Name: a rat model of ecohiv brain infection
Uploaded: 2021-01-21T14:30:00
Description: Watch this Scientific Journal Video about A Rat Model of EcoHIV Brain Infection at JoVE.com

Bu çalışma NIH tarafından HD043680, MH106392, DA013137 ve NS100624 hibeleri tarafından finanse edildi.

Tüm hayvan protokolleri Güney Carolina Üniversitesi Hayvan Bakım ve Kullanım Komitesi tarafından gözden geçirildi ve onaylandı (federal güvence numarası: D16-00028). Altı yetişkin erkek F344/N sıçanı, 12/12 ışık altında kontrollü bir ortamda barındırıldı: yiyecek ve suya ad libitum erişimi olan karanlık döngü. Tüm hayvanlara, Laboratuvar Hayvanlarının Bakımı ve Kullanımı Rehberi'nde Ulusal Sağlık Enstitüleri tarafından oluşturulan yönergeler kullanılarak bakıldı.
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(158), (2020).</li><li>Ko, 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=Macrophages+but+not+Astrocytes+Harbor+HIV+DNA+in+the+Brains+of+HIV-1-Infected+Aviremic+Individuals+on+Suppressive+Antiretroviral+Therapy.">Macrophages but not Astrocytes Harbor HIV DNA in the Brains of HIV-1-Infected Aviremic Individuals on Suppressive Antiretroviral Therapy.</a> Journal of Neuroimmune Pharmacology. 14 (1), 110-119 (2019).</li><li>Sopper, 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=The+effect+of+simian+immunodeficiency+virus+infection+in+vitro+and+in+vivo+on+the+cytokine+production+of+isolated+microglia+and+peripheral+macrophages+from+rhesus+monkey.">The effect of simian immunodeficiency virus infection in vitro and in vivo on the cytokine production of isolated microglia and peripheral macrophages from rhesus monkey.</a> Virology. 220 (2), 320-329 (1996).</li><li>Llewellyn, G. 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Bu protokolde sıçanlarda EcoHIV kaynaklı HIV enfeksiyon modeli oluşturduk. Özellikle, enjeksiyondan 7 gün sonra sıçan beyninde aktif HIV enfeksiyonunu başarıyla indükleyen kortekse bilateral stereotaksik ecohiv enjeksiyonu tanımladık. Ayrıca, sıçanlarda EcoHIV enfeksiyonunun HAND'in temel yönlerini incelemek için iyi bir biyolojik sistem olabileceğini gösteriyoruz. Sekiz hafta sonra- EcoHIV enfeksiyonu, sıçanlar NAc MSN'lerinde zamansal işleme ve sinaptik disfonksiyondaki değişiklikleri içeren ?...

