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Development of pigment-cup eyes in the polychaete Platynereis dumerilii and evolutionary conservation of larval eyes in Bilateria.

The role of Pax6 in eye development in insects and vertebrates supports the view that their eyes evolved from simple pigment-cup ocelli present in their last common ancestors (Urbilateria). The cerebral eyes in errant polychaetes represent prototype invertebrate pigment-cup ocelli and thus resemble the presumed ancestral eyes. We have analysed expression of conserved eye specification genes in the early development of larval and adult pigment-cup eyes in Platynereis dumerilii (Polychaeta, Annelida, Lophotrochozoa). Both larval and adult eyes form in close vicinity of the optic anlagen on both sides of the developing brain ganglia. While pax6 is expressed in the larval, but not in the developing, adult eyes, expression of six1/2 from trochophora stages onwards specifically outlines the optic anlagen and thus covers both the developing larval and adult eyes. Using Platynereis rhabdomeric opsin as differentiation marker, we show that the first pair of adult eye photoreceptor cells is detected within bilateral clusters that transitorily express ath, the Platynereis atonal orthologue, thus resembling proneural sensory clusters. Our data indicate that--similar to insects, but different from the vertebrates--polychaete six1/2 expression outlines the entire visual system from early developmental stages onwards and ath-positive clusters generate the first photoreceptor cells to appear. We propose that pax6-, six1/2- and ath-positive larval eyes, as found in today's trochophora, were present already in Urbilateria.

Amino Acid Sequence↗

Chronic hypoxia alters calbindin D-28k immunoreactivity in lingual and laryngeal taste buds in the rat.

The distribution and abundance of the calcium binding protein, calbindin D-28k (CB) immunoreactivity in the taste buds of the circumvallate papillae and larynx were compared between normoxic and chronically hypoxic rats (10% O2 for 8 weeks). In the normoxic rats, CB immunoreactivity was observed in some cells and fibers of the intragemmal region of the taste buds in the circumvallate papillae. In contrast, in the subgemmal region of the laryngeal taste buds, fibers but not cells were immunoreactive for CB. In chronically hypoxic rats, CB immunoreactive cells and fibers in the taste buds were decreased in the circumvallate papillae. In the laryngeal taste buds, the density of the subgemmal CB immunoreactive fibers in chronically hypoxic rats was greater than in normoxic rats. It is considered that function of the laryngeal taste buds is different from that of the lingual taste buds, so that laryngeal taste buds may be involved in chemosensation other than taste. The altered density of CB immunoreactive cells and fibers in the lingual and laryngeal taste buds is a predominant feature of hypoxic adaptation, and chronic hypoxic exposure might change the chemical sensitivity of the circumvallate papillae and larynx through the regulation of intracellular Ca2+.

Adaptation, Physiological↗

[Study of the chemoreception in mollusca].

Recording of the mollusc osphardium nerve activity revealed the osphardium sensitivity to hyperosmotic pressure, sodium chloride, and aminoacidosis. The osphardium was found to detect quality of the water where the animals were kept. Pont snail osphradium preserves ancestors' multisensority which unites perception of different chemical and physical signals. Patch-clamp method revealed membrane currents in certain ganglia and receptor cells which are sensitive to Na+ and L-aspartate increase in the solution around the osphardium. Inward components of these currents are, probably, of the sodium and/or calcium nature, whereas the outward components--of the potassium one.

Action Potentials↗

A cofactor, TIP30, specifically enhances HIV-1 Tat-activated transcription.

Replication of HIV-1 requires the viral Tat protein, which increases the extent of transcription elongation by RNA polymerase II after activation at the single viral long terminal repeat (LTR) promoter. This effect of Tat on transcription requires Tat interactions with a 5' region (TAR) in nascent transcripts as well as Tat-specific cofactors. The present study identifies a cellular protein, TIP30, that interacts with Tat and with an SRB-containing RNA polymerase II complex both in vivo and in vitro. Coexpression of TIP30 specifically enhances transactivation by Tat in transfected cells, and immunodepletion of TIP30 from nuclear extracts abolishes Tat-activated transcription without affecting Tat-independent transcription. These results implicate TIP30 as a specific coactivator that may enhance formation of a Tat-RNA polymerase II holoenzyme complex.

Acetyltransferases↗

Dependency of hypoxic chemotransduction in cat carotid body on voltage-gated calcium channels.

