Bu yayında bildirilen araştırma diş Ulusal Enstitüsü ve Fasiyal araştırma (NIDCR) ve ulusal sağlık Enstitüleri Ödülü numarası R56 DE025098, UG3 DE027695 ve F30 CA206296 altında Ulusal Kanser Enstitüsü (ncı) tarafından desteklenmiştir. İçeriği yalnızca yazarlar sorumludur ve mutlaka Ulusal Sağlık Enstitüleri resmi görüşlerini temsil etmiyor. Bu eser de NSF DMR 1206219 ve IADR yenilik Oral bakım Ödülü (2016) tarafından desteklenmiştir.
Dr. Eri Maruyama ve Andrew Hollomon tükürük koleksiyonu ile yardım için teşekkür etmek istiyorum. Pei-Lun Weng bezi diseksiyon ile yaptığı yardım için teşekkür etmek istiyorum. Matthew Ingalls şekil hazırlık yaptığı yardım için teşekkür etmek istiyorum. Dr. Elaine Smolock ve Emily Wu bu el yazması eleştirel okuma için teşekkür etmek istiyorum.

Aşağıda verilen tüm vivo içinde yordamlar hayvan kaynakları, Rochester Üniversitesi, Rochester Üniversitesi Komitesi tarafından kabul edildi. NY.
1. hazırlık
<ol><li>Bir analitik denge kullanarak, 20 mg Pilokarpin tartın. 2 ml steril serum microcentrifuge tüp içinde çözülür. Not: Pilokarpin ışığa duyarlı olduğunu ve zaman içinde etkinliğini kaybeder çünkü bu çözüm enjeksiyon gününün hazırlıklı olup yönetilen kadar ışıktan korunan.</li>
	<li>Bir analitik denge kullanarak, tartmak ve tüm koleksiyon tüpler ve cam kılcal tanımlayın. Koleksiyon tüpler ve cam kılcal fareler sayısı eşleşmelidir.</li>
</ol>2. trakeostomi
<ol><li>C57/BL6 fareler bir analitik denge kullanarak tartın.</li>
	<li>0.5 mL 29G x ½" iğne ile kullanarak, fareler 100 mg/kg ketamin ve 10 mg/kg xylazine intraperitoneally enjekte bir steril serum fizyolojik solüsyonu ile anestezi. Fareyi artık genellikle 5-10 dk içinde enjeksiyon takip oluşur (Yani yokluğu pedal çekilme refleks), uyaranlara yanıt verdiğinde aşağıdaki adıma geçin.</li>
	<li>Pamuk uçlu aplikatör üzerinde yağ dağıtımı sonra yavaşça gözlerini uygulamak ve fare sırtüstü pozisyonda Sahne Alanı'nda yer. İmzalat hayvan sıcak yüzey üzerinde tutulmalıdır. Not: Oda sıcaklığında tüm adımlar gerçekleştirilir.</li>
	<li>Burun, bacaklarda ve kuyruk cerrahi bant kullanarak sahne için güvenli.</li>
	<li>Temiz ve boyun bir alkol bezle ıslak. Not: Bu kürk sonraki insizyon girmesini önlemeye yardımcı olur.</li>
	<li>Ventral orta hat boyun deri Forseps ile yükselterek, diseksiyon makası kullanarak bir küçük, yüzeysel kesim yapmak. Makas içine açılış kılavuz ve subkutan doku uçaklar ayırmak için bıçaklar yavaş açılıyor.</li>
	<li>Kaldırdı cilt tutulması, dikkatle ağız aşağıda 1 cm kesin.</li>
	<li>İki yanal insizyon ilk kesim düşük kaliteli ve üstün yönleri olun. Forseps kullanarak, yavaşça uzakta baş ve boyun yapılarına erişim sağlamak için cilt yansıtmak.</li>
	<li>8 X büyütme oranında diseksiyon mikroskop kullanarak görselleştirmek submandibular bezleri (orta hat: şekil 1).</li>
	<li>İyi forseps kullanarak, bezleri Trakea örten dört infrahyoid (Askı) kasları ortaya çıkarmak için yavaşça kaldır. Yırtılma veya çevre damarlara veya boşaltım kanallarını engellemeden kaçının.</li>
	<li>Birden fazla fare tükürük toplama, adımları 2.1-2,10 her fare ile devam etmeden önce işlemini yineleyin. Not: Bu yordam güvenli bir şekilde en fazla 5 fareler aynı anda gerçekleştirilebilir.</li>
	<li>Küçük parçalara makasla kayış kasları medial kısmı kalırken orta hat gibi mümkün olduğunca yakın kaldırın. Sadece nefes borusu görselleştirmek ve kayış kasları yoldan işlem sırasında tutmak gerektiği kadar kesti. Önemli kanama için ikincil birim tükenmesi ise Pilokarpin salgı inducing etkili olmayacağı için herhangi bir yakın gemi nicking kaçının.</li>
	<li>Trakea uzak kayış kasları yansıtır.</li>
	<li>Larinks, trakea ve tiroid bezi görselleştirin. Trakea doku örten açık olduğundan emin olun.</li>
	<li>Yatay bir kesi borusunda inferior/arka küçük diseksiyon makası kullanarak tiroid olun. Hava yolu patent ve sıvı açık olduğundan emin olun.</li>
</ol>3. tükürük toplama
<ol><li>Ağız ve açı diseksiyon sahne aşağı yakınındaki bandı çıkarın (45°, cranially) tükürük akışı ile yardımcı olmak için.</li>
	<li>0.5 mL 29G x ½" iğne ile kullanarak, 10 µL/g vücut ağırlığı Pilokarpin mayi enjeksiyon toplam doz 100 mg/kg için gerçekleştirin.</li>
	<li>Bir sayacı başlatmak. Birden çok fareler dozaj hızlı hareket ve aynı anda kalan fareler enjekte bir asistan sahip sağlama süresini azaltın.</li>
	<li>Standart forseps kullanarak, kılcal tüp erişim için ağzını aç.</li>
	<li>Tükürük bir boncuk ağızda görülmektedir kez proksimal sonuna kılcal tüp distal ucu diseksiyon sahne (önceden tartılır her iki tüpler) altında konumlandırılmış bir toplama tüp içine yerleştirilmiş olan sıvı yerleştirin. Not: C57/BL6 dişi farelerde 6-10 hafta-in yaş, brüt salya Pilokarpin enjeksiyon 2 dk içinde oluşur.</li>
	<li>Tükürük ağız bina ancak tüpten akan değil, önceki adımı deneyin. Tüp giriş engel doğrudan bir Mukozal yüzey karşı yerleştirilmez emin olun.</li>
	<li>Tükürük toplam 12 dk Pilokarpin stimülasyon takip toplamak. Ağzının açık kalmasını sağlamak. Not: Bu dönemde artan gelgit hacmi ve salgıları (tükürük, idrar, dışkı) normaldir.</li>
	<li>Fareler tükürük işlevi depresif Eğer (örn., yaralanma veya müdahale nedeniyle), kalan sıvı P200 micropipette kullanarak toplama tüp ağzından aktarmak. Birden çok fareler toplama ipuçları değiştirilir olun.</li>
	<li>Kayıt toplanan tükürük artı borular Pilokarpin enjeksiyon sonra 12 dk ağırlığı.</li>
</ol>4. bezi diseksiyon
<ol><li>Fareler CO2 ötenazi tarafından ötenazi (ötenazi hayvanlar için travma kurallarda açıklandığı gibi: 2013 Edition) baş ve boyun yapıları korumak için dikkat çekici Torakotomi tarafından takip. Bu nedenle, ötenazi servikal çıkık bir yöntem olarak kaçının.</li>
	<li>Fare, Sahne Alanı'nda bir sırtüstü pozisyonda yerleştirin. Bezleri özgün sitedeki Trakea (şekil 1) yeniden konumlandırın.</li>
	<li>Diseksiyon mikroskop altında 8 X büyütme, kullanarak görselleştirmek büyük bezleri: PG, SMG ve SLG. Bezlerinin diseksiyon sırasında ayırt etmek için SMG ve SLG yanal yalan PG diffüz, tanımlayın. Not: az tanımlanmış ve parçalanmış olması büyük olasılıkla çünkü PG anatomi, önce yapılması gerekir.</li>
	<li>Forseps, temel yapılarını uzak çekerek ile PG kuyruğunun kavramak ve yavaşça gerginlik makas diseksiyon ile PG kafa kesme önce uygulayın.</li>
	<li>İhlali forseps kullanarak SMGs çevreleyen kapsül. Yavaşça sol ve sağ SMG ayırın.</li>
	<li>Forseps kullanarak yapıştırılan nonglandular dokusundan SMG dorsal yönünü ücretsiz.</li>
	<li>SMG boynundan Wharton'ın kanal gergin Forseps ile çizerek ve makas diseksiyon ile kanal kesme kaldırın.</li>
	<li>Yavaşça SLG forseps kullanarak SMG ayırın. Not: SMG superolateral yönü SLG var.</li>
	<li>Analitik bir denge, kayıt SMG ağırlıkları kullanarak.</li>
</ol>

