Name: handling and treatment of male european eels (anguilla anguilla) for hormonal maturation and sperm cryopreservation
Uploaded: 2018-01-28T15:00:00
Description: Watch this Scientific Journal Video about Handling and Treatment of Male European Eels Anguilla anguilla for Hormonal Maturation and Sperm Cryopreservation at JoVE.com

Bu yayın Avrupa Birliği'nin ufuk 2020 araştırma tarafından finanse edildi ve yenilik programı Marie Sklodowska-Curie hibe sözleşmesi kapsamında hiçbir 642893 (IMPRESS), maliyet Office (maliyet eylem SK 1205, AQUAGAMETE) ve mükemmellik Araştırma Merkezi- 1476-4/2016/FEKUT.

Bu protokol için açıklanan Avrupa yılan balığı ile çalışmak için tüm yordamları hayvan deney Etik Komitesi, İspanyol yasa ve yönetmeliklere deneyler kontrol takip Universitat Politècnica de València tarafından kabul edildi ve canlı hayvan işlemler.
1. balık bakım
<ol><li>Avrupa yılan balığı bir araştırma tesisine getir ve onları bir devridaim sistemi ile 200 L aquaria koyardım. Termostatlar ve soğutucular bir sabit 20 ° C sıcaklık sürdürmek için kullanır...