The hypothesis that the entry of extracellular calcium ions into some compartment, quite possibly the type I cells, through voltage-gated calcium channels (VGCC) is essential for hypoxic chemotransduction in the cat carotid body was tested using an in situ perfusion technique. The neural output of the carotid body of anesthetized, paralyzed, and artificially ventilated cats in response to perfusions with Krebs-Ringer bicarbonate solution (KRB), calcium-free KRB, KRB containing calcium channel blockers, or KRB containing BAY K 8644 was recorded. Selective perfusion of the carotid body with hypoxic calcium-free KRB significantly decreased carotid chemoreceptor activity, suggesting that extracellular calcium is essential for hypoxic chemotransduction. Selective perfusion of the carotid body with hypoxic KRB containing verapamil (10-100 microM), diltiazem (10-100 microM), or nifedipine (10-100 microM) dose dependently attenuated the increase in chemoreceptor activity produced by hypoxia, suggesting that VGCC need to be activated for hypoxic chemotransduction. The carotid body response to hyperoxic KRB containing the calcium channel agonist BAY K 8644 (10 microM) was 267 +/- 87% of hyperoxic control KRB, suggesting that an enhanced influx of calcium ions through VGCC stimulates carotid chemoreceptor activity. Selective perfusion of the carotid body with severely hypoxic KRB containing BAY K 8644 did not increase chemoreceptor activity above that produced by severe hypoxia alone. This suggests that severe hypoxia increases intracellular calcium in some compartment of the carotid body to achieve stimulatory maximum response and that further increase in intracellular calcium does not produce further elevation of neural activity.(ABSTRACT TRUNCATED AT 250 WORDS)

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗

Simultaneous recording of [Ca2+]i increases in isolated olfactory receptor neurons retaining their original spatial relationship in intact tissue.

A new method is described for isolating olfactory receptor neurons suitable for simultaneous recording of odorant responses in several cells. This method, called "tissue printing" by Cassab and Varner, was used to isolate cells for measurement of odorant-induced increases in cytosolic-free calcium concentration ([Ca2+]i) using the Ca2+ indicator dye fura-2. A large number of receptors could be isolated from a piece of olfactory epithelium (about 300 microns square), preserving their normal morphology and relative local topology to that in the intact olfactory tissue. The probability that there are one or more receptor cells with odorant-induced responses in [Ca2+]i per preparation was 4 times higher with cells isolated by the tissue printing than with those obtained by the pipetting method. The responses of 2 receptor cells separated by 28 microns in the recording chamber differed for 2 odorants: isoamyl acetate and citralva. The method was useful for isolating receptor neurons without losing their morphological features and for investigating the spatial distribution of odorant responsiveness of each receptor over the olfactory epithelium.

Animals↗

K+-current modulated by PO2 in type I cells in rat carotid body is not a chemosensor.

According to the membrane channel hypothesis of carotid body O2 chemoreception, hypoxia suppresses K+ currents leading to cell depolarization, [Ca2+]i rise, neurosecretion, increased neural discharge from the carotid body. We show here that tetraethylammonium (TEA) plus 4-aminopyridine (4-AP) which suppressed the Ca2+ sensitive and other K+ currents in rat carotid body type I cells, with and without low [Ca2+]o plus high [Mg2+]o, did not essentially influence low PO2 effects on [Ca2+]i and chemosensory discharge. Thus, hypoxia may suppress the K+ currents in glomus cells but K+ current suppression of itself does not lead to chemosensory excitation. Therefore, the hypothesis that K+-O2 current is linked to events in chemoreception is not substantiated. K+-O2 current is an epiphemenon which is not directly linked with O2 chemoreception.

4-Aminopyridine↗

Unexplained peculiarities of the dorsal root ganglion.

The cell soma of primary afferent neurons in the dorsal root ganglion (DRG) is assigned by classical neurophysiology the role of a metabolic depot, charged with supporting the peripheral sensory ending, the conducting axon, and the central synaptic terminals. However, certain peculiarities of DRG morphology and physiology do not sit well with this being its only role. For example, why are DRG cell somata electrically excitable, why are some able to fire repetitively on sustained depolarization, and why does the DRG lack a blood-nerve barrier? Consideration of these and related questions leads to several intriguing hypotheses: (1) Electrical excitability of the soma may be required to insure the reliable propagation of impulses past the DRG T-junction and into the spinal cord. (2) Invasion of the afferent spike into the cell soma may provide an essential feedback signal necessary for the cell soma to regulate the excitability of the sensory ending. 3) The subpopulation of DRG neurons that have repetitive firing capability may be responsible for generating the background sensation that we feel as our body schema. Moreover, these neurons may be chemical sensors that provide essential information about our body's internal milieu.

Animals↗

A gene affecting the specificity of the chemosensory neurons of Drosophila.