<ol>
	<li>Bradley, P., O'Hara, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diseases+of+the+salivary+glands.">Diseases of the salivary glands.</a> Surgery (Oxford). 33 (12), 614-619 (2015).</li><li>Fox, P. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Salivary+enhancement+therapies.">Salivary enhancement therapies.</a> Caries Research. 38 (3), 241-246 (2004).</li><li>Konings, A. W. T., Coppes, R. P., Vissink, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=On+the+mechanism+of+salivary+gland+radiosensitivity.">On the mechanism of salivary gland radiosensitivity.</a> International Journal of Radiation Oncology Biology Physics. 62 (4), 1187-1194 (2005).</li><li>Burlage, F. R., Coppes, R. P., Meertens, H., Stokman, M. A., Vissink, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Parotid+and+submandibular/sublingual+salivary+flow+during+high+dose+radiotherapy.">Parotid and submandibular/sublingual salivary flow during high dose radiotherapy.</a> Radiotherapy and Oncology. 61 (3), 271-274 (2001).</li><li>Aure, M. H., Konieczny, S. F., Ovitt, C. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Salivary+gland+homeostasis+is+maintained+through+acinar+cell+self-duplication.">Salivary gland homeostasis is maintained through acinar cell self-duplication.</a> Developmental Cell. 33 (2), 231-237 (2015).</li><li>Aure, M. H., Arany, S., Ovitt, C. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Salivary+Glands:+Stem+Cells,+Self-duplication,+or+Both?.">Salivary Glands: Stem Cells, Self-duplication, or Both?.</a> Journal of Dental Research. 94 (11), 1502-1507 (2015).</li><li>Ono, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Distinct+effects+of+cevimeline+and+pilocarpine+on+salivary+mechanisms,+cardiovascular+response+and+thirst+sensation+in+rats.">Distinct effects of cevimeline and pilocarpine on salivary mechanisms, cardiovascular response and thirst sensation in rats.</a> Archives of Oral Biology. 57 (4), 421-428 (2012).</li><li>Proctor, G. B., Carpenter, G. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulation+of+salivary+gland+function+by+autonomic+nerves.">Regulation of salivary gland function by autonomic nerves.</a> Auton Neuroscience. 133 (1), 3-18 (2007).</li><li>Nezu, A., Morita, T., Tojyo, Y., Nagai, T., Tanimura, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Partial+agonistic+effects+of+pilocarpine+on+Ca(2+)+responses+and+salivary+secretion+in+the+submandibular+glands+of+live+animals.">Partial agonistic effects of pilocarpine on Ca(2+) responses and salivary secretion in the submandibular glands of live animals.</a> Experimental Physiology. 100 (6), 640-651 (2015).</li><li>Urita, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rebamipide+and+mosapride+enhance+pilocarpine-induced+salivation.">Rebamipide and mosapride enhance pilocarpine-induced salivation.</a> North American Journal of Medical Sciences. 1 (3), 121-124 (2009).</li><li>Arany, S., Benoit, D. S., Dewhurst, S., Ovitt, C. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nanoparticle-mediated+gene+silencing+confers+radioprotection+to+salivary+glands+in+vivo.">Nanoparticle-mediated gene silencing confers radioprotection to salivary glands in vivo.</a> Molecular Therapy. 21 (6), 1182-1194 (2013).</li><li>Kondo, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+differences+in+the+acinar+cells+of+the+murine+major+salivary+glands.">Functional differences in the acinar cells of the murine major salivary glands.</a> Journal of Dental Research. 94 (5), 715-721 (2015).</li><li>Evans, R. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Severe+impairment+of+salivation+in+Na+/K+/2Cl-+cotransporter+(NKCC1)-deficient+mice.">Severe impairment of salivation in Na+/K+/2Cl- cotransporter (NKCC1)-deficient mice.</a> Journal of Biological Chemistry. 275 (35), 26720-26726 (2000).</li><li>Delanian, S., Lefaix, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+radiation-induced+fibroatrophic+process:+therapeutic+perspective+via+the+antioxidant+pathway.">The radiation-induced fibroatrophic process: therapeutic perspective via the antioxidant pathway.</a> Radiotherapy and Oncology. 73 (2), 119-131 (2004).</li><li>Guchelaar, H. J., Vermes, A., Meerwaldt, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Radiation-induced+xerostomia:+pathophysiology,+clinical+course+and+supportive+treatment.">Radiation-induced xerostomia: pathophysiology, clinical course and supportive treatment.</a> Support Care Cancer. 5 (4), 281-288 (1997).</li><li>Lin, A. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measuring+short-term+&#947;-irradiation+effects+on+mouse+salivary+gland+function+using+a+new+saliva+collection+device.">Measuring short-term &#947;-irradiation effects on mouse salivary gland function using a new saliva collection device.</a> Archives of Oral Biology. 46 (11), 1085-1089 (2001).</li><li>Montenegro, M. F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Profound+differences+between+humans+and+rodents+in+the+ability+to+concentrate+salivary+nitrate:+Implications+for+translational+research.">Profound differences between humans and rodents in the ability to concentrate salivary nitrate: Implications for translational research.