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1.
Cell Mol Neurobiol ; 41(3): 525-536, 2021 Apr.
Article in English | MEDLINE | ID: mdl-32415577

ABSTRACT

Olfactory ensheathing cells (OECs) are specialized glial cells of the olfactory system, believed to play a role in the continuous production of olfactory neurons and ensheathment of their axons. Although OECs are used in therapeutic applications, little is known about the cellular mechanisms underlying their migratory behavior. Recently, we showed that OEC migration is sensitive to ganglioside blockage through A2B5 and Jones antibody in OEC culture. Gangliosides are common components of lipid rafts, where they participate in several cellular mechanisms, including cell migration. Here, we characterized OEC lipid rafts, analyzing the presence of specific proteins and gangliosides that are commonly expressed in motile neural cells, such as young neurons, oligodendrocyte progenitors, and glioma cells. Our results showed that lipid rafts isolated from OECs were enriched in cholesterol, sphingolipids, phosphatidylcholine, caveolin-1, flotillin-1, gangliosides GM1 and 9-O-acetyl GD3, A2B5-recognized gangliosides, CNPase, α-actinin, and ß1-integrin. Analysis of the actin cytoskeleton of OECs revealed stress fibers, membrane spikes, ruffled membranes and lamellipodia during cell migration, as well as the distribution of α-actinin in membrane projections. This is the first description of α-actinin and flotillin-1 in lipid rafts isolated from OECs and suggests that, together with ß1-integrin and gangliosides, membrane lipid rafts play a role during OEC migration. This study provides new information on the molecular composition of OEC membrane microdomains that can impact on our understanding of the role of OEC lipid rafts under physiological and pathological conditions of the nervous system, including inflammation, hypoxia, aging, neurodegenerative diseases, head trauma, brain tumor, and infection.


Subject(s)
Membrane Microdomains/metabolism , Olfactory Bulb/cytology , 2',3'-Cyclic-Nucleotide Phosphodiesterases/metabolism , Animals , Biomarkers/metabolism , Cells, Cultured , Cholesterol/metabolism , Cytoskeletal Proteins/metabolism , Gangliosides/metabolism , Membrane Microdomains/ultrastructure , Rats, Wistar , S100 Proteins/metabolism
2.
Int. j. morphol ; 38(5): 1463-1472, oct. 2020. graf
Article in English | LILACS | ID: biblio-1134463

ABSTRACT

SUMMARY: The vomeronasal organ (VNO) is an accessory organ involved on the olfactory pathway, that detects pheromones and emits signals in order to modulate social and reproductive behavior. The VNO stem cells replace neurons throughout life. The aim of this study was to isolate and characterize cells derived from the vomeronasal organ from New Zealand rabbits. Five male rabbits with 120 days were used for cell isolation and culture. Results: VNO-derived cells presented labelling for proliferation (PCNA), undifferentiated profile (Nanog), neuronal (GFAP), mesenchymal stem cells (CD73, CD90 and CD105 and Stro-1). Also, presence of cytoskeletal (Vimentin, b-tubulin and CK-18) and absence of hematopoietic markers (CD34, CD117 and CD45) both by immunofluorescence and flow cytometry. By PCR it was possible to verify the expression of some undifferentiated profile (Oct-4), neuronal (Nestin) and mesenchymal (CD73, CD105 and Vimentin) genes. Functionally, VNO-derived cells differentiate in vitro into adipocytes, osteocytes and chondrocytes, and presented no tumorigenic potential when injected to Balb/c nu/nu mice. In conclusion, the rabbit VNO-derived cells have a profile that could be supportive to VNO olfactory/neuroreceptor epithelium by delivering factors to epithelial turnover or even by differentiation into epithelial cells to replacement of commissural epithelium.


RESUMEN: El órgano vomeronasal (OVN) es un órgano accesorio de la vía olfatoria, que detecta feromonas y emite señales que afectan la modulación del comportamiento social y reproductivo. Las células madre OVN reemplazan las neuronas durante toda la vida. El objetivo de este estudio fue aislar y caracterizar células derivadas del órgano vomeronasal de conejos raza Nueva Zelanda. Para el aislamiento y el cultivo celular se utilizaron cinco conejos machos con una edad de 120 días. Las células del OVN presentaron etiquetado para la proliferación (PCNA), un perfil indiferenciado (Nanog), neuronal (GFAP), células madre mesenquimales (CD73, CD90 y CD105 y Stro-1). Además, se ob- servó presencia de citoesqueleto (Vimentina, β-tubulina y CK-18) y ausencia de marcadores hematopoyéticos (CD34, CD117 y CD45) tanto por inmunofluorescencia como por citometría de flujo. Me- diante PCR fue posible verificar la expresión de algunos genes de perfil indiferenciado (Oct-4), neuronal (Nestin) y mesenquimatoso (CD73, CD105 y Vimentin). Las células derivadas del OVN se diferencian in vitro en adipocitos, osteocitos y condrocitos, y no presentan un potencial tumorigénico al ser infiltrados en ratones Balb / c nu / nu. En conclusión, las células derivadas de OVN de conejo tienen un perfil que podría ser compatible con el epitelio olfatorio / neurorreceptor de OVN transmitiendo factores al recambio epitelial o incluso mediante la diferenciación en células epiteliales para reemplazar el epitelio comisural.