The sensilla on the proboscis and tarsi of Drosophila contain five neurons, four chemosensory and one mechanosensory. The sugar-sensitive neuron, designated S, carries independent acceptor sites for pyranose, furanose and trehalose. Two others, L1 and L2, respond to salts. The fourth neuron, W, is inhibited by salts and sugars, and is believed to mediate detection of water. We describe here a gene in which mutations alter the neurons in such a way that the S cell is excited by salts. As a result, the mutant flies are strongly attracted by NaCl at concentrations which are repellent to the wild type. To our knowledge, this is the first instance of a mutation which changes the specificity of the chemosensory neurons.

Alleles↗

Human olfactory contrast changes during the menstrual cycle.

Several lines of studies have reported that olfactory perception is influenced by physical and hormonal conditions. In the present study, we investigated changes of olfactory perception during the menstrual cycle of the human. Cyclopentadecanolide vapor was used and its perception intensity was measured by 6-point category scale methods. We especially focused on a novel concept termed "olfactory contrast" that has been just very recently derived from the patch clamp experiment that uses the solitary olfactory receptor cell. The results obtained from 18 trials (15 subjects) showed that olfactory contrast was significantly enhanced at the ovulatory and/or menstrual phases. It is suggested that olfactory contrast, which we defined as a new parameter, provides a useful tool in many kinds of studies exploring the olfactory perceived ability.

Adult↗

Oxygen-sensing neurons in the caudal hypothalamus and their role in cardiorespiratory control.

Work from this laboratory has shown that the caudal hypothalamus modulates the cardiorespiratory responses to hypoxia. The purpose of this review is to describe the modulation of respiratory output by the caudal hypothalamus during hypoxia and how neurons in this area respond to hypoxia. The diaphragmatic activity response to hypoxia was significantly attenuated following microinjection of either cobalt chloride or kynurenic acid into the caudal hypothalamus of rats. In addition, caudal hypothalamic neurons in anesthetized rats and cats responded to hypoxia with an increased firing frequency. This response was maintained in the absence of input from the vagus and carotid sinus nerves in the cat. When recorded extracellularly or by whole-cell patch clamp in vitro, these neurons responded to hypoxia with an increase in firing frequency, membrane potential and inward current. These results suggest that the caudal hypothalamus exerts excitatory influence on respiration during hypoxia, that may originate from the ability of these neurons to sense changes in oxygen levels.

Animals↗

Regulation of olfactory signalling via cGMP-dependent protein kinase.

Strong odor stimuli elicit a slow and sustained increase of the cGMP concentration in isolated rat olfactory cilia. Elevated cGMP levels appear to attenuate the primary response to odorant stimulation. Incubating cilia with membrane-permeable cGMP derivates caused a significantly reduced cAMP signal in response to odorant stimulation. This inhibitory effect was mimicked by 8-(4-chlorophenlythio)-cGMP, a selective activator of cGMP-activated protein kinases; in contrast, a selective inhibitor, [8-(4-chlorophenylthio)-guanosine-3',5'-cyclic monophosphorothioate] of cGMP kinases enhanced the reactivity to odorant stimulation. The data suggest that the responsiveness of olfactory sensory cells is governed by a cGMP-dependent protein kinase. Western-blot analysis using subtype-specific antibodies indicated that cytosolic type-I cGMP kinase, but not the membrane-associated type-II cGMP kinase, is expressed in olfactory sensory neurons.

Animals↗

Pituitary and adrenals are required for hyperglycemic reflex initiated by stimulation of CBR with cyanide.

We have previously shown that stimulation of carotid body receptors (CBR) with sodium cyanide (NaCN) elicits a rapid hyperglycemic reflex. Here we explore whether the pituitary and adrenals, two glands involved in glucose homeostasis, are necessary for this reflex. Experiments were performed on anesthetized rats that were artificially ventilated. Measurements of hepatic venous-arterial glucose difference indicated that CBR stimulation with a bolus of 5 micrograms/100 g NaCN produced an immediate increase in the output of glucose by the liver. The same dose of NaCN failed to increase hepatic output of glucose in rats with bilateral adrenalectomy or in rats 1 wk after surgical removal of neurohypophysis. Reflex glucose output by the liver was maintained after adenohypophysectomy or in adrenalectomized rats after adrenal autotransplantation to epiploon. Measurements of epinephrine in plasma and in the grafted adrenal tissue showed that the adrenal autograft can store and secrete catecholamines Immunocytochemical observations indicated that the grafted adrenals retain medullary cells. These results indicate that neurohypophysis and adrenals are necessary for the hyperglycemic reflex initiated by CBR stimulation with NaCN and that the participation of these two organs in this reflex is probably humoral.

Adrenal Glands↗

High-resolution scanning electron microscopy of the murine olfactory receptor.

