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J Ngai

Publications and source records attributed to J Ngai.

At least 19 recordsLinked to original sources
PubMed2001

Onset of odorant receptor gene expression during olfactory sensory neuron regeneration.

Individual olfactory sensory neurons are thought to express only one odorant receptor gene from a repertoire of hundreds to thousands of genes. How do these sensory neurons choose just one specific odorant receptor to express during their differentiation? As an initial attempt toward understanding the process of odorant receptor gene regulation, we studied when odorant receptor expression is activated during sensory neuron regeneration. We find that receptor gene expression is activated in postmitotic neurons and can occur in the absence of the olfactory bulb. These results suggest that receptor expression is restricted to the terminal stages of olfactory neuron differentiation, and sensory neurons do not simply inherit the odorant receptor that is already expressed in mitotic precursor cells. Our results also support a model in which odorant receptor gene expression occurs independent of the olfactory bulb.

Animals↗
PubMed2001

Analysis and characterization of an odorant receptor gene cluster in the zebrafish genome.

A 140.7-kb segment of zebrafish genomic DNA known to contain odorant receptor (OR) genes was fully sequenced to characterize more completely the organization of this gene cluster. A total of 20 OR genes were identified in this region. The most highly related genes are grouped in closest proximity to one another and in the same transcriptional orientation, indicating that a series of tandem duplications was responsible for the expansion of the OR gene family in teleost fish. Our analysis also revealed sequences that may be involved in the transcriptional regulation of OR genes within the cluster.

Amino Acid Sequence↗
PubMed2000

Formation of precise connections in the olfactory bulb occurs in the absence of odorant-evoked neuronal activity.

Olfactory neurons expressing the same odorant receptor converge to a small number of glomeruli in the olfactory bulb. In turn, mitral and tufted cells receive and relay this information to higher cortical regions. In other sensory systems, correlated neuronal activity is thought to refine synaptic connections during development. We asked whether the pattern of connections between olfactory sensory axons and mitral cell dendrites is affected when odor-evoked signaling is eliminated in mice lacking functional olfactory cyclic nucleotide-gated (CNG) channels. We demonstrate that olfactory sensory axons converge normally in the CNG channel mutant background. We further show that the pruning of mitral cell dendrites, although slowed during development, is ultimately unperturbed in mutant animals. Thus, the olfactory CNG channel-and by inference correlated neural activity--is not required for generating synaptic specificity in the olfactory bulb.

Animals↗
PubMed1999

Functional identification of a goldfish odorant receptor.

The vertebrate olfactory system utilizes odorant receptors to receive and discriminate thousands of different chemical stimuli. An understanding of how these receptors encode information about an odorant's molecular structure requires a characterization of their ligand specificities. We employed an expression cloning strategy to identify a goldfish odorant receptor that is activated by amino acids-potent odorants for fish. Structure-activity analysis indicates that the receptor is preferentially tuned to recognize basic amino acids. The receptor is a member of a multigene family of G protein-coupled receptors, sharing sequence similarities with the calcium sensing, metabotropic glutamate, and V2R class of vomeronasal receptors. The ligand tuning properties of the goldfish amino acid odorant receptor provide information for unraveling the molecular mechanisms underlying olfactory coding.

Animals↗
PubMed1999

Development of the vertebrate main olfactory system.

Olfactory receptor neurons project from the sensory epithelium to stereotyped targets within the olfactory bulb. Recent studies suggest that the generation of this precise spatial map probably involves a hierarchy of guidance events, as receptor neurons integrate information present in the epithelium and bulb to reach their target.

Animals↗
PubMed1998

Pathfinding of olfactory neuron axons to stereotyped glomerular targets revealed by dynamic imaging in living zebrafish embryos.

In the vertebrate olfactory system, sensory neurons with common odorant specificities project to specific glomeruli in the olfactory bulb. How do olfactory sensory neurons find their glomerular targets? To address this question, we have visualized the genesis of the peripheral olfactory system in living zebrafish embryos. Dye labelings reveal that a primordial yet stereotyped map of glomeruli is apparent during embryogenesis. By labeling a small number of cells with an ectopically expressed green fluorescent protein reporter, we can observe the dynamic growth behaviors of individual olfactory neuron growth cones as they project to their glomeruli. We find that olfactory axons extend directly to their partner glomeruli, suggesting that these cells' growth cones rely upon pathfinding cues to reach their targets.

Afferent Pathways↗
PubMed1997

Noncoordinate expression of odorant receptor genes tightly linked in the zebrafish genome.

We have characterized the organization and expression of odorant receptor genes clustered within approximately 100 kb of the zebrafish genome. Physical analysis of this genomic region reveals that the receptor genes are tightly linked in tandem arrays. The expression patterns of these genes were evaluated during development as well as in the adult olfactory epithelium. Highly related genes from this array are expressed individually in different olfactory neurons, suggesting that the discriminatory capacity of the vertebrate olfactory system has been maximized by segregating the most similar receptors into distinct cellular pathways. Furthermore, genes from this cluster are activated at different times of development. Together, these results indicate that genomically linked odorant receptor genes are not coordinately regulated.