Subject(s)
Animals , Rabbits/anatomy & histology , Vomeronasal Organ/cytology , Mesenchymal Stem Cells/physiology , Olfactory Bulb/cytology , Stem Cells/physiology , Olfactory Mucosa/cytology , Polymerase Chain Reaction , Fluorescent Antibody Technique , Flow Cytometry , Neurons/physiology
3.
J Anat ; 235(5): 977-983, 2019 11.
Article in English | MEDLINE | ID: mdl-31373393

ABSTRACT

Olfactory ensheathing glia (OEG) are found in the olfactory mucosa, nerve and bulb, and provide in vivo ensheathment for the unmyelinated olfactory axons within the central and peripheral nervous system domains. OEG cells are able to migrate long distances within the neuropil of the central nervous system. Because gangliosides such as 9-O-acetyl GD3 have crucial regulatory roles in neuronal migration during development, we analyzed whether OEG in organotypical cultures are revealed by anti-9-O-acetyl GD3 and/or gangliosides are recognized by the A2B5 antibody (G-A2B5), and whether these gangliosides are involved in OEG migration. Our results showed that all OEG migrating out of a section of olfactory bulb onto a laminin substrate bound to the 9-O-acetyl GD3 and A2B5 antibodies, and that 2',3'-cyclic nucleotide phosphodiesterase (CNPase) colocalized with 9-O-acetyl GD3 and with G-A2B5. Additionally, we showed that the immune blockade of 9-O-acetyl GD3 or G-A2B5 reduced the migration of OEG on laminin, and that 9-O-acetyl GD3 and G-A2B5 colocalized with the ß1-integrin subunit. We also confirmed the phenotype of in-vitro-grown OEG cells derived from adult rats, showing that they express CNPase, and also α-smooth muscle actin, which is not expressed by Schwann cells. Our data showed that the gangliosides 9-O-acetyl GD3 and G-A2B5 participate in the migratory activity of OEG cells, and that the ß1-integrin subunit colocalizes with these gangliosides. These results suggest a new role for ß1-integrin and gangliosides in the polarized migration of OEG cells, and provide new information on the molecules controlling OEG motility and behavior.


Subject(s)
Cell Movement/physiology , Gangliosides/metabolism , Integrin beta1/metabolism , Neuroglia/metabolism , Olfactory Bulb/metabolism , Animals , Neuroglia/cytology , Olfactory Bulb/cytology , Olfactory Mucosa/cytology , Olfactory Mucosa/metabolism , Rats , Rats, Wistar , Schwann Cells/cytology , Schwann Cells/metabolism
4.
J Comp Neurol ; 527(7): 1278-1289, 2019 05 01.
Article in English | MEDLINE | ID: mdl-30628061

ABSTRACT

Many functions of glial cells depend on the formation of selective glial networks mediated by gap junctions formed by members of the connexin family. Olfactory ensheathing cells (OECs) are specialized glia associated with olfactory sensory neuron axons. Like other glia, they form selective networks, however, the connexins that support OEC connectivity in vivo have not been identified. We used an in vivo mouse model to selectively delete candidate connexin genes with temporal control from OECs and address the physiological consequences. Using this model, we effectively abolished the expression of connexin 43 (Cx43) in OECs in both juvenile and adult mice. Cx43-deleted OECs exhibited features consistent with the loss of gap junctions including reduced membrane conductance, largely reduced sensitivity to the gap junction blocker meclofenamic acid and loss of dye coupling. This indicates that Cx43, a typically astrocytic connexin, is the main connexin forming functional channels in OECs. Despite these changes in functional properties, the deletion of Cx43 deletion did not alter the density of OECs. The strategy used here may prove useful to delete other candidate genes to better understand the functional roles of OECs in vivo.


Subject(s)
Connexin 43/physiology , Gap Junctions/physiology , Gene Knockout Techniques , Neuroglia/physiology , Olfactory Bulb/cytology , Aging/metabolism , Animals , Connexin 43/deficiency , Connexin 43/genetics , Crosses, Genetic , Female , Gap Junctions/drug effects , Genes, Reporter , Genes, Synthetic , Integrases/genetics , Male , Meclofenamic Acid/pharmacology , Mice , Mice, Knockout , Myelin Proteolipid Protein/genetics , Olfactory Bulb/metabolism , Patch-Clamp Techniques , Tamoxifen/pharmacology
5.
Anat Rec (Hoboken) ; 301(9): 1570-1584, 2018 09.
Article in English | MEDLINE | ID: mdl-29752870

