长爪沙鼠脑底动脉Willis环变异缺失遗传特性分析及脑缺血模型高发群体的初步培育

目的研究长爪沙鼠脑底动脉Willis环遗传特性,并定向培育脑缺血高发种群。方法通过对5代定向培育的长爪沙鼠高发群动物共398只动物的右侧颈总动脉结扎模型进行观察,比较长爪沙鼠亲代与子代间脑底动脉Willis环的变异缺失类型,探求其遗传特性,并根据该遗传特性将Willis环后交通支缺失且前交通支缺失或细小的同类型的长爪沙鼠父母所生的子代雌雄个体配对繁殖,定向培育长爪沙鼠半脑缺血模型高发生率种群。结果当双亲的Willis环类型一致时,其子代大部分与其父母一致;而当双亲的Willis环类型不一致时,Willis环前交通支与母亲一致率为60.4%,前交通支与父亲的一致率为48.2%,两者的差异有显著性意义（P=0.015）。长爪沙鼠半脑缺血模型高发生率种群定向培育5代后,行单侧颈总动脉结扎时,一侧脑缺血造模成功率由F1代的40%提高到F5代的75%。结论长爪沙鼠脑底动脉Willis环变异有明显的遗传性,初步培育出了长爪沙鼠半脑缺血模型高发生率种群。     

长爪沙鼠脑缺血模型近交系的培育及其发病机制研究进展

长爪沙鼠具有脑血管变异缺失的特性,是脑缺血研究良好的模型动物。普通长爪沙鼠群体中模型成功率低,影响研究结果稳定性,不符合实验动物福利要求。我们团队在证实脑血管Willis环(circle of Willis,COW)变异缺失具有遗传性的基础上,首先通过定向选育建立了脑缺血高发群体,进而通过全同胞近亲繁殖的方式,建立了脑缺血模型近交系,其COW缺失率达76.62%,模型成功率达到88.89%。此外,建立了近交系微卫星DNA和生化位点遗传检测方法,分析了长爪沙鼠脑缺血模型近交系动物的生长发育和血液生理生化等指标,为该模型的推广应用奠定了基础。我们还发现,VEGFA基因、AKT/PI3K和Notch信号通路在长爪沙鼠脑血管发育中发挥重要作用。通过抑制消减杂交方法筛选获得4个与长爪沙鼠脑血管发育相关的基因。长爪沙鼠脑缺血模型近交系的培育成功解决了模型发生率低的问题,减少了动物使用量,提高了研究结果可信度和稳定性,促进了相关机制研究。     

树qu（Tupaia belangeri chinensis）的脑底动脉环

用25只树qu,从升主动脉灌注带色的橡胶乳液,在解剖显微镜下进行解剖观察,用目测微尺进行测量。大多数树qu（22只）有完整的脑底动脉环。由左、右大脑前动脉向内侧各发一前交通动脉组成大脑前总动脉。前交通动脉口径为大脑前动脉的75-85%。后交通动脉口径与大脑后动脉相近,连于颈内动脉与大脑后动脉（基底动脉的分支）之间。测量了组成脑底动脉环有关动脉的口径。由于后交通动脉足够粗大,只有中断左、右颈总动脉和左、右椎动脉,才能造成全脑缺血。     

树鼩(Tupaia belangeri chinensis)的脑底动脉环

用25只树鼩,从升主动脉灌注带色的橡胶乳液,在解剖显微镜下进行解剖观察,用目测微尺进行测量。大多数树鼩(22只)有完整的脑底动脉环。由左、右大脑前动脉向内侧各发一前交通动脉组成大脑前总动脉。前交通动脉口径为大脑前动脉的75～85％。后交通动脉口径与大脑后动脉相近,连于颈内动脉与大脑后动脉(基底动脉的分支)之间。测量了组成脑底动脉环有关动脉的口径。由于后交通动脉足够粗大,只有中断左、右颈总动脉和左、右椎动脉,才能造成全脑缺血。     

