Name: saliva, salivary gland, and hemolymph collection from ixodes scapularis ticks
Uploaded: 2012-02-21T13:00:00
Description: Watch this Scientific Journal Video about Saliva, Salivary Gland, and Hemolymph Collection from Ixodes scapularis Ticks at JoVE.com

Forfatterne vil gerne takke afdelingen for vektorbårne sygdomme Animal Resources Branch, især Andrea Peterson, Lisa Massoudi, Verna O&#39;Brien, og John Liddell for deres pleje og vedligeholdelse af mus og kaniner. Vi vil også gerne takke Amy Ullmann, Theresa Russell, og Barbara J. Johnson for deres bidrag til at dette manuskript. Endelig vil vi gerne anerkende, Alissa Eckert i kontoret for Associate Director for Kommunikation på CDC for at producere de grafiske illustrationer og Judy Lavelle for at lede alle de Betegnelser i forbindelse med optagelserne af dette manuskript.

1. Hæmolymfe kollektion for præparatet (Movie 1) <ol><li> Forsigtigt fjernes aktivt fodring blodmider fra et dyr og placeres i 3% topisk hydrogenperoxid i 5 minutter og derefter i 70% ethanol i 10 minutter til overfladen sterilisere. </li><li> Med en pap pen tegne en cirkel på en silan belagt objektglas og placere kryds i pap pen cirkel. <ol><li> Silan coatede objektglas anvendes til bedst vedhæftning af hæmolymfe til mikroskopobjektglasset. </li></ol></li><li> Se kryds under et disse...

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Indsamlingen af tick hæmolymfe, spytkirtler, og spyt er vigtig i studiet af skovflåtbåren patogen transmission, udbredelse, formidling, spredning og persistens i både kryds og værten 6,11-13,20,23,29. Der er flere måder at dissekere en flåt 30,31. Men i forbindelse med indsamling spytkirtler er det afgørende at dissekere den skovflåten ordentligt, så spytkirtlerne ikke brydes eller tabt i skovflåten resterne. Når de spytkirtlerne fjernes fra blodmiden de skal vaskes adskillige gange for...

<table border="1"><tbody><tr><td> Reagens </td><td> Firma </td><td> Katalognr </td></tr><tr><td> Hydrogenperoxid </td><td> Fisher </td><td> H312-500 </td></tr><tr><td> Ethanol </td><td> Acros </td><td> 61509-5000 </td></tr><tr><td> PBS </td><td> Boston Bioproducts </td><td> BM-2205 </td></tr><tr><td> Dumont fine tænger (3C) </td><td> Fisher </td><td> NC9906085 </td></tr><tr><td> Silanbehandlede mikroskopobjektglas </td><td> Bioworld </td><td> 42763007-1 </td></tr><tr><td> Pap pen </td><td> Bioworld </td><td> 21750008-1 </td></tr><tr><td> Super frost plus mikroskopobjektglas </td><td> Fisher </td><td> 12-550-18 </td></tr><tr><td> Pilocarpin </td><td> Sigma </td><td> P6503-5G </td></tr><tr><td> Proteaseinhibitor-cocktail </td><td> Sigma </td><td> P2714 </td></tr><tr><td> # 11 engangs skalpel </td><td> Feather </td><td> 2975 # 11 </td></tr><tr><td> Ugiftige modellervoks </td><td&gt; Fisher </td><td> S17307 </td></tr><tr><td> Kapillarrør </td><td> Chase videnskabelige Glas, inc </td><td> 40A502 </td></tr></tbody></table>

saliva, salivary gland, and hemolymph collection from ixodes scapularis ticks

Flåter findes over hele verden og plage mennesker med mange kryds-bårne sygdomme. Flåter er vektorer for patogener, der forårsager borreliose og skovflåtbåren recidiverende feber (Borrelia spp.), Rocky Mountain spotted fever (Rickettsia rickettsii), ehrlichiosis (Ehrlichia chaffeensis og E. equi), anaplasmose (Anaplasma phagocytophilum), encephalitis (tick- borne encephalitis virus), babesiose (Babesia spp.), Colorado tick fever (Coltivirus) og tularæmi (Francisella tularensis) 1-8. For at blive ordentligt overføres til værten disse smitstoffer differentielt regulerer genekspression, interagere med tick proteiner, og vandrer gennem kryds 3,9-13. For eksempel, tilpasser Lyme sygdomsagensen, Borrelia burgdorferi, gennem differentiel genekspression til festen og hungersnød stadier af skovflåten har enzootisk cyklus 14,15. Som en Ixodes kryds forbruger enblodmel Borrelia replikere og migrere fra mellemtarmen i hemocoel, hvor de rejser til spytkirtlerne og transmitteres i værten med den udstødte spyt 9,16-19. Som et flueben feeds værten typisk reagerer med en stærk hæmostatisk og medfødte immunrespons 11,13,20-22. Til trods for disse værtsresponser, I. scapularis kan foder til flere dage, fordi tick spyt indeholder proteiner, der er immunmodulerende, lytiske agenter, antikoagulanter og fibrinolysins at hjælpe skovflåten fodring 3,11,20,21,23. De immunmodulerende aktiviteter besat af tick spyt eller spytkirtel ekstrakt (SGE) lette fremsendelsen, spredning og formidling af talrige tick patogener 3,20,24-27. For yderligere at forstå, hvordan skovflåtbåren smitstoffer forårsage sygdom er det vigtigt at dissekere aktivt fodring flåter og indsamle tick spyt. Videoen protokol viser dissektion teknikker tilindsamling af hæmolymfe og fjernelse af spytkirtler fra aktivt at fodre I. scapularis nymfer efter 48 og 72 timer efter mus placering. Vi har også demonstrere spyt samling fra en voksen kvinde I. scapularis tick.

