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Neurogenesis and neuron regeneration in the olfactory system of mammals. II. Degeneration and reconstitution of the olfactory sensory neurons after axotomy.

This report describes the retrograde degeneration affecting olfactory sensory neurons of rats after severance of their axons and illustrates the reconstitution of new neurons originating from stem cells located at the base of the olfactory neuroepithelium. Degeneration of the mature, axotomized neurons, signalled by an increased electron density of their cytoplasmic matrix and by the appearance of lipofuscin-like granules, can be detected in the neuroepithelium as early as 24 h after surgery and becomes conspicuous between the second and the third day. Degenerating neurons can be observed in decreasing number up to the tenth post-operative day. They are removed by macrophages which invade the epithelium. The reconstitution of new neurons begins to occur after eight days, when the stem cells undergo vigorous mitotic activity and differentiate into neurons. The morphology of the reconstituted neurons has been described in detail at different stages of their maturation. After 30 days, the olfactory epithelium appears similar to controls. On the basis of both morphological (in rats) and autoradiographic ( in mice) observations, the basal cells have been recognized as stem cells of the olfactory neurons.

Animals

Evolutionary Process Underlying Receptor Gene Expansion and Cellular Divergence of Olfactory Sensory Neurons in Honeybees.

Olfaction is crucial for animals' survival and adaptation. Unlike the strict singular expression of odorant receptor (OR) genes in vertebrate olfactory sensory neurons (OSNs), insects exhibit complex OR gene expression patterns. In honeybees (Apis mellifera), a significant expansion of OR genes implies a selection preference for the olfactory demands of social insects. However, the mechanisms underlying receptor expression specificity and their contribution to OSN divergence remain unclear. In this study, we used single-nucleus multiomics profiling to investigate the transcriptional regulation of OR genes and the cellular identity of OSNs in A. mellifera. We identified three distinct OR expression patterns, singular OR expression, co-expression of multiple OR genes with a single active promoter, and co-expression of multiple OR genes with multiple active promoters. Notably, ∼50% of OSNs co-expressed multiple OR genes, driven by polycistronic transcription of tandemly duplicated OR genes via a single active promoter. In these OSNs, their identity was determined by the first transcribed receptor. The divergent activation of the promoter for duplicated OR genes ensures the coordinated increased divergence of OSN population. By integrating multiomics data with genomic architecture, we illustrate how fundamental genetic mechanisms drive OR gene expansion and influence flanking regulatory elements, ultimately contributing to the cellular divergence of OSNs. Our findings highlight the interplay between gene duplication and regulatory evolution in shaping OSN diversity, providing new insights into the evolution and adaptation of olfaction in social insects. This study also sheds light on how genetic innovations contribute to the evolution of complex traits.

Animals

Neurogenesis and neuron regeneration in the olfactory system of mammals. I. Morphological aspects of differentiation and structural organization of the olfactory sensory neurons.

The neurogenetic process leading to the formation of primary sensory neurons persists into adult life in the olfactory epithelium of mammals. The morphological stages of maturation and ageing of this exceptional neuron have been described both at light and electron microscopical levels. For descriptive purposes the neural elements have been classified as: (1) basal cells proper, (2) globose basal cells, and (3) neurons. Intermediate stages, however, have been identified. Autoradiographic observations complement the morphological studies and provide a time sequence of the morphological stages leading to the mature neurons. A typical columnar arrangement of the sensory neurons has been described. Furthermore, active and quiescent zones have been recognized in the neuroepithelium. In the active zones the neurogenetic process is vigorous, and the zones are characterized by the presence of immature elements. However, in the quiescent zones there exists a population of mature elements while immature neurons are sparse.

Animals

Determinants of odorant receptor transcription and gene choice.

The mammalian olfactory system enables the detection of a wide variety of chemical compounds via the expression of a repertoire of olfactory receptors comprising the largest gene family in the mammalian genome. Olfactory sensory neurons (OSNs) each express only 1 odorant receptor (OR) gene. In mice, this requires activation of 1 OR gene and repression of over 1,400 other OR genes. In this review, we describe the mechanisms that support the transcription of OR genes and how these mechanisms impact which OR is expressed in each neuron. First, we discuss what is currently known about the role of transcription in OR choice. We then describe the role of specific features of OR genes and enhancers in the regulation of OR transcription. Finally, we discuss characteristics of OSNs which specify transcription of some OR genes while restricting the transcription of others.

