Functional Characterization of GABAA Receptors in Neonatal Hypothalamic Brain Slice

2002 ◽  
Vol 88 (4) ◽  
pp. 1655-1663 ◽  
Author(s):  
Ren-Qi Huang ◽  
Glenn H. Dillon
Keyword(s):  
Functional Characterization ◽  
Minor Component ◽  
Dehydroepiandrosterone Sulfate ◽  
Receptor Subtypes ◽  
Hypothalamic Neurons ◽  
Inhibitory Neurotransmitter ◽  
Old Rats ◽  
A Minor ◽  
Whole Cell Recordings

The hypothalamus influences a number of autonomic functions. The activity of hypothalamic neurons is modulated in part by release of the inhibitory neurotransmitter GABA onto these neurons. GABAA receptors are formed from a number of distinct subunits, designated α, β, γ, δ, ε, and θ, many of which have multiple isoforms. Little data exist, however, on the functional characteristics of the GABAA receptors present on hypothalamic neurons. To gain insight into which GABAA receptor subunits are functionally expressed in the hypothalamus, we used an array of pharmacologic assessments. Whole cell recordings were made from thin hypothalamic slices obtained from 1- to 14-day-old rats. GABAA receptor-mediated currents were detected in all neurons tested and had an average EC50 of 20 ± 1.6 μM. Hypothalamic GABAA receptors were modulated by diazepam (EC50 = 0.060 μM), zolpidem (EC50 = 0.19 μM), loreclezole (EC50 = 4.4 μM), methyl-6,7-dimethoxy-4-ethyl-β-carboline (EC50= 7.7 μM), and 5α-pregnan-3α-hydroxy-20-one (3α-OH-DHP). Conversely, these receptors were inhibited by Zn2+ (IC50 = 70.5 μM), dehydroepiandrosterone sulfate (IC50 = 16.7 μM), and picrotoxin (IC50 = 2.6 μM). The α4/6-selective antagonist furosemide (10–1,000 μM) was ineffective in all hypothalamic neurons tested. The results of our pharmacological analysis suggest that hypothalamic neurons express functional GABAA receptor subtypes that incorporate α1 and/or α2 subunits, β2 and/or β3 subunits, and the γ2 subunit. Our results suggest receptors expressing α3–α6, β1, γ1, and δ, if present, represent a minor component of functional hypothalamic GABAA receptors.

scholarly journals The alpha/B.1.1.7 SARS-CoV-2 variant exhibits significantly higher affinity for ACE-2 and requires lower inoculation doses to cause disease in K18-hACE2 mice

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Rafael Bayarri-Olmos ◽  
Laust Bruun Johnsen ◽  
Manja Idorn ◽  
Line S Reinert ◽  
Anne Rosbjerg ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
United Kingdom ◽  
Disease Progression ◽  
Low Dose ◽  
Transmission Rate ◽  
Functional Characterization ◽  
Fitness Advantage ◽  
The United Kingdom ◽  
A Minor

The alpha/B.1.1.7 SARS-CoV-2 lineage emerged in autumn 2020 in the United Kingdom and transmitted rapidly until winter 2021 when it was responsible for most new COVID-19 cases in many European countries. The incidence domination was likely due to a fitness advantage that could be driven by the RBD residue change (N501Y), which also emerged independently in other Variants of Concern such as the beta/B.1.351 and gamma/P.1 strains. Here we present a functional characterization of the alpha/B.1.1.7 variant and show an eight-fold affinity increase towards human ACE-2. In accordance with this, transgenic hACE-2 mice showed a faster disease progression and severity after infection with a low dose of B.1.1.7, compared to an early 2020 SARS-CoV-2 isolate. When challenged with sera from convalescent individuals or anti-RBD monoclonal antibodies, the N501Y variant showed a minor, but significant elevated evasion potential of ACE-2/RBD antibody neutralization. The data suggest that the single asparagine to tyrosine substitution remarkable rise in affinity may be responsible for the higher transmission rate and severity of the B.1.1.7 variant.

Functional characterization of basolateral and luminal dopamine receptors in rabbit CCD

2001 ◽  
Vol 281 (1) ◽  
pp. F114-F122 ◽  
Author(s):  
Osamu Saito ◽  
Yasuhiro Ando ◽  
Eiji Kusano ◽  
Yasushi Asano
Keyword(s):  
Receptor Agonist ◽  
Collecting Duct ◽  
Functional Characterization ◽  
Receptor Subtypes ◽  
Cortical Collecting Duct ◽  
Basolateral Side ◽  
Skf 81297 ◽  
Transepithelial Voltage ◽  
Specific Antagonist

