073/pnas.101304298.PubMed CentralView ArticlePubMedGoogle Scholar
- Pannebakker BA, Loppin B, Elemans CPH, Humblot L, Vavre F: Parasitic inhibition of cell death facilitates symbiosis. Proc Natl Acad Sci U S A. 2007, 104 (1): 213-215. 10.1073/pnas.0607845104.PubMed CentralView ArticlePubMedGoogle Scholar
- Frydman HM, Li JM, Robson DN, Wieschaus E: Somatic stem cell niche tropism in Wolbachia. Nature. 2006, 441: 509-512. 10.1038/nature04756.View ArticlePubMedGoogle Scholar
- King RC, Rubinson AC, Smith AF: Oogenesis in adult Drosophila melanogaster. Growth. 1956, 20: 121-157.PubMedGoogle Scholar
- Dansereau DA, McKearin D, Lasko P: Oogenesis. Comprehensive Molecular Insect Science. Edited by: Gilbert LI, Iatrou K, Gill SS. 2004, Oxford, Pergamon, 1: Reproduction and Development: 39-85.Google Scholar
- Smith JE, Cummings CA, Cronmiller C: daughterless coordinates somatic cell proliferation, differentiation and germline cyst survival during follicle formation in Drosophila. Development. 2002, 129: 3255-3267.PubMedGoogle Scholar
- D'Herde K, De Prest B, Mussche S, Schotte P, Beyaert R, Coster RV, Roels F: Ultrastructural localization of cytochrome c in apoptosis demonstrates mitochondrial heterogeneity. Cell Death Differ. 2000, 7: 331-337. 10.1038/sj.cdd.4400655.View ArticlePubMedGoogle Scholar
- Brajušković GR, Škaro-Milić AB, Marjanović SA, Cerović SJ, Knežević-Ušaj SF: The ultrastructural investigation of mitochondria in B-CLL cells during apoptosis. Arch Oncol. 2004, 12 (3): 139-141. 10.2298/AOO0403139B.View ArticleGoogle Scholar
- Houwerzijl EJ, Blom NR, van der Want JJ, Esselink MT, Koornstra JJ, Smit JW, Louwes H, Vellenga E, de Wolf JT: Ultrastructural study shows morphologic features of apoptosis and para-apoptosis in megakaryocytes from patients with idiopathic thrombocytopenic purpura. Blood. 2004, 103 (2): 500-506. 10.1182/blood-2003-01-0275.View ArticlePubMedGoogle Scholar
- Reed JC, Green DR: Remodeling for demolition: changes in mitochondrial ultrastructure during apoptosis. Mol Cell. 2002, 9 (1): 1-3. 10.1016/S1097-2765(02)00437-9.View ArticlePubMedGoogle Scholar
- Dudkina NV, Voronin DA, Kiseleva EV: Structural organization and distribution of symbiotic bacteria Wolbachia in early embryos and ovaries of Drosophila melanogaster and D. simulans. Tsitologiia. 2004, 46 (3): 208-220.PubMedGoogle Scholar
- Zhukova MV, Voronin DA, Kiseleva EV: High temperature initiates changes in Wolbachia ultrastructure in ovaries and early embryos of Drosophila melanogaster. Cell and Tissue Biology. 2008, 2 (5): 546-556. 10.1134/S1990519X08050131.View ArticleGoogle Scholar
- Ghedin E, Hailemariam T, DePasse J, Zhang X, Oksov Y, Unnasch TR, Lustigman S: Brugia malayi gene expression in response to the targeting of the Wolbachia endosymbiont by tetracycline treatment. PLoS Negl Trop Dis. 2009, 3 (10): e525-10.1371/journal.pntd.0000525.PubMed CentralView ArticlePubMedGoogle Scholar
- Wright JD, Barr AR: The ultrastructure and symbiotic relationships of Wolbachia of mosquitoes of the Aedes scutellaris group. J Ultrastruct Res. 1980, 72: 52-64. 10.1016/S0022-5320(80)90135-5.View ArticlePubMedGoogle Scholar
- Raben N, Shea L, Hill V, Plotz P: Monitoring autophagy in lysosomal storage disorders. Methods Enzymol. 2009, 453: 417-449.PubMed CentralView ArticlePubMedGoogle Scholar
- Mahowald AP, Strassheim JM: Intercellular migration of centrioles in the germarium of Drosophila melanogaster. An electron microscopic study. J Cell Biol. 1970, 45 (2): 306-20. 10.1083/jcb.45.2.306.PubMed CentralView ArticlePubMedGoogle Scholar
- Megraw TL, Kaufman TC: The centrosome in Drosophila oocyte development. Curr Top Dev Biol. 2000, 49: 385-407.View ArticlePubMedGoogle Scholar
- Ferree PM, Frydman HM, Li JM, Cao J, Wieschaus E, Sullivan W: Wolbachia utilizes host microtubules and Dynein for anterior localization in the Drosophila oocyte. PLoS Pathog. 2005, 1 (2): e14-10.1371/journal.ppat.0010014.PubMed CentralView ArticlePubMedGoogle Scholar
- Li L, Xie T: Stem cell niche: structure and function. Annu Rev Cell Dev Biol. 2005, 21: 605-631. 10.1146/annurev.cellbio.21.012704.131525.View ArticlePubMedGoogle Scholar
- Reynolds KT, Thomson LJ, Hoffmann AA: The effects of host age, host nuclear background and temperature on phenotypic effects of the virulent Wolbachia strain popcorn in Drosophila melanogaster. Genetics. 2003, 164: 1027-1034.PubMed CentralPubMedGoogle Scholar
- Voronin DA, Bocherikov AM, Baricheva EM, Zakharov IK, Kiseleva EV: Action of genotypical surrounding of host Drosophila melanogaster on biological effects of endosymbiont Wolbachia (strain wMelPop). Cell and Tissue Biology. 2009, 3 (3): 263-273. 10.1134/S1990519X09030080.View ArticleGoogle Scholar
- Braig HR, Zhou W, Dobson SL, O'Neill SL: Cloning and characterization of a gene encoding the major surface protein of the bacterial endosymbiont Wolbachia pipientis. J Bacteriol. 1998, 180 (9): 2373-2378.PubMed CentralPubMedGoogle Scholar
- Mpoke SS, Wolfe J: Differential staining of apoptotic nuclei in living cells: application to macronuclear elimination in Tetrahymena. J Histochem Cytochem. 1997, 45 (5): 675-683. 10.1177/002215549704500505.View ArticlePubMedGoogle Scholar
- Abrams JM, White K, Fessler LI, Steller H: Programmed cell death during Drosophila embryogenesis. Development. 1993, 117: 29-43.PubMedGoogle Scholar
- Gold R, Schmied M, Giegerich G, Breitschopf H, Hartung HP, Toyka KV, Lassmann H: Differentiation between cellular apoptosis and necrosis by the combined use of in situ tailing and nick translation techniques. Lab Invest. 1994, 71: 219-225.PubMedGoogle Scholar
- Terasaki M, Runft L, Hand AR: Changes in organization of the endoplasmic reticulum during Xenopus oocyte maturation and activation. Mol Biol Cell. 2001, 12: 1103-1116.PubMed CentralView ArticlePubMedGoogle Scholar
This article is published under license to BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.