The anatomy of the murine olfactory receptor was analyzed using high-resolution scanning electron microscopy with the aldehyde-osmium-dimethyl sulfoxide-osmium method. A good three-dimensional image of the receptor cell architecture was obtained. Within the olfactory vesicle, a complex tubular network was found that seemed to be physically associated with adjacent organelles, the basal aspect of cilia, and the lateral plasmalemma. It is assumed that these structures may be of significance during the olfactory transduction process.

Animals↗

A novel family of mammalian taste receptors.

In mammals, taste perception is a major mode of sensory input. We have identified a novel family of 40-80 human and rodent G protein-coupled receptors expressed in subsets of taste receptor cells of the tongue and palate epithelia. These candidate taste receptors (T2Rs) are organized in the genome in clusters and are genetically linked to loci that influence bitter perception in mice and humans. Notably, a single taste receptor cell expresses a large repertoire of T2Rs, suggesting that each cell may be capable of recognizing multiple tastants. T2Rs are exclusively expressed in taste receptor cells that contain the G protein alpha subunit gustducin, implying that they function as gustducin-linked receptors. In the accompanying paper, we demonstrate that T2Rs couple to gustducin in vitro, and respond to bitter tastants in a functional expression assay.

Animals↗

Multiple sites of oxygen sensing and their contributions to hypoxic pulmonary vasoconstriction.

Oxygen sensing by the pulmonary vasculature is important for the regulation of vessel tone and the matching of lung perfusion to ventilation. Airways hypoxia is a major stimulus for vasoconstriction, which diverts blood from hypoxic alveoli to better ventilated areas of the lung. Several hypotheses have emerged to explain how pulmonary arteries sense a decrease in oxygen and mediate hypoxic pulmonary vasoconstriction (HPV). They differ mainly in where they place the main site of HPV: in the endothelial or smooth muscle cells of the artery wall. HPV probably results from synergistic actions on both cell types, but it can proceed in the absence of endothelium, suggesting that the primary oxygen sensor is the smooth muscle cell and endothelium-derived agents modulate the muscle response. Several oxygen-sensing targets have been identified in smooth muscle, including potassium channels, Ca(2+) stores in the sarcoplasmic reticulum (SR) and the Ca(2+) sensitivity of the contractile proteins. The evidence for different oxygen-sensing mechanisms in pulmonary vessels is discussed.

Animals↗

Modulation of Cl-, K+, and nonselective cation conductances by taurine in olfactory receptor neurons of the mudpuppy Necturus maculosus.

Odors are transduced by processes that modulate the membrane conductance of olfactory receptor neurons. Olfactory neurons from the aquatic salamander, Necturus maculosus, were acutely isolated without enzymes and studied with a resistive whole-cell method to minimize loss of soluble intracellular constituents. 55 of 224 neurons responded to the test compound taurine at concentrations between 10 nM and 100 microM. Four different conductance changes were elicited by taurine: an increased Cl- conductance (33%), an increased nonselective cation conductance (15%), a decreased Cl- conductance (15%), and a decreased K+ conductance (15%); in addition, responses too small to be characterized were elicited in some neurons. In most cases, taurine appeared to modulate only a single conductance in any particular cell. Modulation of each conductance was dose dependent, and each response ran down quickly in the normal whole-cell mode, presumably due to washout of a diffusible component in the transduction pathway. Modulation of taurine-sensitive conductances caused either inhibitory or excitatory responses. A similar diversity of responses in vivo would produce a complex pattern of electrical activity that could encode the identity and characteristics of an odor.

Animals↗

Co-expression of putative pheromone receptors in the sensory neurons of the vomeronasal organ.

Two large and divergent families of G-protein-coupled receptors (V1Rs and V2Rs) are expressed in subsets of neurons in the vomeronasal organ. These receptors are likely to mediate pheromone responses, but it appears that many V2R genes may encode expressed pseudogenes rather than functional proteins. Therefore we have raised antibodies to representative V2Rs and show labeling of vomeronasal neurons demonstrating that V2R genes encode expressed receptors. V2R immunoreactivity was detected at the sensory surface of the vomeronasal organ in dendritic terminals, indicating that these receptors are capable of directly interacting with pheromones and mediating physiological responses. Immunohistochemistry confirmed that three V2R receptors are expressed in small subsets of sensory neurons. However, surprisingly we found that a subfamily of V2R genes is broadly expressed in the Goalpha-layer of the vomeronasal organ and are coexpressed in the same cells as other V2Rs. This is in direct contrast to the main olfactory epithelium where sensory neurons express only a single receptor. Thus, our results suggest that different modes of the information processing may occur in the main and accessory olfactory systems.

Animals↗