Previous studies reported the existence of both D1- and D2-like receptors in the cortical collecting duct (CCD). However, especially with regard to natriuresis, it remains controversial. In the present study, rabbit CCD was perfused to characterize the receptor subtypes responsible for the tubular actions. Basolateral dopamine (DA) induced a dose-dependent depolarization of transepithelial voltage. Basolateral domperidone, a D2-like receptor antagonist, abolished depolarization, whereas SKF-81297, a D1-like receptor agonist, showed no significant change. In addition, bromocriptine, a D2-like receptor agonist, also caused depolarization, whereas SKF-81297, a D1-like receptor agonist, did not depolarize significantly. Moreover, RBI-257, a D4-specific antagonist, reversed the basolateral DA-induced depolarization. In contrast to the basolateral side, luminal DA caused depolarization via a D1-like receptor; however the change was less than that for basolateral DA. For further evaluation, 22Na+ flux ( J Na) was measured to confirm the effect of DA on Na+ transport. Basolateral DA also caused a suppression of J Na, and this reaction was abolished by domperidone. These results suggested that the basolateral D2-like receptor is mainly responsible for the natriuretic action of DA in rabbit CCD.

Phenotypical and functional characterization of Fey receptor I (CD64)-negative monocytes, a minor human monocyte subpopulation with high accessory and antiviral activity

1993 ◽  
Vol 23 (12) ◽  
pp. 3126-3135 ◽  
Author(s):  
Evelin Grage-Griebenow ◽  
Dirk Lorenzen ◽  
Rudolf Fetting ◽  
Hans-Dieter Flad ◽  
Martin Ernst
Keyword(s):  
Antiviral Activity ◽  
Functional Characterization ◽  
Human Monocyte ◽  
A Minor

scholarly journals Biochemical studies on the isolation and characterization of human spleen haemosiderin

1984 ◽  
Vol 223 (1) ◽  
pp. 31-38 ◽  
Author(s):  
M P Weir ◽  
J F Gibson ◽  
T J Peters
Keyword(s):  
Minor Component ◽  
Iron Oxyhydroxide ◽  
Human Spleen ◽  
Major Band ◽  
Isolation And Characterization ◽  
Biochemical Studies ◽  
Characteristic Group ◽  
A Minor ◽  
Proteolytic Product

Haemosiderin was isolated from thalassaemic human spleens by centrifugation through concentrated KI solutions. A method for solubilizing haemosiderin was developed which leaves the iron oxyhydroxide cores and constituent polypeptides intact, facilitating further purification and analysis. Purified haemosiderin contained no detectable haem, trace amounts of carbohydrate, and iron and phosphorus in a molar ration of 6:1; much of the phosphate may be present as core-adsorbed. Several lipids were present, but it is not certain whether these are contaminants or components of the haemosiderin granules. In all preparations examined, a characteristic group of six to seven peptides of apparent Mr 12 900-17 800 were found, with a major band at Mr 14 500 and, in addition, a minor component of Mr 42 000; these peptides co-chromatographed with the cores. Negatively stained electron micrographs suggest that these peptides form an incomplete shell about the cores, consistent with the view that haemosiderin is a proteolytic product of ferritin.

scholarly journals Functional Characterization of Muscarinic Receptors in Human Schwann Cells

2020 ◽  
Vol 21 (18) ◽  
pp. 6666
Author(s):  
Roberta Piovesana ◽  
Alessandro Faroni ◽  
Ada Maria Tata ◽  
Adam J. Reid
Keyword(s):  
Cell Proliferation ◽  
Schwann Cells ◽  
Muscarinic Receptor ◽  
Muscarinic Receptors ◽  
Functional Characterization ◽  
Receptor Subtypes ◽  
Muscarinic Receptor Subtypes ◽  
Selective Stimulation ◽  
Stimulation Of

Functional characterization of muscarinic cholinergic receptors in myelinating glial cells has been well described both in central and peripheral nervous system. Rat Schwann cells (SCs) express different muscarinic receptor subtypes with the prevalence of the M2 subtype. The selective stimulation of this receptor subtype inhibits SC proliferation, improving their differentiation towards myelinating phenotype. In this work, we describe for the first time that human SCs are cholinoceptive as they express several muscarinic receptor subtypes and, as for rat SCs, M2 receptor is one of the most abundant. Human SCs, isolated from adult nerves, were cultured in vitro and stimulated with M2 muscarinic agonist arecaidine propargyl ester (APE). Similarly to that observed in rat, M2 receptor activation causes a decreased cell proliferation and promotes SC differentiation as suggested by increased Egr2 expression with an improved spindle-like shape cell morphology. Conversely, the non-selective stimulation of muscarinic receptors appears to promote cell proliferation with a reduction of SC average cell diameter. The data obtained demonstrate that human SCs are cholinoceptive and that human cultured SCs may represent an interesting tool to understand their physiology and increase the knowledge on how the cholinergic stimulation may contribute to address human SC development in normal and pathological conditions.