ABSTRACT

The existence of neurogenesis in the adult brain is a widely recognized phenomenon, occurring in the subventricular zone (SVZ) of the lateral ventricles and the subgranular zone of the dentate gyrus in several vertebrate species. Neural precursors originated in the SVZ migrate to the main olfactory bulb (MOB), originating the rostral migratory stream (RMS) in the process. To better understand the formation of the adult neurogenic niches in dogs, we investigated the cellular composition and morphological organization of these areas in 57 days-old dog fetuses. Using multiple immunohistochemical markers, we demonstrated that the SVZ in the canine fetus is remarkably similar to the adult SVZ, with glial GFAP-immunoreactive (-ir) cells, DCX-ir neuroblasts and SOX2-ir neuronal progenitors tangentially organized along the dorsal lateral ventricle. The fetal RMS has all the features of its adult counterpart and closely resembles the RMS of other mammalian species. The late-development canine MOB has most of the neurochemical features of the adult MOB, including an early-developed TH-ir population and maturing CALR-ir interneurons, but CALB-ir neurons in the granule cell layer will only appear in the post-partum period. Taken together, our results suggest that the canine fetal development of adult neurogenic niches closely resembles those of primates, and dogs may be suitable models of human adult neurogenesis. Anat Rec, 301:1570-1584, 2018. © 2018 Wiley Periodicals, Inc.


Subject(s)
Cell Movement/physiology , Lateral Ventricles/embryology , Neural Stem Cells/cytology , Neurogenesis/physiology , Olfactory Bulb/embryology , Animals , Cell Proliferation/physiology , Dogs , Lateral Ventricles/cytology , Neurons/cytology , Olfactory Bulb/cytology
6.
Glia ; 66(7): 1267-1301, 2018 07.
Article in English | MEDLINE | ID: mdl-29330870

ABSTRACT

The prospects of achieving regeneration in the central nervous system (CNS) have changed, as most recent findings indicate that several species, including humans, can produce neurons in adulthood. Studies targeting this property may be considered as potential therapeutic strategies to respond to injury or the effects of demyelinating diseases in the CNS. While CNS trauma may interrupt the axonal tracts that connect neurons with their targets, some neurons remain alive, as seen in optic nerve and spinal cord (SC) injuries (SCIs). The devastating consequences of SCIs are due to the immediate and significant disruption of the ascending and descending spinal pathways, which result in varying degrees of motor and sensory impairment. Recent therapeutic studies for SCI have focused on cell transplantation in animal models, using cells capable of inducing axon regeneration like Schwann cells (SchCs), astrocytes, genetically modified fibroblasts and olfactory ensheathing glia cells (OECs). Nevertheless, and despite the improvements in such cell-based therapeutic strategies, there is still little information regarding the mechanisms underlying the success of transplantation and regarding any secondary effects. Therefore, further studies are needed to clarify these issues. In this review, we highlight the properties of OECs that make them suitable to achieve neuroplasticity/neuroregeneration in SCI. OECs can interact with the glial scar, stimulate angiogenesis, axon outgrowth and remyelination, improving functional outcomes following lesion. Furthermore, we present evidence of the utility of cell therapy with OECs to treat SCI, both from animal models and clinical studies performed on SCI patients, providing promising results for future treatments.


Subject(s)
Cell Transplantation , Neuroglia/transplantation , Spinal Cord Injuries/therapy , Animals , Humans , Neuroglia/cytology , Olfactory Bulb/cytology , Olfactory Mucosa/cytology
7.
eNeuro ; 4(5)2017.
Article in English | MEDLINE | ID: mdl-28955723

ABSTRACT

Early sensory experience shapes the anatomy and function of sensory circuits. In the mouse olfactory bulb (OB), prenatal and early postnatal odorant exposure through odorized food (food/odorant pairing) not only increases the volume of activated glomeruli but also increases the number of mitral and tufted cells (M/TCs) connected to activated glomeruli. Given the importance of M/TCs in OB output and in mediating lateral inhibitory networks, increasing the number of M/TCs connected to a single glomerulus may significantly change odorant representation by increasing the total output of that glomerulus and/or by increasing the strength of lateral inhibition mediated by cells connected to the affected glomerulus. Here, we seek to understand the functional impact of this long-term odorant exposure paradigm on the population activity of mitral cells (MCs). We use viral expression of GCaMP6s to examine odor-evoked responses of MCs following prenatal and early postnatal odorant exposure to two dissimilar odorants, methyl salicylate (MS) and hexanal, which are both strong activators of glomeruli on the dorsal OB surface. Previous work suggests that odor familiarity may decrease odor-evoked MC response in rodents. However, we find that early food-based odorant exposure significantly changes MC responses in an unexpected way, resulting in broad increases in the amplitude, number, and reliability of excitatory MC responses across the dorsal OB.


Subject(s)
Odorants , Olfactory Bulb/cytology , Prenatal Exposure Delayed Effects/physiopathology , Sensory Receptor Cells/physiology , Smell/physiology , Action Potentials/drug effects , Action Potentials/physiology , Aldehydes/pharmacology , Animals , Animals, Newborn , Calcium/metabolism , Dose-Response Relationship, Drug , Female , Green Fluorescent Proteins/genetics , Green Fluorescent Proteins/metabolism , Insecticides/pharmacology , Male , Mice , Mice, Transgenic , Olfactory Pathways/physiology , Pregnancy , Sensory Receptor Cells/drug effects , Time Factors , tau Proteins/genetics , tau Proteins/metabolism
8.
Sci Rep ; 6: 36813, 2016 11 09.
Article in English | MEDLINE | ID: mdl-27827453

