The effect of adipokinetic hormone (Pyrap-AKH) in stimulating the function of insect salivary glands (SGs) in extra-oral digestive processes was studied in the firebug, Pyrrhocoris apterus L. (Heteroptera). The analyses were performed on samples of SGs and extracts of linden seeds, a natural source of the bug's food. The SGs from 3-day old P. apterus females (when the food ingestion culminates), primarily contained polygalacturonase (PG) enzyme activity, whereas the level of lipase, peptidase, amylase and α-glucosidase was negligible. The transcription of PG mRNA and enzymatic activity were significantly increased in SGs after Pyrap-AKH treatment. The piercing and sucking of linden seeds by the bugs stimulated the intrinsic enzymatic cocktail of seeds (lipase, peptidase, amylase, glucosidase), and moreover the activity of these enzymes was significantly enhanced when the seeds were fed on by the Pyrap-AKH treated bugs. Similarly, a significant increase in PG activity was recorded in linden seeds fed on by hormonally-treated bugs or when injected by SG extract from hormonally treated ones as compared to untreated controls. The mechanism of AKH action in SGs is unknown, but likely involves cAMP (and excludes cGMP) as a second messenger, since the content of this compound doubled in SGs after Pyrap-AKH treatment. This new and as yet undescribed function of AKH in SGs is compared with the effect of this hormone on digestive processes in the midgut elucidated earlier.
- MeSH
- AMP cyklický metabolismus MeSH
- guanosinmonofosfát cyklický metabolismus MeSH
- Heteroptera fyziologie MeSH
- hmyzí hormony fyziologie MeSH
- kyselina pyrrolidonkarboxylová analogy a deriváty MeSH
- modely u zvířat * MeSH
- oligopeptidy fyziologie MeSH
- slinné žlázy enzymologie MeSH
- trávení MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
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- MeSH
- aspartátové proteasy genetika sekrece izolace a purifikace MeSH
- debridement * metody MeSH
- Diptera MeSH
- genová knihovna MeSH
- helmintoterapie MeSH
- hojení ran MeSH
- hybridizace in situ MeSH
- komplementární DNA genetika MeSH
- larva * enzymologie genetika růst a vývoj MeSH
- lidé MeSH
- metaloproteasy genetika izolace a purifikace sekrece MeSH
- nekróza terapie MeSH
- proteasy * genetika izolace a purifikace sekrece MeSH
- sekvenční analýza DNA MeSH
- serinové proteasy genetika sekrece izolace a purifikace MeSH
- slinné žlázy enzymologie MeSH
- trávicí systém enzymologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
BACKGROUND: In East Africa, Phlebotomus orientalis serves as the main vector of Leishmania donovani, the causative agent of visceral leishmaniasis (VL). Phlebotomus orientalis is present at two distant localities in Ethiopia; Addis Zemen where VL is endemic and Melka Werer where transmission of VL does not occur. To find out whether the difference in epidemiology of VL is due to distant compositions of P. orientalis saliva we established colonies from Addis Zemen and Melka Werer, analyzed and compared the transcriptomes, proteomes and enzymatic activity of the salivary glands. METHODOLOGY/PRINCIPAL FINDINGS: Two cDNA libraries were constructed from the female salivary glands of P. orientalis from Addis Zemen and Melka Werer. Clones of each P. orientalis library were randomly selected, sequenced and analyzed. In P. orientalis transcriptomes, we identified members of 13 main protein families. Phylogenetic analysis and multiple sequence alignments were performed to evaluate differences between the P. orientalis colonies and to show the relationship with other sand fly species from the subgenus Larroussius. To further compare both colonies, we investigated the humoral antigenicity and cross-reactivity of the salivary proteins and the activity of salivary apyrase and hyaluronidase. CONCLUSIONS: This is the first report of the salivary components of P. orientalis, an important vector sand fly. Our study expanded the knowledge of salivary gland compounds of sand fly species in the subgenus Larroussius. Based on the phylogenetic analysis, we showed that P. orientalis is closely related to Phlebotomus tobbi and Phlebotomus perniciosus, whereas Phlebotomus ariasi is evolutionarily more distinct species. We also demonstrated that there is no significant difference between the transcriptomes, proteomes or enzymatic properties of the salivary components of Addis Zemen (endemic area) and Melka Werer (non-endemic area) P. orientalis colonies. Thus, the different epidemiology of VL in these Ethiopian foci cannot be attributed to the salivary gland composition.
