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Method for determination of calcium concentration in rat spinal dorsal horn neuron

An assay method, neuron technology, applied in material excitation analysis, fluorescence/phosphorescence, etc., can solve problems such as influence

Inactive Publication Date: 2015-11-18
ZUNYI MEDICAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Can GC Regulate Ca in Spinal Cord Dorsal Horn Neurons? 2+ Concentration affects its function, yet to be proved experimentally

Method used

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  • Method for determination of calcium concentration in rat spinal dorsal horn neuron
  • Method for determination of calcium concentration in rat spinal dorsal horn neuron
  • Method for determination of calcium concentration in rat spinal dorsal horn neuron

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Embodiment

[0028] The rat spinal cord horn neuron calcium concentration determination method of the present invention comprises the following steps:

[0029] 1 Culture of spinal cord dorsal horn neurons

[0030] Sprague-Dawley rats (3 ), transferred to a centrifuge tube, added 0.125% trypsin and placed at 37°C, 5% CO 2 Digest in a cell culture incubator for 30 minutes; then add DMEM / F12 medium containing 10% fetal bovine serum to terminate the digestion; pipette into a cell suspension, put it in a centrifuge for 5 minutes at 1000r / min, and discard the supernatant; add NB / Resuspend the cells in 200 μl of B27 medium, inoculate them in a laser confocal dish double-coated with polylysine and laminin, and let them stand in the incubator for 3 hours; add 1.5ml of NB / B27 medium (containing penicillin, streptomycin).

[0031] 2 Detection of neuronal Ca by laser confocal technique 2+ concentration change

[0032] Spinal cord dorsal horn neurons were seeded in laser confocal dishes, and the e...

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Abstract

The invention discloses a method for determination of calcium concentration in a rat spinal dorsal horn neuron. The method includes: taking a SD neonatal rat, conducting disinfection and then executing it, separating the spinal dorsal horn tissue, stripping a spinal cord outer membrane, cutting the tissue into small pieces, transferring the pieces into a centrifuge tube, and adding trypsin to conduct digestion; then adding a DMEM / F12 medium containing 10% fetal calf serum to terminate the digestion; conducting blowing to obtain a cell suspension, performing centrifugation and discarding the supernatant; adding an NB / B27 medium to resuspend cells, inoculating the cells into a special laser confocal dish, and performing standing in an incubation box; adding an NB / B27 medium containing penicillin and streptomycin; performing loading with a fluorescent indicator, putting the cells into the incubation box to perform incubation, rinsing the NB medium, then covering the cells with the NB medium, and carrying out scanning detection with a laser scanning confocal microscope. The method for determination of calcium concentration in the rat spinal dorsal horn neuron provided by the invention can be used for observing the influence of corticosterone on the Ca<2+> concentration of the cultured spinal dorsal horn neuron, and creates conditions for studying the regulation of corticosterone on spinal dorsal horn sensory neuron functions and the action mechanism thereof.

Description

technical field [0001] The invention belongs to the field of biomedicine detection, in particular to a method for measuring the calcium concentration of rat spinal cord horn neurons. Background technique [0002] Glucocorticoid (GC) has a wide range of physiological effects and regulates various functions of the body. It is commonly used clinically for anti-inflammation, suppression of immune response and membrane stabilization. GC plays a role through genomic and non-genomic pathways. The non-genomic role has only been gradually recognized and valued in the past 30 years. Its effect occurs rapidly and does not involve gene expression and protein synthesis. However, its molecular mechanism is still unclear . [0003] Recent studies have shown that GC plays an important role in the transmission of nociceptive information and the occurrence of pathological pain. Literature data show that GC is involved in the modulation of nociceptive information in both peripheral and cent...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
Inventor 刘晓红曾俊伟
Owner ZUNYI MEDICAL UNIVERSITY
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