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

이 작품은 NIH 보조금 HD043680, MH106392, DA013137 및 NS100624에 의해 지원되었다.

모든 동물 프로토콜은 사우스 캐롤라이나 대학의 동물 관리 및 사용 위원회에 의해 검토되고 승인되었습니다 (연방 보증 번호 : D16-00028). 6 명의 성인 남성 F344 /N 쥐는 12/12 빛 아래 통제 된 환경에 수용 되었습니다: 음식과 물에 광고 리비툼 액세스 와 어두운 주기. 모든 동물은 실험실 동물의 치료 및 사용에 대한 가이드에서 건강의 국립 연구소에 의해 설립 된 지침을 사용하여 돌보았습니다.
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<ol>
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이 프로토콜에서, 우리는 쥐에 있는 EcoHIV 유도한 HIV 감염 모형을 설치했습니다. 구체적으로, 우리는 성공적으로 쥐 뇌에서 활성 HIV 감염을 유도 피질에 에코 HIV의 양측 스테레오테스테레오테시 주입을 설명 7 주사 후 일. Futhermore, 우리는 쥐에 있는 EcoHIV 감염이 HAND의 중요한 양상을 공부하는 좋은 생물학 시스템이 될 수 있었다는 것을 보여줍니다. 8 주 후- EcoHIV 감염, 쥐는 중요한 신경 인지 장애를...

생물학적 시스템은 HIV-1 관련 신경인지 장애(HAND)와 그 근본적인 신경 메커니즘2에대한 우리의 이해를 향상시켰습니다. 어떤 생물학적 시스템이 어떤 연구에 가장 적합한지 결정하는 것은 종종 관심2의문제에 달려 있습니다. 숙주 동물 모델의 범위의 제한은 HIV-1 질병 발달의 연구에 도전합니다. HIV-1 바이러스 복제 및 병인을 조사하기 위해, Potash 외3은 활성 HIV-1 감염의 마우스 모델을 생성하여 HIV 표면 봉투 당단백질, gp120의 코딩 영역을 에코트로픽 MLV gp80으로 대체하여 마우스4에서성공적인 바이러스 복제를 이끌어 냈다. 키메라 HIV(EcoHIV) 마우스에서 꼬리 정맥 주사 후, HIV-1 혈청 양성 개인(예를 들어, 감염된 림프구 및 대식세포, 항바이러스 면역 반응, 염증3,5,6)과유사한 많은 특성이 관찰되었다.
마우스와 쥐는 모두 무리대의 구성원이지만, 근본적인 종 차이는 특정 실험 질문에 대한 적합성에 영향을 미칠 수 있습니다7. 따라서, 쥐에 에코 HIV 감염 모델의 확장 (일반적으로 약물 남용 및 신경 인지 장애의 연구에 사용) neuroHIV의 연구에서 유리 할 것 이다. 예를 들어, 그들의 큰 크기는 약물 자가 투여 절차에 대한 경정맥 카테터 이식을 보다 실용적으로8. 쥐의 약물 자가 투여 기술은 HIV-19의동기 부여를 평가하기 위해 활용되었습니다. 또한, 많은 신경 인지/행동 작업은 처음에 쥐를 위해 설계 되었다10. 여기에서, 우리는 EcoHIV 감염 모형을 확장하고 neuroHIV 및 HAND와 관련있는 새로운 질문을 해결하기 위하여 중요한 기회를 마련하기 위하여 쥐에 있는 EcoHIV의 입체 주사의 활용을 보고합니다.

<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 감염 마우스 모델은 HIV 관련 신경 학적 합병증을 조사하는 데 중요한 유틸리티라는 것을 잘 연구되었습니다. 약물 남용 및 신경 인지 장애의 연구에 대 한 EcoHIV 감염 된 쥐 모델의 설립, neuroHIV와 HIV-1 관련 된 신경 인지 장애의 연구에 도움이 될 것 이다 (HAND). 본 연구에서는, 우리는 키메라 HIV (EcoHIV)를 사용하여 활성 HIV 감염의 쥐 모델의 성공적인 생성을 보여줍니다. 첫째, 에코HIV의 렌티바이러스 구조는 48시간 동안 배양된 293FT 세포로 포장되었다. 이어서, 조건부 배지는 농축되고 흐트러졌다. 다음으로, 우리는 F344/N 쥐 뇌 조직에 EcoHIV-EGFP의 양측 스테레오택시 주사를 수행했습니다. 감염 후 1주일 후, EGFP 형광 신호가 감염된 뇌 조직에서 검출되어 EcoHIV가 쥐에서 활성 HIV 감염을 성공적으로 유발한다는 것을 나타냅니다. 또한, 마이크로글리아세포 마커인 Iba1에 대한 면역염색이 수행되었다. 결과는 microglia가 EcoHIV를 품고 있는 우세한 세포 모형이었다는 것을 표시했습니다. 더욱이, EcoHIV 쥐는 감염 후 8 주 동안 HAND의 잠재적 기본 신경 행동 메커니즘뿐만 아니라 시간 적 처리에 변화를 나타냈다. 종합적으로, 본 연구는 뇌의 HIV-1 바이러스 저장소뿐만 아니라 HAND 및 약물 남용과 같은 관련 comorbidities를 연구하는 귀중한 생물학적 시스템을 제공하는 쥐에 HIV-1 감염의 EcoHIV 모델을 확장합니다.

