Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 12 de 12
Filtrar
1.
Cell ; 178(6): 1375-1386.e11, 2019 09 05.
Artículo en Inglés | MEDLINE | ID: mdl-31474366

RESUMEN

In search of the molecular identities of cold-sensing receptors, we carried out an unbiased genetic screen for cold-sensing mutants in C. elegans and isolated a mutant allele of glr-3 gene that encodes a kainate-type glutamate receptor. While glutamate receptors are best known to transmit chemical synaptic signals in the CNS, we show that GLR-3 senses cold in the peripheral sensory neuron ASER to trigger cold-avoidance behavior. GLR-3 transmits cold signals via G protein signaling independently of its glutamate-gated channel function, suggesting GLR-3 as a metabotropic cold receptor. The vertebrate GLR-3 homolog GluK2 from zebrafish, mouse, and human can all function as a cold receptor in heterologous systems. Mouse DRG sensory neurons express GluK2, and GluK2 knockdown in these neurons suppresses their sensitivity to cold but not cool temperatures. Our study identifies an evolutionarily conserved cold receptor, revealing that a central chemical receptor unexpectedly functions as a thermal receptor in the periphery.


Asunto(s)
Proteínas de Caenorhabditis elegans/fisiología , Caenorhabditis elegans/genética , Receptores de Glutamato/fisiología , Receptores de Ácido Kaínico/fisiología , Receptores de Glutamato Metabotrópico/fisiología , Sensación Térmica/fisiología , Animales , Células CHO , Proteínas de Caenorhabditis elegans/genética , Frío , Cricetulus , Humanos , Ratones , Neuronas/metabolismo , Receptores de Glutamato/genética , Receptores de Ácido Kaínico/genética , Receptores de Glutamato Metabotrópico/genética , Sensación Térmica/genética
2.
RNA Biol ; 16(1): 13-24, 2019 01.
Artículo en Inglés | MEDLINE | ID: mdl-30574830

RESUMEN

Long non-coding RNAs have emerged as an important regulatory layer in biological systems. Of the various types of lncRNAs, one class (designated as divergent RNAs/XH), which is in head-to-head overlap with the coding genes, has emerged as a critical biotype that regulates development and cellular differentiation. This work aimed to analyze previously published data on differential expression, epigenetic and network analysis in order to demonstrate the association of divergent lncRNAs, a specific biotype with the differentiation of human neural progenitor cells (hNPCs). We have analyzed various available RNAseq databases that address the neuronal and astrocytic differentiation of hNPCs and identified differentially expressed lncRNAs (DELs) during cell-fate determination. Key DELs identified from the databases were experimentally verified by us in our in-vitro hNPC differentiation system. We also analyzed the change in promoter activity using ChIP-seq datasets of the histone markers H3K4me3 (activation) and H3K27me3 (inactivation) of these DELs. Additionally, we explored the change in the euchromatinization state of DELs (by analyzing DNase-seq data) during lineage-specific differentiation of hNPCs and performed their network analysis. We were able to identify differences between neuronal and astrocytic differentiation of hNPCs at the level of divergent DELs epigenetic markers, DNAase hypersensitive sites and gene expression network. Divergent lncRNAs are more involved in neuronal rather than astrocytic differentiation, while the sense downstream lncRNA biotype appears to be more involved in astrocytic differentiation. By studying the lncRNA involvement of distinct biotypes, we have been able to indicate the preferential role of a particular biotype during lineage-specific differentiation.


Asunto(s)
Diferenciación Celular/genética , Linaje de la Célula/genética , Epigénesis Genética , Células-Madre Neurales/citología , Células-Madre Neurales/metabolismo , ARN Largo no Codificante/genética , Astrocitos/citología , Astrocitos/metabolismo , Inmunoprecipitación de Cromatina , Biología Computacional/métodos , Perfilación de la Expresión Génica , Regulación del Desarrollo de la Expresión Génica , Redes Reguladoras de Genes , Secuenciación de Nucleótidos de Alto Rendimiento , Histonas/metabolismo , Humanos
3.
Glia ; 65(2): 250-263, 2017 02.
Artículo en Inglés | MEDLINE | ID: mdl-27761954