Biyolojik sistemler HIV-1 ilişkili nöroklasitif bozukluklar (EL) ve bunların altında kalan sinirsel mekanizmalar hakkındaki anlayışımızı geliştirmiştir2. Herhangi bir çalışma için hangi biyolojik sistemin en uygun olduğunu belirlemek genellikle ilgi sorusuna bağlıdır2. Konak hayvan modellerinin çeşitliliğinin sınırlandırılması HIV-1 hastalığı gelişimi çalışmalarına meydan okuyor. HIV-1 viral replikasyonu ve patogenezini araştırmak için Potash ve ark.3, HIV yüzey zarfı glikoprotein kodlama bölgesi gp120'yi farelerde başarılı viral replikasyona yol açan ekotropik MLV gp80 ile değiştirerek aktifHIV-1enfeksiyonunun bir fare modelini oluşturdu 4 . Kimerik HIV (EcoHIV) farelerinde kuyruk damarı enjeksiyonlarından sonra, HIV-1 seropositive bireylerinkine benzeyen birçok özellik gözlenmiştir (örneğin, antiviral immün yanıtlar için hedeflenen enfekte lenfositler ve makrofajlar ve iltihap3,5,6).
Fareler ve sıçanlar Muridae'nin her ikisi de üyesi olmasına rağmen, temel tür farklılıkları belirli deneysel sorulara uygunluklarını etkileyebilir7. Bu nedenle, EcoHIV enfeksiyon modelinin sıçanlara uzatılması (yaygın olarak ilaç kötüye kullanımı ve nöroknatif bozukluk çalışmalarında kullanılır) nöroHIV çalışmasında avantajlı olacaktır. Örneğin, daha büyük boyutları, ilaç kendi kendine uygulama prosedürleri için juguler kateter implantasyonunu daha pratik hale getirir8. HIV-19'da motivasyonun değerlendirilmesi için sıçanlarda ilaçkendi kendine uygulama teknikleri kullanılmıştır. Ayrıca, birçok nöroknatif / davranışsal görev başlangıçta sıçanlar için tasarlanmıştır10. Burada, EcoHIV enfeksiyon modelini genişletmek ve neuroHIV ve HAND ile ilgili yeni soruları ele almak için önemli bir fırsat sağlamak için sıçanlarda EcoHIV'in stereotaksik enjeksiyonlarının kullanımını rapor ediyoruz.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>293FT cells</td>
			<td>ThermoFisher Scientific</td>
			<td>R70007</td>
			<td></td>
		</tr>
		<tr>
			<td>Antibiotic-Antimycotic solution</td>
			<td>Cellgro</td>
			<td>30004CI</td>
			<td>100X</td>
		</tr>
		<tr>
			<td>Corning BioCoatGelatin 75cm&#178; Rectangular Canted Neck Cell Culture Flask with Vented Cap</td>
			<td>Life Technologies</td>
			<td>354488</td>
			<td></td>
		</tr>
		<tr>
			<td>Corning DMEM with L-Glutamine, 4.5 g/L Glucose and Sodium Pyruvate</td>
			<td>Life Technologies</td>
			<td>10013CV</td>
			<td></td>
		</tr>
		<tr>
			<td>Cover glass</td>
			<td>VWR</td>
			<td>637-137</td>
			<td></td>
		</tr>
		<tr>
			<td>drill</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Dumont #5 Forceps</td>
			<td>World Precision Instruments</td>
			<td>14095</td>
			<td></td>
		</tr>
		<tr>
			<td>Dumont #7 Forceps</td>
			<td>World Precision Instruments</td>
			<td>14097</td>
			<td></td>
		</tr>
		<tr>
			<td>Eppendorf Snap-Cap Microcentrifuge Biopur Safe-Lock Tubes</td>
			<td>Life Technologies</td>
			<td>22600028</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethicon Vicryl Plus Antibacterial, 4-0 Polyglactin 910 Suture, 27in. FS-2</td>
			<td>Med Vet International</td>
			<td>VCP422H</td>
			<td></td>
		</tr>
		<tr>
			<td>Hamilton syringe</td>
			<td>Hamilton</td>
			<td>1701</td>
			<td></td>
		</tr>
		<tr>
			<td>Invitrogen Lipofectamine 3000 Transfection Reagent</td>
			<td>Life Technologies</td>
			<td>L3000015</td>
			<td></td>
		</tr>
		<tr>
			<td>Iris Forceps</td>
			<td>World Precision Instruments</td>
			<td>15914</td>
			<td></td>
		</tr>
		<tr>
			<td>Iris Scissors</td>
			<td>World Precision Instruments</td>
			<td>500216</td>
			<td></td>
		</tr>
		<tr>
			<td>Lentivirus-Associated p24 ELISA Kit</td>
			<td>Cell Biolabs, inc.</td>
			<td>VPK-107-5</td>
			<td></td>
		</tr>
		<tr>
			<td>Lenti-X Concentrator</td>
			<td>Takara</td>
			<td>PT4421-2</td>
			<td></td>
		</tr>
		<tr>
			<td>Opti-MEM I Reduced Serum Medium</td>
			<td>Life Technologies</td>
			<td>11058021</td>
			<td></td>
		</tr>
		<tr>
			<td>Paraformaldehyde</td>
			<td>Sigma-Aldrich</td>
			<td>158127-500G</td>
			<td></td>
		</tr>
		<tr>
			<td>Paraformaldehyde</td>
			<td>Sigma</td>
			<td>P6148</td>
			<td></td>
		</tr>
		<tr>
			<td>ProLong Gold</td>
			<td>Fisher Scientific</td>
			<td>P36930</td>
			<td></td>
		</tr>
		<tr>
			<td>Sevoflurane</td>
			<td>Merritt Veterinary Supply</td>
			<td>347075</td>
			<td></td>
		</tr>
		<tr>
			<td>stereotaxic apparatus</td>
			<td>Kopf Instruments</td>
			<td>Model 900</td>
			<td></td>
		</tr>
		<tr>
			<td>SuperFrost Plus Slides</td>
			<td>Fisher Scientific</td>
			<td>12-550-154%</td>
			<td></td>
		</tr>
		<tr>
			<td>Vannas Scissors</td>
			<td>World Precision Instruments</td>
			<td>500086</td>
			<td></td>
		</tr>
	</tbody>
</table>

