scholarly journals Neuronal Chloride Accumulation in Olfactory Epithelium of Mice Lacking NKCC1

2006 ◽  
Vol 95 (3) ◽  
pp. 2003-2006 ◽  
Author(s):  
William T. Nickell ◽  
Nancy K. Kleene ◽  
Robert C. Gesteland ◽  
Steven J. Kleene
Keyword(s):  
Olfactory Epithelium ◽  
Electrical Response ◽  
Niflumic Acid ◽  
Olfactory Receptor Neurons ◽  
Flufenamic Acid ◽  
Wild Type ◽  
In The Wild ◽  
Receptor Neurons ◽  
Chloride Accumulation

When stimulated with odorants, olfactory receptor neurons (ORNs) produce a depolarizing receptor current. In isolated ORNs, much of this current is caused by an efflux of Cl−. This implies that the neurons have one or more mechanisms for accumulating cytoplasmic Cl− at rest. Whether odors activate an efflux of Cl− in intact olfactory epithelium, where the ionic environment is poorly characterized, has not been previously determined. In mouse olfactory epithelium, we found that >80% of the summated electrical response to odors is blocked by niflumic acid or flufenamic acid, each of which inhibits Ca2+-activated Cl− channels in ORNs. This indicates that ORNs accumulate Cl− in situ. Recent evidence has shown that NKCC1, a Na+-K+-2Cl− cotransporter, contributes to Cl− accumulation in mammalian ORNs. However, we find that the epithelial response to odors is only reduced by 39% in mice carrying a null mutation in Nkcc1. As in the wild-type, most of the response is blocked by niflumic acid or flufenamic acid, indicating that the underlying current is carried by Cl−. We conclude that ORNs effectively accumulate Cl− in situ even in the absence of NKCC1. The Cl−-transport mechanism underlying this accumulation has not yet been identified.

Neuronal nitric oxide synthase-like immunoreactivity in olfactory epithelium throughout the life cycle of the sea lamprey, Petromyzon marinus L.

2000 ◽  
Vol 78 (3) ◽  
pp. 346-351 ◽  
Author(s):  
Hong N Hua ◽  
Aliya U Zaidi ◽  
Barbara S Zielinski
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Olfactory Epithelium ◽  
Olfactory Receptor ◽  
Petromyzon Marinus ◽  
Neuronal Nitric Oxide Synthase ◽  
Sea Lamprey ◽  
Olfactory Receptor Neurons ◽  
Receptor Neurons ◽  
Oxide Synthase

This study is the first to show that neuronal nitric oxide synthase-like immunoreactivity is located in the olfactory epithelium at all developmental stages of a vertebrate. Western immunoblotting of sea lamprey (Petromyzon marinus L.) olfactory mucosa with a monoclonal antibody against the NADPH-binding epitope of neuronal nitric oxide synthase showed that the molecular mass of this protein was 200 kDa. In the larval stage, neuronal nitric oxide synthase-like immunoreactivity was strongest in the basal region of the olfactory epithelium, the site of proliferating olfactory receptor neurons. This staining gradually diminished as the life cycle progressed. In the juvenile stage, the intensity of neuronal nitric oxide synthase-like immunoreactivity was striking in the wide cell bodies and dendrites on olfactory receptor neurons. These results confirm previous evidence that nitric oxide modulates development in the olfactory epithelium.

scholarly journals Improving 3-methylphenol (m-cresol) production in yeast via in vivo glycosylation or methylation

2020 ◽  
Vol 20 (8) ◽  
Author(s):  
Julia Hitschler ◽  
Eckhard Boles
Keyword(s):  
Saccharomyces Cerevisiae ◽  
De Novo ◽  
Wild Type ◽  
Biotechnological Production ◽  
Yeast Saccharomyces Cerevisiae ◽  
In Situ Extraction ◽  
Methylsalicylic Acid Synthase ◽  
In The Wild

