Mechanistic understanding about the nature of cellular cryoinjury and mechanisms by which some animals survive freezing while others do not is currently lacking. Here, we exploited the broadly manipulable freeze tolerance of larval malt flies (Chymomyza costata) to uncover cell and tissue morphological changes associated with freeze mortality. Diapause induction, cold acclimation and dietary proline supplementation generate malt fly variants ranging from weakly to extremely freeze tolerant. Using confocal microscopy and immunostaining of the fat body, Malpighian tubules and anterior midgut, we described tissue and cytoskeletal (F-actin and α-tubulin) morphologies among these variants after exposure to various cold stresses (from chilling at -5°C to extreme freezing at -196°C), and upon recovery from cold exposure. Fat body tissue appeared to be the most susceptible to cryoinjury: freezing caused coalescence of lipid droplets, loss of α-tubulin structure and apparent aggregation of F-actin. A combination of diapause and cold acclimation substantially lowered the temperature at which these morphological disruptions occurred. Larvae that recovered from a freezing challenge repaired F-actin aggregation but not lipid droplet coalescence or α-tubulin structure. Our observations indicate that lipid coalescence and damage to α-tubulin are non-lethal forms of freeze injury, and suggest that repair or removal (rather than protection) of actin proteins is a potential mechanism of acquired freeze tolerance.
- Klíčová slova
- Actin, Chymomyza, Cytoskeleton, Diapause, Drosophilid, Freeze tolerance,
- MeSH
- aklimatizace * MeSH
- cytoskelet fyziologie MeSH
- Drosophilidae cytologie růst a vývoj fyziologie MeSH
- gastrointestinální trakt cytologie fyziologie MeSH
- larva cytologie růst a vývoj fyziologie MeSH
- malpighické trubice cytologie fyziologie MeSH
- nízká teplota škodlivé účinky MeSH
- tukové těleso cytologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Blastocrithidia papi is a unique trypanosomatid in that its life cycle is synchronized with that of its host, and includes an obligate stage of development in Malpighian tubules (MTs). This occurs in firebugs, which exited the winter diapause. In the short period, preceding the mating of overwintered insects, the flagellates penetrate MTs of the host, multiply attached to the epithelial surface with their flagella, and start forming cyst-like amastigotes (CLAs) in large agglomerates. By the moment of oviposition, a large number of CLAs are already available in the rectum. They are discharged on the eggs' surface with feces, used for transmission of bugs' symbiotic bacteria, which are compulsorily engulfed by the newly hatched nymphs along with the CLAs. The obligate development of B. papi in MTs is definitely linked to the life cycle synchronization. The absence of peristalsis allow the trypanosomatids to accumulate and form dense CLA-forming subpopulations, whereas the lack of peritrophic structures facilitates the extensive discharge of CLAs directly into the hindgut lumen. The massive release of CLAs associated with oviposition is indispensable for maximization of the infection efficiency at the most favorable time point.
- MeSH
- epitelové buňky parazitologie ultrastruktura MeSH
- feces parazitologie MeSH
- Hemiptera parazitologie ultrastruktura MeSH
- interakce hostitele a patogenu * MeSH
- kladení vajíček MeSH
- malpighické trubice parazitologie ultrastruktura MeSH
- stadia vývoje MeSH
- střeva parazitologie ultrastruktura MeSH
- Trypanosomatina růst a vývoj ultrastruktura MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
We describe the detection of sialylated N-linked glycans in partially fed Ixodes ricinus tick females using matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry. Sialylated glycans were detected in salivary glands as well as in tick guts and we propose the host origin of these structures. In addition, we mapped the transport of sialylated structures from the blood meal through the gut to the salivary glands using electron microscopy. Specific localization of sialylated glycans to basement membranes of salivary glands was observed. Finally, the influence of the sample preparation methods for electron microscopy on ultrastructure and immunogold labeling was evaluated.
- MeSH
- epitopy MeSH
- gastrointestinální trakt metabolismus MeSH
- imunohistochemie MeSH
- klíště metabolismus MeSH
- kyselina N-acetylneuraminová metabolismus MeSH
- malpighické trubice metabolismus MeSH
- polysacharidy metabolismus MeSH
- slinné žlázy metabolismus MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
- transmisní elektronová mikroskopie MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
- Názvy látek
- epitopy MeSH
- kyselina N-acetylneuraminová MeSH
- polysacharidy MeSH
The existing set of methods for assessing toxicity of noxas, based on experiments with whole animals (subclinical toxicity, toxicokinetics, carcinogenity, teratogenity, neurotoxicology etc.) does not provide much information about cellular and subcellular effects such compounds may exert. We suggest to complement the current methodology by combining a traditional morphological observation in an electron microscope with a spectroscopic method of electron microprobe X-ray analysis (or X-ray microanalysis). The latter makes it possible to measure concentrations of chemical elements in individual cells and organelles and effects of noxas can thus be assessed (i) at subcellular level, (ii) directly in situ and (iii) quantitatively. Concentrations of biologically important elements such as phosphorus, sulfur or zinc were measured in individual organelles in both intact and noxa-treated tissues, thus offering a possibility of comparing the effects of various noxious species at subcellular level (with the noxa previously applied to whole tissue or animal). The suggested correlation of analytical and morphological information may also provide new insights into cellular targeting of noxas (and potentially also drugs) as some organelles appear to be much more susceptible to damage than others.