</a> Redox biology. 10, 206-210 (2016).</li><li>Choi, J. S., Park, I. S., Kim, S. K., Lim, J. Y., Kim, Y. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Morphometric+and+Functional+Changes+of+Salivary+Gland+Dysfunction+After+Radioactive+Iodine+Ablation+in+a+Murine+Model.">Morphometric and Functional Changes of Salivary Gland Dysfunction After Radioactive Iodine Ablation in a Murine Model.</a> Thyroid. 23 (11), 1445-1451 (2013).</li><li>Imamura, T. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhibition+of+pilocarpine-induced+saliva+secretion+by+adrenergic+agonists+in+ICR+mice.">Inhibition of pilocarpine-induced saliva secretion by adrenergic agonists in ICR mice.</a> Clinical and Experimental Pharmacology and Physiology. 39 (12), 1038-1043 (2012).</li><li>Ma, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Defective+Secretion+of+Saliva+in+Transgenic+Mice+Lacking+Aquaporin-5+Water+Channels.">Defective Secretion of Saliva in Transgenic Mice Lacking Aquaporin-5 Water Channels.</a> Journal of Biological Chemistry. 274 (29), 20071-20074 (1999).</li><li>Parkes, M. W., Parks, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Supersensitivity+of+salivation+in+response+to+pilocarpine+after+withdrawal+of+chronically+administered+hyoscine+in+the+mouse.">Supersensitivity of salivation in response to pilocarpine after withdrawal of chronically administered hyoscine in the mouse.</a> British Journal of Pharmacology. 46 (2), 315-323 (1972).</li><li>Nishiyama, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Up-Regulated+PAR-2-Mediated+Salivary+Secretion+in+Mice+Deficient+in+Muscarinic+Acetylcholine+Receptor+Subtypes.">Up-Regulated PAR-2-Mediated Salivary Secretion in Mice Deficient in Muscarinic Acetylcholine Receptor Subtypes.</a> Journal of Pharmacology and Experimental Therapeutics. 320 (2), 516 (2007).</li><li>Yang, B., Song, Y., Zhao, D., Verkman, A. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phenotype+analysis+of+aquaporin-8+null+mice.">Phenotype analysis of aquaporin-8 null mice.</a> American Journal of Physiology-Cell Physiology. 288 (5), C1161-C1170 (2005).</li><li>Kamiya, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=X-Ray-Induced+Damage+to+the+Submandibular+Salivary+Glands+in+Mice:+An+Analysis+of+Strain-Specific+Responses.">X-Ray-Induced Damage to the Submandibular Salivary Glands in Mice: An Analysis of Strain-Specific Responses.</a> BioResearch Open Access. 4 (1), 307-318 (2015).</li><li>Patel, R. M., Varma, S., Suragimath, G., Zope, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Estimation+and+Comparison+of+Salivary+Calcium,+Phosphorous,+Alkaline+Phosphatase+and+pH+Levels+in+Periodontal+Health+and+Disease:+A+Cross-sectional+Biochemical+Study.">Estimation and Comparison of Salivary Calcium, Phosphorous, Alkaline Phosphatase and pH Levels in Periodontal Health and Disease: A Cross-sectional Biochemical Study.</a> Journal of Clinical and Diagnostic Research. 10 (7), ZC58-ZC61 (2016).</li><li>Droebner, K., Sandner, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modification+of+the+salivary+secretion+assay+in+F508del+mice--the+murine+equivalent+of+the+human+sweat+test.">Modification of the salivary secretion assay in F508del mice--the murine equivalent of the human sweat test.</a> Journal of Cystic Fibrosis. 12 (6), 630-637 (2013).</li><li>Lamy, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Changes+in+mouse+whole+saliva+soluble+proteome+induced+by+tannin-enriched+diet.">Changes in mouse whole saliva soluble proteome induced by tannin-enriched diet.</a> Proteome Science. 8 (1), 65 (2010).</li><li>Mahomed, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recent+advances+in+mucin+immunohistochemistry+in+salivary+gland+tumors+and+head+and+neck+squamous+cell+carcinoma.">Recent advances in mucin immunohistochemistry in salivary gland tumors and head and neck squamous cell carcinoma.</a> Oral Oncology. 47 (9), 797-803 (2011).</li><li>Kohlgraf, K. G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitation+of+SPLUNC1+in+saliva+with+an+xMAP+particle-based+antibody+capture+and+detection+immunoassay.">Quantitation of SPLUNC1 in saliva with an xMAP particle-based antibody capture and detection immunoassay.</a> Archives of Oral Biology. 57 (2), 197-204 (2012).</li><li>Maimets, M., Bron, R., de Haan, G., van Os, R., Coppes, R. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Similar+ex+vivo+expansion+and+post-irradiation+regenerative+potential+of+juvenile+and+aged+salivary+gland+stem+cells.">Similar ex vivo expansion and post-irradiation regenerative potential of juvenile and aged salivary gland stem cells.</a> Radiotherapy and Oncology. , (2015).</li><li>Lombaert, I. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rescue+of+salivary+gland+function+after+stem+cell+transplantation+in+irradiated+glands.">Rescue of salivary gland function after stem cell transplantation in irradiated glands.</a> PLoS One. 3 (4), e2063 (2008).</li></ol>

