Building of extracorporal ischemia model for oxygen-glucose deprivation on hippocampal section

A technology of glucose deprivation and hippocampus, applied in the field of biomedicine, can solve problems of different severity

Inactive Publication Date: 2017-05-31
宁波美丽人生医药生物科技发展有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although rollingNagoya and leaner mutant mice exhibit normal lifespans, their a

Method used

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  • Building of extracorporal ischemia model for oxygen-glucose deprivation on hippocampal section
  • Building of extracorporal ischemia model for oxygen-glucose deprivation on hippocampal section
  • Building of extracorporal ischemia model for oxygen-glucose deprivation on hippocampal section

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Experimental program
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Effect test

Embodiment 1

[0037] Expression pattern of Cav2.1α1 mRNA

[0038] We used in situ hybridization to evaluate the distribution of Cav2.1α1 mRNA in wild-type mice (n=6), rollingNagoya (n=5) and leanermice (n=5); the distribution in the three mice was the same (Data not shown). We detected a widespread expression of α1 subunit in the brains of all three mice using antisense probes, especially in the olfactory bulb neurons, cerebral cortex neurons, hippocampus neurons and cerebellum neurons There are strong expressions. However, no signal was detected after using the sense probe. We used real-timeqRT-PCR to detect the expression levels of CaV2.1α1mRNA in the olfactory bulb, cerebral cortex, caudate putamen, hippocampus, cerebellum and liver in these three mice. In the olfactory bulb, cerebral cortex, caudate putamen, hippocampus, and cerebellum of these three mice, the relative expression levels of total CaV2.1α1 were not significantly different (wild-type mice, rolling Nagoya and leaner mice...

Embodiment 2

[0040] CaV2.1α1 mutant mice have reduced infarct volume in an in vivo ischemia model

[0041]Wild-type mice (n=12), rollingNagoya (n=11) and leanermice (n=12) were treated with middle cerebral artery occlusion. We distinguish cerebral infarction from the color, which is a white unstained area surrounded by red living tissue. Fig.1A is a set of representative experimental results, which are the results obtained after staining for 24 hours after the infarct was permanently occluded with TTC. After treatment with middle cerebral artery occlusion, the infarct area was significantly different in the three mice. The proportion of the infarct region to the total brain volume was 27.1±3.5% in rollingNagoya, 20.2±3.5% in leanermice, and 42.9±4.5% in wild-type mice.

Embodiment 3

[0043] CaV2.1α1 gene mutant mice in in vitro ischemia model [Ca 2+ ]i reduction

[0044] To investigate whether the reduction in infarct volume in mutant mice is related to the neuronal calcium pathway, we used hippocampal slices to examine [Ca 2+ ]i changes, OGD treatment can make local ischemia. Ca V The 2.1 channel is strongly expressed in the hippocampus. Before OGD treatment, there was no significant difference in the basal ratio of the pyramidal cell layer in the CA1 region among the three mice (wild type: 0.847±0.011; rollingNagoya: 0.827±0.022; leanermice: 0.803±0.013). Once OGD starts processing, [Ca 2+ ]i appeared increasing, and within 4 minutes after treatment, the normalized index of wild-type mice was much higher than that of the other two groups of mice. The maximum rate of increase was also different among the three mice. The rate of increase was 0.083±0.007 / min (p<0.01) in rollingNagoya, 0.062±0.006 / min (p<0.01) in leanermice, and 0.105±0.008 / min in wild...

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Abstract

The invention relates to the field of biological medicines, in particular to building of an extracorporal ischemia model for oxygen-glucose deprivation on a hippocampal section. Cell deaths and ischemia caused by cerebral injury mainly result from dysregulation of Ca2+ ions. Depolarization of neuron cytomembrane results in activation of voltage gate Ca2+ control aisle and internal flow of Ca2+. In experiments, two kinds of Cav2.1 aisle alpha 1 subunit mutant mice (rolling Nagoya and leaner mice) are utilized to study the physiological effect of CaV2.1 (P-Q type) in ischemic neuron injury. As partial ischemia experiments show, the brain tissue embolism areas of the two kinds of mutant models of rolling Nagoya and leaner mice are obviously reduced compared with those of wild type mice. In the study of extracorporal Ca2+ imaging, the growth rate of Ca2+ concentration in the two kinds of Cav2.1 aisle mutant mice is very low compared with that of the wild type mice. These study results show that it is necessary to build the extracorporal ischemia model for oxygen-glucose deprivation on the hippocampal section.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to the establishment of an in vitro ischemia model of oxygen-glucose deprivation on hippocampal slices. Background technique [0002] During membrane depolarization, voltage-gated Ca 2+ (Ca V ) channel allows Ca 2+ into the cell. In the nervous system, Ca V Channels play an important role in the regulation of diverse neuronal functions due to intracellular Ca 2+ increase in concentration [1] . Ca V A channel is a complex molecular complex composed of α1, β, α2-δ and γ subunits [2] . The α1 subunit is essential for proper channel function and for determining the fundamental properties of the channel [1] . At presynaptic terminals, there are three major Ca V2 Channel type, Ca V2.1 (P / Q type), Ca V2.2 (N-type) and Ca V2.3 (R type), these three types are widely expressed in the central nervous system [3] and participate in Ca 2+ dependent exocytosis release of neurotransmitte...

Claims

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Application Information

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IPC IPC(8): A61D1/00C12Q1/68
CPCA61D1/00C12Q1/6888C12Q2600/158
Inventor 田晓丽
Owner 宁波美丽人生医药生物科技发展有限公司
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