scholarly journals Functional characterization of two melanocortin (MC) receptors in lamprey showing orthology to the MC1 and MC4 receptor subtypes

2007 ◽  
Vol 7 (1) ◽  
pp. 101 ◽  
Author(s):  
Tatjana Haitina ◽  
Janis Klovins ◽  
Akiyoshi Takahashi ◽  
Maja Löwgren ◽  
Aneta Ringholm ◽  
...  
Keyword(s):  
Functional Characterization ◽  
Receptor Subtypes

scholarly journals Purification and Characterization of Thin Pili of IncI1 Plasmids ColIb-P9 and R64: Formation of PilV-Specific Cell Aggregates by Type IV Pili

1998 ◽  
Vol 180 (11) ◽  
pp. 2842-2848 ◽  
Author(s):  
Tetsu Yoshida ◽  
Nobuhisa Furuya ◽  
Masayuki Ishikura ◽  
Toshiaki Isobe ◽  
Kazu Haino-Fukushima ◽  
...  
Keyword(s):  
Gradient Centrifugation ◽  
Minor Component ◽  
Electrophoretic Analysis ◽  
Type Iv Pili ◽  
Specific Cell ◽  
Cell Aggregates ◽  
Type Iv ◽  
Cscl Density Gradient ◽  
A Minor

ABSTRACT Thin pili of the closely related IncI1 plasmids ColIb-P9 and R64 are required only for liquid mating and belong to the type IV family of pili. They were sedimented by ultracentrifugation from culture medium in which Escherichia coli cells harboring ColIb-P9- or R64-derived plasmids had been grown, and then the pili were purified by CsCl density gradient centrifugation. In negatively stained thin pilus samples, long rods with a diameter of 6 nm, characteristic of type IV pili, were observed under an electron microscope. Gel electrophoretic analysis of purified ColIb-P9 thin pili indicated that thin pili consist of two kinds of proteins, pilin and the PilV protein. Pilin was demonstrated to be the product of the pilS gene. Pilin was first synthesized as a 22-kDa prepilin from the pilS gene and subsequently processed to a 19-kDa protein by the function of thepilU product. The N-terminal amino group of the processed protein was shown to be modified. The C-terminal segments of thepilV products vary among six or seven different types, as a result of shufflon DNA rearrangements of the pilV gene. These PilV proteins were revealed to comprise a minor component of thin pili. Formation of PilV-specific cell aggregates by ColIb-P9 and R64 thin pili was demonstrated and may play an important role in liquid mating.

Analysis of plant genomes. IV. Isolation and characterization of satellite DNA components from two dicotyledons cucumber (Cucumis sativus) and radish (Raphanus sativus)

1978 ◽  
Vol 56 (8) ◽  
pp. 808-815 ◽  
Author(s):  
P. K. Ranjekar ◽  
D. Pallotta ◽  
J. G. Lafontaine
Keyword(s):  
Satellite Dna ◽  
Raphanus Sativus ◽  
Nuclear Dna ◽  
Minor Component ◽  
Isolation And Characterization ◽  
Dna Contents ◽  
A Minor ◽  
Dna Components ◽  
Denaturation Profile

Satellite DNA fractions from cucumber and radish, two plants having low DNA contents and relatively small chromosomes, were isolated and characterized. Reassociation studies of satellite and total nuclear DNA showed that the satellite fractions in these two plants contain most of the rapidly reassociating DNA. Cucumber satellite I was found to contain one major component (70% of the total satellite) having a density of 1.706 g/cm3 and a Tm of 90.5 °C and a minor component with a density of 1.712 g/cm3 and a Tm of 93.5 °C. The complexity of the major component was estimated to be 3.8 × 105 daltons while that of the minor one was 12.9 × 107 daltons. Although cucumber satellite II banded as a single peak at adensity of 1.700 g/cm3 in neutral CsCl gradients, it was observed to have a rather broad denaturation profile with a Tm of 86.5 °C. Its Cot curve was also broader than that of satellite I and one of its components (40% of the total) had a complexity of 5.8 × 105 daltons. Two satellite fractions were also observed in the case of radish DNA but only satellite I was isolated in a pure form and characterized. This radish satellite formed a sharp, symmetrical peak at a density of 1.698 g/cm3 in neutral CsCl gradients and underwent denaturation in a narrow temperature range of 6 to 7 °C. An analysis of the optical reassociation kinetics showed that this satellite contained a major and a minor component. The major component, which comprised 80% of the satellite, had a complexity of 12.9 × 105 daltons. Hybridization experiments revealed that the ribosomal DNA was present in satellite II.

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