<ol>
	<li>ICES_Working_Group_on_Eels. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Report of the 2011 Session of the Joint EIFAAC/ICES Working Group on Eels. , 246 (2011).</li><li>Moriarty, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=European+Catches+of+Elver+of+1928-1988.">European Catches of Elver of 1928-1988.</a> Int. Rev. Hydrobiol. 75 (6), 701-706 (1990).</li><li>Dekker, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+fractal+geometry+of+the+European+eel+stock.">The fractal geometry of the European eel stock.</a> ICES J. Mar. Sci. 57 (1), 109-121 (2000).</li><li>Jacoby, D., Gollock, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Anguilla anguilla. The IUCN red list of threatened species. Version 2014.2. , (2014).</li><li>Asturiano, J. 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=Effects+of+hCG+as+spermiation+inducer+on+European+eel+semen+quality.">Effects of hCG as spermiation inducer on European eel semen quality.</a> Theriogenology. 66 (4), 1012-1020 (2006).</li><li>P&#233;rez, 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=Induction+of+maturation+and+spermiation+in+the+male+European+eel:+assessment+of+sperm+quality+throughout+treatment.">Induction of maturation and spermiation in the male European eel: assessment of sperm quality throughout treatment.</a> J. Fish Biol. 57 (6), 1488-1504 (2000).</li><li>Gallego, V., 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=Study+of+the+effects+of+thermal+regime+and+alternative+hormonal+treatments+on+the+reproductive+performance+of+European+eel+males+(Anguilla+anguilla)+during+induced+sexual+maturation.">Study of the effects of thermal regime and alternative hormonal treatments on the reproductive performance of European eel males (Anguilla anguilla) during induced sexual maturation.</a> Aquaculture. 354, 7-16 (2012).</li><li>Asturiano, J. F., S&#248;rensen, S. R., P&#233;rez, L., Lauesen, P., Tomkiewicz, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=First+Production+of+Larvae+Using+Cryopreserved+Sperm:+Effects+of+Preservation+Temperature+and+Cryopreservation+on+European+Eel+Sperm+Fertilization+Capacity.">First Production of Larvae Using Cryopreserved Sperm: Effects of Preservation Temperature and Cryopreservation on European Eel Sperm Fertilization Capacity.</a> Reprod. Domestic Anim. 51 (4), 485-491 (2016).</li><li>Di Biase, A., 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=Co-treatment+with+androgens+during+artificial+induction+of+maturation+in+female+eel,+Anguilla+anguilla:+Effects+on+egg+production+and+early+development.">Co-treatment with androgens during artificial induction of maturation in female eel, Anguilla anguilla: Effects on egg production and early development.</a> Aquaculture. 479, 508-515 (2017).</li><li>Lokman, P., Wylie, M. J., Downes, M., Di Biase, A., Damsteegt, E. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Artificial+induction+of+maturation+in+female+silver+eels,+Anguilla+australis:+The+benefits+of+androgen+pre-treatment.">Artificial induction of maturation in female silver eels, Anguilla australis: The benefits of androgen pre-treatment.</a> Aquaculture. 437, 111-119 (2015).</li><li>Mylonas, C. C., Duncan, N. J., Asturiano, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hormonal+manipulations+for+the+enhancement+of+sperm+production+in+cultured+fish+and+evaluation+of+sperm+quality.">Hormonal manipulations for the enhancement of sperm production in cultured fish and evaluation of sperm quality.</a> Aquaculture. , (2016).</li><li>Ohta, H., Izawa, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diluent+for+cool+storage+of+the+Japanese+eel+(Anguilla+japonica)+spermatozoa.">Diluent for cool storage of the Japanese eel (Anguilla japonica) spermatozoa.</a> Aquaculture. 142 (1-2), 107-118 (1996).</li><li>Pe&#241;aranda, D. S., 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=European+Eel+Sperm+Diluent+for+Short-term+Storage.">European Eel Sperm Diluent for Short-term Storage.</a> Reprod. Domestic Anim. 45 (3), 407-415 (2010).</li><li>Ohta, H., Kagawa, H., Tanaka, H., Unuma, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Control+by+the+environmental+concentration+of+ions+of+the+potential+for+motility+in+Japanese+eel+spermatozoa.">Control by the environmental concentration of ions of the potential for motility in Japanese eel spermatozoa.</a> Aquaculture. 198 (3), 339-351 (2001).</li><li>Asturiano, J., 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=Media+and+methods+for+the+cryopreservation+of+European+eel+(Anguilla+anguilla)+sperm.">Media and methods for the cryopreservation of European eel (Anguilla anguilla) sperm.</a> Fish Physiol. Biochem. 28 (1), 501-502 (2003).</li><li>Asturiano, J., 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=Physio-chemical+characteristics+of+seminal+plasma+and+development+of+media+and+methods+for+the+cryopreservation+of+European+eel+sperm.">Physio-chemical characteristics of seminal plasma and development of media and methods for the cryopreservation of European eel sperm.</a> Fish Physiol. Biochem. 30 (3), 283-293 (2004).</li><li>Szab&#243;, 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=Cryopreservation+of+European+eel+(Anguilla+anguilla)+sperm+using+different+extenders+and+cryoprotectants.">Cryopreservation of European eel (Anguilla anguilla) sperm using different extenders and cryoprotectants.</a> Acta. Biol. Hung. 56 (1-2), 173-175 (2005).