Amino Acid Sequence↗
PubMed1997

Comparison of peripheral blood CD34+ concentration, colony-forming units granulocyte-macrophage and mononuclear cells in leukapheresed product for the prediction of peripheral blood CD34+ cell yield harvest.

The aim of this study was to evaluate reliability of parameters which may be used to guide peripheral stem cell harvests in cancer patients prior to myeloablative chemotherapy. Each leukapheresed product was analysed for CD34-positive (CD34+) cell count, mononuclear cell (MNC) count and the number of colony-forming units granulocyte-macrophage (CFU-GM). Each patient's peripheral blood (PB) taken before leukapheresis was analysed for CD34+ concentration. We evaluated whether the CD34+ yield from leukapheresis correlated with any of the three parameters. A total of 119 procedures were performed in 33 patients. The yield of CD34+ cells by leukapheresis correlated weakly but significantly with the peripheral blood CD34+ cell count (R = 0.4 P < 0.05), the MNC cells (R = 0.4, P < 0.05), and CFU-GM (R = 0.4, P < 0.05). When a PB CD34+ count of 50 x 10(6)/L was used as a cut-off value, the accuracy for prediction of adequate leukapheresis (> 1 x 10(6) CD34+ cells/kg) was 78%.

Adolescent↗
PubMed1996

Asynchronous onset of odorant receptor expression in the developing zebrafish olfactory system.

The functional identity of an olfactory neuron is determined in large part by the odorant receptors it expresses. As an approach toward understanding the events that underlie the specification of olfactory neurons, we have examined the patterns of odorant receptor gene expression in the developing zebrafish. Surprisingly, we find that the onset of specific odorant receptor expression occurs asynchronously in the developing olfactory placode. Our results suggest that odorant receptor expression is not strictly stochastic, but rather is governed by temporally regulated cues during development. Moreover, by restricting the number of receptor genes competent for transcription at different times of development, temporal waves of expression may provide a mechanism for simplifying the regulation of the large odorant receptor gene family.

Age Factors↗
PubMed1996

General anosmia caused by a targeted disruption of the mouse olfactory cyclic nucleotide-gated cation channel.

Olfactory neurons transduce the binding of odorants into membrane depolarization. Two intracellular messengers, cyclic AMP (cAMP) and inositol trisphosphate (IP3), are thought to mediate this process, with cAMP generating responses to some odorants and IP3 mediating responses to others. cAMP causes membrane depolarization by activating a cation-selective cyclic nucleotide-gated (CNG) channel. We created a mutant "knockout" mouse lacking functional olfactory CNG channels to assess the roles of different second messenger pathways in olfactory transduction. Using an electrophysiological assay, we find that excitatory responses to both cAMP- and IP3-producing odorants are undetectable in knockout mice. Our results provide direct evidence that the CNG channel subserves excitatory olfactory signal transduction, and further suggest that cAMP is the sole second messenger mediating this process.

Animals↗
PubMed1993

Spatial segregation of odorant receptor expression in the mammalian olfactory epithelium.

The signal elicited by the interaction of odorous ligands with receptors on olfactory sensory neurons must be decoded by the brain to determine which of the numerous receptors have been activated. We have examined the patterns of odorant receptor expression in the rat olfactory epithelium to determine whether the mammalian olfactory system employs spatial segregation of sensory input to encode the identity of an odorant stimulus. In situ hybridization experiments with probes for 11 different odorant receptors demonstrate that sensory neurons expressing distinct receptors are topologically segregated into a small number of broad, yet circumscribed, zones within the olfactory epithelium. Within a given zone, however, olfactory neurons expressing a specific receptor appear to be randomly distributed, rather than spatially localized. The complex mammalian olfactory system may therefore compartmentalize the epithelium into anatomically and functionally discrete units, such that each zone expresses only a subset of the entire receptor repertoire.

Animals↗
PubMed1993

The family of genes encoding odorant receptors in the channel catfish.

The anatomical and numerical simplicity of the fish olfactory system has led us to examine the family of olfactory receptors expressed in the catfish. We have identified a family of genes encoding seven transmembrane domain receptors that share considerable homology with the odorant receptors of the rat. The size of the catfish receptor repertoire appears to be far smaller than in mammals. Analysis of the nucleotide sequences suggests that these receptor genes have undergone positive Darwinian selection to generate enhanced diversity within the putative odorant-binding domains. Individual receptor clones anneal with 0.5%-2% of the olfactory neurons, suggesting that a single cell expresses only a small subset of distinct odorant receptors. Each cell, therefore, possesses a unique identity defined by the receptors it expresses. These data suggest that the brain may discriminate among odors by determining which neurons have been activated.

Amino Acid Sequence↗
PubMed1993

Coding of olfactory information: topography of odorant receptor expression in the catfish olfactory epithelium.