Receptors, Odorant

Transcriptional interference gates monogenic odorant receptor expression in ants.

Communication is crucial to social life, and in ants, it is mediated primarily through olfaction. Ants have more odorant receptor (OR) genes than any other group of insects, generated through tandem duplications that produce large genomic arrays of related genes. The mechanism by which olfactory sensory neurons (OSNs) produce a single functional OR from these arrays remains unclear. In ant OSNs, only mRNA from one OR in an array is exported into the cytoplasm, while upstream genes are silent and transcripts from downstream genes remain nuclear. Here, we show that readthrough transcription in the downstream direction generates non-translated transcripts. We also find that OR promoters are bidirectional, producing antisense long non-coding RNAs. We suspect that neither readthrough nor antisense transcription produces functional RNA but that bidirectional transcription alone is critical to suppressing the expression of all other OR genes in a tandem array. Finally, we present evidence that this regulatory architecture is conserved across ants and bees, suggesting that this mechanism for functionally monogenic OR expression is widespread in insects with expanded OR repertoires.

Animals

NAD activates olfactory receptor 1386 to regulate type I interferon responses in Plasmodium yoelii YM infection.

Olfactory receptors (Olfr) are G protein-coupled receptors that are normally expressed on olfactory sensory neurons to detect volatile chemicals or odorants. Interestingly, many Olfrs are also expressed in diverse tissues and function in cell-cell recognition, migration, and proliferation as well as immune responses and disease processes. Here, we showed that many Olfr genes were expressed in the mouse spleen, linked to Plasmodium yoelii genetic loci significantly, and/or had genome-wide patterns of LOD scores (GPLSs) similar to those of host Toll-like receptor genes. Expression of specific Olfr genes such as Olfr1386 in HEK293T cells significantly increased luciferase signals driven by IFN-β and NF-κB promoters, with elevated levels of phosphorylated TBK1, IRF3, P38, and JNK. Mice without Olfr1386 were generated using the CRISPR/Cas9 method, and the Olfr1386-/- mice showed significantly lower IFN-α/β levels and longer survival than wild-type (WT) littermates after infection with P. yoelii YM parasites. Inhibition of G protein signaling and P38 activity could affect cyclic AMP-responsive element promoter-driven luciferase signals and IFN-β mRNA levels in HEK293T cells expressing the Olfr1386 gene, respectively. Screening of malaria parasite metabolites identified nicotinamide adenine dinucleotide (NAD) as a potential ligand for Olfr1386, and NAD could stimulate IFN-β responses and phosphorylation of TBK1 and STAT1/2 in RAW264.7 cells. Additionally, parasite RNA (pRNA) could significantly increase Olfr1386 mRNA levels. This study links multiple Olfrs to host immune response pathways, identifies a candidate ligand for Olfr1386, and demonstrates the important roles of Olfr1386 in regulating type I interferon (IFN-I) responses during malaria parasite infections.

Animals

Neuronal degeneration and regeneration in the olfactory epithelium of pigeon following transection of the first cranial nerve.

The pigeon olfactory nerve has been sectioned to explore the course of retrograde degeneration of the sensory neurons' perikarya, which are located in the olfactory neuroepithelium. Both light- and electron-microscopic observations have shown that from 3 to 8 days after axotomy the sensory neurons undergo retrograde, irreversible degeneration. Following disappearance of the mature neurons, the basal cells of the neuroepithelium actively divide and differentiate into mature olfactory sensory neurons. Consequently, the basal cells represent true stem cells of the olfactory sensory neurons. The olfactory mucosa regains a structural organization close to normal in a period of 30-50 days after axotomy. These observations indicate that, when the primary olfactory neurons degenerate as a consequence of the experimental section of their axons, restitutio ad integrum of the sensory olfactory connections can be reestablished by new elements which differentiate from basal cells of the olfactory neuroepithelium.20

Animals

EEG analysis gives model of neuronal template-matching mechanism for sensory search with olfactory bulb.

