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1.
J Biol Chem ; 291(18): 9762-75, 2016 Apr 29.
Article in English | MEDLINE | ID: mdl-26957543

ABSTRACT

Formyl peptide receptor 3 (Fpr3, also known as Fpr-rs1) is a G protein-coupled receptor expressed in subsets of sensory neurons of the mouse vomeronasal organ, an olfactory substructure essential for social recognition. Fpr3 has been implicated in the sensing of infection-associated olfactory cues, but its expression pattern and function are incompletely understood. To facilitate visualization of Fpr3-expressing cells, we generated and validated two new anti-Fpr3 antibodies enabling us to analyze acute Fpr3 protein expression. Fpr3 is not only expressed in murine vomeronasal sensory neurons but also in bone marrow cells, the primary source for immune cell renewal, and in mature neutrophils. Consistent with the notion that Fpr3 functions as a pathogen sensor, Fpr3 expression in the immune system is up-regulated after stimulation with a bacterial endotoxin (lipopolysaccharide). These results strongly support a dual role for Fpr3 in both vomeronasal sensory neurons and immune cells. We also identify a large panel of mouse strains with severely altered expression and function of Fpr3, thus establishing the existence of natural Fpr3 knock-out strains. We attribute distinct Fpr3 expression in these strains to the presence or absence of a 12-nucleotide in-frame deletion (Fpr3Δ424-435). In vitro calcium imaging and immunofluorescence analyses demonstrate that the lack of four amino acids leads to an unstable, truncated, and non-functional receptor protein. The genome of at least 19 strains encodes a non-functional Fpr3 variant, whereas at least 13 other strains express an intact receptor. These results provide a foundation for understanding the in vivo function of Fpr3.


Subject(s)
Gene Expression Regulation/immunology , Receptors, Formyl Peptide/immunology , Sensory Receptor Cells/immunology , Vomeronasal Organ/immunology , Animals , Bone Marrow Cells/cytology , Bone Marrow Cells/immunology , HEK293 Cells , Humans , Mice , Mice, Knockout , Receptors, Formyl Peptide/genetics , Sensory Receptor Cells/cytology , Species Specificity , Vomeronasal Organ/cytology
2.
São Paulo; s.n; 5 nov. 2008. [116] p. ilus, tab, graf.
Thesis in Portuguese | LILACS | ID: lil-508081

ABSTRACT

No genoma de camundongo existem por volta de 1000 genes que codificam para receptores olfatórios (ORs) e 150 genes que codificam para receptores de feromônios do tipo 1 (V1Rs) distribuídos em vários cromossomos. Cada neurônio olfatório e vomeronasal seleciona um único alelo de um único gene de receptor OR ou de V1R, respectivamente, para expressar enquanto que o restante do repertório é mantido silenciado. Os mecanismos que regulam esse padrão de expressão não são conhecidos. As similaridades no padrão de expressão dos genes de ORs e de V1Rs sugerem que o mecanismo de regulação possa ser comum. Até então poucas regiões promotoras de genes de ORs e de genes de V1Rs haviam sido experimentalmente determinadas e pesquisadas. Realizamos uma análise na qual regiões a montante de um grande número de diferentes genes de ORs e de genes de V1Rs foram comparadas...


Subject(s)
Animals , Mice , Gene Expression/genetics , Pheromones/genetics , In Vitro Techniques , Molecular Biology , Olfactory Receptor Neurons , Smell/genetics , Smell/immunology , Vomeronasal Organ/immunology , Receptors, Odorant/analysis , Receptors, Odorant/biosynthesis , Dissection , Electrophoresis , Spectrophotometry , Polymerase Chain Reaction/methods , Polymerase Chain Reaction
3.
Curr Biol ; 13(5): 394-400, 2003 Mar 04.
Article in English | MEDLINE | ID: mdl-12620187