ABSTRACT

Olfactory ensheathing cells (OECs) are a type of specialized glial cell currently considered as having a double function in the nervous system: one regenerative, and another immune. Streptococcus pneumoniae is a major agent of severe infections in humans, including meningitis. It is commonly found in the nasopharynx of asymptomatic carriers, and, under certain still unknown conditions, can invade the brain. We evaluated whether pneumococcal cells recovered from lysed OECs and microglia are able to survive by manipulating the host cell activation. An intracellular-survival assay of S. pneumoniae in OECs showed a significant number of bacterial CFU recovered after 3 h of infection. In contrast, microglia assays resulted in a reduced number of CFU. Electron-microscopy analysis revealed a large number of pneumococci with apparently intact morphology. However, microglia cells showed endocytic vesicles containing only bacterial cell debris. Infection of OEC cultures resulted in continuous NF-κB activation. The IFN-γ-induced increase of iNOS expression was reversed in infected OECs. OECs are susceptible to S. pneumoniae infection, which can suppress their cytotoxic mechanisms in order to survive. We suggest that, in contrast to microglia, OECs might serve as safe targets for pneumococci, providing a more stable environment for evasion of the immune system.


Subject(s)
Microglia/cytology , Olfactory Bulb/cytology , Streptococcus pneumoniae/growth & development , Animals , Cells, Cultured , Colony Count, Microbial , Interferon-gamma/metabolism , Microglia/metabolism , Microglia/microbiology , Microscopy, Electron , NF-kappa B/metabolism , Nitric Oxide Synthase Type II/metabolism , Olfactory Bulb/metabolism , Olfactory Bulb/microbiology , Rats
9.
Genet Mol Res ; 14(4): 13437-55, 2015 Oct 28.
Article in English | MEDLINE | ID: mdl-26535658

ABSTRACT

We examined the effect of muscle basal lamina (MBL) with neural stem cells (NSCs) and olfactory ensheathing cells (OECs) on spinal cord injury repair. Seventy-two Sprague-Dawley rats were subjected to spinal cord hemisection and divided into 6 groups. In blank control group (group A), the ends of the spinal cord hemisection model were flushed with physiological saline. In NSC transplantation group (B), OEC transplantation group (C), MBL with NSC transplantation group (D), MBL with OEC transplantation group (E), and MBL with NSC and OEC transplantation group (F), NSCs, OECs, MBL with NSCs, MBL with OECs, and MBL with NSCs and OECs were implanted into the ends of the hemisection model. Survival and migration of transplanted cells were detected by immunohistochemistry and immunofluorescence after 4 and 8 weeks. Hind limb function repair was evaluated by Bundle branch block score at various time points before and after surgery. MBL could promote NSC growth along its lumen and promote host cell advancement in the lumen, reducing local inflammatory responses. Using MBL with NSCs and/or OECs for spinal cord repair shows advantages over simple cell transplantation. Group F contained more nerve cells in muscle basal lamina than group E. This method is useful for forming more axons, synaptic connections, and signal transduction pathways. However, these new axons showed nerve demyelination, which may greatly limit nerve signal conduction. In group F, OECs could induce neural stem cells, axonal growth, and synaptic connection formation, but its role is limited.


Subject(s)
Neural Stem Cells/cytology , Animals , Cell Transplantation , Female , Male , Olfactory Bulb/cytology , Rats , Rats, Sprague-Dawley , Spinal Cord Injuries , Spinal Cord Regeneration/physiology
10.
PLoS One ; 9(11): e111733, 2014.
Article in English | MEDLINE | ID: mdl-25372872

ABSTRACT

Sex differences in the human olfactory function reportedly exist for olfactory sensitivity, odorant identification and memory, and tasks in which odors are rated based on psychological features such as familiarity, intensity, pleasantness, and others. Which might be the neural bases for these behavioral differences? The number of cells in olfactory regions, and especially the number of neurons, may represent a more accurate indicator of the neural machinery than volume or weight, but besides gross volume measures of the human olfactory bulb, no systematic study of sex differences in the absolute number of cells has yet been undertaken. In this work, we investigate a possible sexual dimorphism in the olfactory bulb, by quantifying postmortem material from 7 men and 11 women (ages 55-94 years) with the isotropic fractionator, an unbiased and accurate method to estimate absolute cell numbers in brain regions. Female bulbs weighed 0.132 g in average, while male bulbs weighed 0.137 g, a non-significant difference; however, the total number of cells was 16.2 million in females, and 9.2 million in males, a significant difference of 43.2%. The number of neurons in females reached 6.9 million, being no more than 3.5 million in males, a difference of 49.3%. The number of non-neuronal cells also proved higher in women than in men: 9.3 million and 5.7 million, respectively, a significant difference of 38.7%. The same differences remained when corrected for mass. Results demonstrate a sex-related difference in the absolute number of total, neuronal and non-neuronal cells, favoring women by 40-50%. It is conceivable that these differences in quantitative cellularity may have functional impact, albeit difficult to infer how exactly this would be, without knowing the specific circuits cells make. However, the reported advantage of women as compared to men may stimulate future work on sex dimorphism of synaptic microcircuitry in the olfactory bulb.