a rat model of ecohiv brain infection

EcoHIV enfekte fare modelinin HIV ile ilişkili nörolojik komplikasyonların araştırılmasında önemli bir yararı olduğu iyi incelenmiştir. İlaç kötüye kullanımı ve nöroknasitif bozukluklar çalışmaları için EcoHIV enfekte sıçan modelinin kurulması, nöroHIV ve HIV-1 ilişkili nörokinetif bozuklukların (EL) çalışmasında faydalı olacaktır. Bu çalışmada, kimerik HIV (EcoHIV) kullanılarak aktif HIV enfeksiyonunun sıçan modelinin başarılı bir şekilde oluşturulduğunu göstermektedir. İlk olarak, EcoHIV'in lentiviral yapısı 48 saat boyunca kültürlü 293 FT hücrelerde paketlendi. Daha sonra, koşullu ortam yoğunlaştı ve titretildi. Daha sonra, EcoHIV-EGFP'nin F344/N sıçan beyin dokusuna bilateral stereotaksik enjeksiyonlarını gerçekleştirdik. Enfeksiyondan bir hafta sonra, enfekte beyin dokusunda EGFP floresan sinyalleri tespit edildi ve bu da EcoHIV'in sıçanlarda aktif bir HIV enfeksiyonunu başarıyla teşvik ettiğini gösterdi. Ayrıca Iba1 mikroglial hücre belirteci için immünostaining yapıldı. Sonuçlar, mikroglianın EcoHIV'i barındıran baskın hücre tipi olduğunu gösterdi. Ayrıca, EcoHIV sıçanları, HAND'in potansiyel bir nörobehavioral mekanizması olan zamansal işlemde ve enfeksiyondan sekiz hafta sonra sinaptik disfonksiyonda değişiklikler gösterdi. Toplu olarak, bu çalışma HIV-1 enfeksiyonunun EcoHIV modelini, beyindeki HIV-1 viral rezervuarlarını ve EL'i ve uyuşturucu kullanımı gibi ilişkili komorbiditeleri incelemek için değerli bir biyolojik sistem sunan sıçana genişletmektedir.

Şartlandırılmış ortam, EcoHIV-EGFP enfekte 293FT hücrelerinin lentivirüslerinden toplanmıştır. Daha sonra konsantre edildi ve titretildi, daha sonra stereotaksik olarak F344/N sıçanlarının beynine (kortikal bölge) enjekte edildi. Enjeksiyondan yedi gün sonra sıçanlar kurban edildi ve bregma 5.64 mm'den bregma -4.68 mm'ye kadar değişen koronal beyin dilimlerinden görüntüler alındı. Şekil 1A'da, beyinde, özellikle korteks ve hipokampal dentat girusunda önemli EcoHIV...

Watch this Scientific Journal Video about A Rat Model of EcoHIV Brain Infection at JoVE.com

A Rat Model of EcoHIV Brain Infection

Burada, beyindeki HIV-1 viral rezervuarlarını anlamamızı geliştirmek ve HIV ile ilişkili nöroknaktif bozuklukları ve ilişkili komorbiditeleri (yani uyuşturucu bağımlılığı) incelemek için bir sistem sunmak için kritik öneme sahip olan kimerik HIV (EcoHIV) kullanarak aktif HIV enfeksiyonunun yeni bir sıçan modelini oluşturmak için bir protokol sunuyoruz.

EcoHIV Beyin Enfeksiyonunun Bir Sıçan Modeli

It has been well studied that the EcoHIV infected mouse model is of significant utility in investigating HIV associated neurological complications. ...