ABSTRACT

The vomeronasal organ (VNO) is a chemosensory organ specialized in the detection of pheromones in higher vertebrates. In mouse and rat, two gene superfamilies, V1r and V2r vomeronasal receptor genes, are expressed in sensory neurons whose cell bodies are located in, respectively, the apical and basal layers of the VNO epithelium. Here, we report that neurons of the basal layer express another multigene family, termed H2-Mv, representing nonclassical class I genes of the major histocompatibility complex. The nine H2-Mv genes are expressed differentially in subsets of neurons. More than one H2-Mv gene can be expressed in an individual neuron. In situ hybridization with probes for H2-Mv and V2r genes reveals complex and nonrandom combinations of coexpression. While neural expression of Mhc class I molecules is increasingly being appreciated, the H2-Mv family is distinguished by variegated expression across seemingly similar neurons and coexpression with a distinct multigene family encoding neural receptors. Our findings suggest that basal vomeronasal sensory neurons may consist of multiple lineages or compartments, defined by particular combinations of V2r and H2-Mv gene expression.


Subject(s)
Histocompatibility Antigens Class II/metabolism , Multigene Family , Neurons, Afferent/immunology , Neurons, Afferent/metabolism , Receptors, Vasopressin/metabolism , Vomeronasal Organ/immunology , Vomeronasal Organ/metabolism , Amino Acid Sequence , Animals , Histocompatibility Antigens Class II/genetics , Histocompatibility Antigens Class II/immunology , Mice , Molecular Sequence Data
4.
Brain Behav Evol ; 57(1): 18-32, 2001 Jan.
Article in English | MEDLINE | ID: mdl-11359045

ABSTRACT

Neuroanatomical distribution of FMRFamide-like immunoreactivity was investigated in the brain and olfactory system of the viviparous skink, Chalcides chalcides. In the adult brain FMRFamide immunoreactive (ir) perikarya were observed in the diagonal band of Broca, medial septal nucleus, accumbens nucleus, bed nucleus of the anterior commissure, periventricular hypothalamic nucleus, lateral forebrain bundle, and lateral preoptic, subcommissural, suprachiasmatic and lateral hypothalamic areas. This pattern was seen in both male and female brains. Though all major brain areas showed FMRFamide-ir innervation, the densest ir fiber network was observed in the hypothalamus. During development, ir elements were observed for the first time in embryos at mid-pregnancy. FMRFamide perikarya were located along the ventral surface of the vomeronasal nerve, in the olfactory peduncle mediobasally, as well as in the anterior olfactory nucleus and olfactory tubercle. Furthermore, some ir neurons were observed in the rhombencephalic reticular substance; however, the ir fiber network was poorly developed. Later in development FMRFamide-ir neurons appeared also in the bed nucleus of the anterior commissure as well as the rhombencephalic nucleus of solitary tract and the dorsal motor nucleus of vagus nerve. In juveniles, the distribution profile of FMRFamide immunoreactivity was substantially similar to that of the adults, with a less widespread neuronal distribution and a more developed fiber network. Ontogenetic presence of FMRFamide immunoreactivity in the nasal area has been linked to the presence of a nervus terminalis in this reptile.


Subject(s)
Brain/immunology , FMRFamide/immunology , Reptiles/anatomy & histology , Animals , Brain/metabolism , FMRFamide/metabolism , Female , Immunohistochemistry , Male , Nerve Fibers/immunology , Nerve Fibers/metabolism , Olfactory Pathways/immunology , Olfactory Pathways/metabolism , Vomeronasal Organ/immunology , Vomeronasal Organ/metabolism
5.
Chem Senses ; 25(4): 369-80, 2000 Aug.
Article in English | MEDLINE | ID: mdl-10944499