Subject(s)
Olfactory Bulb/cytology , Sex Characteristics , Aged , Aged, 80 and over , Cell Count , Female , Humans , Male , Middle Aged , Neuroglia/cytology , Neurons/cytology , Olfactory Bulb/anatomy & histology , Organ Size , Sex Factors
11.
Dev Dyn ; 243(12): 1619-31, 2014 Dec.
Article in English | MEDLINE | ID: mdl-25255735

ABSTRACT

BACKGROUND: The peripheral olfactory sensory system arises from morphologically identifiable structures called placodes. Placodes are relatively late developing structures, evident only well after the initiation of somitogenesis. Placodes are generally described as being induced from the ectoderm suggesting that their development is separate from the coordinated cell movements generating the central nervous system. RESULTS: With the advent of modern techniques it is possible to follow the development of the neurectoderm giving rise to the anterior neural tube, including the olfactory placodes. The cell movements giving rise to the optic cup are coordinated with those generating the olfactory placodes and adjacent telencephalon. The formation of the basal lamina separating the placode from the neural tube is coincident with the anterior migration of cranial neural crest. CONCLUSIONS: Olfactory placodes are transient morphological structures arising from a continuous sheet of neurectoderm that gives rise to the peripheral and central nervous system. This field of cells is specified at the end of gastrulation and not secondarily induced from ectoderm. The separation of olfactory placodes and telencephalon occurs through complex cell movements within the developing neural plate similar to that observed for the developing optic cup.


Subject(s)
Neural Plate/embryology , Neural Tube/embryology , Olfactory Bulb/embryology , Organogenesis/physiology , Zebrafish/embryology , Animals , Neural Plate/cytology , Neural Tube/cytology , Olfactory Bulb/cytology
12.
Genet Mol Res ; 13(2): 4124-9, 2014 May 30.
Article in English | MEDLINE | ID: mdl-24938704

ABSTRACT

The aim of this study was to determine the effectiveness and safety of transplantation of olfactory ensheathing cells for functional repair of the spinal cord. An olfactory bulb was obtained from a 4- to 5-month-old aborted fetus, and it was digested into single olfactory ensheathing cells and then cultured and purified for 1 to 2 weeks. Under general anesthesia, these single-cell suspensions of olfactory ensheathing cells were injected into the corresponding spinal injury site with 0.45-mm-diameter injections. The American Spinal Injury Association (ASIA) Impairment Scale was used to evaluate spinal function. A total of 15 patients (12 men, 3 women; age range, 18-56 years; mean age, 40) were admitted for obsolete spinal injuries. Spinal functions of the 15 patients were observed and followed postoperatively for a period ranging from 2 weeks to 1 month. All the 15 patients exhibited improvements in spinal function, and the improvement tendencies continued. Twelve patients had obvious spinal function improvement, and three had slight improvement according to the ASIA scale, with an obvious difference between preoperation and postoperation measures (P < 0.05). No fevers, infections, functional deteriorations, or deaths were seen. Thus, transplantation of olfactory ensheathing cells promoted spinal and neurofunctional recovery in patients with malignant spinal injuries, and this therapeutic method was safe.


Subject(s)
Cell Transplantation , Nerve Regeneration , Olfactory Bulb/transplantation , Spinal Cord Injuries/surgery , Adolescent , Adult , Female , Humans , Male , Middle Aged , Olfactory Bulb/cytology , Olfactory Bulb/surgery , Smell/genetics , Spinal Cord/pathology , Spinal Cord/surgery , Spinal Cord Injuries/physiopathology , Young Adult
13.
Neurosci Lett ; 575: 101-6, 2014 Jul 11.
Article in English | MEDLINE | ID: mdl-24882722

ABSTRACT

Abuse of toluene-containing inhalants is associated to various cognitive impairments that have been partly associated to deviation of the hippocampal neurogenesis processes during adulthood. In the present study we analyzed the effect of chronic toluene exposure (6000ppm) on cell proliferation and migration in the other selected area of the rodent brain where neurogenesis persist throughout adulthood, the subventricular zone of the lateral ventricle (SVZ). We used an anti-Ki67 antibody to evaluate SVZ cell proliferation, BrdU to evaluate cell survival and double-staining with BrdU and the migration marker doublecortin (DCX) to evaluate migration, by immunofluorescence 2h, 1, 5, 10 or 15 days after 20 sessions of toluene exposure. We found that toluene induced an initial burst of cell proliferation in the SVZ but not a significant increase in migration toward the rostral migratory stream (RMS) or the number of cells that migrate to the olfactory bulb. In addition, we detected a small number of new migrating cells in the corpus callosum and striatum of control mice that was similar in toluene-exposed brains. These results may underline the homeostatic capabilities of the populations of dividing cells, previously demonstrated using other drugs of abuse and demonstrate that toluene misuse can alter cellular proliferation in the postnatal brain.