The spatial pattern of EEG activity at the surface of the olfactory bulb tends to be invariant with respect to input and to change to a new pattern whenever an animal is trained to expect or search for a particular odor. It is postulated here that the spatial EEG pattern is dependent on a neural template for that odor that is formed during training. This hypothesis is expressed in the form of a model consisting of an array of interconnected elements (1 X 10 or 6 X 6). Each element represents 2 excitatory and 2 inhibitory subsets of neurons with 3 types of internal feedback: negative, mutually excitatory, and mutually inhibitory. The elements are interconnected only by mutual excitation and mutual inhibition. Each neural subset is represented by a nonlinear differential equation; the connections are represented by modifiable coupling coefficients. With appropriate values of the time, coupling, and gain coefficients, and with input that is modelled on olfactory input, the set of 40 or 144 equations gives output that simulates the time and space patterns of the EEG. In the naive state the coefficients are uniform. A template is formed by giving input to selected elements, cross-correlating the outputs, and weighting the mutually excitatory coupling coefficient between each pair of elements by the corresponding correlation coefficient. When a template has been formed, input to nontemplate elements is treated as noise. Optionally a matched filter is made to simulate habituation by reducing the synaptic gain coefficients of those excitatory subsets that receive the noise. The model is tested by giving input to nontemplate elements and to none, part or all of the template elements. There are two outputs of the model. One is the spatial pattern Vj of the root mean square (rms) amplitudes of the individual outputs v(j, t) of the elements. The other output is the rms amplitude Erms of the ensemble average E(t) over v(j, t). The results show that Vj depends on the template and is relatively insensitive to input, whether or not input is given to template elements. However, Erms increases in proportion to the number of "hits" on the template. If the number of elements receiving noise does not exceed the number of elements in a template, or if the noise is matched with a habituation filter, then Erms rises above the noise level for a "hit" on any one or more template elements irrespective of location or combination. Vj conforms to the performance of the surface EEG. Erms is not yet accessible to physiological measurement.

Animals

Scanning electron microscopy of the channel catfish olfactory lamellae.

The olfactory lamellae of the channel catfish (Ictalurus punctatus) are composed of sensory and indifferent (non-sensory) epithelia organized into two distinct regions on both surfaces of each lamella. The smaller sensory region located adjacent to the midline raphe has fewer cilia per unit surface area than the indifferent epithelium and contains the olfactory neurons whose ciliated dendritic terminals occur at the epithelial surface. The indifferent epithelium, comprising the greater surface area of the olfactory lamella, is covered with a dense mat of non-sensory cilia. Fractured carbon dioxide critical point dried lamellar tissue revealed the underlying cellular structure. The lamellae are composed of two layers of epithelium enclosing a thin stromal layer. Olfactory receptors were observed in the fractured tissue only within the sensory epithelium.

Animals

[Ultrastructure of the olfactory epithelium of the rat: polymorphism of the receptors].

The olfactory epithelium of the Rat is studied by transmission electron microscopy. Two types of bipolar sensory cells are described. One (A type) is homologous to the receptors already described in vertebrates. The other (B type) is less frequently found and shows different morphological characteristics. Another peculiar epithelial cell-type (C type) was occasionally observed.

Animals

Vomeronasal organ: critical role in mediating sexual behavior of the male hamster.

Sexual behavior in male hamsters is totally abolished by bilateral removal of the olfactory bulbs. This operation eliminates sensory input from both the olfactory and the vomeronasal systems. We previously demonstrated that peripheral destruction of the olfactory receptors caused anosmia but did not impair male hamster mating behavior. Here we demonstrate that peripheral deafferentation of the vomeronasal system produces severe sexual behavior deficits in approximately one-third of the treated animals. Combined deafferentation of both the vomeronasal and the olfactory systems eliminates copulation in 100 percent of the animals. This is the first experimental demonstration of a functional role for the vomeronasal organ in a mammalian species.

Animals

An integrated anatomical, functional and evolutionary view of the Drosophila olfactory system.