Sıçanlar kullanılarak, bu protokol kimerik HIV kullanarak aktif HIV enfeksiyonunun yeni bir modelini oluşturmak için kullanılabilir. sıçanlar ile, ilaç kullanımı ve nörobilişsel bozukluklar çalışmaları için EcoHIV enfeksiyon modelinin kurulması nöro HIV ve HIV-1 ilişkili nörobilişsel bozuklukların incelenmesine faydalı olabilir. Prosedürü gösteren Hailong Li olacak, laboratuvarımdan bir araştırma görevlisi.Virüsün 293 T hücresine paketlenmesi için, DMEM mililitresi başına beş çarpı 10 ila beşinci 293 T hücresi, iki jelatin kaplı 75 santimetrekare şişenin her birine% 10 FBS ile desteklenmiştir ve kültürler% 50 izne ulaşana kadar hücreleri 37 santigrat derecede kuluçkaya yatırmaktadır. Transfeksiyon gününde, 22,5 mikrolitre transfeksiyon reaktifini 1,5 mililitrelik bir mikrosantrifüj tüpünde şişe başına 750 mikrolitre orta derecede seyreltin ve çözeltiyi üç saniye boyunca girdaplayın. Chimeric EcoHIV plazmid DNA'sının 20 mikrogramını 750 mikrolitre orta derecede, şişe başına ikinci bir 1,5 mililitre mikrosantrifüj tüpünde iyice karıştırarak seyreltin.Seyreltilmiş DNA'yı seyreltilmiş transfeksiyon reaktifi ile nazik karıştırma ile birleştirin ve elde edilen çözeltiyi oda sıcaklığında 15 dakika kuluçkaya bırakın. Kuluçkanın sonunda, her virüs süspansiyonunu 10 mililitrelik savaş öncesi DMEM ortamına ekleyin ve her virüs takviyeli çözeltiyi 293 T hücresinden oluşan bir şişeye ekleyin. İki günlük hücre kültüründen sonra, şişelerden gelen süpernatantı santrifüjleme için tek bir 50 mililitre konik tüpte bir araya getirmek ve netleştirilmiş süpernatantı yeni bir 50 mililitrelik tüpe aktarın.Netleştirilmiş süpernatanta sekiz mililitre Lenti-X Konsantratör ekleyin ve yumuşak inversiyonla karıştırın. Dört santigrat derecede iki günlük bir inkübasyondan sonra, karışımı santrifüjleyin ve süpernatantı dikkatlice çıkarın, ardından peleti 100 milimoler PBS'nin 100 mikrolitresi ile hafifçe yeniden kullanın ve virüs konsantrasyonunu titretmek için bir P24 ELISA kiti kullanın. EcoHIV-EGFP enjeksiyonu için, pedal refleksine yanıt eksikliğini onayladıktan sonra, saçları beyin bölgesinden tıraş edin ve maruz kalan cildi% 70 etanol ve klorheksidin bazlı bir ovalama ile iki kez sterilize edin.Sıçanı stereotaksik bir aparatta eğilimli bir pozisyonda sabitleyin ve kafa derisi orta çizgisi boyunca deriden beş ila altı santimetrelik bir kesi yapın. Bregma'ya 0,8 milimetre yanal ve bir 1,2 milimetre rostralde bir delme pozisyonu işaretleyin ve her kafatası pozisyonunda 0,4 milimetre çapında bir delik açın. Titrek EcoHIV Lentivirus çözeltisinin mililitre başına altıncı transdüksiyon ünitelerine 1.04 kez 10 kez 10 mikrolitre enjeksiyon şırıngasına yükleyin ve şırıngayı stereotaksik aparatlara sabitleyin.İğneyi bir delme deliğinin yüzeyine yaklaştırın ve iğneyi deliğe 2,5 mililitre yerleştirin. Dakikada 0,2 mikrolitre virüs oranında bir mikrolitre virüs çözeltisi demleyin. Tüm virüs enjekte edildiğinde, iğneyi fare kafatasının dışına çıkıncaya kadar yavaşça geri çekmeden önce iğneyi enjeksiyon bölgesinin içinde beş dakika bekletin.Cilt kesisini 4-0 ipek iplik dikişi ile kapatın ve çözeltiyi% 70 etanol ile sterilize edin, ardından sıçanı görev süresine kadar izlemeli bir ısıtma yastığı ile bir geri kazanım odasına yerleştirin. Viral infüzyondan bir ila sekiz hafta sonra, pedal refleksine yanıt eksikliğini doğruladıktan sonra, sıçanı bir duman kaputunun içinde bir supine pozisyonunda sabitlayın ve cildi torasik orta hat boyunca inzivaya getirin. Torasik boşluğu açmak için diyaframı kesin ve sol ventrikül içine 25 milimetrelik bir iğne ile 20 ölçer yerleştirin.Hemen sağ kulakçığı makasla açın ve dakikada beş mililitre debide 50 mililitre önceden soğutulmuş 100 milimoler PBS'yi perfüzyona maruz bırakın. Tüm PBS teslim edildiğinde, 100 mililitre soğuk% 4 paraformaldehit ile perfüzyon. Tüm fiksatif teslim edildiğinde, beyni kafatasından çıkarın ve dokuyu bir gecede taze% 4 paraformaldehitte sabitleyin.Ertesi sabah, örneği 100 milimolar PBS'de% 30 sakkarozun 40 mililitresine, beyin metil butanoldeki beyin dokusunu eksi 80 santigrat derecede iki dakika dondurmadan önce yaklaşık üç gün boyunca 50 mililitrelik bir tüpe yerleştirin. Donduktan sonra, 50 mikron kalınlığında koronal bölüm elde etmek için eksi 20 santigrat bir kriyostat ve ince bir boya fırçası kullanın. Tüm bölümler elde edildiğinde, numuneleri 300 mikrolitre anti-solmaya karşı orta ile örtün ve 22 adet 50 mililitrelik kapakla monte edin.Ortam kuruduğunda, bölümleri konfokal mikroskopi ile görüntüleyin. Lentivirüs enjeksiyonundan yedi gün sonra, sıçan koroner beyin dilimi görüntüleri, beyin dokusu boyunca, özellikle korteks ve hipokampal dentat girus içinde Önemli miktarda EcoHIV-EGFP sinyali varlığını ortaya koymaktadır. Iba1 ve EcoHIV-EGFP probları ile çift etiketleme, mikroglianın beyinde EcoHIV ekspresyonını barındıran baskın hücre tipi olduğuna dair güçlü kanıtlar sağlar.Enfeksiyondan sekiz hafta sonra, EcoHIV enjekte edilen hayvanlar, tuzlu su kontrollerine kıyasla nispeten daha düz bir aralık fonksiyonu ile kanıtlandıkça, uyaranlar arası aralığın manipülasyonuna görece bir duyarsızlık gösterir. Ek olarak, EcoHIV enjekte edilen sıçanlar, kontrol hayvanlarına göre artan baş ve boyun çaplarına sahip daha kısa dendritik dikenlerin artan göreceli sıklığı ile kanıtlandıkça, dendritik omurga morfolojilerinde derin değişiklikler gösterir. Stereotaksik enjeksiyondan önce durum ortamının konsantrasyonu ve titratingi, deneyler arasında tutarlı ve tekrarlanabilir sonuçların sağlanması için kritik öneme sahiptir.EcoHIV'in bu spesifik bölgesel stereotaksik enjeksiyonu beyinde etkili bir HIV enfeksiyonu sağlar ve sıçanlarda zamansal işlem açıkları üretir. EcoHIV enfeksiyon modelini genişletmek için sıçanlarda stereotaksik HIV enjeksiyonlarının kullanılması, eldeki nöro HIV ile ilgili yeni soruları ele almak için önemli bir fırsat sürmektedir.