Yazarlar ifşa gerek yok.

Biz bezi yaralanma ve tedavi uygulanan Tükürük bezi işlevi değerlendirmek için multistep bir yöntem mevcut. Bizim yordamı trakeostomi, tükürük toplama ve bezi diseksiyon, her biri bir tümleşik çalışma Tükürük bezi Biyoloji destekleyen deneysel uygulamaları sahip içerir. Örneğin, fare trakeostomi ağız boşluğu tıkayan işlemler sırasında genel hava yolu yönetimi için kullanılmıştır.
Uygun diseksiyon ve trakeal kesi Pilokarpin için gerekli teşvik tükürük s...

Tükürük bezi bozuklukları sendromlar dysregulated veya Engelli salgı aşırı üretimi (sialorrhea) veya tükürük1eksik (xerostomia ve hyposalivation) içerir. Her iki durumda da, terapötik geliştirme2sonu hedefi doğru Tükürük bezi biyoloji anlayışımız iyileştirilmesi bir ilgi vardır.
Tükrük bezleri son derece radiosensitive organları ve genellikle kalıcı ağız kuruluğu (xerostomia)3,4' e lider baş ve boyun kanser radyoterapi sırasında hasar. Radiosensitive diğer dokular, aksine Ancak, Tükürük bezi Ciro oranı nispeten düşüktür ve salgı kaybı kötü anlaşılan5,6mekanizmadır. Bu benzersiz yaralanma ayarında doku rejenerasyonu ve radioprotection stratejileri tükürük işlevsel değerlendirme gerektirir. Deneysel, fare tükürük toplama radyasyon ve terapötik ajanlar bezi cevaben değerlendirilmesinde özellikle değerli bir araçtır.
Burada, performans ve uyarılmış tükürük salgısı Pilokarpin, güçlü muscarinic agonist7kullanarak miktarının bir yöntem mevcut. Pilokarpin bezi salgısının8,9ikna etmek için otonom sinir sistemini uyarır. Bu test uygun şekilde tamamlamak için bir trakeostomi fareyi bir patent hava yolu yordamı boyunca tutar emin olmak için ve boğulma ve aspirasyon biriken salgıları ağız boşluğu10üzerinden riskleri azaltmak için gereklidir.
Bu üç büyük tükürük bezleri kaldırılmasına sonuçlanan bir terminal, bir uygulamadır: parotid (PG), çene (SMG) ve hapı (SLG). Fonksiyonel çalışmalar, bezi ağırlık kaydedilir ve tükürük ölçüm11,12,13normalleştirmek için sıklıkla kullanılır. Bu veriler, neyin bezi Atrofi beklenen sonucu14,15 yaşında radyasyon çalışmalarda, özellikle önemlidir
Edebiyat nasıl uyarılmış tükürük salgısı açısından değişkenlik gerçekleştirilir ve16bildirdi. Örneğin, edebiyat yayılma alanı en az üç büyüklük17,18,19,20,21,22,23içinde doz Pilokarpin. Burada, bir en iyi duruma getirilmiş yüksek doz Pilokarpin protokolü ile yöntem yürütme hem teknikleri olarak adapte edilebilir (trakeostomi, tükürük toplama ve bezi diseksiyon) modüler bir platform sağlayan geliştirilmiş tekrarlanabilirlik amacıyla mevcut ihtiyacım vardı.
İletişim kuralı gösteri ek olarak 2 hafta fraksiyonlara radyasyon (4 doz 6.85 Gy) SMG bölgesine takip tükürük akışı temsilcisi fonksiyonel verileri içerir.