The Drosophila melanogaster olfactory system is one of the most intensively studied parts of the nervous system in any animal. Composed of ~60 independent olfactory neuron classes, with several associated hygrosensory and thermosensory pathways, it has been subject to diverse types of experimental analyses. However, synthesizing the available data is limited by the incompleteness and inconsistent nomenclature found in the literature. In this work, we first "complete" the peripheral sensory map through the identification of a previously uncharacterized antennal sensory neuron population expressing Or46aB, and the definition of an exceptional "hybrid" olfactory neuron class comprising functional Or and Ir receptors. Second, we survey developmental, anatomical, connectomic, functional and evolutionary studies to generate an integrated dataset of these sensory neuron pathways - and associated visualizations - creating an unprecedented comprehensive resource. Third, we illustrate the utility of the dataset to reveal relationships between different organizational properties of this sensory system, and the new questions these stimulate. These examples emphasize the power of this resource to promote further understanding of the construction, function and evolution of these neural circuits.

Journal Article

Olfactory epithelium of Necturus maculosus and Ambystoma tigrinum.

The morphological study presented here provides a general description of the elements of the olfactory epithelium in the mud puppy and tiger salamander,, and gives evidence about their dynamic activity and interrelationships. There are morphological indications of local bursts of reduplication and a continual line of differentiation of receptor cells from basal cell progenitors through stages of mature development to senescence (indicated by the accumulation of pigment granules) and cell death and disposal (by expulsion of pycnotic cell nuclei and by phagocytosis by macrophages). The supporting cells probably play several roles: a secretory role which supplements the activity of Bowman's glands, a minor insulating role in which some dendrites are shielded from the surrounding milieu, and a skeletal role in which they facilitate the efficient displacement of dendrites. The dendrites are regularly arranged in organized relationships with one another and are for the most part in direct apposition, separated only by a 200 A intercellular gap, thus suggesting the possibility of functional interrelationships. This study emphasizes the fact that efficient planning of experimental investigations must include knowledge and consideration of the thickness of the particular olfactory epithelium under study. It also suggests that because of the large receptor-cell size, the mud puppy and/or tiger salamander would make good model systems for single cell recording. Further, the olfactory epithelia of these species are suggested as favorable targets for studies of the aging process in nerve cells.

Ambystoma

Olfactory receptor response characteristics: a factor analysis.

The responses to odor stimulation of 22 single olfactory units in the frog olfactory mucosa were recorded with metal filled micropipettes. Seven purified odorants matched in concentration and a pure air stimulus were administered. The change in firing frequency for each unit to each chemical was evaluated to determine: (1) whether the response patterns developed across all units are different from chemical to chemical and, (2) whether individual units can be grouped in terms of their similarity of their responses to all odorants. The results of a Bartlett Sphericity test suggest that each odorant produces an independent pattern of responses across units. To answer the second question, a factor analysis was employed. It examined the responses of each unit to all chemicals and yielded 7 out of a possible 22 independent factors, suggesting that there are 7 ways by which the units studied look at the odorant stimuli and an air control employed in this study. These analyses were evaluated and discussed in terms of previously published data suggesting receptor site specificity.

Animals

The fine structure of the olfactory and vomeronasal organs of a lizard (Tiliqua scincoides scincoides).

Olfactory epithelium in Tiliqua scinoides is of a loosely packed pseudostratified type. It receives secretion from the supporting cells and the underlying glands of Bowman. Its surface bears microvilli and cilia from sensory cells and microvilli from supporting cells. The vomeronasal epithelium is also pseudostratified but higher and more closely packed. Its surface carries microville from sensory and supporting cells but lacks cilia. Vascular connective tissue penetrates it almost to the epithelial surface but is always outlined by basal cell processes and a basal lamina. There are no secretory cells in or under the sensory epithelium but some cells in the epithelium of the mushroom body contain secretion granules. Sensory cells of both epithelia are bipolar neurons. The perikarya of the vomeronasal cells are more neuronal in character. Axonic processes are similar in both, dendrites are distinctive. Olfactory dendrites end in rounded rods bearing microvilli and cilia of an unusual type. Microville with filamentous cores occur on vomeronasal dendrites. There are no cilia, but 2-6 centrioles appear below the cell surface. Basal cells are structurally similar in both epithelia, but axonic processes of olfactory cells are surrounded by supporting cell processes, while vomeronasal axonic processes are surrounded by basal cells before they leave the epithelium. The presence of cilia and microville on the surface of the sensory cells is discussed in relation to the physical conditions surrounding them.

Animals