</li><li>M&#252;ller, 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=Cryopreservation+of+sperm+of+farmed+European+eel+Anguilla+anguilla.">Cryopreservation of sperm of farmed European eel Anguilla anguilla.</a> J World Aquac Soc. 35 (2), 225-231 (2004).</li><li>Pe&#241;aranda, D. S., P&#233;rez, L., Gallego, V., Jover, M., Asturiano, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improvement+of+European+eel+sperm+cryopreservation+method+by+preventing+spermatozoa+movement+activation+caused+by+cryoprotectants.">Improvement of European eel sperm cryopreservation method by preventing spermatozoa movement activation caused by cryoprotectants.</a> Cryobiology. 59 (2), 119-126 (2009).</li><li>Marco-Jim&#233;nez, 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=Cryopreservation+of+European+eel+(Anguilla+anguilla)+spermatozoa:+effect+of+dilution+ratio,+foetal+bovine+serum+supplementation,+and+cryoprotectants.">Cryopreservation of European eel (Anguilla anguilla) spermatozoa: effect of dilution ratio, foetal bovine serum supplementation, and cryoprotectants.</a> Cryobiology. 53 (1), 51-57 (2006).</li><li>Asturiano, J. 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=Effect+of+sperm+cryopreservation+on+the+European+eel+sperm+viability+and+spermatozoa+morphology.">Effect of sperm cryopreservation on the European eel sperm viability and spermatozoa morphology.</a> Reprod. Domestic Anim. 42 (2), 162-166 (2007).</li><li>Mordenti, M., Di Biase, A., Sirri, R., Modugno, S., Tasselli, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+Sexual+Maturation+in+Wild+Female+European+Eels+(Anguilla+anguilla)+in+Darkness+and+Light.">Induction of Sexual Maturation in Wild Female European Eels (Anguilla anguilla) in Darkness and Light.</a> Isr. J. Aquac. 64, (2012).</li><li>Ohta, H., 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=Artificial+induction+of+maturation+and+fertilization+in+the+Japanese+eel,+Anguilla+japonica.">Artificial induction of maturation and fertilization in the Japanese eel, Anguilla japonica.</a> Fish Physiol. Biochem. 17 (1), 163-169 (1997).</li><li>Tanaka, S., 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=Long-term+cryopreservation+of+sperm+of+Japanese+eel.">Long-term cryopreservation of sperm of Japanese eel.</a> J. Fish Biol. 60 (1), 139-146 (2002).</li><li>Gallego, V., 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=Subpopulation+pattern+of+eel+spermatozoa+is+affected+by+post-activation+time,+hormonal+treatment+and+the+thermal+regimen.">Subpopulation pattern of eel spermatozoa is affected by post-activation time, hormonal treatment and the thermal regimen.</a> Reprod. Fertil. Dev. 27 (3), 529-543 (2015).</li><li>Asturiano, J. F., Cabrita, E., Horv&#225;th, &. #. 1. 9. 3. ;. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Progress,+challenges+and+perspectives+on+fish+gamete+cryopreservation:+A+mini-review.">Progress, challenges and perspectives on fish gamete cryopreservation: A mini-review.</a> Gen. Comp. Endocrinol. , (2016).</li><li>Kime, D. 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=Computer-assisted+sperm+analysis+(CASA)+as+a+tool+for+monitoring+sperm+quality+in+fish.">Computer-assisted sperm analysis (CASA) as a tool for monitoring sperm quality in fish.</a> Comp. Biochem. Physiol. C Toxicol. Pharmacol. 130 (4), 425-433 (2001).</li><li>Rurangwa, E., Kime, D. E., Ollevier, F., Nash, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+measurement+of+sperm+motility+and+factors+affecting+sperm+quality+in+cultured+fish.">The measurement of sperm motility and factors affecting sperm quality in cultured fish.</a> Aquaculture. 234 (1-4), 1-28 (2004).</li><li>Verstegen, J., Iguer-Ouada, M., Onclin, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Computer+assisted+semen+analyzers+in+andrology+research+and+veterinary+practice.">Computer assisted semen analyzers in andrology research and veterinary practice.</a> Theriogenology. 57 (1), 149-179 (2002).</li><li>Magnotti, C., 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=Cryopreservation+and+vitrification+of+fish+semen:+a+review+with+special+emphasis+on+marine+species.">Cryopreservation and vitrification of fish semen: a review with special emphasis on marine species.</a> Rev. Aquacult. , (2016).</li><li>Dziewulska, K., Rzemieniecki, A., Czerniawski, R., Domaga&#322;a, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Post-thawed+motility+and+fertility+from+Atlantic+salmon+(Salmo+salar+L.)+sperm+frozen+with+four+cryodiluents+in+straws+or+pellets.">Post-thawed motility and fertility from Atlantic salmon (Salmo salar L.) sperm frozen with four cryodiluents in straws or pellets.</a> Theriogenology. 76 (2), 300-311 (2011).</li><li>Babiak, I., Bolla, S., Ottesen, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Suitable+methods+for+cryopreservation+of+semen+from+Atlantic+halibut,+Hippoglossus+hippoglossus+L.">Suitable methods for cryopreservation of semen from Atlantic halibut, Hippoglossus hippoglossus L.</a> Aquac. Int. 16 (6), 561-572 (2008).</li><li>M&#252;ller, 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=Japanese+eel+(Anguilla+japonica+Temminck+&amp;+Schlegel,+1846)+propagation+using+cryopreserved+sperm+samples.">Japanese eel (Anguilla japonica Temminck &amp; Schlegel, 1846) propagation using cryopreserved sperm samples.</a> J. Appl. Ichthyol. 33 (3), 550-552 (2017).</li></ol>