正常成人脑主要供血动脉的MSCT血管成像研究

目的:观测正常脑动脉多层螺旋CT血管成像(multi-slice spiral CT angiography,MSCTA)主要动脉的起始部内径、走行、形态及其分支的显示情况,为脑血管疾病的诊断提供参考资料.方法:选择40例健康体检者行全脑血管64层MSCTA检查,采用ADW4.2工作站测量各主要动脉起始部内径,观测其走行、形态和变异,并统计各级分支的显示率.结果:正常成人脑动脉常见的变异发生在组成脑底动脉环的大脑前动脉A1段、前后交通动脉及大脑后动脉起始段,以后交通动脉变异最大.颈内动脉末段、大脑前动脉A1段、大脑中动脉、大脑后动脉、眼动脉、前交通动脉、后交通动脉及脉络膜前动脉在MSCTA上的显示率分别为100%、95%、100%、100%、100%、92.5%、58.8%和72.5%.主要脑动脉起始部内径分别是:颈内动脉末段4.013±0.770 mm、大脑前动脉A1段2.709±0.877 mm、大脑中动脉3.498±0.640 mm、大脑后动脉2.025±0.608 mm、眼动脉1.640±0.334 mm、后交通动脉1.491±0.697mm及脉络膜前动脉1.460±0.483mm.结论:正常成人脑血管变异主要发生于脑底动脉环,脑血管直径与其供血范围及分布区血流动力学因素相关.     

炔雌醚对内蒙古农牧交错带长爪沙鼠(Meriones unguiculatus)家群大小和巢域行为的影响

为检验施用炔雌醚对农牧交错带长爪沙鼠家群大小与巢域行为的影响,于2006年5月至9月对内蒙古锡林郭勒盟锡林浩特市农牧交错区域分布的长爪沙鼠家群使用炔雌醚进行种群不育控制实验。分设投药区和对照区两个组别,分别于5月、7月与9月份采用1/4样圆面积有效洞口计数法,随机选取24个长爪沙鼠家群,调查对照区域和投药区内沙鼠的家群洞口数量以及家群大小。另在对照区和投药区,随机测定了30个沙鼠的家群活动范围,对比使用炔雌醚前后长爪沙鼠巢域平均半径和活动面积的影响。结果为:对照区家群平均洞口数58个,投药区仅23个,在炔雌醚投药区长爪沙鼠家群受炔雌醚影响,洞口数仅为23个,表明其对长爪沙鼠家群大小控制效果显著。投药区内长爪沙鼠家群密度在投药后连续下降,最大下降幅度较对照区家群密度低70%,表明炔雌醚对长爪沙鼠的家群密度影响较大,可显著降低长爪沙鼠家群密度(P<0.05)。炔雌醚对长爪沙鼠家群平均半径和活动面积的研究结果显示,施用炔雌醚后家群巢域半径与活动面积均显著缩小(P<0.05),与对照区巢域平均面积相比缩小15%、平均活动半径缩小30%。通过以上结果可得出:单独施用炔雌醚对控制野外长爪沙鼠家群以及降低长爪沙鼠巢域、活动面积效果显著。炔雌醚可有效降低农牧交错带长爪沙鼠种群数量和有效活动范围,这对于农牧交错带的鼠害防控,同时降低鼠源性疾病,包括鼠疫的传播都有一定的意义。     

长爪沙鼠脑缺血后大脑皮质Parvalbumin(PV)的表达变化

目的:观察长爪沙鼠大鼠前脑短暂缺血后前额皮质Parvalbumin(PV)的表达变化,为进一步研究其功能及其对脑缺血损伤的治疗作用提供形态学依据。方法:24只健康雄性长爪鼠随机分为时照组(6只)和缺血组(18只),夹闭长爪沙鼠双测颈总动脉10min诱导前脑缺血后,动物分别存活1天、3天或7天。用免疫组化方法、阳性细胞计数和相对平均灰度值检测了前额皮质中PV的表达情况。结果:与正常组相比,各缺血组中前额皮质中PV表达均减少。缺血再灌1天时,PV表达最弱(P＜0．01);3天时PV表达开始恢复(P＜0．05);7天时PV表达进一步恢复,但仍低于正常组(P＞0．05)。结论:前脑短暂缺血后可造成长爪沙鼠前额皮质PV的表达在短时间内急剧下降;但随着缺血消失后时间的延长,机体可进行部分的自行修复。     