RESUMEN

Astroglia are indispensable component of the tripartite synapse ensheathing innumerous soma and synapses. Its proximity to neurons aids the regulation of neuronal functions, health and survival through dynamic neuroglia crosstalk. Susceptibility of astrocyte to HIV-1 infection and subsequent latency culminates in compromised neuronal health. The viral protein HIV-1 transactivator of transcription (Tat) is neurotoxic. HIV-1 Tat is detected in brain of AIDS patients even in cases where viral load is non-detectable due to successful HAART therapy. Recently, we demonstrated that HIV-1 Tat triggers excess ATP release from astrocytes that causes neuronal death by activating purinergic receptor system. Using well-characterized model system of human primary astrocytes and neurons, we probed into the molecular mechanism for enhanced ATP release in HIV-1 Tat affected astrocytes. HIV-1 Tat modulated the miRNA machinery in astrocytes and perturbed the levels of voltage dependent anion channel 1 (VDAC1), a channel present in the outer mitochondrial membrane and plasma membrane that regulates extracellular ATP release. Our studies with autopsy tissue sections also showed concordantly dysregulated VDAC1 and miR-320a levels in HIV-1 patients suffering from mild cognitive impairment (MCI). We report a novel molecular cascade of miRNA-mediated ATP release through regulation of VDAC1. Downregulation of VDAC1 either with miR-320a mimic or VDAC1 siRNA in HIV-1 Tat-affected astroglia could rescue the neurons from glia-mediated indirect death. Our findings reveal a novel upstream therapeutic target that could be employed to thwart the astroglia-mediated neurotoxicity in HIV-1 neuropathogenesis. GLIA 2017;65:250-263.


Asunto(s)
Astrocitos/metabolismo , Regulación de la Expresión Génica/genética , MicroARNs/metabolismo , Neuronas/patología , Canal Aniónico 1 Dependiente del Voltaje/metabolismo , Regiones no Traducidas 3'/fisiología , Apoptosis/efectos de los fármacos , Apoptosis/genética , Astrocitos/efectos de los fármacos , Astrocitos/virología , Autopsia , Encéfalo/patología , Encéfalo/virología , Células Cultivadas , Disfunción Cognitiva/etiología , Disfunción Cognitiva/virología , Feto , Regulación de la Expresión Génica/efectos de los fármacos , Infecciones por VIH/complicaciones , Infecciones por VIH/patología , Humanos , Etiquetado Corte-Fin in Situ , MicroARNs/genética , Células Madre Neoplásicas/efectos de los fármacos , Neuronas/metabolismo , Neuronas/virología , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/metabolismo , Telencéfalo/citología , Factores de Tiempo , Canal Aniónico 1 Dependiente del Voltaje/genética , Productos del Gen tat del Virus de la Inmunodeficiencia Humana/genética , Productos del Gen tat del Virus de la Inmunodeficiencia Humana/metabolismo , Productos del Gen tat del Virus de la Inmunodeficiencia Humana/farmacología
4.
Nat Neurosci ; 27(4): 679-688, 2024 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-38467901

RESUMEN

Thermosensors expressed in peripheral somatosensory neurons sense a wide range of environmental temperatures. While thermosensors detecting cool, warm and hot temperatures have all been extensively characterized, little is known about those sensing cold temperatures. Though several candidate cold sensors have been proposed, none has been demonstrated to mediate cold sensing in somatosensory neurons in vivo, leaving a knowledge gap in thermosensation. Here we characterized mice lacking the kainate-type glutamate receptor GluK2, a mammalian homolog of the Caenorhabditis elegans cold sensor GLR-3. While GluK2 knockout mice respond normally to heat and mechanical stimuli, they exhibit a specific deficit in sensing cold but not cool temperatures. Further analysis supports a key role for GluK2 in sensing cold temperatures in somatosensory DRG neurons in the periphery. Our results reveal that GluK2-a glutamate-sensing chemoreceptor mediating synaptic transmission in the central nervous system-is co-opted as a cold-sensing thermoreceptor in the periphery.