조건부 배지는 293FT 세포에 감염된 EcoHIV-EGFP의 렌티바이러스로부터 수집되었다. 다음으로, 그것은 집중 하 고 titered, 다음 스테레오로 뇌에 주입 (피 질 영역) F344/N 쥐의. 7일 후 주사 후, 쥐를 희생하고 브레그마 -4.68 mm에 이르는 관상 동맥 뇌 슬라이스에서 촬영되었다. 도 1A에서,특히 피질과 해마 규명 자이루스에서 뇌 전체에 걸쳐 중요한 EcoHIV-EGFP 신호가 있습니다. 더욱이, ?...

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

A Rat Model of EcoHIV Brain Infection

여기에서, 우리는 키메라 HIV (EcoHIV)를 사용하여 활성 HIV 감염의 새로운 쥐 모형을 설치하는 프로토콜을 제시합니다, 두뇌에 있는 HIV-1 바이러스 성 저수지의 우리의 이해를 강화하고 HIV 관련 신경 인지 장애 및 관련 comorbidities를 공부하는 시스템을 제공하는 것을 위해 중요합니다 (즉, 약물 남용).

에코HIV 뇌 감염의 쥐 모델

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

쥐를 사용하여, 이 프로토콜은 키메라 HIV를 사용하여 활성 HIV 감염의 새로운 모형을 확립하기 위하여 이용될 수 있습니다. 쥐와 함께, 약물 남용 및 신경 인지 장애의 연구에 대 한 EcoHIV 감염 모델의 설립 신경 HIV와 HIV-1 관련 된 신경 인지 장애의 연구에 도움이 될 수 있습니다. 절차를 시연하는 것은 내 실험실에서 연구 동료 인 하이롱 리 (Hailong Li)가 될 것입니다.293 T 세포로 바이러스의 포장을 위해, DMEM의 밀리리터 당 다섯 번째 293 T 세포에 5 번 10 을 종자 10% FBS로 보충 두 젤라틴 코팅 75 평방 센티미터 플라스크와 배양이 도달 할 때까지 섭씨 37도에서 세포를 배양하 50% 수렴성. 트랜스펙트 당일, 플라스크당 750마이크로리터의 미약재 22.5마이크로리터를 1.5밀리리터 마이크로센트심분리기 튜브로 희석하고 3초 동안 용액을 소용돌이시다. 플라스크당 1.5 밀리리터 마이크로센심분리기 튜브에 750마이크로리터의 키메라 에코HIV 플라스미드 DNA 20마이크로그램을 철저한 혼합으로 희석시킵니다.희석된 DNA와 희석된 형질 전환 시약을 부드러운 혼합과 결합하고 실온에서 15분 동안 생성된 용액을 배양합니다. 인큐베이션의 끝에서, prewarm DMEM 배지의 10 밀리리터에 각 바이러스 현탁액을 추가하고 293 T 세포의 한 플라스크에 각 바이러스 보충 솔루션을 추가합니다. 세포 배양 이틀 후, 원심분리를 위해 플라스크에서 상체를 풀고 명확한 상체를 새로운 50 밀리리터 튜브로 옮긴다.렌티-X 농축기 8밀리리터를 명확히 된 상체에 넣고 부드러운 반전과 섞으세요. 섭씨 4도에서 2일 간 배양한 후 혼합물을 원심분리하고 상체를 조심스럽게 제거한 다음, 100밀리머 PBS의 100 마이크로리터로 펠릿을 부드럽게 재연하고 P24 ELISA 키트를 사용하여 바이러스 농도를 자극합니다. EcoHIV-EGFP 주사의 경우 페달 반사에 대한 반응부족을 확인한 후 뇌 부위에서 모발을 면도하고 노출된 피부를 70%에탄올과 클로르헨시딘 기반 스크럽으로 두 번 살균합니다.스테레오탁스 장치에서 쥐를 수습된 자세로 고정하고 두피 미드라인을 따라 피부를 통해 5~6센티미터 절개를 합니다. 1개의 드릴링 위치를 측면 0.8밀리미터, 1.2밀리미터 로스트랄로 표시하여 각 두개골 위치에서 직경 0.4mm의 구멍을 뚫습니다. 10 마이크로리터 주사 주사기에 titered EcoHIV Lentivirus 용액의 밀리리터 당 1.04 배 10 ~ 여섯 번째 트랜스듀싱 유닛을 로드하고 스테레오탁스 장치에 주사기를 고정시킵니다.바늘을 하나의 드릴링 구멍의 표면에 가깝게 이동하고 구멍에 바늘 2.5 밀리리터를 삽입합니다. 분당 0.2 마이크로리터의 바이러스 속도로 하나의 마이크로리터의 바이러스 용액을 주입합니다. 모든 바이러스가 주입되었을 때, 쥐 두개골 이밖에있을 때까지 천천히 바늘을 철회하기 전에 5 분 동안 주사 영역 내부에 바늘을 둡니다.4-0 실크 실 봉합사로 피부 절개를 닫고 70 %에탄올로 용액을 살균 한 다음 쥐를 가열 패드에 놓고 방수까지 모니터링하십시오. 바이러스 주입 후 1~8주 후, 페달 반사에 대한 반응이 부족하다는 것을 확인한 후, 연기 후드 내부의 척추 자세로 쥐를 고정하고 흉부 미드라인을 따라 피부를 절개합니다. 격막을 잘라 흉부 구멍을 열고 왼쪽 심실에 20 게이지를 삽입합니다.즉시 가위로 오른쪽 아트리움을 열고 분당 5 밀리리터로 미리 냉각된 100 밀리리터 PBS의 50밀리리터를 견딜 수 있습니다. 모든 PBS가 전달되면 100 밀리리터의 차가운 4 % 파라 포름알데히드가 제공됩니다. 모든 고정이 전달되면 두개골에서 뇌를 제거하고 신선한 4 %의 파라 포름 알데히드에서 하룻밤 조직을 수정합니다.다음 날 아침, 뇌가 메틸 부타놀에서 뇌 조직을 영하 80도에서 2분 동안 동결하기 전에 약 3일 동안 50 밀리리터 튜브에 100 밀리미터 PBS의 30%의 30%의 자당40밀리리터에 샘플을 놓습니다. 동결 후 영하 20도의 냉동 저온과 미세 한 페인트 브러시를 사용하여 50 미크론 두께의 관상 섹션을 획득하십시오. 모든 단면을 얻을 때, 300 마이크로리터의 페이드 방지 매체로 샘플을 덮고 22 50 밀리리터 커버립으로 장착하십시오.매체가 건조되면, 공초점 현미경검사법에 의해 단면을 이미지. 렌티바이러스 주입 후 7일 후, 쥐 관상 동맥 뇌 슬라이스 이미지는 뇌 조직 전반에 걸쳐 EcoHIV-EGFP 신호의 상당한 존재를 드러냅니다, 특히 피질과 해마 의 장축 자이루스 내에서. 이중 라벨링 및 이바1 및 EcoHIV-EGFP 프로브는 마이크로글리아가 뇌에서 에코HIV 발현을 수용하는 주요 세포 유형이라는 강력한 증거를 제공합니다.8 주 감염 후, EcoHIV 주입 동물은 식염수 대조군에 비해 상대적으로 아첨 간격 기능에 의해 입증 된 바와 같이 상호 자극 간격의 조작에 상대적 무감각을 나타낸다. 또한, EcoHIV 주입 쥐는 대조동물에 비해 머리와 목 직경이 증가하는 짧은 수지상 척추의 상대적 빈도증가에 의해 입증된 바와 같이 수지상 척추 형태에 심오한 변화를 표시합니다. 스테레오탁스 주입 전에 조건 매체의 농도 및 적정은 실험 전반에 걸쳐 일관되고 복제 가능한 결과를 보장하는 데 중요합니다.EcoHIV의 이 특정 지역 입체 주사는 두뇌 내의 능률적인 HIV 감염을 제공하고 쥐에 있는 시간 적 처리 적자를 일으킵니다. 에코 HIV 감염 모델을 확장 하기 위해 쥐에 스테레오 탁스 HIV 주사의 활용은 손에 신경 HIV와 관련 된 새로운 질문을 해결 하기 위한 중요 한 기회를 제공.

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.

비 심실 플라스미드 주입 및 Electroporation에 의해 출생 후의 쥐 뇌에 유전자 전달

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

연구

신경과학

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.

성인 쥐의 특정 두뇌 영역의 Excitotoxic 병변에 대한 Stereotaxic 외과

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 협와 가벼운 자궁내 Hypoperfusion의 쥐 모델

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.

허 혈 성 뇌졸중의 쥐 모델에서 뇌혈관 장애의 비보에 평가

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).

쥐 모델에 수정 사용 하 여 중앙 피로의 여러 플랫폼 메서드

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.

Neurodegeneration Ex Vivo 쥐 두뇌 분할 영역을 사용 하 여 기본 이벤트를 공부 하는 양자 택일 접근

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.

쥐 모델에서 집중 초음파를 사용하여 위치 특정 혈액 뇌 장벽 개방에 대한 벤치 탑 접근

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.

쥐 뇌 조직을 위한 소수성 조직 청산 방법

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.

파킨슨 병의 쥐 모델에서 레보도파 유도 운동 장애의 유도 및 평가

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

mTBI 쥐 모델에서 부상 반응 및 행동 평가 검증

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.

저자 스포트라이트: 외상성 뇌 손상 연구의 발전 - 정확한 복제 및 신속한 평가를 위한 폐쇄형 헤드 모델 

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.

폐쇄 머리 경미한 외상성 손상의 쥐 모델 및 그 검증

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