Bu iletişim kuralı için Avrupa yılan balığı olgunlaşma, işleme ve sperm dondurma tam işlemini açıklar. Burada açıklanan yetiştirme koşulları hızlı olgunlaşma ve yüksek kaliteli sperm bu tür 6,7,25yüksek hacimli üretim için en iyi durumda. Başarısı bu dondurma Protokolü ve potansiyel kullanımı çözdürme sonra gübreleme için taze sperm 26kalitesini büyük ölçüde bağl?...

Son 25 yılda Avrupa sahilde gelen Avrupa yılan balığı (Anguilla anguilla) sayısı % 90 1,2,3tarafından giderek düştü. Kirliliği, enfeksiyonlar, aşırı avlanma ve habitat imha da dahil olmak üzere bu köklü damla açıklamak çeşitli faktörler vardır. Bunların hepsi Uluslararası Birliği için doğa koruma (IUCN) listesi olarak kritik tehlikedeki 4Tarih Avrupa yılan balığı eklenmesi için önde gelen bu tür üzerinde derin bir etkisi olmuştur. Sonuç olarak, geliştirme teknikleri ve protokolleri esaret üreme için gerekli.
Avrupa yılan balığı olgunlaşma tutsak hormonal tedavi 5,6,7 tarafından elde edilir ama her iki cinsiyette gamet üretim 8eşitlemek zordur. Yılan balıkları 9,oogenesis hızlandırmak için yeni androjen implantlar geliştirilmesi göstermiştir10, dişilerde son olgunlaşma zamanlaması hala son derece değişken ve 11kontrol etmek zor olsa bile. Bu nedenle, kısa süreli depolama sperm 12,13,14 ve dondurma teknikleri gamet eşitleme gereksiz 8yapma üreme yönetimi için gerekli.
Avrupa yılan balığı sperm dondurma 2003 15,16beri geliştirmiştir. Çeşitli araştırmacılar Dimetil sülfoksit (DMSO) veya metanol cryoprotectants 16,17,18,19,20kullanarak başarılı iletişim kuralları tasarlanmış. Her ne kadar her iki iletişim kurallarını başarıyla kullanmış, DMSO ile cryopreserved çözdürülen sperm elde edilen hücre canlılığı metanol 20,21ile daha düşüktür. Ayrıca, yılan balığı sperm DMSO ile temas harekete geçirmek ve daha sıkıcı sperm manipülasyon 19gerektirir, bu nedenle metanol DMSO daha Avrupa yılan balığı sperm için daha uygun bir cryoprotectant.
Burada, en uygun kullanım ve Avrupa yılan balığı hormonal tedavi için protokol aşağıda açıklanacaktır. Buna ek olarak, metanol bir cryoprotectant ve bir iletişim kuralı bu türler içinde sperm kalitesinin değerlendirilmesi için en iyi Avrupa yılan balığı sperm dondurma iletişim kuralı'nı da açıklanan olacaktır.