Subject(s)
Lateral Ventricles/drug effects , Neural Stem Cells/drug effects , Olfactory Bulb/drug effects , Psychotropic Drugs/toxicity , Toluene/toxicity , Animals , Cell Movement/drug effects , Cell Proliferation/drug effects , Doublecortin Protein , Lateral Ventricles/cytology , Male , Mice , Neural Stem Cells/cytology , Neural Stem Cells/physiology , Olfactory Bulb/cytology
14.
PLoS One ; 9(1): e85826, 2014.
Article in English | MEDLINE | ID: mdl-24465729

ABSTRACT

The cortical amygdala receives direct olfactory inputs and is thought to participate in processing and learning of biologically relevant olfactory cues. As for other brain structures implicated in learning, the principal neurons of the anterior cortical nucleus (ACo) exhibit intrinsic subthreshold membrane potential oscillations in the θ-frequency range. Here we show that nearly 50% of ACo layer II neurons also display electrical resonance, consisting of selective responsiveness to stimuli of a preferential frequency (2-6 Hz). Their impedance profile resembles an electrical band-pass filter with a peak at the preferred frequency, in contrast to the low-pass filter properties of other neurons. Most ACo resonant neurons displayed frequency preference along the whole subthreshold voltage range. We used pharmacological tools to identify the voltage-dependent conductances implicated in resonance. A hyperpolarization-activated cationic current depending on HCN channels underlies resonance at resting and hyperpolarized potentials; notably, this current also participates in resonance at depolarized subthreshold voltages. KV7/KCNQ K+ channels also contribute to resonant behavior at depolarized potentials, but not in all resonant cells. Moreover, resonance was strongly attenuated after blockade of voltage-dependent persistent Na+ channels, suggesting an amplifying role. Remarkably, resonant neurons presented a higher firing probability for stimuli of the preferred frequency. To fully understand the mechanisms underlying resonance in these neurons, we developed a comprehensive conductance-based model including the aforementioned and leak conductances, as well as Hodgkin and Huxley-type channels. The model reproduces the resonant impedance profile and our pharmacological results, allowing a quantitative evaluation of the contribution of each conductance to resonance. It also replicates selective spiking at the resonant frequency and allows a prediction of the temperature-dependent shift in resonance frequency. Our results provide a complete characterization of the resonant behavior of olfactory amygdala neurons and shed light on a putative mechanism for network activity coordination in the intact brain.


Subject(s)
Amygdala/cytology , Electrophysiological Phenomena , Neurons/physiology , Olfactory Bulb/cytology , Theta Rhythm/physiology , Animals , Cerebral Cortex/physiology , Computer Simulation , Ion Channel Gating , Ions , Kinetics , Male , Membrane Potentials/physiology , Models, Neurological , Rats , Rats, Sprague-Dawley , Sodium Channels/metabolism
15.
J Neurosci Res ; 91(12): 1572-80, 2013 Dec.
Article in English | MEDLINE | ID: mdl-24105692

ABSTRACT

Complex carbohydrate structures are essential molecules of infectious bacteria, parasites, and host cells and are involved in cell signaling associated with immune responses, glycoprotein homeostasis, and cell migration. The uptake of mannose-tailed glycans is usually carried out by professional phagocytes to trigger MHC class I- and MHC class II-restricted antigen presentation or, alternatively, to end inflammation. We have detected the mannose receptor (MR) in cultured olfactory ensheathing cells (OECs), so we investigated by flow cytometry whether recently dissociated cells of the olfactory bulb (OB) nerve fiber layer (ONL) could bind a mannosylated ligand (fluorescein conjugate of mannosyl bovine serum albumin; Man/BSA-FITC) in a specific manner. In addition, we estimated the relative proportion of ONL OECs, microglia, and astrocytes, tagged by 2'3'-cyclic nucleotide 3'-phosphodiesterase (CNPase), by the B4 isolectin of Griffonia simplicifonia (IB4), and by glial fibrillary acidic protein (GFAP), respectively, that were Man/BSA-FITC(+) . We also determined by histochemistry and/or immunohistochemistry whether Man/BSA-FITC or an anti-MR antibody (anti-C-terminal MR peptide; anti-cMR) labeled OECs and/or parenchymal microglia. In addition, we confirmed by Western blot with the K1K2 (against the entire MR molecule) antibody that a band of about 180 kDA is expressed in the OB. Our findings are compatible with a prospective sentinel role of OECs against pathogens of the upper airways and/or damage-associated glycidic patterns as well as with homeostasis of OB mannosylated glycoproteins.


Subject(s)
Lectins, C-Type/biosynthesis , Mannose-Binding Lectins/biosynthesis , Neuroglia/metabolism , Olfactory Bulb/cytology , Olfactory Bulb/metabolism , Receptors, Cell Surface/biosynthesis , Animals , Blotting, Western , Flow Cytometry , Immunohistochemistry , Mannose Receptor , Rats , Rats, Wistar
16.
Rev Neurol ; 56(10): 521-31, 2013 May 16.
Article in Spanish | MEDLINE | ID: mdl-23658035

ABSTRACT

INTRODUCTION: Spinal cord injury is a catastrophic event with permanent consequences during the all life. Treatment research has been based in the development of therapies that reduce the discapacity, but since the nineties there has been an important advance and several cellular transplants have been tested in spinal cord animal models, like Schwann cells, astrocytes and olfactory and olfactory ensheathing cells (OEC). AIM: Detailed account of spinal cord injury pathogeny, primary and secondary, and the OEC mechanisms for the regeneration effects that have been described in the literature. DEVELOPMENT: After the trauma, spinal cord injury develops in two phases, the primary injury with characteristics compression lesions, and the secondary produce for several factors that occur in parallel and include: vascular, cellular and molecular factors, and glial scar formation. The most of spinal cord models and OEC transplants have been reported functional recovery, remielinization and axonal regeneration. These cells exert their action in a direct way by producing grow factors and in an indirect way inducing directly neuronal an axonal regeneration and remielinization. CONCLUSIONS: OEC are a therapeutic option in patients with spinal cord injury, because they induce in a direct or indirect way, neuronal and axonal regeneration, remielinization, decrease the glial scar and produce other effects that conduce a functional recovery.