Research

JoVE Journal

Neuroscience

Gene Delivery to Postnatal Rat Brain by Non-ventricular Plasmid Injection and Electroporation

Creation of transgenic animals is a standard approach in studying functions of a gene of interest in vivo. However, many knockout or transgenic animals are not viable in those cases where the modified gene is expressed or deleted in the whole organism. Moreover, a variety of compensatory mechanisms often make it difficult to interpret the results. The compensatory effects can be alleviated by either timing the gene expression or limiting the amount of transfected cells.
The method of postnatal non-ventricular microinjection and in vivo electroporation allows targeted delivery of genes, siRNA or dye molecules directly to a small region of interest in the newborn rodent brain. In contrast to conventional ventricular injection technique, this method allows transfection of non-migratory cell types. Animals transfected by means of the method described here can be used, for example, for two-photon in vivo imaging or in electrophysiological experiments on acute brain slices.

This protocol describes a non-viral method of delivery of genetic constructs to a certain area of living rodent brain. The method consists of plasmid preparation, micropipette fabrication, neonatal rat pup surgery, microinjection of the construct, and in vivo electroporation.

Non-ventriküler Plazmid Enjeksiyon ve Elektroporasyon Postnatal Rat Beyin Gen Teslimat

Creation of transgenic animals is a standard approach in studying functions of a gene of interest in vivo. However, many knockout or transgenic animals ...

Nörobilim

Stereotaxic Surgery for Excitotoxic Lesion of Specific Brain Areas in the Adult Rat

Many behavioral functions in mammals, including rodents and humans, are mediated principally by discrete brain regions. A common method for discerning the function of various brain regions for behavior or other experimental outcomes is to implement a localized ablation of function. In humans, patient populations with localized brain lesions are often studied for deficits, in hopes of revealing the underlying function of the damaged area. In rodents, one can experimentally induce lesions of specific brain regions.
Lesion can be accomplished in several ways. Electrolytic lesions can cause localized damage but will damage a variety of cell types as well as traversing fibers from other brain regions that happen to be near the lesion site. Inducible genetic techniques using cell-type specific promoters may also enable site-specific targeting. These techniques are complex and not always practical depending on the target brain area. Excitotoxic lesion using stereotaxic surgery, by contrast, is one of the most reliable and practical methods of lesioning excitatory neurons without damaging local glial cells or traversing fibers.
Here, we present a protocol for stereotaxic infusion of the excitotoxin, N-methyl-D-aspartate (NMDA), into the basolateral amygdala complex. Using anatomical indications, we apply stereotaxic coordinates to determine the location of our target brain region and lower an injection needle in place just above the target. We then infuse our excitotoxin into the brain, resulting in excitotoxic death of nearby neurons. While our experimental subject of choice is a rat, the same methods can be applied to other mammals, with the appropriate adjustments in equipment and coordinates.
This method can be used on a variety of brain regions, including the basolateral amygdala1-6, other amygdala nuclei6, 7, hippocampus8, entorhinal cortex9 and prefrontal cortex10. It can also be used to infuse biological compounds such as viral vectors1, 11. The basic stereotaxic technique could also be adapted for implantation of more permanent osmotic pumps, allowing more prolonged exposure to a compound of interest.