<table border="0" cellpadding="0" cellspacing="0" width="538">
	<tbody>
		<tr height="19">
			<td height="19" style="height:19px;width:151px;">AQUACEN BENZOCA&#205;NA +D2A2:D24</td>
			<td style="width:128px;">AQUACEN</td>
			<td style="width:119px;">3394ESP</td>
			<td style="width:141px;">Benzocaine</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">NaCl</td>
			<td>Avantor&#160;</td>
			<td>3624-19</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Syringe</td>
			<td>BD Plastipak</td>
			<td>305501</td>
			<td>Syringe 1 mL</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">FC500</td>
			<td>Beckman Coulter</td>
			<td></td>
			<td>Flow cytometer</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Air pump 100</td>
			<td>EHEIM</td>
			<td>4011708370032</td>
			<td>Modified for suction</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Eppendorf 1.5</td>
			<td>Eppendorf</td>
			<td>175508</td>
			<td>Plastic tubes 1.5 mL</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Falcon tubes</td>
			<td>Falcon</td>
			<td>175747</td>
			<td>Plastic tubes 15 mL</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Flexible bucket</td>
			<td>Fiel</td>
			<td>1040101</td>
			<td>40 L, Black color.</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Ethanol</td>
			<td>Guinama SL</td>
			<td>Mg96270</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Cyopreservation straws&#160;</td>
			<td>IMV Technologies</td>
			<td>14550</td>
			<td>500 &#181;L straws</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Live/Dead Sperm Viability Kit</td>
			<td>Invitrogen</td>
			<td>L7011</td>
			<td>Cytometer Kit</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Modelling clay</td>
			<td>JOVI</td>
			<td>Art 30</td>
			<td>4 different colors</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Precision scale PCB</td>
			<td>KERN &#38; Sohn GmbH</td>
			<td>PCB35002</td>
			<td>Digital scale</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Saline solution Vitulia 0.9%</td>
			<td>Laboratorios Ern, S.A.</td>
			<td>C.N. 999790.8</td>
			<td>Saline solution</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Forceps</td>
			<td>Levantina de Lab. SL</td>
			<td>3710025</td>
			<td>30 cm metal forceps</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Scissors</td>
			<td>Levantina de Lab. SL</td>
			<td>3700014</td>
			<td>Scissors 14cm</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Ovitrelle (r-hCG)</td>
			<td>Merck SL</td>
			<td>EMEA/H/C/000320</td>
			<td>Human chorionic gonadotropin</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Nikon Eclipse 80i</td>
			<td>Nikon</td>
			<td></td>
			<td>Microscope</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">CaCl2</td>
			<td>Panreac Quimica SAU</td>
			<td>2112211210</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Na2SO4</td>
			<td>Panreac Quimica SAU</td>
			<td>3257091611</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">NaHCO3</td>
			<td>Panreac Quimica SAU</td>
			<td>1416381210</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">782M Camera</td>
			<td>Proiser</td>
			<td>782M</td>
			<td>Camera for CASA</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">ISAS v1</td>
			<td>Proiser</td>
			<td>ISAS v1</td>
			<td>CASA software</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">SpermTrack-10</td>
			<td>Proiser</td>
			<td>SpermTrack-10</td>
			<td>Countig chamber for CASA</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Beaker Pyrex 3L</td>
			<td>PYREX</td>
			<td>2110668</td>
			<td>Glass beaker 3L</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Flask Pyrex 250 GL-45</td>
			<td>PYREX</td>
			<td>21801365</td>
			<td>Glass flask 250 mL</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">KCl</td>
			<td>Scharlab SL</td>
			<td>PO02000500</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Methanol</td>
			<td>Scharlab SL</td>
			<td>ME03061000</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">MgCl2</td>
			<td>Scharlab SL</td>
			<td>MA00370500</td>
			<td></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">BSA</td>
			<td>Sigma Aldrich</td>
			<td>05482</td>
			<td>Bovine serum albumina</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Styrofoam box</td>
			<td></td>
			<td></td>
			<td>34x34x30 cm and 5cm thick</td>
		</tr>
	</tbody>
</table>

handling and treatment of male european eels (anguilla anguilla) for hormonal maturation and sperm cryopreservation