TITLE: Patogenia de la lesion medular y mecanismos de reparacion inducidos por las celulas de la glia envolvente olfatoria.Introduccion. La lesion medular es un evento catastrofico, cuyas consecuencias persisten durante toda la vida del paciente. La investigacion en tratamiento se ha basado principalmente en el desarrollo de terapias que reduzcan la discapacidad, pero desde los anos noventa hay un avance significativo y se han probado varios trasplantes celulares en modelos animales de lesion medular, celulas de Schwann, astrocitos y celulas de la glia envolvente olfatoria (CGEO). Objetivo. Hacer un recuento detallado de la patogenia de la lesion medular primaria y secundaria y de los mecanismos por los cuales las CGEO inducirian sus posibles efectos regenerativos descritos en la bibliografia. Desarrollo. Despues del traumatismo, la lesion se desarrolla en dos fases, la primaria se caracteriza por las lesiones de compresion y la secundaria se produce por una serie de factores que se dan en paralelo y que incluyen factores vasculares, celulares, moleculares y formacion de cicatriz glial. La mayoria de los modelos de lesion medular y trasplante con CGEO han comunicado recuperacion funcional, remielinizacion y regeneracion axonal. Estas celulas ejercen su accion de manera indirecta a traves de la produccion de factores de crecimiento y de manera directa induciendo regeneracion neuronal, axonal y remielinizacion. Conclusiones. Las CGEO son una opcion terapeutica en pacientes con lesion medular debido a que inducen de modo directo o indirecto regeneracion neuronal, axonal, remielinizacion de axones, disminucion de cicatriz glial y otros efectos que conducen a la recuperacion funcional.


Subject(s)
Cell Transplantation , Olfactory Bulb/cytology , Regeneration/physiology , Spinal Cord Injuries/etiology , Wound Healing/physiology , Animals , Astrocytes/physiology , Cicatrix/pathology , Cytokines/physiology , Demyelinating Diseases/etiology , Demyelinating Diseases/physiopathology , Edema/etiology , Humans , Ischemia/etiology , Lymphocytes/physiology , Macrophages/physiology , Mice , Microcirculation , Microglia/physiology , Nerve Growth Factors/physiology , Nerve Growth Factors/therapeutic use , Nerve Tissue Proteins/physiology , Neurogenesis , Neutrophils/physiology , Rats , Retrograde Degeneration/physiopathology , Spinal Cord/blood supply , Spinal Cord Compression/complications , Spinal Cord Injuries/pathology , Spinal Cord Injuries/physiopathology , Spinal Cord Injuries/surgery , Stem Cells/physiology
17.
PLoS One ; 8(2): e56148, 2013.
Article in English | MEDLINE | ID: mdl-23405261

ABSTRACT

Interneurons in the olfactory bulb are key elements of odor processing but their roles have not yet being fully understood. Two types of inhibitory interneurons, periglomerular and granule cells, act at two different levels within the olfactory bulb and may have different roles in coordinating the spiking of mitral cells, which are the principal output neurons of the olfactory bulb. In this work we introduce a reduced compartmental model of the periglomerular cell and use it to investigate its role on mitral cell spiking in a model of an elementary cell triad composed of these two cell types plus a granule cell. Our simulation results show that the periglomerular cell is more effective in inhibiting the mitral cell than the granule cell. Based on our results we predict that periglomerular and granule cells have different roles in the control of mitral cell spiking. The periglomerular cell would be the only one capable of completely inhibiting the mitral cell, and the activity decrease of the mitral cell through this inhibitory action would occur in a stepwise fashion depending on parameters of the periglomerular and granule cells as well as on the relative times of arrival of external stimuli to the three cells. The major role of the granule cell would be to facilitate the inhibitory action of the periglomerular cell by enlarging the range of parameters of the periglomerular cell which correspond to complete inhibition of the mitral cell. The combined action of the two interneurons would thus provide an efficient way of controling the instantaneous value of the firing rate of the mitral cell.