长爪沙鼠对植物血凝素的反应及能量学代价的性别和季节差异

以野生长爪沙鼠为对象,通过外源注射植物血凝素(Phytohemagglutinin,PHA)和磷酸盐缓冲液,分别测定了夏季和冬季沙鼠注射前、注射24 h 和48 h 后体重和静止代谢率(Resting metabolic rate,RMR)的变化,以及48 h 后沙鼠足增重量和白细胞总数(White blood cell counts,WBCs)的变化,以检测沙鼠的免疫功能和能量代价的性别和季节差异。结果显示:1)注射PHA 能显著增加沙鼠足重和白细胞的总数,夏季白细胞的总数高
于冬季;2)冬季沙鼠体重高于夏季,雄鼠大于雌鼠,但PHA 处理对注射前后沙鼠的体重无影响;3)冬季RMR高于夏季,雄鼠大于雌鼠,但PHA 处理对沙鼠的RMR 无影响,PHA 处理对注射前后沙鼠的RMR 也无影响。这些结果表明,长爪沙鼠对PHA 的反应具有季节差异,但无性别差异,也没有发现明显的能量学代价,这可能与野外环境条件的大幅度波动(如环境温度和食物条件等)和沙鼠的繁殖状态有关。     

种间转移扩增法筛选长爪沙鼠微卫星位点

目的筛选长爪沙鼠新的微卫星位点,为长爪沙鼠遗传分析提供遗传标记物。方法从GenBank中随机选取小鼠微卫星位点引物536对,用这些引物对长爪沙鼠基因组DNA扩增,将阳性目的条带进行序列分析,找出符合微卫星序列特征的短串联重复序列。结果 536对小鼠微卫星引物在长爪沙鼠基因组中扩增出了313个阳性条带,经序列分析,确定130个长爪沙鼠微卫星位点;其中完美型位点占80.77%(105/130),不完美型位点占19.23%(25/130),与小鼠同源性为24.3%(130/536)。将筛选出的微卫星位点在GenBank中注册,注册号从GU562694到GU562823。结论小鼠和沙鼠的微卫星位点具有较高的同源性,用小鼠的微卫星位点引物直接扩增长爪沙鼠基因组DNA可有效地筛选出长爪沙鼠微卫星位点。     

长爪沙鼠资源开发利用进展

长爪沙鼠是源于我国的＂多功能＂实验动物资源,在某些研究领域发挥重要作用。随着研究深入,越来越多的长爪沙鼠生物学特性将被发现,这些将使长爪沙鼠资源利用更加多元化。本文对长爪沙鼠的开发研究历程及在分类学、寄生虫学、脑缺血、脂质代谢、神经性疾病和肿瘤学等研究中的应用作简要概述。     

Characteristics of circle of Willis variations in the mongolian gerbil and a newly established ischemia-prone gerbil group

Mongolian gerbils (Meriones unguiculatus) have an incomplete circle of Willis (CoW), as a result of which approximately 30-40% of these animals develop focal cerebral ischemia after unilateral carotid occlusion (UCO). There are four types of patterns of the anterior and posterior communicating arteries (ACoAs and PCoAs, respectively) of the CoW and they determine the severity of the ischemic symptoms. We used 398 gerbils from five generations, including a selectively bred ischemia-prone group, to investigate post-UCO ischemic symptoms and possible correlations of ACoA and PCoA patterns between parents and their progeny. We observed that if the parents had complete ACoAs, their progeny also had complete ACoAs, and we found significant differences when the parents' ACoAs were incomplete: in 60.4% of offspring the type of ACoA was consistent with that of the mother and in 48.2% it was consistent with that of the father. The severity of the neurological symptoms after UCO was significantly related to the patterns of the ACoAs when PCoAs were absent. The proportion of UCO ischemia in gerbils with incomplete ACoAs was significantly higher than in those with complete ACoAs. After selectively breeding five generations, the proportion of UCO ischemia increased from 40% in the F1 animals to 75% in the F5 animals. Our results suggest that variations in the CoW are genetic and demonstrate that we successfully established an ischemia-prone group of gerbils.     