ABSTRACT Heterologous expression of 6-methylsalicylic acid synthase (MSAS) together with 6-MSA decarboxylase enables de novo production of the platform chemical and antiseptic additive 3-methylphenol (3-MP) in the yeast Saccharomyces cerevisiae. However, toxicity of 3-MP prevents higher production levels. In this study, we evaluated in vivo detoxification strategies to overcome limitations of 3-MP production. An orcinol-O-methyltransferase from Chinese rose hybrids (OOMT2) was expressed in the 3-MP producing yeast strain to convert 3-MP to 3-methylanisole (3-MA). Together with in situ extraction by dodecane of the highly volatile 3-MA this resulted in up to 211 mg/L 3-MA (1.7 mM) accumulation. Expression of a UDP-glycosyltransferase (UGT72B27) from Vitis vinifera led to the synthesis of up to 533 mg/L 3-MP as glucoside (4.9 mM). Conversion of 3-MP to 3-MA and 3-MP glucoside was not complete. Finally, deletion of phosphoglucose isomerase PGI1 together with methylation or glycosylation and feeding a fructose/glucose mixture to redirect carbon fluxes resulted in strongly increased product titers, with up to 897 mg/L 3-MA/3-MP (9 mM) and 873 mg/L 3-MP/3-MP as glucoside (8.1 mM) compared to less than 313 mg/L (2.9 mM) product titers in the wild type controls. The results show that methylation or glycosylation are promising tools to overcome limitations in further enhancing the biotechnological production of 3-MP.

scholarly journals Comparison of the Nasal Olfactory Organs of Various Species of Lizardfishes (Teleostei: Aulopiformes: Synodontidae) with Additional Remarks on the Brain

2010 ◽  
Vol 2010 ◽  
pp. 1-8 ◽  
Author(s):  
L. Fishelson ◽  
D. Golani ◽  
B. Galil ◽  
M. Goren
Keyword(s):  
Olfactory Epithelium ◽  
Olfactory Receptor ◽  
Olfactory Receptor Neurons ◽  
Adult Fish ◽  
Supporting Cells ◽  
Receptor Neurons ◽  
Olfactory Organs ◽  
Species Specific ◽  
The Brain

The olfactory organs of lizardfishes (Synodontidae) are situated in two capsules connected to the outside by incurrent and excurrent openings. The olfactory epithelium is in form of petal rosettes each composed of lamellae and a rephe, and bear olfactory receptor neurons, supporting cells and cells with kinocillia. The dimension of rosettes and lamellae, as well as the number of lamellae, increase with growth of the fish; until in adult fish these parameters remaine constant, species specific. In adultSynodusspp. andTrachinocephalus myopsthe rosettes are 3.5–4.0 mm long, with 5–8 lamellae, whereas inSauridaspp. they are 8.0 mm and possess up tp 22 lamellae. The number of ORN ranges from 2,600 on the smaller lamellae to 20,000 on the largest ones. The number of ORN/m of olfactory is ca. 30,000 inSauridaspp. Thus the rosettes ofS. macrolepiswith 20 lamellae possess a total of ca. 170,000 ORN, whereas those ofSy. variegatusandT. myopswith the average of six lamellae possess only ca. 50,000–65,000 ORN. The olfactory nerves lead from the rosettes to the olfactory balbs situated on the olfactory lobes. The differences among the species in olfactory organs are discussed in correlation with their distribution.

scholarly journals Calcium-activated chloride channels clamp odor-evoked spike activity in olfactory receptor neurons

2018 ◽  
Author(s):  
Joseph D. Zak ◽  
Julien Grimaud ◽  
Rong-Chang Li ◽  
Chih-Chun Lin ◽  
Venkatesh N. Murthy
Keyword(s):  
Olfactory Receptor ◽  
Chloride Channels ◽  
Feedback Mechanism ◽  
Olfactory Receptor Neurons ◽  
Odor Source ◽  
Wild Type ◽  
Behavioral Deficits ◽  
Receptor Neurons

AbstractThe calcium-activated chloride channel anoctamin-2 (Ano2) is thought to amplify transduction currents in ORNs, a hypothesis supported by previous studies in dissociated neurons from Ano2-/- mice. Paradoxically, despite a reduction in transduction currents in Ano2-/- ORNs, their spike output for odor stimuli may be higher. We examined the role of Ano2 in ORNs in their native environment in freely breathing mice by imaging activity in ORN axons as they arrive in the olfactory bulb glomeruli. Odor-evoked responses in ORN axons of Ano2-/- mice were consistently larger for a variety of odorants and concentrations. In an open arena, Ano2-/- mice took longer to approach a localized odor source than wild-type mice, revealing clear olfactory behavioral deficits. Our studies provide the first in vivo evidence toward an alternative role for Ano2 in the olfactory transduction cascade, where it may serve as a feedback mechanism to clamp ORN spike output.