Subject(s)
Computer Simulation , Interneurons/physiology , Olfactory Bulb/physiology , Olfactory Nerve/physiology , Animals , Electrophysiology , Evoked Potentials , Interneurons/cytology , Olfactory Bulb/cytology , Olfactory Nerve/cytology , Rats
18.
Neuroscience ; 232: 151-60, 2013 Mar 01.
Article in English | MEDLINE | ID: mdl-23262235

ABSTRACT

We have previously shown that the first-paced mating encounter increases the number of newborn cells in the granule cell layer (Gra; also known as internal cell layer, ICL) of the accessory olfactory bulb (AOB) in the adult female rat (Corona et al., 2011). In the present study we evaluated if repetition of the stimulus (paced mating) could increase the arrival of more newborn neurons in the olfactory bulb generated during the first session of paced sexual contact. Sexually naive female rats were bilaterally ovariectomized, hormonally supplemented with estradiol (E2) and progesterone (P) and randomly assigned to one of four groups: (1) without sexual contact, (2) one session of paced mating, (3) four sessions of paced mating, and (4) four sessions of non-paced mating. We also included a group of gonadally intact females. On the first day of the experiment, all females were i.p. injected with the marker of DNA synthesis bromodeoxyuridine and were killed 16 days later. Blood was collected at sacrifice to determine the plasma levels of E2 and P. The number of newborn neurons that arrived at the ICL of the AOB and the Gra of the main olfactory bulb (MOB) increased, relative to all other groups, only in the group that repeatedly mated under pacing conditions. No differences were found in E2 and P levels between supplemented groups indicating that our results are not influenced by changes in hormone concentrations. We suggest that repeated paced mating promotes the arrival of more newborn neurons in the AOB and MOB.


Subject(s)
Neurogenesis/physiology , Neurons/physiology , Olfactory Bulb/physiology , Animals , Antigens, Nuclear/metabolism , Bromodeoxyuridine , Cell Count , Estradiol/administration & dosage , Estradiol/metabolism , Female , Microscopy, Confocal , Nerve Tissue Proteins/metabolism , Neurons/cytology , Olfactory Bulb/cytology , Ovariectomy , Progesterone/administration & dosage , Progesterone/metabolism , Random Allocation , Rats, Wistar , Sexual Behavior, Animal/physiology
19.
J Chem Neuroanat ; 46(1-2): 30-4, 2012 Dec.
Article in English | MEDLINE | ID: mdl-23000948

ABSTRACT

The mammalian brain preserves the ability to replace olfactory periglomerular cells (PGC) throughout life. Even though we have detailed a great deal the mechanisms underlying stem and amplifying cells maintenance and proliferation, as well as those modulating migration and differentiation, our knowledge on PGC phenotypic plasticity is at best fragmented and controversial. Here we explored whether chronically reinforced olfactory conditioning influences the phenotype of newborn PGC. Accordingly, olfactory conditioned rats showed increased numbers of GAD 65/67 positive PGC. Because such phenotypic change was not accompanied neither by increments in the total number of PGC, or periglomerular cell nuclei labeled with bromodeoxyuridine, nor by reductions in the number of tyrosine hydroxylase (TH), calbindin (CB) or calretinin (CR) immunoreactive PGC, we speculate that increments in the number of GABAergic PGC occur at the expense of other PGC phenotypes. In any event, these results support that adult newborn PGC phenotype may be subjected to phenotypic plasticity influenced by sensory stimulation.


Subject(s)
Conditioning, Operant/physiology , GABAergic Neurons/cytology , GABAergic Neurons/physiology , Olfactory Bulb/cytology , Olfactory Bulb/physiology , Reinforcement, Psychology , Animals , Male , Neurogenesis/physiology , Neuronal Plasticity/physiology , Rats , Rats, Wistar
20.
Neurosci Lett ; 522(1): 6-11, 2012 Jul 26.
Article in English | MEDLINE | ID: mdl-22634627

ABSTRACT

The olfactory bulb (OB) is rich in the number and variety of neurotransmitter and neuropeptide containing cells, in particular in the glomerular layer. Several reports suggest that numbers of some periglomerular phenotypes could change depending on age. However, it is unclear whether the different classes of periglomerular interneurons are modified or are maintained stable throughout life. Thus, our first objective was to obtain the absolute number of cells belonging to the different periglomerular phenotypes at adulthood. On the other hand, the olfactory bulb is continously supplied with newly generated periglomerular neurons produced by stem cells located in the subventricular zone (SVZ) and rostral migratory stream. Previously, we demonstrated that the implantation of a physical barrier completely prevents SVZ neuroblast migration towards the OB. Then, another objective of this study was to evaluate whether stopping the continuous supply of SVZ neuroblasts modified the different periglomerular populations throughout time. In summary, we estimated the total number of TH-IR, CalB-IR, CalR-IR and GAD-IR cells in the OB glomerular layer at several time points in control and barrier implanted adult rats. In addition, we estimated the volume of glomerular, granular and complete OB. Our main finding was that the number of the four main periglomerular populations is age-dependent, even after impairment of subventricular neuroblast migration. Furthermore, we established that these changes do not correlate with changes in the volume of glomerular layer.


Subject(s)
Interneurons/cytology , Olfactory Bulb/cytology , Age Factors , Animals , Calbindin 2 , Calbindins , Cell Count , Cell Division , Cell Movement , Cerebral Ventricles , Glutamic Acid/metabolism , Immunohistochemistry , Interneurons/physiology , Male , Olfactory Bulb/growth & development , Rats , Rats, Wistar , S100 Calcium Binding Protein G/metabolism , Tyrosine 3-Monooxygenase/metabolism
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