Experimental study of hemodynamics in the circle of willis

Background

The Circle of Willis (CoW) is an important collateral pathway of the cerebral blood flow. An experimental study of the cerebral blood flow (CBF) distribution in different anatomical variations may help to a better understanding of the collateral mechanism of the CoW.

Methods

An in-vitro test rig was developed to simulate the physiological cerebral blood flow in the CoW. Ten anatomical variations were considered in this study, include a set of different degrees of stenosis in L-ICA and L-ICA occlusion coexist with common anatomical variations. Volume flow rates of efferent arteries and pressure signals at the end of communicating arteries of each case were recorded. Physiological pressure waveforms were applied as inlet boundary condition.

Results

In the development of L-ICA stenosis, the total CBF decreases with the increase of stenosis degree. The blood supply of ipsilateral middle cerebral artery (MCA) was affected most by the stenosis of L-ICA. Anterior communicating artery (ACoA) and ipsilateral posterior communicating artery (PCoA) function as important collateral pathways of cerebral collateral circulation when unilateral stenosis occurred. The blood supply of anterior cerebral circulation was compensated by the posterior cerebral circulation through ipsilateral PCoA when L-ICA stenosis degree is greater than 40% and the affected side was compensated immediately by the unaffected side through ACoA. Blood flow of the anterior circulation and the total CBF reached the minimum among all cases studied when L-ICA occlusion coexist with the absence of PCoA.

Conclusion

The results demonstrated the flow distribution patterns of the CoW under anatomical variations and clarified the collateral mechanism of the CoW. The flow ACoA is the most sensitive indexes to the morphology change of ipsilateral ICA. The relative independence of the circulation in anterior and posterior sections of the CoW is not broken and the function of ipsilateral PCoA is not activated until a severe stenosis of unilateral ICA occurs. PCoA is the most important collateral pathway of the collateral circulation and the missing of PCoA has the highest risk of stroke when the ipsilateral ICA has severe stenosis. These findings may provide the basis for future therapeutic and diagnosis applications.

     

Species Differences in the Immunoreactive Expression of Oxytocin,Vasopressin, Tyrosine Hydroxylase and Estrogen Receptor Alpha in the Brain of Mongolian Gerbils (Meriones unguiculatus) and Chinese Striped Hamsters (Cricetulus barabensis)

Species differences in neurochemical expression and activity in the brain may play an important role in species-specific patterns of social behavior. In the present study, we used immunoreactive (ir) labeling to compare the regional density of cells containing oxytocin (OT), vasopressin (AVP), tyrosine hydroxylase (TH), or estrogen receptor alpha (ERα) staining in the brains of social Mongolian gerbils (Meriones unguiculatus) and solitary Chinese striped hamsters (Cricetulus barabensis). Multiple region- and neurochemical-specific species differences were found. In the anterior hypothalamus (AH), Mongolian gerbils had higher densities of AVP-ir and ERα-ir cells than Chinese striped hamsters. In the lateral hypothalamus (LH), Mongolian gerbils also had higher densities of AVP-ir and TH-ir cells, but a lower density of OT-ir cells, than Chinese striped hamsters. Furthermore, in the anterior nucleus of the medial preoptic area (MPOAa), Mongolian gerbils had higher densities of OT-ir and AVP-ir cells than Chinese striped hamsters, and an opposite pattern was found in the posterior nucleus of the MPOA (MPOAp). Some sex differences were also observed. Females of both species had higher densities of TH-ir cells in the MPOAa and of OT-ir cells in the intermediate nucleus of the MPOA (MPOAi) than males. Given the role of these neurochemicals in social behaviors, our data provide additional evidence to support the notion that species-specific patterns of neurochemical expression in the brain may be involved in species differences in social behaviors associated with different life strategies.     

