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大鼠胚胎第16天腹侧中脑的差异解剖以及醛脱氢酶阳性TH神经元亚群对6-羟基多巴胺损伤纹状体的存活和再支配

Differential dissection of the rat E16 ventral mesencephalon and survival and reinnervation of the 6-OHDA-lesioned striatum by a subset of aldehyde dehydrogenase-positive TH neurons.

作者信息

Haque N S, LeBlanc C J, Isacson O

机构信息

Neuroregeneration Laboratory, Harvard Medical School/McLean Hospital, Belmont, MA 02178, USA.

出版信息

Cell Transplant. 1997 May-Jun;6(3):239-48. doi: 10.1177/096368979700600307.

Abstract

The retinoic acid-generating enzyme, aldehyde dehydrogenase (AHD), is expressed in a subpopulation of dopaminergic neurons found in the substantia nigra. Using AHD and tyrosine hydroxylase (TH) as immunohistochemical markers, we determined whether differential dissection of the embryonic (E16) ventral mesencephalon (VM) into its lateral and medial portions contributed equally to the number of TH cells surviving transplantation, if grafted AHD/TH neurons reinnervate the host striatum according to their normal projection patterns, and examined the functional recovery caused by the implanted cells as assessed by amphetamine-induced rotation in a 6-OHDA-lesioned model of Parkinson's disease. The embryonic tissue was transplanted as solid pieces injected via a 20-gauge lumbar puncture needle into the center of the deafferented striatum. Groups received either one complete ventral mesencephalic piece (VM), two medial pieces of ventral mesencephalic tissue (MVM), or two lateral pieces of ventral mesencephalic tissue (LVM). Both VM and MVM groups showed a significant decrease in amphetamine-induced rotation over time and, there was no difference in the degree of reduction observed between the two groups. Histological evaluation of the transplants revealed a much larger total number of surviving TH cells in grafts from the VM and MVM groups compared to the LVM group. Surviving AHD/TH neurons were found in all groups. Whereas TH staining of the transplanted striatum displayed a halo of graft-derived fibers all around the transplant and integration of these fibers into the host neuropil, AHD staining showed a preferential reinnervation of the dorsolateral striatum corresponding to the normal projection pattern of AHD/TH neurons. In summary, selective dissection of the embryonic ventral mesencephalon is possible, functional recovery as assessed by amphetamine-induced rotation in animals transplanted with MVM is similar to that seen in animals grafted with VM, and AHD/TH neurons have a selective reinnervation pattern in the PD transplantation paradigm. These findings may have implications for the grafting of fetal mesencephalic tissue in PD patients.

摘要

维甲酸生成酶醛脱氢酶(AHD)在黑质中的多巴胺能神经元亚群中表达。我们以AHD和酪氨酸羟化酶(TH)作为免疫组织化学标记物,确定将胚胎期(E16)腹侧中脑(VM)分别分离为外侧和内侧部分后,移植后存活的TH细胞数量是否相同;移植的AHD/TH神经元是否按照其正常投射模式重新支配宿主纹状体;并通过帕金森病6-OHDA损伤模型中苯丙胺诱导的旋转来评估植入细胞引起的功能恢复情况。胚胎组织以固体块的形式通过20号腰穿针注射到去传入神经的纹状体中心。各组分别接受一块完整的腹侧中脑组织(VM)、两块内侧腹侧中脑组织(MVM)或两块外侧腹侧中脑组织(LVM)。VM组和MVM组的苯丙胺诱导旋转均随时间显著减少,两组间减少程度无差异。对移植物的组织学评估显示,与LVM组相比,VM组和MVM组移植物中存活的TH细胞总数要多得多。所有组均发现有存活的AHD/TH神经元。移植纹状体的TH染色显示移植物周围有一圈源自移植物的纤维,且这些纤维整合到宿主神经毡中,而AHD染色显示背外侧纹状体有优先的再支配,这与AHD/TH神经元的正常投射模式一致。总之,胚胎腹侧中脑的选择性分离是可行的,MVM移植动物中通过苯丙胺诱导旋转评估的功能恢复与VM移植动物相似,且在帕金森病移植模型中AHD/TH神经元具有选择性再支配模式。这些发现可能对帕金森病患者胎儿中脑组织移植有重要意义。

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