Method for detecting cortical neuron axonal growth guidance and application thereof

A technology of neurons and axons, applied in the field of neuroscience, to achieve the effect of excellent optical transparency and flexible methods

Inactive Publication Date: 2015-07-15
SHANGHAI TENTH PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this method needs to select neurons with larger growth cones (such as Xenopus spinal cord neurons),

Method used

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  • Method for detecting cortical neuron axonal growth guidance and application thereof
  • Method for detecting cortical neuron axonal growth guidance and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0036] 1. Preparation of PDMS stamp

[0037] Raw materials and instruments: mold (the surface of the mold is engraved with parallel fine grooves at equal intervals, the groove width is 60 μm, the groove height is 100 μm, and the interval between each groove is 90 μm.), polydimethylsiloxane ( PDMS) (Momentive RTV 615A solution, B solution, Suzhou Wenhao Chip Technology Co., Ltd.), thick aluminum foil, petri dish (150mm); vacuum box, oven, scalpel.

[0038] process:

[0039] 1. Spread aluminum foil on a large petri dish, weigh liquid A and liquid B (respectively prepolymer and coupling agent), stir well, turn up the edge of the aluminum foil, and vacuum in a vacuum box for 10 minutes (Note 1: pump When vacuuming, it can be pumped all the time; it can also be pumped and put at the same time);

[0040] 2. Spread aluminum foil on a new petri dish, and flatten it around. Put the mold on the aluminum foil, and pour the vacuum-pumped glue in 1 into the groove of the mold (Note 2: Be...

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PUM

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Abstract

The invention relates to a method for detecting cortical neuron axonal growth guidance. The method comprises the following steps: preparing a stamp with a microscopy channel on a mold through polydimentylsiloxane, buckling the stamp on a glass slide for coating poly-L-lysine, adding a solution containing guidance factors and fluorescent molecules from one side of the channel so as to form parallel stripes with equal intervals after entering ducts; and performing neuron cultivation on the glass slide coated with the fluorescent stripe, observing whether newborn axon enters the stripe containing the guidance factors, and computing proportion of the newborn axon and the stripe, statistically analyzing. The method disclosed by the invention is not only in favor of understanding the forming mechanism of the neural circuit in a brain, but also has potential treatment value to the axonal degeneration and axonal injury diseases. On the basis of the method for detecting the cortical neuron axonal growth guidance, a disease model is established, the specificity expression of the secretion factor in the brain in the time and the space is simulated through exogenously giving cell secretion factor, the functional reconstruction of the neutron is induced, and the experimental basis is established for clinical translational medicine for treating above diseases.

Description

technical field [0001] The invention relates to the technical field of neuroscience, in particular to a method for detecting the axon growth orientation of cortical neurons and its application. Background technique [0002] During brain development, neurons differentiate into axons and dendrites, two regions with different functions, and form synaptic connections with other neurons for information transmission. During axon growth, long-distance pathfinding is required to establish connections with distant neurons. This process needs to be strictly regulated in order to form delicate neural circuits and complete the advanced functions of the brain. After the newborn neurons of the cerebral cortex are produced in the ventricle wall (Ventricular zone, VZ), they differentiate into two protruding cells (bipolar cells) at the front and back, and migrate outward along the radial glial fibers. During migration, leading processes differentiate into apical dendrites, and caudal proc...

Claims

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Application Information

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IPC IPC(8): G01N33/53
CPCG01N33/5058
Inventor 张敏赖思强宋瑷宏许东升
Owner SHANGHAI TENTH PEOPLES HOSPITAL
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