Son yıllarda birkaç araştırma grubu geliştirme ve iyileştirme Avrupa yılan balığı kullanım ve olgunlaşma için yeni iletişim kuralları üzerinde çalışıyoruz. Derecede-in henüz, insan Korionik gonadotropin (hCG) haftalık enjeksiyonlari tedavi sırasında birkaç hafta yüksek miktarlar yüksek kaliteli Sperm üreten, sadece 5-6 hafta sonra erkek maturate kanıtladılar. Buna ek olarak, sperm dondurma farklı Extender kullanarak iletişim kuralları, cryoprotectants ve soğutma ve kez çözme daha önce için Avrupa yılan balığı tarif edilmistir. İşte, Tanaka´s extender çözüm doğrudan veya dondurma, gübreleme için kullanılabilir çözümler ve dilutions farklı kullanımı gereksiz yapma göstermektedir. Gibi metanol düşük toksisite ve sperm etkinleştirmez Ayrıca, bir cryoprotectant olarak metanol kullanımı bu iletişim kuralı kullanımını kolaylaştırır. Sperm cryoprotectant eklenmesinden sonra hemen cryopreserved gerekmez ve uzun çözdürülen sonra kullanılabilir. Ayrıca, sperm motilitesi taze sperm daha düşük olmasına rağmen çözdürme sonra hala yüksektir. Bu çalışmanın amacı, Avrupa yılan balığı için en iyi kullanılabilir iletişim kuralı işleme, olgunlaşma ve sperm dondurma göstermektir.

Spermi ile % 70 veya daha yüksek, sperm motilitesi 18 yılan balıkları bu çalışma için seçildi. Çözdürme için taze sperm (Tablo 1 ve Şekil 2) insanlardan karşılaştırıldığında sonra sonuçları tüm kalite parametreleri bir azalma gösterdi. Hareketliliği sonuçları (yani ± S.E.M., n = 18) daha yüksek toplam hareketliliği ve ilerici bir hareketliliği toplam hareketliliği ve ilerici hareket sonrası tezcan sperm örn...

Watch this Scientific Journal Video about Handling and Treatment of Male European Eels Anguilla anguilla for Hormonal Maturation and Sperm Cryopreservation at JoVE.com

Handling and Treatment of Male European Eels Anguilla anguilla for Hormonal Maturation and Sperm Cryopreservation

İletişim kuralları Avrupa yılan balığı olgunlaşma ve sperm dondurma için son yıl içinde geliştirilmiştir. Bu makalede, en iyi Protokolü olgunlaşma ve cryoprotectant olarak metanol inducing için insan Korionik gonadotropin (hCG) kullanarak kullanılabilir.

Kullanım ve tedavi, erkek Avrupa yılan balığı (Anguilla anguilla) Hormonal olgunlaşma ve Sperm dondurma için

During the last years, several research groups have been working on the development and improvement of new protocols for the European eel handling and ...

Hi, my name is Juan Asturiano from the Polytechnic University of Valencia, and we are with some of our colleagues from the Szent Istv

Research

JoVE Journal

Developmental Biology

Handling and Treatment of Male European Eels (Anguilla anguilla) for Hormonal Maturation and Sperm Cryopreservation

Manipulation and In Vitro Maturation of Xenopus laevis Oocytes, Followed by Intracytoplasmic Sperm Injection, to Study Embryonic Development

Manipulation and In Vitro Maturation of Xenopus laevis Oocytes, Followed by Intracytoplasmic Sperm Injection, to Study Embryonic Development

Manipülasyon ve<em&gt; İn Vitro</emVe&gt; Olgunlaşma<em&gt; Xenopus laevis</emIntrasitoplazmik sperm enjeksiyonu izledi&gt; oositler, Embriyonik Geliştirme Çalışması

Amphibian eggs have been widely used to study embryonic development. Early embryonic development is driven by maternally stored factors accumulated during ...

