68 results about "GABAergic neuron" patented technology

Transcriptomes of individual neurons provide rich information about cell types and dynamic states. However, it is difficult to capture rare dynamic processes, such as adult neurogenesis, because isolation from dense adult tissue is challenging, and markers for each phase are limited. Here, Applicants developed Nuc-seq, Div-Seq, and Dronc-Seq. Div-seq combines Nuc-Seq, a scalable single nucleus RNA-Seq method, with EdU-mediated labeling of proliferating cells. Nuc-Seq can sensitively identify closely related cell types within the adult hippocampus. Div-Seq can track transcriptional dynamics of newborn neurons in an adult neurogenic region in the hippocampus. Dronc-Seq uses a microfluidic device to co-encapsulate individual nuclei in reverse emulsion aqueous droplets in an oil medium together with one uniquely barcoded mRNA-capture bead. Finally, Applicants found rare adult newborn GABAergic neurons in the spinal cord, a non-canonical neurogenic region. Taken together, Nuc-Seq, Div-Seq and Dronc-Seq allow for unbiased analysis of any complex tissue.

The present invention provides a method of more efficiently producing a high-quality dopaminergic neuron from neural progenitor cells, specifically, a production method of a dopaminergic neuron including a step of culturing neural progenitor cells in a medium containing (i) a cAMP analogue and (ii) a MEK inhibitor. Moreover, the present invention also provides a medicament containing a dopaminergic neuron obtained by the method, and a reagent and a kit to be used for the method.

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 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.

This invention discloses the uses of IL-22 in the treatment and prevention of a nerve damage disease or a neurodegenerative disease. In particular, the invention discloses the uses of IL-22 or IL-22 dimers as follows: (i) can protect neurons to recover the functions of injured neurons after ischemic nerve damage in animals in vivo, thus enabling effective treatment of nerve damage diseases, (ii) can significantly inhibit the loss of dopaminergic neurons in substantia nigra in PD model animal, enhance the functions of dopaminergic neurons, significantly reduce neuronal apoptosis in hippocampus, improve learning and memory capacity of AD model rats, and effectively prevent neuronal loss, thereby enabling more effective treatment of neurodegenerative diseases.

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.

This invention relates to compositions and methods for the transplantation of GABAergic neurons. GABAergic neurons and compositions comprising the same according to the present invention may be used as cell-based therapies for restoring or reinforcing central inhibition in the nervous system of a subject and for the treatment of neurological conditions, diseases and disorders associated with impaired or aberrant neural function. In a preferred embodiment, the transplant composition comprise of GABAergic neurons, a GFR-alpha agonist, and at least one cell death inhibitor, and that the GABAergicneurons are generated by differentiating pluripotent stem cells, multipotent stem cells, or progenitor cells.

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.

A cell composition includes neural stem cells and neural stem cells derived from mesenchymal stem cells, and at least one selected from the group consisting of immature neural stem cells and mesenchymal stem cells. The number of the neural stem cells is at least 70% of the number of the cells included in the cell composition, and the total number of the cells of the immature neural stem cells and the mesenchymal stem cells is at most 30% of the number of the cells included in the cell composition. The cell composition has a very low probability of being developed into a cancer cell and thus can be used as a cell therapeutic.

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 relates to a GABAergic neuron conditional knockout gene PGC-1 alpha mouse model and a construction method thereof and belongs to the field of animal models and application thereof. The construction method includes the steps that a PGC-1 alpha <flox / flox> mouse is obtained in the first place; then, a PGC-1 alpha <flox / +> is mated with a C57BL / 6J, and the PGC-1 alpha <flox / +> is obtained, then the PGC-1 alpha <flox / +> and the C57BL / 6J are mated with a Dlx5 / 6<Cre-IRES-EGFP> mouse, and the full knockout D1x5 / 6< Cre-IRES-EGFP >, PGC-1 alpha <flox / flox>, semi knockout D1x5 / 6< Cre-IRES-EGFP > and a PGC-1 alpha <flox> mouse are obtained; as a result, a gene knockout mouse on GABAergic internuncial neuron conditional knockout PGC-1 alpha is prepared, and a credible animal model is provided for mechanism research on neurodevelopment and neurodegenerative diseases.

A local stimulation method of a predetermined region of brain tissue, the brain tissue including at least GABAergic neuron type. The method includes at least one activation step of a set of bipolar electrodes, according to a predetermined electric intensity and a predetermined biphasic pulse frequency, delivering a predetermined electric field. The predetermined electric field induces an activation of the GABAergic type neurons and a virtual absence of activation of other types of neurons.

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.

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 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.

The invention discloses application of baicalein(chemical name:5,6,7-trihydroxy flavone;English name: baicalein) shown as a general formula (I) to preparation of a medicament for preventing and treating Parkinson's disease. The baicalein can lower the tremorfrequency and amplitude of the Parkinson's disease, and improve and treat Parkinson's disease symptoms; the baicalein can protect dopaminergic neurons from being injured, and can effectively prevent and treat pathological change of the Parkinson's disease and the development of the Parkinson's disease; through the anti-apoptosis, anti-inflammation and anti-oxidation functions of the baicalein, the effect of protecting thedopaminergic neurons and the effect of restraining the symptoms of the Parkinson's disease are achieved. The invention relates to separate use of the baicalein to preparation of the medicament for preventing and treating the Parkinson's disease and preparation of the baicalein into various preparations in combination with other substances. The general formula (I) is show in the description.

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.

An object of the present invention is to provide a probe, a primer, a primer set and an antibody for use in the detection or selection of a dopaminergic neuron progenitor cell. The present invention provides a probe, a primer and a primer set for use in the detection or selection of a mesencephalon dopaminergic neuron progenitor cell, and preferably a dopaminergic neuron proliferative progenitor cell, which can hybridize with a nucleotide sequence of a 187A5 gene, or a complementary sequence thereto, and an antibody for use in the detection or selection of a mesencephalon dopaminergic neuron progenitor cell, and preferably a dopaminergic neuron progenitor cell, which is capable of binding to a 187A5 protein.

The present invention relates to a method for producing cholinergic neurons comprising obtaining neural progenitor cells from stem cells so as to continuously produce cholinergic neural cells with high purity and the same traits, followed by differentiating the neural progenitor cells into the cholinergic neurons, and cholinergic neurons produced therefrom. Since the method of preparing the cholinergic neurons provided in the present invention enables not only production of the cholinergic neurons with high purity, but also rapid production of the cholinergic neurons with the same traits, it can be widely used for effectively treating degenerative cranial nerve diseases such as Alzheimer's disease.

The present invention discloses an application of acetylcholinesterase inhibitor in the preparation of medicine for curing Parkinson's disease. The described inhibitor includes the compounds which can inhibit activity of acetylcholinesterase and have the action of curing Parkinson's disease. Said inhibitor can effectively improve motion defect resulted from Parkinson's disease and can protect dopaminergic neuron from injury.