67 results about "GABAergic neuron" patented technology

The present disclosure is directed to improved methods for efficiently producing neuroprogenitor cells and differentiated neural cells such as GABAergic neurons from pluripotent stem cells, for example embryonic stem cells. Using the disclosed methods, cell populations containing a high proportion of GABAergic neurons have been isolated. The neuroprogenitor cells and terminally differentiated cells of the present disclosure can be generated in large quantities, and therefore may serve as an excellent source for cell replacement therapy in neurodegenerative disorders and neuronal diseases such as stroke, ischemia, epilepsy, and Huntington's disease.

This document provides methods and materials for generating GABAergic neurons in brains. For example, methods and materials for using nucleic acid encoding a NeuroD1 polypeptide and nucleic acid encoding a Dlx2 polypeptide to trigger glial cells (e.g., NG2 glial cells or astrocytes) within the brain (e.g., striatum) into forming GABAergic neurons (e.g., neurons resembling medium spiny neurons such as DARPP32-positive GABAergic neurons) that are functionally integrated into the brain of a living mammal (e.g., a human) are provided.

The invention relates to a method for constructing an animal model of epilepsy, belonging to the technical field of construction of animal models. The method comprises the steps of mating FGF9fl/fl mice with GABAergic neurons Cre mice to obtain FGF9fl/+VGAT-Cre hybrid knockout mice, and then mating the obtained hybrid knockout mice with FGF9fl/fl homozygous mice to obtain FGF9 homozygous knockoutmice (FGF9fl/flVGAT-Cre), namely, the animal model of epilepsy. The animal model of epilepsy is closer to the symptoms and mechanisms of clinical epilepsy, and provides a good research tool for the study of the etiology of clinical epilepsy as well as further diagnosis and treatment.

The present disclosure is directed to improved methods for efficiently producing neuroprogenitor cells and differentiated neural cells such as GABAergic neurons from pluripotent stem cells, for example embryonic stem cells. Using the disclosed methods, cell populations containing a high proportion of GABAergic neurons have been isolated. The neuroprogenitor cells and terminally differentiated cells of the present disclosure can be generated in large quantities, and therefore may serve as an excellent source for cell replacement therapy in neurodegenerative disorders and neuronal diseases such as stroke, ischemia, epilepsy, and Huntington's disease.

The invention provides a composition capable of specifically exciting GABAergic neurons in nucleus accumbens and application thereof in improving schizophrenia abnormal behaviors. The composition comprises a light-sensation-gene-carrying virus vector for infecting GABAergic neurons in nucleus accumbens and an irradiation device capable of generating blue light, wherein the vector comprises a promoter, a light sensation gene and a green fluorescence labeling gene; the promoter is an Syn promoter; the irradiation device is used for carrying out irradiation control on the infected nucleus accumbens to excite the nucleus accumbens. The composition can accurately and specifically excite GABAergic neurons in nucleus accumbens, and can effectively improve the schizophrenia abnormal behaviors.

To treat epilepsy or schizophrenia by transplanting precursor cells of GABAergic neurons into a region where GABAergic neurons are lost or decreased in the brain of a patient suffering from the disease, it is intended to provide a method for separating a precursor cell of GABAergic neuron in an adult or a fetal nerve tissue or a precursor cell of GABAergic neuron derived from an embryo stem cell. The invention of this application comprises the step of preparing a cell population containing a precursor cell of GABAergic neuron, the step of introducing a DNA, in which a reporter gene emitting fluorescence detectable even in vivo is attached to the downstream of a promoter of GAD67 gene or GAD65 gene that is gene of an inhibitory neurotransmitter GABA synthase, into dispersed cells, the step of isolating a GABAergic neuron and a precursor cell of GABAergic neuron based on the presence/absence of the fluorescence from the reporter protein, and the step of isolating the precursor cell of GABAergic neuron based on the possession of proliferative capability.

The invention discloses a method for preparing dopaminergic neurons and/or dopaminergic neural precursor cells and modified stem cells used by the method. The method comprises the following step that:the modified stem cells are induced and differentiated into the dopaminergic neurons and/or the dopaminergic neural precursor cells. The modified stem cells are in-vitro stem cells for expressing Nurr1 or expressing protein with the same function as the Nurr1. According to the method, Nurr1 genes are introduced into human pluripotent stem cells to obtain the modified stem cells, the Nurr1 genes are used for inducing the human pluripotent stem cells to differentiate towards a dopaminergic neuron subtype direction, and finally dopaminergic neural precursor cells and dopaminergic neurons are obtained. Compared with the prior method, the method can obviously enhance the efficiency of an inducing and differentiating process and obtaining of dopaminergic neuron subtype cells. The method can beused for treatment of diseases caused by dopaminergic neuron degeneration and/or damage, cell technology application, establishment of an in-vitro disease model, screening and development of drugs andthe like.

The present invention discloses a GABAergic neuron-specific promoter and an application thereof. The promoter is a brain neuron-specific promoter obtained from a mouse genome of a C57BL/6J breed and can regulate specific expression of genes in GABAergic neurons. The promoter is loaded into an adeno-associated virus vector, infects a mouse brain in vivo and can mediate the specific expression of the target gene in the GABAergic neuron, thereby marking or manipulating GABAergic neuron groups existing in the brain. The promoter has a wide range of application value and market prospects in structure and function analysis of the GABAergic neuron groups in specific brain regions, disease model establishment, gene therapy, etc.