Asunto(s)
Receptor de Ácido Kaínico GluK2 , Receptores de Ácido Kaínico , Animales , Ratones , Caenorhabditis elegans/metabolismo , Frío , Receptor de Ácido Kaínico GluK2/metabolismo , Ácido Glutámico , Mamíferos/metabolismo , Neuronas/metabolismo , Receptores de Ácido Kaínico/genética , Receptores de Ácido Kaínico/metabolismo , Transmisión Sináptica
5.
Front Mol Neurosci ; 14: 790435, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-35058747

RESUMEN

Thermosensitive transient receptor potential V3 (TRPV3) is a polymodal receptor implicated in nociceptive, thermoceptive, pruritoceptive, and inflammatory pathways. Reports focused on understanding the role of TRPV3 in thermoception or nociception are not conclusive. Previous studies also show that aberrant hyperactivity of TRPV3 channels results in spontaneous itch and dermatitis-like symptoms, but the resultant behavior is highly dependent on the background of the animal and the skin microbiome. To determine the function of hyperactive TRPV3 channels in somatosensory sensations, we tested different somatosensory behaviors using a genetic mouse model that carries a gain-of-function point mutation G573S in the Trpv3 gene (Trpv3 G573S ). Here we report that Trpv3 G573S mutants show reduced perception of cold, acetone-induced cooling, punctate, and sharp mechanical pain. By contrast, locomotion, noxious heat, touch, and mechanical itch are unaffected in Trpv3 G573S mice. We fail to observe any spontaneous itch responses and/or dermatitis in Trpv3 G573S mutants under specific pathogen (Staphylococcus aureus)-free conditions. However, we find that the scratching events in response to various pruritogens are dramatically decreased in Trpv3 G573S mice in comparison to wild-type littermates. Interestingly, we observe sensory hypoinnervation of the epidermis in Trpv3 G573S mutants, which might contribute to the deficits in acute mechanical pain, cool, cold, and itch sensations.

6.
Genome Biol Evol ; 13(1)2021 01 07.
Artículo en Inglés | MEDLINE | ID: mdl-33434274

RESUMEN

Alu repeats contribute to phylogenetic novelties in conserved regulatory networks in primates. Our study highlights how exonized Alus could nucleate large-scale mRNA-miRNA interactions. Using a functional genomics approach, we characterize a transcript isoform of an orphan gene, CYP20A1 (CYP20A1_Alu-LT) that has exonization of 23 Alus in its 3'UTR. CYP20A1_Alu-LT, confirmed by 3'RACE, is an outlier in length (9 kb 3'UTR) and widely expressed. Using publically available data sets, we demonstrate its expression in higher primates and presence in single nucleus RNA-seq of 15,928 human cortical neurons. miRanda predicts ∼4,700 miRNA recognition elements (MREs) for ∼1,000 miRNAs, primarily originated within these 3'UTR-Alus. CYP20A1_Alu-LT could be a potential multi-miRNA sponge as it harbors ≥10 MREs for 140 miRNAs and has cytosolic localization. We further tested whether expression of CYP20A1_Alu-LT correlates with mRNAs harboring similar MRE targets. RNA-seq with conjoint miRNA-seq analysis was done in primary human neurons where we observed CYP20A1_Alu-LT to be downregulated during heat shock response and upregulated in HIV1-Tat treatment. In total, 380 genes were positively correlated with its expression (significantly downregulated in heat shock and upregulated in Tat) and they harbored MREs for nine expressed miRNAs which were also enriched in CYP20A1_Alu-LT. MREs were significantly enriched in these 380 genes compared with random sets of differentially expressed genes (P = 8.134e-12). Gene ontology suggested involvement of these genes in neuronal development and hemostasis pathways thus proposing a novel component of Alu-miRNA-mediated transcriptional modulation that could govern specific physiological outcomes in higher primates.


Asunto(s)
Regiones no Traducidas 3'/genética , Elementos Alu/genética , Sistema Enzimático del Citocromo P-450/genética , Exones , MicroARNs/genética , Poríferos/genética , Primates/genética , Isoformas de Proteínas/genética , Animales , Regulación de la Expresión Génica , Respuesta al Choque Térmico/genética , Humanos , Neuronas , Filogenia , ARN Mensajero , Regulación hacia Arriba
7.
Cell Death Discov ; 6: 28, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32351715