- MeSH
- buňky účinky léků MeSH
- hmyz cytologie účinky léků MeSH
- insekticidy toxicita MeSH
- malpighické trubice účinky léků patologie MeSH
- mikroanalýza elektronovou sondou metody MeSH
- pyrany toxicita MeSH
- testy toxicity metody MeSH
- tukové těleso účinky léků patologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- insekticidy MeSH
- pyrany MeSH
- tetranactin MeSH Prohlížeč
Proteins of the Malpighian tubules (MT), midgut tissue (MG), salivary glands (SG), internal reproductive organs (RO), epidermis (EP), cerebral ganglion (CG), rectal ampulla (RA) and larval homogenate (LA) of Argas (Argas) polonicus were studied for their antigenicity and lecin affinity using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), immunoblotting, lectin affinoblotting and enzyme-linked lectin sorbentassay (ELLSA) techniques. A glycoprotein of 305 kDA was found in all tissues studied. All low molecular weight antigenic proteins recognized by anti-larval immune pigeon serum, except for one of 35 kDA, i.e. the 19-, 21-, 23-, 27-, 34-, and 46- kDa proteins, were shown to be glycoproteins. The glycosylation was shown to be N-linked in all of these antigens, but O-type glycosylation was also demonstrated in the 34-kDa glycoprotein. The correlation between the glycosylation and antigenicity of these proteins is also discussed.
- MeSH
- antigeny analýza MeSH
- epidermis chemie MeSH
- glykoproteiny analýza MeSH
- klíšťata analýza MeSH
- larva analýza MeSH
- lektiny analýza MeSH
- malpighické trubice chemie MeSH
- oligosacharidy analýza MeSH
- pohlavní orgány chemie MeSH
- slinné žlázy chemie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antigeny MeSH
- glykoproteiny MeSH
- lektiny MeSH
- oligosacharidy MeSH
Rickettsiella phytoseiuli naturally occurring in Phytoseiulus persimilis mites was cultivated in adult female Dermacentor reticulatus ticks. It demonstrates all six known developmental stages: dense, intermediate, bacterial, giant, crystal-forming and small dark particles. These stages of rickettsiae were found in salivary glands, Malpighian tubules, synganglion, ovaries, tracheal complex, haemolymph, fat body and alimentary tract. Rickettsiella phytoseiuli did not infect the Gené's organ. It multiplied in female ticks in a manner similar to that in the typical host mite, P. persimilis.
- MeSH
- Dermacentor mikrobiologie ultrastruktura MeSH
- elektronová mikroskopie MeSH
- hemocyty mikrobiologie MeSH
- klíšťata mikrobiologie MeSH
- malpighické trubice mikrobiologie MeSH
- nervový systém mikrobiologie MeSH
- ovarium mikrobiologie MeSH
- Rickettsiaceae růst a vývoj ultrastruktura MeSH
- slinné žlázy mikrobiologie MeSH
- střeva mikrobiologie MeSH
- trachea mikrobiologie MeSH
- tukové těleso mikrobiologie MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
(S)-9-(2,3-dihydroxypropyl) adenine (DHPA), D-eritadenine and some other open-chain nucleoside analogues, which exhibit adverse biological effects in microorganisms, plants and animals, cause pronounced inhibition of intestinal phosphatases in the hemipteran insect Pyrrhocoris apterus. The rate of p-nitrophenylphosphate hydrolysis by homogenates from intestinal epithelium and Malpighian tubules was inhibited up to 94% by 2-10 millimolar concentrations of these drugs. This effect is stronger than that of sodium fluoride, which is recognized as a common inhibitor of phosphatase. We conclude that inhibition of phosphatase activity in the digestive and excretory organs may be responsible for the previously reported massive excretion of phosphorylated derivatives of the nucleoside analogues after their oral administration to insects.
- MeSH
- adenin analogy a deriváty farmakologie MeSH
- alkalická fosfatasa antagonisté a inhibitory MeSH
- fosfatasy antagonisté a inhibitory MeSH
- Hemiptera enzymologie MeSH
- kyselá fosfatasa antagonisté a inhibitory MeSH
- malpighické trubice enzymologie MeSH
- nitrofenoly metabolismus MeSH
- organofosforové sloučeniny metabolismus MeSH
- střeva enzymologie MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- 9-(2,3-dihydroxypropyl)adenine MeSH Prohlížeč
- adenin MeSH
- alkalická fosfatasa MeSH
- eritadenine MeSH Prohlížeč
- fosfatasy MeSH
- kyselá fosfatasa MeSH
- nitrofenoly MeSH
- nitrophenylphosphate MeSH Prohlížeč
- organofosforové sloučeniny MeSH
The larva of Ch. costulata possesses seven paired glands which can be differentiated according to their position and histological structure. In five of them, the dynamics of their secretory or excretory activity may be studied histologically during the feeding process, when not only the deposition of secretion in the gland cells, but also the size of the whole gland is changed. The morphology of the remaining glands is not affected by feeding.
- MeSH
- larva cytologie MeSH
- malpighické trubice embryologie MeSH
- roztoči embryologie MeSH
- slinné žlázy embryologie MeSH
- střeva embryologie MeSH
- Trombiculidae embryologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- MeSH
- členovci - vektory * MeSH
- Coxiella růst a vývoj izolace a purifikace MeSH
- ganglia mikrobiologie MeSH
- hemolymfa mikrobiologie MeSH
- klíšťata * MeSH
- kůže mikrobiologie MeSH
- malpighické trubice mikrobiologie MeSH
- morčata MeSH
- ovarium mikrobiologie MeSH
- Q-horečka epidemiologie MeSH
- slinné žlázy mikrobiologie MeSH
- trávicí systém mikrobiologie MeSH
- zvířata MeSH
- Check Tag
- morčata MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