Targeted ablation of specific brain region(s) by infusion of an excitotoxin using stereotaxic coordinates is described. This technique could also be adapted for infusion of other chemicals into the rat brain.

Yetişkin Rat spesifik Beyin Alanlarının eksitotoksik Lezyon için Stereotaksik Cerrahi

Many behavioral functions in mammals, including rodents and humans, are mediated principally by discrete brain regions. A common method for discerning the ...

A Rat Model of Mild Intrauterine Hypoperfusion with Microcoil Stenosis

Intrauterine hypoperfusion/ischemia is one of the major causes of intrauterine/fetal growth restriction, preterm birth, and low birth weight. Most studies of this phenomenon have been performed in either models with severe intrauterine ischemia or models with gradient degree of intrauterine hypoperfusion. No study has been performed in a model on uniform mild intrauterine hypoperfusion (MIUH). Two models have been used for studies of MIUH: a model based on suture ligation of either side of the arterial arcade formed with the uterine and ovarian arteries, and a transient model based on clipping the bilateral ovarian arteries and aorta having patency. Those two rodent models of MIUH have some limitations, e.g., not all fetuses are subjected to MIUH, depending on their position in the uterine horn. In our MIUH model, all fetuses are subjected to a comparable level of intrauterine hypoperfusion. MIUH was achieved by mild stenosis of all four arteries feeding the uterus, i.e., the bilateral uterine and ovarian arteries.
Arterial stenosis was induced by metal microcoils wrapped around the feeding arteries. Producing arterial stenosis with microcoils allowed us to control, optimize, and reproduce decreased blood flow with very little inter-animal variability and a low mortality rate, thus enabling accurate evaluation. When microcoils with an inner diameter of 0.24 mm were used, the blood flow in both the placenta and fetus was mildly decreased (approximately 30% from the pre-stenosis level in the placenta). The offspring of our MIUH model clearly demonstrates long-lasting alterations in neurological, neuroanatomical and behavioral test results.

Mild intrauterine hypoperfusion was produced by artery stenosis with metal microcoils wrapped around the uterine and ovarian arteries in rats at embryonic day 17. This procedure produced prenatal hypoperfusion and intrauterine growth restriction.

Microcoil stenoz ile hafif intrauterin hipoperfüzyon fare modeli

Intrauterine hypoperfusion/ischemia is one of the major causes of intrauterine/fetal growth restriction, preterm birth, and low birth weight. Most studies ...

An In Vivo Assessment of Blood-Brain Barrier Disruption in a Rat Model of Ischemic Stroke

Ischemic stroke leads to vasogenic cerebral edema and subsequent primary brain injury, which is mediated through destruction of the blood-brain barrier (BBB). Rats with induced ischemic stroke were established and used as in vivo models to investigate the functional integrity of the BBB. Spectrophotometric detection of Evans blue (EB) in the brain samples with ischemic injury could provide reliable justification for the research and development of novel therapeutic modalities. This method generates reproducible results, and is applicable in any laboratory without a need for special equipment. Here, we present a visualized and technical guideline on the detection of the extravasation of EB following induction of ischemic stroke in rats.

An In Vivo Assessment of Blood-Brain Barrier Disruption in a Rat Model of Ischemic Stroke

The overall goal of this procedure is to provide a highly reproducible technique for in vivo assessment of the blood-brain barrier disruption in rat models of ischemic stroke.

Kan - beyin bariyerini bozulma iskemik inme, bir fare modeli Vivo içinde değerlendirilmesi

Ischemic stroke leads to vasogenic cerebral edema and subsequent primary brain injury, which is mediated through destruction of the blood-brain barrier ...

A Rat Model of Central Fatigue Using a Modified Multiple Platform Method

In this article, we introduced a rat model of central fatigue using the modified multiple platform method (MMPM). The Multiple Platform box was designed as a water tank with narrow platforms on the bottom. The model rats were put into the tank and stood on the platforms for 14 h (18:00 - 8:00) per day for a consecutive 21 days, with a blank control group set for contrast. At the end of modeling, rats in the model group showed an obvious fatigued appearance. To assess the model, we performed several behavioral tests, including the open field test (OFT), the elevated plus maze (EPM) test, and the exhaustive swimming (ES) test. The results showed that anxiety, spatial cognition impairment, poor muscle performance, and declined voluntary activity presented in model rats confirm the diagnosis of&#160;central fatigue. Changes of the central neurotransmitters also verified the result. In conclusion, the model successfully simulated central fatigue, and future study with the model may help reveal the pathological mechanism of the disease.