Discrimination among the vast array of odors requires that the brain discern which of the numerous odorant receptors have been activated. If individual olfactory neurons express only a subset of the odorant receptor repertoire, then the nature of a given odorant can be discerned by identifying which cells have been activated. We performed in situ hybridization experiments demonstrating that individual olfactory neurons express different complements of odorant receptors and are therefore functionally distinct. Thus, a topographic map, defining either the positions of specific neurons in the epithelium or the positions of their projections, may be employed to determine the quality of an olfactory stimulus. Neurons expressing specific receptors appear to be randomly distributed within the olfactory epithelium. These data are consistent with a model in which randomly dispersed olfactory neurons with common receptor specificities project to common glomeruli in the olfactory bulb.

Animals↗
PubMed1992

Molecular cloning and single-channel properties of the cyclic nucleotide-gated channel from catfish olfactory neurons.

We have cloned a functional cDNA encoding the cyclic nucleotide-gated channel selectively expressed in catfish olfactory sensory neurons. The cyclic nucleotide-gated channels share sequence and structural features with the family of voltage-gated ion channels. This homology is most evident in transmembrane region S4, the putative voltage sensor domain, and the H5 domain, thought to form the channel pore. We have characterized the single-channel properties of the cloned catfish channel and compared these properties with the channel in native catfish olfactory sensory neurons. The channel is activated equally well by cAMP and cGMP, shows only a slight voltage dependence of gating, and exhibits a pH- and voltage-dependent subconductance state similar to that observed for the voltage-gated L-type calcium channel.

Amino Acid Sequence↗
PubMed1990

Localization of newly synthesized vimentin subunits reveals a novel mechanism of intermediate filament assembly.

We have assessed the mechanism of intermediate filament assembly by assaying the sites of incorporation of chicken vimentin subunits expressed under the control of an inducible promoter in transfected mouse fibroblasts. The localization of newly synthesized vimentin was determined by immunofluorescence and immunoelectron microscopy at short time periods of induced synthesis, using antibodies specific for chicken vimentin. Under conditions where neither the soluble subunit pools nor the steady-state distribution of endogenous filaments are affected, newly synthesized vimentin incorporates into the vimentin filament network at numerous and discrete sites throughout the cell. Over time, the pattern of newly assembled vimentin converts to a continuous array coincident with preexisting vimentin filaments. These results are consistent with a novel mechanism of intermediate filament assembly, whereby growth of intermediate filaments occurs by topographically restricted and localized subunit addition, necessitating a transient disruption of filament integrity.

Animals↗
PubMed1987

Regulated expression of multiple chicken erythroid membrane skeletal protein 4.1 variants is governed by differential RNA processing and translational control.

Protein 4.1 is an extrinsic membrane protein that facilitates the interaction of spectrin and actin in the erythroid membrane skeleton and exists as several structurally related polypeptides in chickens. The ratio of protein 4.1 variants is developmentally regulated during terminal differentiation of chicken erythroid and lenticular cells. To examine the mechanisms by which multiple chicken protein 4.1 variants are differentially expressed, we have isolated cDNA clones specific for chicken erythroid protein 4.1. We show that a single protein 4.1 gene gives rise to multiple 6.6-kilobase mRNAs by differential RNA processing. Furthermore, the ratios of protein 4.1 mRNAs change during chicken embryonic erythropoiesis. We observe a quantitative difference in variant ratios when protein 4.1 is synthesized in vivo or in a rabbit reticulocyte lysate in vitro. Our results show that the expression of multiple protein 4.1 polypeptides is regulated at the levels of translation and RNA processing.

Amino Acid Sequence↗
PubMed1987

Expression of transfected vimentin genes in differentiating murine erythroleukemia cells reveals divergent cis-acting regulation of avian and mammalian vimentin sequences.

We studied the expression of transfected chicken and hamster vimentin genes in murine erythroleukemia (MEL) cells. MEL cells normally repress the levels of endogenous mouse vimentin mRNA during inducermediated differentiation, resulting in a subsequent loss of vimentin filaments. Expression of vimentin in differentiating MEL cells reflects the disappearance of vimentin filaments during mammalian erythropoiesis in vivo. In contrast, chicken erythroid cells express high levels of vimentin mRNA and vimentin filaments during terminal differentiation. We demonstrate here that chicken vimentin mRNA levels increase significantly in differentiating transfected MEL cells, whereas similarly transfected hamster vimentin genes are negatively regulated. In conjunction with in vitro nuclear run-on transcription experiments, these results suggest that the difference in vimentin expression in avian and mammalian erythropoiesis is due to a divergence of cis-linked vimentin sequences that are responsible for transcriptional and posttranscriptional regulation of vimentin gene expression. Transfected chicken vimentin genes produce functional vimentin protein and stable vimentin filaments during MEL cell differentiation, further demonstrating that the accumulation of vimentin filaments is determined by the abundance of newly synthesized vimentin.

Animals↗