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			<td height="42" style="height:42px;width:219px;">Goat polyclonal anti-Nkcc1</td>
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			<td>Nkcc1 Antibody</td>
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			<td height="42" style="height:42px;width:219px;">DAPI (4&#39;,6-Diamidino-2-Phenylindole, Dihydrochloride)</td>
			<td style="width:128px;">Thermo Fisher Scientific</td>
			<td style="width:119px;">D1306</td>
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			<td style="width:128px;">BD</td>
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			<td>Syringe for intraperitoneal injection</td>
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murine salivary functional assessment via pilocarpine stimulation following fractionated radiation

Hyposalivation otoimmün reaksiyon Sjoegren Sendromu veya aşağıdaki radyasyon hasarı büyük tükürük bezleri için yaygın olarak görülmektedir. Bu gibi durumlarda, hastalığın patogenezinde ve etkili müdahaleler ile ilgili sorular kalır. Ekzokrin bezi biyoloji, disfonksiyon ve tedavi soruşturma için çok değerli tükürük bezlerinin işlevsel değerlendirme sağlar en iyi duruma getirilmiş bir tekniktir. Burada, Pilokarpin gerçekleştirmek için adım adım bir yaklaşım mevcut uyarılmış tükürük salgısı, trakeostomi ve üç büyük fare tükürük bezlerinin diseksiyon dahil olmak üzere. Biz de bu teknikler sırasında erişilen uygun fare baş ve boyun anatomisine ayrıntı. Ölçeklenebilir, böylece iş akışı verimliliği artırmak aynı anda, işlenmek üzere birden çok fareler için izin veren bir yaklaşımdır. Bu yöntemlerin her biri daha fazla alan içinde uygulamaları olan tekrarlanabilirlik geliştirmek hedefliyoruz. Tükürük toplama ek olarak, biz miktarının ve bu dokuların fonksiyonel kapasite normalleştirme için ölçümler tartışıyorlar. Temsilcisi veri 2 hafta fraksiyonlara radyasyon (4 doz 6.85 Gy) takip depresif Tükürük bezi fonksiyonu ile çene bezlerinden dahil edilir.

Yüksek doz Pilokarpin gerçekleştirme tükürük toplama uyardığında, aspirasyon önlemek için hava yolu korumak önemli veya oral kavite içinde salgıları den boğulma. Trakeostomi şematik (şekil 1) sağlanır. Trakeal kesi, stoma doku ve sıvı açık kalmalıdır.
Kılcal eylem tükürük toplama sırasında geliştirmek için fare başlarını aşağıya doğru 45 ° açıyla yerleşti...

Watch this Scientific Journal Video about Murine Salivary Functional Assessment via Pilocarpine Stimulation Following Fractionated Radiation at JoVE.com

Murine Salivary Functional Assessment via Pilocarpine Stimulation Following Fractionated Radiation

Fare tükürük işlevsel değerlendirme Pilokarpin stimülasyon aşağıdaki yolu ile radyasyon şeker

Biz tükürük toplama, fare trakeostomi ve üç büyük tükürük bezleri yalıtımı da dahil olmak üzere performans detaylı bir yaklaşım sunuyoruz.

Hyposalivation is commonly observed in the autoimmune reaction of Sj&#246;gren's syndrome or following radiation injury to the major salivary glands. In ...

murine-salivary-functional-assessment-via-pilocarpine-stimulation

Research

JoVE Journal

Medicine

11.0K Görüntüleme.  University of Rochester. Biz tükürük toplama, fare trakeostomi ve üç büyük tükürük bezleri yalıtımı da dahil olmak üzere performans detaylı bir yaklaşım sunuyoruz.

Video: Murine Salivary Functional Assessment via Pilocarpine Stimulation Following Fractionated Radiation

Cannulation of the Mouse Submandibular Salivary Gland via the Wharton's Duct

Severe salivary gland hypofunction is frequently found in patients with Sj&ouml;gren's syndrome and those who receiving therapeutic 
irradiation in their head and neck regions for cancer treatment. Both groups of patients experience symptoms such as xerostomia (dry mouth), dysphagia 
(impaired chewing and swallowing), severe dental caries, altered taste, oro-pharyngeal infections (candidiasis), mucositis, pain and discomfort.
One innovative approach of regenerative medicine for the treatment of salivary gland hypo-function is speculated in RS Redman, E Mezey 
et al. 2009: stem cells can be directly deposited by cannulation into the gland as a potent method in reviving the functions of the impaired organ. Presumably, 
the migrated foreign stem cells will differentiate into glandular cells to function as part of the host salivary gland. Also, this cannulation technique is an 
expedient and effective delivery method for clinical gene transfer application.
Here we illustrate the steps involved in performing the cannulation procedure on the mouse submandibular salivary gland via the Wharton's duct 
(Fig 1). C3H mice (Charles River, Montreal, QC, Canada) are used for this experiment, which have been kept under clean conventional conditions at 
the McGill University animal resource center. All experiments have been approved by the University Animal Care Committee and were in accordance with the 
guidelines of the Canadian Council on Animal Care.
For this experiment, a trypan blue solution is infused into the gland through the opening of the Wharton's duct using a 
insulin syringe with a 29-gauge needle encased inside a polyethylene tube. Subsequently, the mouse is dissected to show that the infusions migrated 
into the gland successfully.