- MeSH
- enzymy chemie klasifikace genetika MeSH
- hmyz - vektory genetika MeSH
- leishmanióza viscerální imunologie MeSH
- molekulární sekvence - údaje MeSH
- myši inbrední BALB C MeSH
- myši MeSH
- Phlebotomus genetika MeSH
- sekvence aminokyselin MeSH
- sekvenční seřazení MeSH
- slinné proteiny a peptidy chemie klasifikace genetika imunologie MeSH
- slinné žlázy chemie enzymologie MeSH
- transkriptom genetika MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Intramural MeSH
- Geografické názvy
- Etiopie MeSH
To date, a single report has appeared on the use of salivary cortisol for adrenal function testing with a low dose ACTH, although 1 µg has become preferred as a more physiological stimulus than the commonly used 250 µg ACTH test. Our present study was aimed to obtain physiological data on changes of free salivary cortisol after 1 µg ACTH stimulation. This approach was compared with the common method based on the changes of total serum cortisol. Intravenous, lowdose ACTH test was performed in 15 healthy women (aged 22-40 years) with normal body weight, not using hormonal contraceptives, in the follicular phase of the menstrual cycle. Blood and saliva for determination of cortisol were collected before ACTH administration and 30 and 60 min after ACTH administration. Basal concentration of salivary cortisol (mean ± S.E.M., 15.9±1.96 nmol/l) increased after 1 µg ACTH to 29.1±2.01 nmol/l after 30 min, and to 27.4±2.15 nmol/l after 60 min. The differences between basal and stimulated values were highly significant (p<0.0001). The values of salivary cortisol displayed very little interindividual variability (p<0.04) in contrast to total serum cortisol values (p<0.0001) A comparison of areas under the curve (AUC) related to initial values indicated significantly higher AUC values for salivary cortisol than for total serum cortisol (1.89±0.88 vs. 1.22±0.19, p<0.01). Correlation analysis of serum and salivary cortisol levels showed a borderline relationship between basal levels (r=0.5183, p=0.0525); correlations after stimulation were not significant. Low-dose ACTH administration appeared as a sufficient stimulus for increasing salivary cortisol to a range considered as a normal adrenal functional reserve.
- MeSH
- adrenokortikální funkční testy metody využití MeSH
- adrenokortikotropní hormon diagnostické užití krev MeSH
- financování organizované využití MeSH
- hydrokortison aplikace a dávkování diagnostické užití krev MeSH
- menstruační cyklus MeSH
- slinné žlázy enzymologie účinky léků MeSH
- sliny enzymologie účinky léků MeSH
- transkortin diagnostické užití izolace a purifikace MeSH
- ženy MeSH
- MeSH
- finanční podpora výzkumu jako téma MeSH
- klíšťata imunologie parazitologie MeSH
- lidé MeSH
- lymfocyty imunologie MeSH
- slinné žlázy enzymologie imunologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- techniky in vitro MeSH
- MeSH
- druhová specificita MeSH
- hyaluronoglukosaminidasa metabolismus MeSH
- koncentrace vodíkových iontů MeSH
- Phlebotomus enzymologie MeSH
- slinné žlázy enzymologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- srovnávací studie MeSH
- MeSH
- alkalická fosfatasa analýza fyziologie MeSH
- laktace MeSH
- myši MeSH
- pohlavní dimorfismus MeSH
- slinné žlázy cytologie enzymologie růst a vývoj MeSH
- těhotenství MeSH
- vývojová biologie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- srovnávací studie MeSH