RESUMEN

lncRNA genes can be genic or "intergenic". "Genic" RNAs can be further divided into six biotypes. Through genome-wide analysis of a publicly available data set on corticogenesis, we found that the divergent lncRNA (XH) biotype, comprising the lncRNA and the coding gene being in opposite directions in a head-to-head manner, was most prominent during neural commitment. Within this biotype, a coding gene/divergent RNA pair of the BASP1 gene and the uncharacterized RNA loc285696 (hitherto referred as BASP1-AS1) formed a major HUB gene during neuronal differentiation. Experimental validation during the in vitro differentiation of human neural progenitor cells (hNPCs) showed that BASP1-AS1 regulates the expression of its adjacent coding gene, BASP1. Both transcripts increased sharply on the first day of neuronal differentiation of hNPCs, to fall steadily thereafter, reaching very low levels in differentiated neurons. BASP1-AS1 RNA and the BASP1 gene formed a molecular complex that also included the transcription factor TCF12. TCF12 is coded by the DYX1 locus, associated with inherited dyslexia and neurodevelopmental defects. Knockdown of BASP1-AS1, BASP1, or TCF12 impaired the neuronal differentiation of hNPCs, as seen by reduction in DCX and TUJ1-positive cells and by reduced neurite length. There was also increased cell proliferation. A common set of critical genes was affected by the three molecules in the complex. Our study thus identified the role of the XH biotype and a novel mediator of neuronal differentiation-the complex of BASP1-AS1, BASP1, and TCF12. It also linked a neuronal differentiation pathway to inherited dyslexia.

8.
Front Mol Neurosci ; 12: 293, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31920530

RESUMEN

Long non-coding RNAs (lncRNAs) have emerged as an important regulatory control in biological systems. Though the field of lncRNA has been progressing rapidly, a complete understanding of the role of lncRNAs in neuroblastoma pathogenesis is still lacking. To identify the abrogated lncRNAs in primary neuroblastoma and in the metastasized as well as the relapsed form of neuroblastoma, we analyzed an RNA-seq dataset on neuroblastoma that is available online to identify the lncRNAs that could potentially be contributing to the biology of neuroblastoma. The identified lncRNAs were further scrutinized using a publicly available epigenetic dataset of neuroblastoma and a cancer database. After this cross-sectional study, we were able to identify three significant lncRNAs, CASC15, PPP1R26-AS1, and USP3-AS1, which could serve as potential biomarkers in clinical studies of neuroblastoma pathogenesis.

9.
Pain ; 160(5): 1166-1174, 2019 05.
Artículo en Inglés | MEDLINE | ID: mdl-30913166

RESUMEN

Recent studies have made significant progress in identifying distinct populations of peripheral neurons involved in itch transmission, whereas the cellular identity of spinal interneurons that contribute to itch processing is still a debate. Combining genetic and pharmacological ablation of spinal excitatory neuronal subtypes and behavioral assays, we demonstrate that spinal somatostatin-positive (SOM) excitatory interneurons transmit pruritic sensation. We found that the ablation of spinal SOM/Lbx1 (SOM) neurons caused significant attenuation of scratching responses evoked by various chemical pruritogens (chemical itch). In an attempt to identify substrates of spinal itch neural circuit, we observed that spinal SOM neurons partially overlapped with neurons expressing natriuretic peptide receptor A (Npra), the receptor of peripheral itch transmitter B-type natriuretic peptide. Spinal SOM neurons, however, did not show any overlap with itch transmission neurons expressing gastrin-releasing peptide receptor in the dorsal spinal cord, and the gastrin-releasing peptide-triggered scratching responses were intact after ablating spinal SOM neurons. Dual ablation of SOM and Npra neurons in the spinal cord reduced chemical itch responses to a greater extent than ablation of SOM or Npra neurons alone, suggesting the existence of parallel spinal pathways transmitting chemical itch. Furthermore, we showed that SOM peptide modulated itch processing through disinhibition of somatostatin receptor 2A-positive inhibitory interneuron. Together, our findings reveal a novel spinal mechanism for sensory encoding of itch perception.


Asunto(s)
Interneuronas/metabolismo , Prurito/inducido químicamente , Prurito/patología , Somatostatina/metabolismo , Médula Espinal/patología , Potenciales de Acción/efectos de los fármacos , Potenciales de Acción/genética , Inhibidores de la Angiogénesis/farmacología , Animales , Cloroquina/toxicidad , Modelos Animales de Enfermedad , Técnicas In Vitro , Interneuronas/fisiología , Proteínas Luminiscentes/genética , Proteínas Luminiscentes/metabolismo , Lisina/análogos & derivados , Lisina/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Proteínas Musculares/genética , Proteínas Musculares/metabolismo , Nitrobenzoatos/farmacología , Técnicas de Placa-Clamp , Proteínas Proto-Oncogénicas c-fos/metabolismo , Somatostatina/genética , Médula Espinal/efectos de los fármacos , Médula Espinal/metabolismo , p-Metoxi-N-metilfenetilamina/toxicidad , Proteínas tau/genética , Proteínas tau/metabolismo
10.
Neuron ; 103(6): 1135-1149.e6, 2019 09 25.
Artículo en Inglés | MEDLINE | ID: mdl-31324538