A protocol for the cannulation of the mouse submandibular salivary gland via the Wharton's duct is described. For this experiment, the trypan blue solution is used as a dyer to demonstrate how this technique effectively delivers infusions into the targeted gland, and to suggest the reliability of this new approach as a potential clinical drug/cell therapy for the regeneration of salivary glands.

Wharton Kanalı üzerinden Fare Submandibular Tükürük Bezi kanülasyonu

Severe salivary gland hypofunction is frequently found in patients with Sj&ouml;gren's syndrome and those who receiving therapeutic 
irradiation in their ...

Tıp

A Murine Model of Muscle Training by Neuromuscular Electrical Stimulation

Neuromuscular electrical stimulation (NMES) is a common clinical modality that is widely used to restore1, maintain2 or enhance3-5 muscle functional capacity. Transcutaneous surface stimulation of skeletal muscle involves a current flow between a cathode and an anode, thereby inducing excitement of the motor unit and the surrounding muscle fibers. 
NMES is an attractive modality to evaluate skeletal muscle adaptive responses for several reasons. First, it provides a reproducible experimental model in which physiological adaptations, such as myofiber hypertophy and muscle strengthening6, angiogenesis7-9, growth factor secretion9-11, and muscle precursor cell activation12 are well documented. Such physiological responses may be carefully titrated using different parameters of stimulation (for Cochrane review, see 13). In addition, NMES recruits motor units non-selectively, and in a spatially fixed and temporally synchronous manner14, offering the advantage of exerting a treatment effect on all fibers, regardless of fiber type. Although there are specified contraindications to NMES in clinical populations, including peripheral venous disorders or malignancy, for example, NMES is safe and feasible, even for those who are ill and/or bedridden and for populations in which rigorous exercise may be challenging. 
Here, we demonstrate the protocol for adapting commercially available electrodes and performing a NMES protocol using a murine model. This animal model has the advantage of utilizing a clinically available device and providing instant feedback regarding positioning of the electrode to elicit the desired muscle contractile effect. For the purpose of this manuscript, we will describe the protocol for muscle stimulation of the anterior compartment muscles of a mouse hindlimb.

A murine model of neuromuscular electrical stimulation (NMES), a safe and inexpensive clinical modality, to the anterior compartment muscles is described. This model has the advantage of modifying a readily available clinical device for the purpose of eliciting targeted and specific muscle contractions in mice.

Nöromüsküler Elektriksel Stimülasyon tarafından Kas Eğitimi A Mürin Modeli

Neuromuscular electrical stimulation (NMES) is a common clinical modality that is widely used to restore1, maintain2 or enhance3-5 muscle functional ...

Collecting Saliva and Measuring Salivary Cortisol and Alpha-amylase in Frail Community Residing Older Adults via Family Caregivers

Salivary measures have emerged in bio-behavioral research that are easy-to-collect, minimally invasive, and relatively inexpensive biologic markers of stress. This article we present the steps for collection and analysis of two salivary assays in research with frail, community residing older adults-salivary cortisol and salivary alpha amylase. The field of salivary bioscience is rapidly advancing and the purpose of this presentation is to provide an update on the developments for investigators interested in integrating these measures into research on aging. Strategies are presented for instructing family caregivers in collecting saliva in the home, and for conducting laboratory analyses of salivary analytes that have demonstrated feasibility, high compliance, and yield quality specimens. The protocol for sample collection includes: (1) consistent use of collection materials; (2) standardized methods that promote adherence and minimize subject burden; and (3) procedures for controlling certain confounding agents. We also provide strategies for laboratory analyses include: (1) saliva handling and processing; (2) salivary cortisol and salivary alpha amylase assay procedures; and (3) analytic considerations.

We demonstrate: (1) procedures for collection of salivary samples in cognitive impaired older adults by family caregivers in the home setting, (2) procedures for measuring stress activity via salivary cortisol and alpha amylase, and (3) representative profiles. Protocols that allow researchers to study stress-linked processes advance our understanding of biological sensitivity and susceptibility.

Salivary measures have emerged in bio-behavioral research  that are easy-to-collect, minimally invasive, and relatively inexpensive  biologic markers of ...

Myocardial Infarction and Functional Outcome Assessment in Pigs

Introduction of newly discovered cardiovascular therapeutics into first-in-man trials depends on a strictly regulated ethical and legal roadmap. One important prerequisite is a good understanding of all safety and efficacy aspects obtained in a large animal model that validly reflect the human scenario of myocardial infarction (MI). Pigs are widely used in this regard since their cardiac size, hemodynamics, and coronary anatomy are close to that of humans. Here, we present an effective protocol for using the porcine MI model using a closed-chest coronary balloon occlusion of the left anterior descending artery (LAD), followed by reperfusion. This approach is based on 90 min of myocardial ischemia, inducing large left ventricle infarction of the anterior, septal and inferoseptal walls. Furthermore, we present protocols for various measures of outcome that provide a wide range of information on the heart, such as cardiac systolic and diastolic function, hemodynamics, coronary flow velocity, microvascular resistance, and infarct size. This protocol can be easily tailored to meet study specific requirements for the validation of novel cardioregenerative biologics at different stages (i.e. directly after the acute ischemic insult, in the subacute setting or even in the chronic MI once scar formation has been completed). This model therefore provides a useful translational tool to study MI, subsequent adverse remodeling, and the potential of novel cardioregenerative agents.

This protocol describes the porcine myocardial infarction (MI) model using a 90 min closed-chest coronary balloon occlusion of the left anterior descending artery (LAD), followed by reperfusion. Furthermore, the protocol for several outcome parameters, such as cardiac function, hemodynamics, microvascular resistance, and infarct size, are also presented.