ABSTRACT

We observed the surface of the anterior part of the nasal septum of living subjects using an endoscope. In approximately 13% of 1842 patients without pathology of the septum, the vomeronasal pit was clearly observed on each side of the septum, and in 26% it was observed only on one side. The remaining observations indicated either the presence of putative pits or no visible evidence of a pit. However, repetitive observations on 764 subjects depicted changes over time, from nothing visible to well-defined pits and vice versa. Based on 130 subjects observed at least four times, we estimate that approximately 73% of the population exhibits at least one clearly defined pit on some days. By computer tomography, the vomeronasal cavities were located at the base of the most anterior part of the nasal septum. Histological studies indicated that the vomeronasal cavities consisted of a pit generally connected to a duct extending in a posterior direction under the nasal mucosa. Many glands were present around the duct, which contained mucus. There was no sign of the pumping elements found in other mammalian species. Most cells in the vomeronasal epithelium expressed keratin, a protein not expressed by olfactory neurons. Vomeronasal epithelial cells were not stained by an antibody against the olfactory marker protein, a protein expressed in vomeronasal receptor neurons of other mammals. Moreover, an antibody against protein S100, expressed in Schwann cells, failed to reveal the existence of vomeronasal nerve bundles that would indicate a neural connection with the brain. Positive staining was obtained with the same antibodies on specimens of human olfactory epithelium. The lack of neurons and vomeronasal nerve bundles, together with the results of other studies, suggests that the vomeronasal epithelium, unlike in other mammals, is not a sensory organ in adult humans.


Subject(s)
Vomeronasal Organ/anatomy & histology , Adolescent , Adult , Endoscopy , Female , Humans , Immunohistochemistry , Keratins/immunology , Male , Middle Aged , Phosphopyruvate Hydratase/immunology , S100 Proteins/immunology , Tomography, X-Ray Computed , Vomeronasal Organ/diagnostic imaging , Vomeronasal Organ/immunology
6.
Anat Embryol (Berl) ; 199(3): 215-24, 1999 Mar.
Article in English | MEDLINE | ID: mdl-10068087

ABSTRACT

The development of the rat vomeronasal organ was studied morphologically and immunocytochemically, using the monoclonal antibodies (MAbs) VOBM1, VOBM2 and VOM2 that react with the luminal surface of the vomeronasal sensory epithelium. Postnatal day (P) 7, 14, 21, 28, 35 and adult animals were examined. The vomeronasal organ and the blood vessel of the organ markedly increased in size and the vomeronasal glands increased in number between P7 and P14. At P35, the shape of the vomeronasal organ was similar to that of the adult but its size was slightly smaller. Electron microscopy showed that only a few scattered microvilli were present on supporting cells, and receptor cells were immature at P7. At P21, well-branched microvilli of the receptor cells and many microvilli of the supporting cells were observed on the luminal surface of the sensory epithelium. At P35, most apical endings of supporting cells and receptor cells were covered with numerous microvilli. Less developed areas were also present at the luminal surface of the epithelium at P35. At P7, immunoreactivities of the three antibodies were observed as discontinuous thin-layered bands only on the luminal surface of the sensory epithelium and no immunoreactivity was observed in other regions of the vomeronasal organ. Immunoreactivities of the VOBM1, VOBM2 and VOM2 increased with age and were observed as continuous thin-layered bands on the luminal surface of the epithelium by P35. These finding suggest that the development of the vomeronasal organ continues after birth and that the organ may reach maturity just before puberty (P42-49).


Subject(s)
Antibodies, Monoclonal/analysis , Sensory Receptor Cells/growth & development , Vomeronasal Organ/growth & development , Animals , Animals, Newborn , Epithelium/growth & development , Epithelium/immunology , Immunoenzyme Techniques , Mice , Mice, Inbred BALB C , Microscopy, Electron , Microvilli/ultrastructure , Olfactory Mucosa/chemistry , Olfactory Mucosa/growth & development , Rats , Rats, Sprague-Dawley , Sensory Receptor Cells/cytology , Sensory Receptor Cells/immunology , Vomeronasal Organ/cytology , Vomeronasal Organ/immunology
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