Gelişim Biyolojisi

Isolation and Cryopreservation of Neonatal Rat Cardiomyocytes

İzolasyon ve Neonatal Rat kardiyomiyositlerde Kryoprezervasyon

Cell culture has become increasingly important in cardiac research, but due to the limited proliferation of cardiomyocytes, culturing cardiomyocytes is ...

Protocols for Analyzing the Role of Paneth Cells in Regenerating the Murine Intestine using Conditional Cre-lox Mouse Models

Protocols for Analyzing the Role of Paneth Cells in Regenerating the Murine Intestine using Conditional Cre-lox Mouse Models

Şartlı kullanarak Fare İnce Bağırsağında Yenileyici içinde Paneth Hücrelerinin Rolü analiz için protokoller<em&gt; Cre-lox</em&gt; Fare Modelleri

The epithelial surface of the mammalian intestine is a dynamic tissue that renews every 3 - 7 days. Understanding this renewal process identified a ...

Modified MicroSecure Vitrification: A Safe, Simple and Highly Effective Cryopreservation Procedure for Human Blastocysts

Modifiye MicroSecure Vitrifikasyon: İnsan Blastosistlerinin A, Güvenli Basit ve Etkili Kryoprezervasyon Prosedürü

Clinical embryo vitrification evolved with the development of unique vitrification devices in the 21st century and with the misconception that ultra-rapid ...

Functional Manipulation of Maternal Gene Products Using In Vitro Oocyte Maturation in Zebrafish

Functional Manipulation of Maternal Gene Products Using In Vitro Oocyte Maturation in Zebrafish

Kullanılması Anne Gen Ürünlerinin Fonksiyonel Manipülasyonu<em&gt; İn Vitro</emZebra balığı&gt; Oosit Olgunlaştırma

Cellular events that take place during the earliest stages of animal embryonic development are driven by maternally derived gene products deposited into ...

Maturation of Human Stem Cell-derived Cardiomyocytes in Biowires Using Electrical Stimulation

Elektriksel Uyarı Biowires İnsan Kök Hücre kaynaklı kardiyomiyositlerde Olgunlaşma

Human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) have been a promising cell source and have thus encouraged the investigation of their ...

A Standardized Approach for Multispecies Purification of Mammalian Male Germ Cells by Mechanical Tissue Dissociation and Flow Cytometry

Memeli Erkek Germ Hücrelerinin Çok Denekli Saflaştırılmasında Standart Bir Yaklaşım, Mekanik Doku Ayrılma ve Akış Sitometrisi ile

Fluorescence-activated cell sorting (FACS) has been one of the methods of choice to isolate enriched populations of mammalian testicular germ cells. ...

Induction and Validation of Cellular Senescence in Primary Human Cells

İndüksiyon ve birincil insan hücrelerinde hücresel yaşlanma doğrulama

Cellular senescence is a state of permanent cell cycle arrest activated in response to different damaging stimuli. Activation of cellular senescence is a ...

Fluorimetric Techniques for the Assessment of Sperm Membranes

Fluorimetric teknikleri Sperm membran değerlendirilmesi için

Standard spermiograms describing sperm quality are mostly based on the physiological and visual parameters, such as ejaculate volume and concentration, ...

Measuring Sperm Guidance and Motility within the Caenorhabditis elegans Hermaphrodite Reproductive Tract

Measuring Sperm Guidance and Motility within the Caenorhabditis elegans Hermaphrodite Reproductive Tract

Ölçme sperm rehberlik ve motilite içinde Caenorhabditis elegans hermafrodit üreme yolu

Successful fertilization is fundamental to sexual reproduction, yet little is known about the mechanisms that guide sperm to oocytes within the female ...