RESUMEN

Lightly stroking the lips or gently poking some skin regions can evoke mechanical itch in healthy human subjects. Sensitization of mechanical itch and persistent spontaneous itch are intractable symptoms in chronic itch patients. However, the underlying neural circuits are not well defined. We identified a subpopulation of excitatory interneurons expressing Urocortin 3::Cre (Ucn3+) in the dorsal spinal cord as a central node in the pathway that transmits acute mechanical itch and mechanical itch sensitization as well as persistent spontaneous itch under chronic itch conditions. This population receives peripheral inputs from Toll-like receptor 5-positive (TLR5+) Aß low-threshold mechanoreceptors and is directly innervated by inhibitory interneurons expressing neuropeptide Y::Cre (NPY+) in the dorsal spinal cord. Reduced synaptic inhibition and increased intrinsic excitability of Ucn3+ neurons lead to chronic itch sensitization. Our study sheds new light on the neural basis of chronic itch and unveils novel avenues for developing mechanism-specific therapeutic advancements.


Asunto(s)
Interneuronas/fisiología , Mecanorreceptores/fisiología , Vías Nerviosas/fisiopatología , Prurito/fisiopatología , Piel/inervación , Médula Espinal/fisiopatología , Animales , Sensibilización del Sistema Nervioso Central/fisiología , Ácido Glutámico/metabolismo , Interneuronas/metabolismo , Mecanorreceptores/metabolismo , Ratones , Inhibición Neural/fisiología , Neuropéptido Y/metabolismo , Estimulación Física , Médula Espinal/citología , Receptor Toll-Like 5/metabolismo , Urocortinas/metabolismo
11.
Cell Death Dis ; 8(1): e2556, 2017 01 19.
Artículo en Inglés | MEDLINE | ID: mdl-28102850

RESUMEN

Japanese encephalitis virus (JEV), which is a causative agent of sporadic encephalitis, harbours itself inside the neural stem/progenitor cells. It is a well-known fact that JEV infects neural stem/progenitor cells and decreases their proliferation capacity. With mass spectrometry-based quantitative proteomic study, it is possible to reveal the impact of virus on the stem cells at protein level. Our aim was to perceive the stem cell proteomic response upon viral challenge. We performed a two-dimensional gel electrophoresis-based proteomic study of the human neural stem cells (hNS1 cell line) post JEV infection and found that 13 proteins were differentially expressed. The altered proteome profile of hNS1 cell line revealed sustained endoplasmic reticulum stress, which deteriorated normal cellular activities leading to cell apoptosis. The proteomic changes found in hNS1 cell line were validated in vivo in the subventricular zone of JE infected BALB/c mice. Congruent alterations were also witnessed in multipotent neural precursor cells isolated from human foetus and in autopsy samples of human brain clinically diagnosed as cases of JE patients. Endoplasmic reticulum resident chaperone GRP78, mitochondrial protein Prohibitin and heterogeneous nuclear ribonucleoprotein hnRNPC (C1/C2) have been shown to interact with viral RNA. Hence it is proposed that these are the principle candidates governing endoplasmic reticulum stress-induced apoptosis in JEV infection.


Asunto(s)
Encefalitis Japonesa/genética , Proteínas de Choque Térmico/genética , Ribonucleoproteína Heterogénea-Nuclear Grupo C/genética , Proteínas Represoras/genética , Animales , Apoptosis/genética , Proliferación Celular/genética , Electroforesis en Gel Bidimensional , Virus de la Encefalitis Japonesa (Especie)/genética , Virus de la Encefalitis Japonesa (Especie)/patogenicidad , Encefalitis Japonesa/virología , Chaperón BiP del Retículo Endoplásmico , Estrés del Retículo Endoplásmico/genética , Regulación de la Expresión Génica , Proteínas de Choque Térmico/biosíntesis , Ribonucleoproteína Heterogénea-Nuclear Grupo C/biosíntesis , Humanos , Ratones , Prohibitinas , Proteoma/genética , ARN Viral/genética , Proteínas Represoras/biosíntesis
12.
Neurosci Bull ; 37(11): 1639-1641, 2021 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-34273096

Asunto(s)
Neuralgia , Humanos
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA