Name: induction of maternal immune activation in mice at mid-gestation stage with viral mimic poly(i:c)
Uploaded: 2016-03-25T13:00:00
Description: Watch this Scientific Journal Video about Induction of Maternal Immune Activation in Mice at Mid-gestation Stage with Viral Mimic PolyI:C at JoVE.com

我们要纪念已故保罗·帕特森博士对MIA模型，自闭症和精神分裂症研究的进步作出贡献。我们承认萨尔基斯K. Mazmanian就这一协议的大力支持;鲁本·M.巴戎寺，伊薇特·加西亚 - 弗洛雷斯，卡伦C.伦乔尼，和Leslie A.诺伊曼行政援助;阿里Khoshnan和Jan C.柯对拍摄的援助;伊莱恩Y.萧和Natalia Malkova它们对MIA感应意见;杰弗里·科克伦S.，华金·古铁雷斯，关F·李，海梅·罗德里格斯，洛雷娜C.桑多瓦尔，和Natalie A.贝尔杜斯科及专家畜牧业。这项工作是由美国国立卫生研究院康特中心奖（NIH 5P50MH086383-04，保罗·H·帕特森）的支持;自闭症（＃7670，保罗·H·帕特森）;西蒙斯基金会（＃322839，以萨尔基斯K. Mazmanian）;美国国立卫生研究院培训资助（NIH / NRSA T32GM07616到K-HC）;加州理工大学暑期大学生研究奖学金（SURF）（以ZY）;安进学者计划在加州理工学院（ZY到）;和博士后奖学金FROM国家科学委员会，台湾（NSC 101-2917-I-564-039，到W-LW）。

所有协议均技术机构动物护理和使用委员会的加州理工学院（IACUC）的批准下进行的。 1.准备定时交配双<ol><li>使用基因表达分析至少6个水坝的行为实验和3。 注意：使用估计雌性小鼠的双数，如只有50％左右的小鼠将从定时交配堵塞。 </li><li>使用雌性小鼠事先没有怀孕，并在8-16周龄。 注：雌性小鼠有性行为之前接触到雄性小鼠但一直没怀孕是可以接受的。 </li><li...

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MIA感应在不同的时间窗扰乱在啮齿类动物不同脑发育的事件，并因此导致在后代不同行为异常和neuropathologies。这里，我们描述了一种协议以诱导MIA小鼠与聚（I：C）在E12.5注射。 MIA感应的此方法导致行为，神经病学，免疫学，并与自闭症和精神分裂症的后代8,10,11,16有关的胃肠道异常。要注意，这一点很重要，因为MIA是一个环境风险因素，它们的后代的表型，预计将有比来自神经发育障碍...

母体免疫激活（MIA）的概念从母体感染患有自闭症和精神分裂症1的关联流行病学研究起源。由于没有在胎盘或母体病毒感染2,3后胎儿大脑检测复制病毒病原体，感染的对后代的影响是假设由母体免疫系统的激活，而不是病原体本身引起的。 为了阐明MIA和精神障碍，注入化学合成，病毒模拟双链RNA聚肌苷之间的原因和结果关系：胞苷酸：成怀孕啮齿类动物（聚（I C））已被广泛地用作动物模型MIA 4,5。聚（I：C）通过toll样受体3（TLR3），和聚的全身给药确认（I：C）诱导病毒样急性炎症反应。一种机制，通过它的聚（I：C）公关oduces行为异常和neuropathologies的后代是由母体胎盘胎儿轴6使亲和抗炎细胞因子的不平衡。一些研究小组已经通过在MIA模型理解精神障碍7的病因，并且由于研究组之间的不同的利益，免疫激活的不同时间点已被用来实现对脑发育和行为7不同扰动。 在保罗·帕特森实验室技术加州理工学院采用注射聚的策略（I：C）为孕鼠胚胎在12.5天（E12.5），它已经成功地证明了MIA能够诱导行为，神经的，并且在与自闭症和精神分裂症相关8-11后代的免疫学变化。我们之前的工作表明，MIA后代显示行为异常（ 如社会impairmenT，通信逆差，重复的行为，焦虑样行为，以及潜在抑制赤字8,10,12），免疫失调和细胞因子失衡8,13,14，胎儿大脑的基因表达15的改变，在小叶第七浦肯野细胞的损失小脑11，海马突触9性质改变，基因与环境互作13，肠道通透性的改变，以及肠道菌群组成16。此外，治疗和预防策略也从该模型系统13,16,17的发展。通过在E12.5诱导MIA，其他人已经表明MIA产生脑突触分子17的胎儿胶质细胞活化和前脑基底胆碱能18发育改造，株特异性相互作用19，大脑突触脑超微结构异常，脑线粒体呼吸链功能亢进abnormalties，下调，抑郁样行为，认知和海马长时程增强（LTP）障碍，与成人海马神经20的赤字。 在这里，我们提供了如何通过聚诱导MIA在E12.5的详细方法（I：C），以及如何运用这一模型来研究自闭症和精神分裂症的病因学范式。要注意的是MIA为多种病症4的危险因素是很重要的，其结果是对时间和感应的方法，以及在怀孕水坝的饲养极为敏感。这样，实验室之间甚至轻微不一致常常导致低的再现性和/或后代不同的表型。我们的方法是专门为那些有兴趣在学习MIA为自闭症和精神分裂症的环境风险因素设计的，详细的描述中提供将有助于研究人员提高他们的数据的可重复性。

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			<td height="21" style="height:21px;width:283px;">Polyinosinic&#8211;polycytidylic acid potassium salt</td>
			<td style="width:129px;">SIGMA</td>
			<td style="width:167px;">P9582</td>
			<td style="width:133px;"></td>
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		<tr height="21">
			<td height="21" style="height:21px;">0.9% sodium chloride INJ. USP</td>
			<td>HOSPIRA</td>
			<td>NDC 0409-4888-10</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">MONOJECT insuline syrinage 3/10 mL 29G x 1/2&#34;</td>
			<td>COVIDIEN</td>
			<td>8881600145</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">50 ml conical screw cap tubes</td>
			<td>USA SCIENTIFIC</td>
			<td>1500-1211</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Nanodrop 1000 spectrophotometer</td>
			<td>THERMO SCIENTIFIC</td>
			<td>1000</td>
			<td>Optional</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Stereomicroscope</td>
			<td>Wild Heerbrugg</td>
			<td>M5A</td>
			<td>Optional</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dumont #5 Forceps Inox Tip Size .10 X .06mm</td>
			<td>Roboz</td>
			<td>RS-5045</td>
			<td>Optional</td>
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		<tr height="21">
			<td height="21" style="height:21px;">RNAlater RNA stabilization reagent</td>
			<td>Qiagen</td>
			<td>76104</td>
			<td>Optional</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">TRIzol reagent</td>
			<td>Life Technologies</td>
			<td>15596-026&#160;</td>
			<td>Optional</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">RQ1 Rnase-free DNase</td>
			<td>Promega</td>
			<td>M610A</td>
			<td>Optional</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">iScript cDNA synthesis kit</td>
			<td>Bio-Rad</td>
			<td>170-8891</td>
			<td>Optional</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">FastStart universal SYBR green master mix with ROX&#160;</td>
			<td>Roche</td>
			<td>4913922001</td>
			<td>Optional</td>
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		<tr height="21">
			<td height="21" style="height:21px;">Real-time PCR</td>
			<td>ABI</td>
			<td>7300</td>
			<td>Optional</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Primer: Il6 forward</td>
			<td>Life Technologies</td>
			<td>TAGTCCTTCCTACCCCAATTTCC</td>
			<td>Optional</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Primer: Il6 Reverse</td>
			<td>Life Technologies</td>
			<td>TTGGTCCTTAGCCACTCCTTC</td>
			<td>Optional</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Primer: beta-actin forward</td>
			<td>Life Technologies</td>
			<td>AGAGGGAAATCGTGCGTGAC</td>
			<td>Optional</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Primer: beta-actin Reverse</td>
			<td>Life Technologies</td>
			<td>CAATAGTGATGACCTGGCCGT</td>
			<td>Optional</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">MicroAmp optical 96-well reaction plate</td>
			<td>Life Technologies</td>
			<td>4306737</td>
			<td>Optionl</td>
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			<td height="21" style="height:21px;">MicroAmp optical adhesive film&#160;</td>
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			<td>4311971</td>
			<td>Optionl</td>
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			<td height="21" style="height:21px;">EthoVision</td>
			<td>Noldus</td>
			<td>EthoVision</td>
			<td>Optionl</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">SR-LAB apparatus (PPI)</td>
			<td>San Diego Instruments&#160;</td>
			<td>SR-LAB</td>
			<td>Optionl</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Marbles</td>
			<td>PENN-PLAX</td>
			<td>Blue gem stones marbles</td>
			<td>Optionl</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dulbecco&#39;s Phosphate-Buffered Saline (DPBS)</td>
			<td>Life Technologies</td>
			<td>21600-069</td>
			<td>Optionl</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Paraformaldehyde</td>
			<td>MACRON</td>
			<td>2621-59</td>
			<td>Optionl</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Vibratome</td>
			<td>Leica</td>
			<td>VT1000 S</td>
			<td>Optionl</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sodium azide</td>
			<td>Sigma</td>
			<td>S2002</td>
			<td>Optionl</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Triton x-100</td>
			<td>Sigma</td>
			<td>X100</td>
			<td>Optionl</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hydrogen peroxide solution</td>
			<td>Sigma</td>
			<td>18312</td>
			<td>Optionl</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Goat serum</td>
			<td>Vector Laboratories</td>
			<td>S-1000</td>
			<td>Optionl</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Rabbit anti-calbindin antibody</td>
			<td>Abcam</td>
			<td>ab11426</td>
			<td>Optionl</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Biotinlyated goat anti-rabbit IgGantibody</td>
			<td>Vector Laboratories</td>
			<td>BA-1000</td>
			<td>Optionl</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">VECTASTAIN ABC Kit</td>
			<td>Vector Laboratories</td>
			<td>PK-4000</td>
			<td>Optionl</td>
		</tr>
	</tbody>
</table>

induction of maternal immune activation in mice at mid-gestation stage with viral mimic poly(i:c)

Maternal immune activation (MIA) model is increasingly well appreciated as a rodent model for the environmental risk factor of various psychiatric disorders. Numerous studies have demonstrated that MIA model is able to show face, construct, and predictive validity that are relevant to autism and schizophrenia. To model MIA, investigators often use viral mimic polyinosinic:polycytidylic acid (poly(I:C)) to activate the immune system in pregnant rodents. Generally, the offspring from immune activated dam exhibit behavioral abnormalities and physiological alterations that are associated with autism and schizophrenia. However, poly(I:C) injection with different dosages and at different time points could lead to different outcomes by perturbing brain development at different stages. Here we provide a detailed method of inducing MIA by intraperitoneal (i.p.) injection of 20 mg/kg poly(I:C) at mid-gestational embryonic 12.5 days (E12.5). This method has been shown to induce acute inflammatory response in the maternal-placental-fetal axis, which ultimately results in the brain perturbations and behavioral phenotypes that are associated with autism and schizophrenia.

<html>的20毫克/公斤的聚注射：在E12.5（I C）可以唤起在母体胎盘胎儿轴急性炎症反应并沉淀到大脑发育和行为表型12,13慢性效果。一个促炎性细胞因子水平升高，白细胞介素（IL）-6，是MIA后急性炎症反应的可靠指标。高峰时间在胎盘IL6的基因表达和胎儿大脑是在3小时后的聚（I：C）注射12,13。我们已经表明，聚（I：C）诱导整个脾脏产妇胎盘，胎儿的...

Watch this Scientific Journal Video about Induction of Maternal Immune Activation in Mice at Mid-gestation Stage with Viral Mimic PolyI:C at JoVE.com

Induction of Maternal Immune Activation in Mice at Mid-gestation Stage with Viral Mimic PolyI:C

母体免疫激活诱导小鼠的病毒性模仿聚妊娠中期阶段（I：C）

Maternal immune activation (MIA) is a model for an environmental risk factor of autism and schizophrenia. The goal of this article is to provide a step-by-step procedure of how to induce MIA in the pregnant mice in order to enhance the reproducibility of this model.

Maternal immune activation (MIA) model is increasingly well appreciated as a rodent model for the environmental risk factor of various psychiatric ...

Research

JoVE Journal

Immunology and Infection

Induction of Maternal Immune Activation in Mice at Mid-gestation Stage with Viral Mimic Poly(I:C)

Induction of Experimental Autoimmune Hypophysitis in SJL Mice

SJL小鼠诱导实验性自身免疫性垂体炎

Autoimmune hypophysitis can be reproduced experimentally by the injection of pituitary proteins mixed with an adjuvant into susceptible mice1. Mouse ...

科研

免疫与感染

Seven Steps to Stellate Cells

星状细胞的七个步骤

Hepatic stellate cells are liver-resident  cells of star-like morphology and are located in the space of Disse between  liver sinusoidal endothelial cells ...

Measuring Bacterial Load and Immune Responses in Mice Infected with Listeria monocytogenes

Measuring Bacterial Load and Immune Responses in Mice Infected with Listeria monocytogenes

测量细菌感染小鼠和免疫反应李斯特菌</em

Listeria monocytogenes (Listeria) is a Gram-positive facultative intracellular pathogen1. Mouse studies typically employ intravenous injection of ...

Right Ventricular Systolic Pressure Measurements in Combination with Harvest of Lung and Immune Tissue Samples in Mice

右心室收缩压测量组合在小鼠的肺和免疫组织样品的收获

The function of the right heart is to pump blood through the lungs, thus linking right heart physiology and pulmonary vascular physiology. Inflammation is ...

Dissecting Innate Immune Signaling in Viral Evasion of Cytokine Production

细胞因子产生的病毒逃避解剖先天免疫信号

In response to a viral infection, the host innate immune response is activated to up-regulate gene expression and production of antiviral cytokines. ...

Induction of Murine Intestinal Inflammation by Adoptive Transfer of Effector CD4+CD45RBhigh T Cells into Immunodeficient Mice

Induction of Murine Intestinal Inflammation by Adoptive Transfer of Effector CD4+CD45RBhigh T Cells into Immunodeficient Mice

诱导小鼠肠道炎症由效应CD4的过继转移<sup&gt; +</sup&gt; CD45RB<sup&gt;高</sup&gt; T细胞免疫缺陷小鼠

There are many different animal models available for studying the pathogenesis of human inflammatory bowel diseases (IBD), each with its own advantages ...

Isolation of Leukocytes from the Murine Tissues at the Maternal-Fetal Interface

从小鼠组织在母胎界面分离白细胞

Immune tolerance in pregnancy requires that the immune system of the mother undergoes distinctive changes in order to accept and nurture the developing ...

Induction of Experimental Autoimmune Encephalomyelitis in Mice and Evaluation of the Disease-dependent Distribution of Immune Cells in Various Tissues

在免疫细胞在不同组织与疾病相关的分布和小鼠实验评价性自身免疫性脑脊髓炎的诱导

Multiple sclerosis is presumed to be an inflammatory autoimmune disease, which is characterized by lesion formation in the central nervous system (CNS) ...

Human Placental and Decidual Organ Cultures to Study Infections at the Maternal-fetal Interface

人胎盘及蜕膜器官培养研究在母胎界面感染

The placenta shows a large degree of interspecies anatomic variability. To best understand biology and pathophysiology of the human placenta, it is ...

Determining Immune System Suppression versus CNS Protection for Pharmacological Interventions in Autoimmune Demyelination

确定免疫系统抑制中枢神经系统对保护药物干预自身免疫性脱髓鞘

A major hallmark of the autoimmune demyelinating disease multiple sclerosis (MS) is immune cell infiltration into the brain and spinal cord resulting in ...