scholarly journals Evidence of SARS-CoV2 entry protein ACE2 in the human nose and olfactory bulb

2020 ◽  
Author(s):  
M. Klingenstein ◽  
S. Klingenstein ◽  
P.H. Neckel ◽  
A. F. Mack ◽  
A. Wagner ◽  
...  
Keyword(s):  
Olfactory Bulb ◽  
Olfactory Epithelium ◽  
Olfactory Receptor ◽  
Respiratory Epithelium ◽  
Olfactory Receptor Neurons ◽  
Basal Cells ◽  
Sustentacular Cells ◽  
Glandular Cells ◽  
Receptor Neurons ◽  
Human Nose

ABSTRACTUsually, pandemic COVID-19 disease, caused by SARS-CoV2, presents with mild respiratory symptoms such as fever, cough but frequently also with anosmia and neurological symptom. Virus-cell fusion is mediated by Angiotensin-Converting Enzyme 2 (ACE2) and Transmembrane Serine Protease 2 (TMPRSS2) with their organ expression pattern determining viral tropism. Clinical presentation suggests rapid viral dissemination to central nervous system leading frequently to severe symptoms including viral meningitis. Here, we provide a comprehensive expression landscape of ACE2 and TMPRSS2 proteins across human, post-mortem nasal and olfactory tissue. Sagittal sections through the human nose complemented with immunolabelling of respective cell types represent different anatomically defined regions including olfactory epithelium, respiratory epithelium of the nasal conchae and the paranasal sinuses along with the hardly accessible human olfactory bulb. ACE2 can be detected in the olfactory epithelium, as well as in the respiratory epithelium of the nasal septum, the nasal conchae and the paranasal sinuses. ACE2 is located in the sustentacular cells and in the glandular cells in the olfactory epithelium, as well as in the basal cells, glandular cells and epithelial cells of the respiratory epithelium. Intriguingly, ACE2 is not expressed in mature or immature olfactory receptor neurons and basal cells in the olfactory epithelium. Similarly ACE2 is not localized in the olfactory receptor neurons albeit the olfactory bulb is positive. Vice versa, TMPRSS2 can also be detected in the sustentacular cells and the glandular cells of the olfactory epithelium.Our findings provide the basic anatomical evidence for the expression of ACE2 and TMPRSS2 in the human nose, olfactory epithelium and olfactory bulb. Thus, they are substantial for future studies that aim to elucidate the symptom of SARS-CoV2 induced anosmia of via the olfactory pathway.

scholarly journals Lineage, Identity, and Fate of Distinct Progenitor Populations in the Embryonic Olfactory Epithelium

2021 ◽  
Author(s):  
Elizabeth M Paronett ◽  
Corey A Bryan ◽  
Thomas M Maynard ◽  
Anthony-S. LaMantia
Keyword(s):  
Olfactory Epithelium ◽  
Olfactory Nerve ◽  
Late Gestation ◽  
Olfactory Receptor Neurons ◽  
Proliferative Capacity ◽  
Temporal Dimension ◽  
Sustentacular Cells ◽  
Distinct Cell ◽  
Receptor Neurons ◽  
Temporal And Spatial

We defined a temporal dimension of precursor diversity and lineage in the developing mouse olfactory epithelium (OE) at mid-gestation that results in genesis of distinct cell classes. Slow, symmetrically dividing Meis1+/ Pax7+ progenitors in the early differentiating lateral OE give rise to small numbers of Ascl1+ precursors in the dorsolateral and ventromedial OE. Few of the initial progeny of the Ascl1+ precursors immediately generate olfactory receptor neurons (ORNs). Instead, most early progeny of this temporally defined precursor cohort, labeled via temporally discreet tamoxifen-dependent Ascl1Cre-driven recombination, populate a dorsomedial OE domain comprised of proliferative Ascl1+ as well as Ascl1- cells from which newly generated ORNs are mostly excluded. The most prominent early progeny of these Ascl1+ OE precursors are migratory mass cells associated with the nascent olfactory nerve (ON) in the frontonasal mesenchyme. These temporal, regional and lineage distinctions are matched by differences in proliferative capacity and modes of division in isolated, molecularly distinct lateral versus medial OE precursors. By late gestation, the progeny of the temporally and spatially defined Ascl1+ precursor cohort include few proliferating precursors. Instead, these cells generate a substantial subset of OE sustentacular cells, spatially restricted ORNs, and ensheathing cells associated with actively growing as well as mature ON axons. Accordingly, from the earliest stages of OE differentiation, distinct temporal and spatial precursor identities provide a template for acquisition of subsequent OE and ON cellular diversity.