Domuzlar miyokard enfarktüsü ve Fonksiyonel Sonuç Değerlendirmesi

Introduction of newly discovered cardiovascular therapeutics into first-in-man trials depends on a strictly regulated ethical and legal roadmap. One ...

Controlling Parkinson's Disease With Adaptive Deep Brain Stimulation

Adaptive deep brain stimulation (aDBS) has the potential to improve the treatment of Parkinson&#39;s disease by optimizing stimulation in real time according to fluctuating disease and medication state. In the present realization of adaptive DBS we record and stimulate from the DBS electrodes implanted in the subthalamic nucleus of patients with Parkinson&#39;s disease in the early post-operative period. Local field potentials are analogue filtered between 3 and 47 Hz before being passed to a data acquisition unit where they are digitally filtered again around the patient specific beta peak, rectified and smoothed to give an online reading of the beta amplitude. A threshold for beta amplitude is set heuristically, which, if crossed, passes a trigger signal to the stimulator. The stimulator then ramps up stimulation to a pre-determined clinically effective voltage over 250 msec and continues to stimulate until the beta amplitude again falls down below threshold. Stimulation continues in this manner with brief episodes of ramped DBS during periods of heightened beta power.
Clinical efficacy is assessed after a minimum period of stabilization (5 min) through the unblinded and blinded video assessment of motor function using a selection of scores from the Unified Parkinson&#39;s Rating Scale (UPDRS). Recent work has demonstrated a reduction in power consumption with aDBS as well as an improvement in clinical scores compared to conventional DBS. Chronic aDBS could now be trialed in Parkinsonism.

Adaptive deep brain stimulation (aDBS) is effective for Parkinson&#8217;s disease, improving symptoms and reducing power consumption compared to conventional deep brain stimulation (cDBS). In aDBS we track a local field potential biomarker (beta oscillatory amplitude) in real time and use this to control the timing of stimulation.

Adaptif Derin Beyin Stimülasyonu İle Parkinson Hastalığı kontrol

Adaptive deep brain stimulation (aDBS) has the potential to improve the treatment of Parkinson&#39;s disease by optimizing stimulation in real time ...

Diffuse Optical Spectroscopy for the Quantitative Assessment of Acute Ionizing Radiation Induced Skin Toxicity Using a Mouse Model

Acute skin toxicities from ionizing radiation (IR) are a common side effect from therapeutic courses of external beam radiation therapy (RT) and negatively impact patient quality of life and long term survival. Advances in the understanding of the biological pathways associated with normal tissue toxicities have allowed for the development of interventional drugs, however, current response studies are limited by a lack of quantitative metrics for assessing the severity of skin reactions. Here we present a diffuse optical spectroscopic (DOS) approach that provides quantitative optical biomarkers of skin response to radiation. We describe the instrumentation design of the DOS system as well as the inversion algorithm for extracting the optical parameters. Finally, to demonstrate clinical utility, we present representative data from a pre-clinical mouse model of radiation induced erythema and compare the results with a commonly employed visual scoring. The described DOS method offers an objective, high through-put evaluation of skin toxicity via functional response that is translatable to the clinical setting.

We present a diffuse optical spectroscopic (DOS) approach that provides quantitative optical biomarkers of skin response to radiation. We describe DOS instrumentation design, optical parameters extraction algorithms and the animal handling procedures required to yield representative data from a pre-clinical mouse model of radiation induced erythema.

Bir fare Modeli Kullanılarak Akut Radyasyondan Bağlı Cilt zehirlilik Kantitatif Değerlendirme optik tayf Diffüz

Acute skin toxicities from ionizing radiation (IR) are a common side effect from therapeutic courses of external beam radiation therapy (RT) and ...

Retroductal Submandibular Gland Instillation and Localized Fractionated Irradiation in a Rat Model of Salivary Hypofunction

Normal tissues that lie within the portals of radiation are inadvertently damaged. Salivary glands are often injured during head and neck radiotherapy. Irreparable cell damage results in a chronic loss of salivary function that impairs basic oral activities, and increases the risk of oral infections and dental caries. Salivary hypofunction and its complications gravely impact a patient's comfort. Current symptomatic management of the condition is ineffective, and newer therapies to assuage the condition are needed. 
Salivary glands are exocrine glands, which expel their secretions into the mouth via excretory ducts. Cannulation of these ducts provides direct access to the glands. Retroductal delivery of a contrast agent to major salivary glands is a routine out-patient procedure for diagnostic imaging. Using a similar procedure, localized treatment of the glands is feasible. However, performing this technique in preclinical studies with small animals poses unique challenges. In this study we describe the technique of retroductal administration in rat submandibular glands, a procedure that was refined in Dr. Bruce Baum's laboratory (NIH)1, and lay out a procedure for local gland irradiation.

Salivary gland hypofunction, a major adverse effect of head and neck radiotherapy diminishes a patient's quality of life. The demonstration of efficacy of new therapies in animal models is a prerequisite before clinical transition. This protocol describes retroductal administration and local irradiation of rat submandibular glands.

Tükürük hipofonksiyonunun bir Sıçan Modelinde Retroductal Submandibuler Bezi damlatma ve yerelleştirilmiş fraksiyonlara Işınlama

Normal tissues that lie within the portals of radiation are inadvertently damaged. Salivary glands are often injured during head and neck radiotherapy. ...