Blood vessel size of circulus arteriosus cerebri (circle of Willis): a statistical research on 100 human subjects

The brain weight of 100 fresh cadavers of Italian subjects (60 males and 40 females), aged between 17 and 84 years, was obtained and the corrected circumference of the following blood vessels was measured: basilar artery, internal carotid arteries, anterior and posterior cerebral arteries, and anterior and posterior communicating arteries. The cerebral 'potential flow' was expressed in each case by adding the circumference of the basilar artery to that of the internal carotid arteries. Moreover, the sides and the perimeter of the circle of Willis as well as the length of the basilar artery were calculated. The statistical analysis of the obtained data yielded the following main results: (1) the brain weight decreases with aging, is lower in females than in males and and is statistically correlated neither with the circumferences of the considered arteries and the 'potential flow' nor with the perimeter of the arterial polygon (circle of Willis); (2) the arteries of the left side appear to be larger than those of the right one; (3) no significant difference exists in the circumference and length of the arteries between males and females; (4) the increase of the perimeter of the arterial polygon is achieved by means of a harmonious increase of all its sides; (5) the anterior and posterior communicating arteries have an anarchic pattern, because of the relatively frequent anomalies and the lack of a correlation between their circumference and that of the vessel of origin or of outlet.     

Development of Taenia asiatica cysticerci to infective stage and adult stage in Mongolian gerbils

The development of metacestodes and adult worms of Taenia asiatica in Mongolian gerbils (Meriones unguiculatus) were observed. Cysticerci were recovered from gerbils subcutaneously injected with hatched oncospheres. The recovery rate ranged from 0.1 to 3.2%. No cysticerci were recovered from the orally inoculated gerbils. The infectivity of the cysticerci recovered at 48 weeks post-infection was evaluated. Tapeworms were recovered on day 14 post-infection from the small intestine of 5 of 11 gerbils, with a recovery rate of 27% (6 worms recovered/22 worms inoculated). Three and four adult worms were recovered from two human volunteers who ingested five cysticerci after 4 months post-infection. In worms recovered from gerbils, segmentation and genital primordia in the posterior proglottids and hooklets in the residual rostellum were observed. The results indicate that gerbils can serve as an alternative intermediate host and that partial development of the adult worm stage occurs in gerbils.     

A 3D numerical study of the collateral capacity of the circle of Willis with anatomical variation in the posterior circulation

Background

The Circle of Willis (CoW) is the most important collateral pathway of the cerebral artery. The present study aims to investigate the collateral capacity of CoW with anatomical variation when unilateral internalcarotid artery (ICA) is occluded.

Methods

Basing on MRI data, we have reconstructed eight 3D models with variations in the posterior circulation of the CoW and set four different degrees of stenosis in the right ICA, namely 24%, 43%, 64% and 79%, respectively. Finally, a total of 40 models are performed with computational fluid dynamics simulations. All of the simulations share the same boundary condition with static pressure and the volume flow rate (VFR) are obtained to evaluate their collateral capacity.

Results

As for the middle cerebral artery (MCA) and the anterior cerebral artery (ACA), the transitional-type model possesses the best collateral capacity. But for the posterior cerebral artery (PCA), unilateral stenosis of ICA has the weakest influence on the unilateral posterior communicating artery (PCoA) absent model. We also find that the full fetal-type posterior circle of Willis is an utmost dangerous variation which must be paid more attention.

Conclusion

The results demonstrate that different models have different collateral capacities in coping stenosis of unilateral ICA and these differences can be reflected by different outlets. The study could be used as a reference for neurosurgeon in choosing the best treatment strategy.

     

Scanning electron microscope study of the communications between the anterior chamber and the sinus venosus sclerae in primates

We have studied the morphology of the anatomical structures that permit communication between the anterior chamber and the sinus venosus sclerae. Examination of the posterior or inner wall of this canal, represented by the sclerocorneal trabecula, in 15 species of primates and 5 adult humans, has enabled us to observe the existence of some small orifices or stomata that are the outermost part of the so-called Sondermann's canals, which in our opinion are made by the successive confluence of the interstices worked in the interior of the sclerocorneal trabecula by means of contraction of the longitudinal portion of the ciliary muscle.