Here, we present a protocol to introduce a rat model of central fatigue using the modified multiple platform method (MMPM).

Bir fare modeli değiştirilmiş bir kullanarak merkezi yorgunluk birden çok Platform yöntemi

In this article, we introduced a rat model of central fatigue using the modified multiple platform method (MMPM). The Multiple Platform box was designed ...

An Alternative Approach to Study Primary Events in Neurodegeneration Using Ex Vivo Rat Brain Slices

Despite&#160;numerous studies that attempt to develop reliable animal models&#160;which reflecting the primary processes underlying neurodegeneration, very few have been widely accepted. Here, we propose&#160;a new procedure&#160;adapted from the well-known ex vivo brain slice technique, which offers a closer in vivo-like scenario than in vitro preparations, for investigating the early events triggering cell degeneration, as observed in Alzheimer&#39;s disease (AD). This variation consists of simple and easily reproducible steps, which enable preservation of the anatomical cytoarchitecture of the selected brain region and its local functionality in a physiological milieu. Different anatomical areas can be obtained from the same brain, providing the opportunity to perform multiple experiments with the treatments in question in a site-, dose-, and time-dependent manner. Potential limitations&#160;which could affect the outcomes related to this methodology are related to the conservation of the tissue, i.e., the maintenance of its anatomical integrity during the slicing and incubation steps and the section thickness, which can influence the biochemical and immunohistochemical analysis. This approach can be employed for different purposes, such as exploring molecular mechanisms involved in physiological or pathological conditions, drug screening, or dose-response assays. Finally, this protocol could also reduce the number of animals employed in behavioral studies. The application reported here has been recently described and tested for the first time on ex vivo rat brain slices containing the basal forebrain (BF), which is one of the cerebral regions primarily affected in AD. Specifically, it has been demonstrated that the administration of a toxic peptide derived from the C-terminus of acetylcholinesterase (AChE) could prompt an AD-like profile, triggering, along the antero-posterior axis of the BF, a differential expression of proteins altered in AD, such as the alpha7 nicotinic receptor (&#945;7-nAChR), phosphorylated Tau (p-Tau), and amyloid beta (A&#946;).

An Alternative Approach to Study Primary Events in Neurodegeneration Using Ex Vivo Rat Brain Slices

We present a method which can&#160;provide further insights into the early events underlying neurodegeneration and based on the established ex-vivo brain technique, combining the advantages of in vivo and in vitro experiments. Moreover, it represents a unique opportunity for direct comparison of treated and untreated group in the same anatomical plane.

Ateş Ex Vivo sıçan beyin dilimleri kullanarak birincil olayları incelemek için alternatif bir yaklaşım

Despite&#160;numerous studies that attempt to develop reliable animal models&#160;which reflecting the primary processes underlying neurodegeneration, ...

A Benchtop Approach to the Location Specific Blood Brain Barrier Opening using Focused Ultrasound in a Rat Model

Stereotaxic surgery is the gold standard for localized drug and gene delivery to the rodent brain. This technique has many advantages over systemic delivery including precise localization to a target brain region and reduction of off target side effects. However, stereotaxic surgery is highly invasive which limits its translational efficacy, requires long recovery times, and provides challenges when targeting multiple brain regions. Focused ultrasound (FUS) can be used in combination with circulating microbubbles to transiently open the blood brain barrier (BBB) in millimeter sized regions. This allows intracranial localization of systemically delivered agents that cannot normally cross the BBB. This technique provides a noninvasive alternative to stereotaxic surgery. However, to date this technique has yet to be widely adopted in neuroscience laboratories due to the limited access to equipment and standardized methods. The overall goal of this protocol is to provide a benchtop approach to FUS BBB opening (BBBO) that is affordable and reproducible and can therefore be easily adopted by any laboratory.

Focused ultrasound with microbubble agents can open the blood brain barrier focally and transiently. This technique has been used to deliver a wide range of agents across the blood brain barrier. This article provides a detailed protocol for the localized delivery to the rodent brain with or without MRI guidance.

Bir Sıçan Modelinde Odaklanmış Ultrason Kullanarak Konuma Özgü Kan Beyin Bariyeri Açılışına Tezgah Üstü Yaklaşım

Stereotaxic surgery is the gold standard for localized drug and gene delivery to the rodent brain. This technique has many advantages over systemic ...