Electrophysiological Assessment of Murine Atria with High-Resolution Optical Mapping

Recent genome-wide association studies targeting atrial fibrillation (AF) have indicated a strong association between the genotype and electrophysiological phenotype in the atria. That encourages us to utilize a genetically-engineered mouse model to elucidate the mechanism of AF. However, it is difficult to evaluate the electrophysiological properties in murine atria due to their small size. This protocol describes the electrophysiological evaluation of atria using an optical mapping system with a high temporal and spatial resolution in Langendorff perfused murine hearts. The optical mapping system is assembled with dual high-speed complementary metal oxide semiconductor cameras and high magnification objective lenses, to detect the fluorescence of a voltage-sensitive dye and Ca2+ indicator. To focus on the assessment of murine atria, optical mapping is performed with an area of 2 mm &#215; 2 mm or 10 mm x 10 mm, with a 100 &#215; 100 resolution (20 &#181;m/pixel or 100 &#181;m/pixel) and sampling rate of up to 10 kHz (0.1 ms) at maximum. A 1-French size quadripolar electrode pacing catheter is placed into the right atrium through the superior vena cava avoiding any mechanical damage to the atrium, and pacing stimulation is delivered through the catheter. An electrophysiological study is performed with programmed stimulation including constant pacing, burst pacing, and up to triple extrastimuli pacing. Under a spontaneous or pacing rhythm, the optical mapping recorded the action potential duration, activation map, conduction velocity, and Ca2+ transient individually in the right and left atria. In addition, the programmed stimulation also determines the inducibility of atrial tachyarrhythmias. Precise activation mapping is performed to identify the propagation of the excitation in the atrium during an induced atrial tachyarrhythmia. Optical mapping with a specialized setting enables a thorough electrophysiological evaluation of the atrium in murine pathological models.

This protocol describes the electrophysiological evaluation of murine atria utilizing an optical mapping system with a high temporal and spatial resolution, including dual recordings of the membrane voltage and Ca2+ transient under programmed stimulation through a specialized electrode catheter.

Yüksek çözünürlüklü optik eşleştirmenin fare Atria Elektrofizyolojik değerlendirme

Recent genome-wide association studies targeting atrial fibrillation (AF) have indicated a strong association between the genotype and ...

Development and Functional Characterization of Murine Tolerogenic Dendritic Cells

The immune system operates by maintaining a tight balance between coordinating responses against foreign antigens and maintaining an unresponsive state against self-antigens as well as antigens derived from commensal organisms. The disruption of this immune homeostasis can lead to chronic inflammation and to the development of autoimmunity. Dendritic cells (DCs) are the professional antigen-presenting cells of the innate immune system involved in activating na&#239;ve T cells to initiate immune responses against foreign antigens. However, DCs can also be differentiated into TolDCs that act to maintain and promote T cell tolerance and to suppress effector cells contributing to the development of either autoimmune or chronic inflammation conditions. The recent advancement in our understanding of TolDCs suggests that DC tolerance can be achieved by modulating their differentiation conditions. This phenomenon has led to tremendous growth in developing TolDC therapies for numerous immune disorders caused due to break in immune tolerance. Successful studies in preclinical autoimmunity murine models have further validated the immunotherapeutic utility of TolDCs in the treatment of autoimmune disorders. Today, TolDCs have become a promising immunotherapeutic tool in the clinic for reinstating immune tolerance in various immune disorders by targeting pathogenic autoimmune responses while leaving protective immunity intact. Although an array of strategies has been proposed by multiple labs to induce TolDCs, there is no consistency in characterizing the cellular and functional phenotype of these cells. This protocol provides a step-by-step guide for the development of bone marrow-derived DCs in large numbers, a unique method used to differentiate them into TolDCs with a synthetic triterpenoid 2-cyano-3,12-dioxooleana-1,9-dien-28-oic acid-difluoro-propyl-amide (CDDO-DFPA), and the techniques used to confirm their phenotype, including analyses of essential molecular signatures of TolDCs. Finally, we show a method to assess TolDC function by testing their immunosuppressive response in vitro and in vivo in a preclinical model of multiple sclerosis.

Here, we present a protocol to develop and characterize tolerogenic dendritic cells (TolDCs) and evaluate their immunotherapeutic utility.

Geliştirme ve işlevsel fare Tolerogenic dendritik hücrelerin karakterizasyonu

The immune system operates by maintaining a tight balance between coordinating responses against foreign antigens and maintaining an unresponsive state ...

Radiation Treatment of Organotypic Cultures from Submandibular and Parotid Salivary Glands Models Key In Vivo Characteristics

Hyposalivation and xerostomia create chronic oral complications that decrease the quality of life in head and neck cancer patients who are treated with radiotherapy. Experimental approaches to understanding mechanisms of salivary gland dysfunction and restoration have focused on in vivo models, which are handicapped by an inability to systematically screen therapeutic candidates and efficiencies in transfection capability to manipulate specific genes. The purpose of this salivary gland organotypic culture protocol is to evaluate maximal time of culture viability and characterize cellular changes following ex vivo radiation treatment. We utilized immunofluorescent staining and confocal microscopy to determine when specific cell populations and markers are present during a 30-day culture period. In addition, cellular markers previously reported in in vivo radiation models are evaluated in cultures that are irradiated ex vivo. Moving forward, this method is an attractive platform for rapid ex vivo assessment of murine and human salivary gland tissue responses to therapeutic agents that improve salivary function.

Using three-dimensional organotypic cultures to visualize morphology and functional markers of salivary glands may provide novel insights into the mechanisms of tissue damage following radiation. Described here is a protocol to section, culture, irradiate, stain, and image 50&#8211;90 &#956;m thick salivary gland sections prior to and following exposure to ionizing radiation.

Submandibular ve parotis tükürük bezlerinden Organotipik kültürlerin radyasyon tedavisi anahtar In vivo özellikleri

Hyposalivation and xerostomia create chronic oral complications that decrease the quality of life in head and neck cancer patients who are treated with ...