The invention discloses a method for inducing differentiation of adipose-derived stem cells into dopaminergic neurons, and belongs to the technical field of biomedicine. The dopaminergic neurons are obtained through separation of the adipose-derived stem cells, subculture of the adipose-derived stem cells and induced culture and conversion, and a large number of the adipose-derived stem cells areobtained through separation and culture of adipose tissue from autologous sources; the adipose-derived stem cells are converted into the dopaminergic neurons through two specific culture media, batches of stable dopaminergic neurons can be obtained safely and effectively, and a high differentiation rate is achieved. It is shown through immunofluorescence and cell counting experiments that the conversion rate of the dopaminergic neurons is 90.4+/-0.76, the survival rate is 98.9+/-0.55, and the average amount of dopamine which is secreted by the converted dopaminergic neurons and is detected byan ELISA kit is 450 pg/ml; and therefore, it is confirmed that the steps of extraction and conversion of the adipose-derived stem cells are gentle and stable, and the extracted cells are suitable formass application.

The invention belongs to the technical field of traditional Chinese medicines, and particularly discloses application of a traditional Chinese medicine composition in preparation of a medicine for preventing or treating neurodegenerative diseases. The traditional Chinese medicine composition is prepared from leeches, ligusticum wallichii, salvia miltiorrhiza and astragalus membranaceus. The traditional Chinese medicine composition has the effects of promoting blood circulation to remove blood stasis and tonifying qi and dredging collaterals, and can effectively improve general behavioral performance, enhance balance coordination ability and autonomous activity ability, relieve motion retardation symptoms, relieve anxiety mood and obviously increase the number of nigra TH + cells; in addition, the traditional Chinese medicine composition can effectively treat the Parkinson's disease, reduces or delays the loss of dopaminergic neurons of the midbrain, has a neuroprotective effect and is free of liver and kidney function damage. The composition has a remarkable effect on treating the Parkinson's disease, is safe and effective, and has a good popularization and application prospect.

The invention discloses a new application of a compound Boropinol-B in preparation of medicines for treating insomnia for the first time. Boropinol-B as shown in a formula I acts on a neuron GABAA receptor, and enhances chloride ion internal flow to hyperpolarize nerve cells, thereby achieving central nervous sedation and inhibiting effects. The Boropinol-B can be used for improving GABA level in the brain by enhancing the activity of GABAergic neurons in a VLPO region in the brain of an insomnia model mouse and thus increasing the release of GABA. The compound has remarkable sedative and hypnotic effects, fast onset of action, quick in-vivo metabolism elimination, short biological half-life period, neither obvious residual effect nor obvious toxic side effects, and is expected to become medicines for treating insomnia with great application prospects.

The invention relates to a method for preparing basal forebrain cholinergic neurons reserving age characteristics through non-nerve cell transformation. According to the preparation method disclosed by the invention, key cell direct transdifferentiation gene combinations and promoters for regulating and controlling the expression level are optimized, viruses capable of efficiently infecting donor-derived cells of various ages are packaged, and adherent growth of the donor-derived cells and transformed neuronal cells is promoted by adopting a proper coating matrix; an induced differentiation culture solution containing small molecular compounds capable of promoting transdifferentiation and growth factors is utilized; a large number of basal forebrain cholinergic neurons are obtained throughtransdifferentiation after the differentiation culture solution is replaced for multiple times within 10-14 days; and finally, the high-purity basal forebrain cholinergic neurons are obtained throughseparation and purification.

The invention discloses a method for constructing a female AD model by utilizing an oxytocin receptor gene silencing technology and an identification method. Constructing and screening are carried outto obtain an AAV9-Oxtr-RNAi vector, AAV9-Oxtr-RNAi viruses are injected into a rat basal nucleus region by utilizing brain stereotaxis, according to a brain infection pathological state, detection and evaluation are carried out in combination with senile dementia symptoms caused by estrogen decline of clinical women, rats are randomly divided into a sham group, a con-shRNA control group and an OXTR-shRNA group, and the learning and memory abilities of the rats are detected by adopting a behavioral Morris water maze experiment and a bright and dark box experiment; an Elisa kit is used for detecting the change of the level of the rat brain tissue cortical estrogen (Estrogen, E2) and the change of the level of a acetylcholine neurotransmitter (acetylcholine, Ach); and in combination with animmunofluorescence technology, the co-localization and expression conditions of incoming nerve fibers of hippocampus in a brain basal nuclear cholinergic neuron fiber projection region, OTR and ChAT are observed, and a basis can be provided for better exploring and researching the pathogenesis and pathological changes of female Alzheimer disease.

A method for co-culturing with nerve stem cells to inducing rASCs to be DA Neurons by Lmx1a mediation, belongs to a method for treating Parkinson's disease by inducing stem cells to replacing defective Neuron cells. The method of the invention comprises steps of: building Lmx1a gene clone, recombining plasmid, building, packaging and recombining adenovirus Ad-Lmx1a, and co-culturing rASCs and NSCs which are infected by Ad-Lmx1a to differentiating them to be DA Neurons; and culturing rASCs and NSCs passage cells. Protein marker for expressing dopaminergic neuron cells can be differentiated from inducted neuron-like cells in the invention, and the inducted cells can transmit electric signal or secrete neurotransmitters, to prove that the inducted cells are the Neuron cells.

The invention relates to an application of a peptide MOTS-c to preparation of a medicine for treating Parkinson's disease. The peptide MOTS-c can reduce the level of oxidative stress injury to the midbrain of a rotenone-induced Parkinson rat animal model, protect rat midbrain nigra striatum dopaminergic neurons and reduce loss of dopaminergic neurons. The dopamine level of striatum is obviously improved, and the neurobehavioral symptoms of rats with Parkinson's disease are effectively improved. Therefore, the peptide MOTS-c has important application significance in preparation of the medicine for resisting the Parkinson's disease.