scholarly journals Effects of Mutations in the Drosophila melanogaster Rif1 Gene on the Replication and Underreplication of Pericentromeric Heterochromatin in Salivary Gland Polytene Chromosomes

Cells ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 1501
Author(s):  
Tatyana D. Kolesnikova ◽  
Alexandra V. Kolodyazhnaya ◽  
Galina V. Pokholkova ◽  
Veit Schubert ◽  
Viktoria V. Dovgan ◽  
...  
Keyword(s):  
Salivary Gland ◽  
Satellite Dna ◽  
Polytene Chromosomes ◽  
Replication Timing ◽  
Wild Type ◽  
Chromosome X ◽  
Substantial Fraction ◽  
In The Wild ◽  
Specific Control

In Drosophila salivary gland polytene chromosomes, a substantial portion of heterochromatin is underreplicated. The combination of mutations SuURES and Su(var)3-906 results in the polytenization of a substantial fraction of unique and moderately repeated sequences but has almost no effect on satellite DNA replication. The Rap1 interacting factor 1 (Rif) protein is a conserved regulator of replication timing, and in Drosophila, it affects underreplication in polytene chromosomes. We compared the morphology of pericentromeric regions and labeling patterns of in situ hybridization of heterochromatin-specific DNA probes between wild-type salivary gland polytene chromosomes and the chromosomes of Rif1 mutants and SuUR Su(var)3-906 double mutants. We show that, despite general similarities, heterochromatin zones exist that are polytenized only in the Rif1 mutants, and that there are zones that are under specific control of Su(var)3-9. In the Rif1 mutants, we found additional polytenization of the largest blocks of satellite DNA (in particular, satellite 1.688 of chromosome X and simple satellites in chromosomes X and 4) as well as partial polytenization of chromosome Y. Data on pulsed incorporation of 5-ethynyl-2′-deoxyuridine (EdU) into polytene chromosomes indicated that in the Rif1 mutants, just as in the wild type, most of the heterochromatin becomes replicated during the late S phase. Nevertheless, a significantly increased number of heterochromatin replicons was noted. These results suggest that Rif1 regulates the activation probability of heterochromatic origins in the satellite DNA region.

scholarly journals Compensatory plasticity in the olfactory epithelium: age, timing, and reversibility

2015 ◽  
Vol 114 (3) ◽  
pp. 2023-2032 ◽  
Author(s):  
Casey N. Barber ◽  
David M. Coppola
Keyword(s):  
Nasal Cavity ◽  
Olfactory Epithelium ◽  
Adult Mouse ◽  
Olfactory Receptor Neurons ◽  
Nasal Airflow ◽  
Stimulus Response ◽  
Open Side ◽  
Receptor Neurons ◽  
Compensatory Plasticity ◽  
Stimulus Environment

Like other biological systems, olfaction responds “homeostatically” to enduring change in the stimulus environment. This adaptive mechanism, referred to as compensatory plasticity, has been studied almost exclusively in developing animals. Thus it is unknown if this phenomenon is limited to ontogenesis and irreversible, characteristics common to some other forms of plasticity. Here we explore the effects of odor deprivation on the adult mouse olfactory epithelium (OE) using nasal plugs to eliminate nasal airflow unilaterally. Plugs were in place for 2–6 wk after which electroolfactograms (EOGs) were recorded from the occluded and open sides of the nasal cavity. Mean EOG amplitudes were significantly greater on the occluded than on the open side. The duration of plugging did not affect the results, suggesting that maximal compensation occurs within 2 wk or less. The magnitude of the EOG difference between the open and occluded side in plugged mice was comparable to adults that had undergone surgical naris occlusion as neonates. When plugs were removed after 4 wk followed by 2 wk of recovery, mean EOG amplitudes were not significantly different between the always-open and previously plugged sides of the nasal cavity suggesting that this form of plasticity is reversible. Taken together, these results suggest that compensatory plasticity is a constitutive mechanism of olfactory receptor neurons that allows these cells to recalibrate their stimulus-response relationship to fit the statistics of their current odor environment.

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