Watch this Scientific Journal Video about Saliva, Salivary Gland, and Hemolymph Collection from Ixodes scapularis Ticks at JoVE.com

Saliva, Salivary Gland, and Hemolymph Collection from Ixodes scapularis Ticks

Indsamlingen af ​​inficeret skovflåt hæmolymfe, spytkirtler, og spyt er vigtigt at undersøge, hvordan skovflåtbåren patogener forårsage sygdom. I denne protokol vi vise, hvordan man indsamler hæmolymfe og spytkirtler fra fodring<em&gt; Ixodes scapularis</em&gt; Nymfer. Vi har også demonstrere spyt samling af kvindelige<em&gt; I. scapularis</em&gt; Voksne.

Spyt, spytkirtel, og hæmolymfe Collection fra Ixodes scapularis Flåter

Ticks are found worldwide and afflict humans with many tick-borne illnesses. Ticks are vectors for pathogens that cause Lyme disease and tick-borne ...

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Immunology and Infection

Saliva, Salivary Gland, and Hemolymph Collection from Ixodes scapularis Ticks

RNA Interference in Ticks

Ticks are obligate hematophagous ectoparasites of wild and domestic animals and humans, and are considered to be second worldwide to mosquitoes as vectors ...

Hybridization in situ of Salivary Glands, Ovaries, and Embryos of Vector Mosquitoes

Hybridization in situ of Salivary Glands, Ovaries, and Embryos of Vector Mosquitoes

Hybridisering<em&gt; In situ</em&gt; Af spytkirtler, æggestokke og embryoner af Vector Myg

Mosquitoes are vectors for a diverse set of pathogens including arboviruses, protozoan parasites and nematodes.  Investigation of transcripts and gene ...

Feeding of Ticks on Animals for Transmission and Xenodiagnosis in Lyme Disease Research

Transmission of the etiologic agent of Lyme disease, Borrelia burgdorferi, occurs by the  attachment and blood feeding of Ixodes species ticks on ...

Developing a Salivary Antibody Multiplex Immunoassay to Measure Human Exposure to Environmental Pathogens

Udvikling af en Spyt Antibody Multiplex Immunoassay at måle menneskelig eksponering for miljømæssige patogener

The etiology and impacts of human exposure to environmental pathogens are of major concern worldwide and, thus, the ability to assess exposure and ...

Rearing Ixodes scapularis, the Black-legged Tick: Feeding Immature Stages on Mice

Rearing Ixodes scapularis, the Black-legged Tick: Feeding Immature Stages on Mice

opdræt<em&gt; Ixodes scapularis,</em&gt; The Black-legged Tick: Feeding Immature Stages på Mus

Ixodes scapularis, the vector of Lyme disease, is one of the most important disease vectors in the eastern and Midwestern United States. This species is a ...

A Method for the Measurement of Salivary Gland Function in Mice

En metode til måling af spytkirtel funktion i mus

Patients with Sj&#246;gren&#39;s syndrome, an autoimmune disease affecting the exocrine glands, develop salivary gland inflammation and have reduced ...

Preparation of Murine Submandibular Salivary Gland for Upright Intravital Microscopy

Forberedelse af Murine submandibulære spytkirtel til opretstående Intravital mikroskopi

The submandibular salivary gland (SMG) is one of the three major salivary glands, and is of interest for many different fields of biological research, ...

Isolation of Salivary Epithelial Cells from Human Salivary Glands for In Vitro Growth as Salispheres or Monolayers

Isolering af spyt Epiteliale celler fra humane spytkirtler til in vitro-vækst som Salisfærer eller Monolayers

The salivary glands are a site of significant interest for researchers interested in multiple aspects of human disease. One goal of researchers is to ...

Purification of High Yield Extracellular Vesicle Preparations Away from Virus

Rensning af ekstracellulære vesikulære præparater med høj udbytte afstand fra virus

One of the major hurdles in the field of extracellular vesicle (EV) research today is the ability to achieve purified EV preparations in a viral infection ...

Disclosing Hemolymph Collection and Inoculation of Metarhizium Blastospores into Rhipicephalus Microplus Ticks Towards Invertebrate Pathology Studies

Disclosing Hemolymph Collection and Inoculation of Metarhizium Blastospores into Rhipicephalus Microplus Ticks Towards Invertebrate Pathology Studies

Videregivelse af hemolymph indsamling og inokulation af Metarhizium blastosporer til Rhipicephalus Microplus flåter mod Invertebrat patologiske studier

Ticks are obligate hematophagous ectoparasites and Rhipicephalus microplus has great importance in veterinary medicine because it causes anemia, weight ...