A Hydrophobic Tissue Clearing Method for Rat Brain Tissue

Hydrophobic tissue clearing methods are easily adjustable, fast, and low-cost procedures that allows for the study of a molecule of interest in unaltered tissue samples. Traditional immunolabeling procedures require cutting the sample into thin sections, which restricts the ability to label and examine intact structures. However, if brain tissue can remain intact during processing, structures and circuits can remain intact for the analysis. Previously established clearing methods take significant time to completely clear the tissue, and the harsh chemicals can often damage sensitive antibodies. The iDISCO method quickly and completely clears tissue, is compatible with many antibodies, and requires no special lab equipment. This technique was initially validated for the use in mice tissue, but the current protocol adapts this method to image hemispheres of control and transgenic rat brains. In addition to this, the present protocol also makes several adjustments to preexisting protocol to provide clearer images with less background staining. Antibodies for Iba-1 and tyrosine hydroxylase were validated in the HIV-1 transgenic rat and in F344/N control rats using the present hydrophobic tissue clearing method. The brain is an interwoven network, where structures work together more often than separately of one another. Analyzing the brain as a whole system as opposed to a combination of individual pieces is the greatest benefit of this whole brain clearing method.

Here we present a hydrophobic tissue clearing method that allows for the viewing of target molecules as part of intact brain structures. This technique has now been validated for F344/N control and HIV-1 transgenic rats of both sexes.

Sıçan Beyin Dokusu için Hidrofobik Doku Temizleme Yöntemi

Hydrophobic tissue clearing methods are easily adjustable, fast, and low-cost procedures that allows for the study of a molecule of interest in unaltered ...

Induction and Assessment of Levodopa-induced Dyskinesias in a Rat Model of Parkinson's Disease

Levodopa (L-DOPA) remains the gold-standard therapy used to treat Parkinson&#39;s disease (PD) motor symptoms. However, unwanted involuntary movements known as L-DOPA-induced dyskinesias (LIDs) develop with prolonged use of this dopamine precursor.&#160;It is estimated that the incidence of LIDs escalates to approximately 90% of individuals with PD within 10&#8211;15 years of treatment. Understanding the mechanisms of this malady and developing both novel and effective anti-dyskinesia treatments requires consistent and accurate modeling for pre-clinical testing of therapeutic interventions. A detailed method for reliable induction and comprehensive rating of LIDs following 6-OHDA-induced nigral lesioning in a rat model of PD is presented here. Dependable LID assessment in rats provides a powerful tool that can be readily utilized across laboratories to test emerging therapies focused on reducing or eliminating this common treatment-induced burden for individuals with PD.

Induction and Assessment of Levodopa-induced Dyskinesias in a Rat Model of Parkinson&#039;s Disease

This article describes methods to induce and evaluate levodopa-induced dyskinesias in a rat model of Parkinson&#39;s disease. The protocol offers detailed information regarding the intensity and frequency of a range of dyskinetic behaviors, both dystonic and hyperkinetic, providing a reliable tool to test treatments targeting this unmet medical need.

Parkinson Hastalığının Sıçan Modelinde Levodopa Kaynaklı Diskinezilerin İndüksiyonu ve Değerlendirilmesi

Levodopa (L-DOPA) remains the gold-standard therapy used to treat Parkinson&#39;s disease (PD) motor symptoms. However, unwanted involuntary movements ...

Bir mTBI Sıçan Modelinde Yaralanma Tepkisinin ve Davranışsal Değerlendirmelerin Doğrulanması

Rat Model of Closed-Head Mild Traumatic Injury and its Validation

Animal models are crucial for advancing our understanding of mild traumatic brain injury (mTBI) and guiding clinical research. To achieve meaningful insights, developing a stable and reproducible animal model is essential. In this study, we report a detailed description of a closed-head mTBI model and a representative validation method using Sprague-Dawley rats to verify the modeling effect. The model involves dropping a 550 g mass weight from a height of 100 cm directly onto the head of a rat on a destructible surface, followed by a 180-degree turn. To assess the injury, rats underwent a series of neurobehavioral assessments 10 min post-injury, including time of loss of consciousness, first seeking-behavior time, escape ability, and beam balance ability test. During the acute and subacute stages following the injury, behavioral tests were conducted to assess motor coordination ability (Beam task), anxiety (Open Field test), and learning and memory abilities (Morris Water Maze test). The closed-head mTBI model produced a consistent injury response with minimal mortality and replicated real-life situations. The validation method effectively verified the model development and ensured the stability and consistency of the model.

Yazar Spotlight: Travmatik Beyin Hasarı Araştırmalarını İlerletmek - Doğru Çoğaltma ve Hızlı Değerlendirme için Kapalı Kafa Modeli 

Here, we present a closed-head mild traumatic brain injury (mTBI) rat model and its validation exhibiting remarkable similarity to human mTBI concerning behavioral manifestations during the acute and subacute stages.

Kapalı Kafa Hafif Travmatik Yaralanmanın Sıçan Modeli ve Validasyonu

Animal models are crucial for advancing our understanding of mild traumatic brain injury (mTBI) and guiding clinical research. To achieve meaningful ...