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Chronic hypoxia-induced morphological and neurochemical changes in the carotid body.

The carotid body (CB) plays an important role in the control of ventilation. Type I cells in CB are considered to be the chemoreceptive element which detects the levels of PO(2), PCO(2), and [H(+)] in the arterial blood. These cells originate from the neural crest and appear to retain some neuronal properties. They are excitable and produce a number of neurochemicals. Some of these neurochemicals, such as dopamine and norepinephrine, are considered to be primarily inhibitory to CB function and others, such as adenosine triphosphate, acetylcholine, and endothelin, are thought to be primarily excitatory. Chronic hypoxia (CH) induces profound morphological as well as neurochemical changes in the CB. CH enlarges the size of CB and causes hypertrophy and mitosis of type I cells. Also, CH changes the vascular structure of CB, including inducing marked vasodilation and the growth of new blood vessels. Moreover, CH upregulates certain neurochemical systems within the CB, e.g., tyrosine hydroxylase and dopaminergic activity in type I cells. There is also evidence that CH induces neurochemical changes within the innervation of the CB, e.g., nitric oxide synthase. During CH the sensitivity of the CB chemoreceptors to hypoxia is increased but the mechanisms by which the many CH-induced structural and neurochemical changes affect the sensitivity of CB to hypoxia remains to be established.

Animals↗

Tormogen cell and receptor-lymph space in insect olfactory sensilla. Fine structure and histochemical properties in Calliphora.

(1) The basiconic sensilla on the antennae of Calliphora resemble other insect epidermal sensilla; one or several bipolar sense cells are surrounded by three non-neural cells. (2) The apical cell membrane of the tormogen cell (one of the three accessory cells) forms microvilli coated internally with particles. (3) In the (extracellular) outer receptor-lymph space hyaluronic acid can be demonstrated histochemically. (4) Demonstration of non-specific alkaline phosphatase, Mg2+-activated ATPase, and the presence of mitochondria in the apical part of the tormogen cell suggest active transport processes through these cells into the outer receptor-lymph space.

Adenosine Triphosphatases↗

Neuronal expression of Fos and Jun protein in the rat medulla and spinal cord after anoxic and hypercapnic stimulations.

After inhalation of 100% N2 (anoxia) or 8% CO2 (hypercapnia), neurons that express Fos protein (FOS) and/or Jun protein (JUN) were immunohistochemically identified in the CNS of rats anesthetized with urethane and alpha-chloralose. JUN-labeled cells were characteristically found in the respiration-related motor nuclei of the medulla and spinal cord, and in the central chemoreceptive area of the ventral medullary surface on which hypercapnia was more effective than anoxia, whereas FOS-labeled cells were dominant in the respiration-related sensory nuclei of the medulla on which anoxia was more effective than hypercapnia. The results suggest that the peripheral chemoreceptor and the central chemoreceptive pathways in the medulla and spinal cord may be traced by a combination of FOS and JUN expression methods.

Animals↗

The vomeronasal (Jacobson's) organ in man: ultrastructure and frequency of occurrence.

These investigations address three major questions: (1) What is the frequency of occurrence of the vomeronasal (Jacobson's) organ (VNO) in man? (2) what is the ultrastructure of the human VNO? and (3) does the VNO contain sensory receptor cells? Macroscopic and microscopic intranasal clinical examinations of over 200 persons revealed paired bilateral vomeronasal pits on the anterior 1/3 of the nasal septum in all cases. Biopsies of the vomeronasal pits and surrounding tissues were examined by light and electron microscopy. These studies showed that the vomeronasal pit leads to a closed tube, 2-8 mm long, lined by a unique pseudostratified columnar epithelium unlike any other in the human body. The anterior end of the tube is lined by tall, columnar cells with a sparse population of short microvilli. The posterior end of the VNO is lined by an epithelium that contains three morphologically distinct cell types: (1) basal cells; (2) "dark cells--tall, slender cells with heterochromatic nuclei and electron-dense cytoplasm that often contain mucigen-like granules; and (3) "light" cells--large, clear cells, extending from the basement membrane to the organ's lumen. Each "light" cell has a round, euchromatic nucleus and a clear cytoplasm that often contains many Golgi stacks and membrane-limited vesicles filled with material of modest electron density. The cell apex is tipped by a few short microvilli. Whether these cells subserve any sensory function awaits further investigation.

Adult↗

Sensitivity of the negative mucosal potential to the trigeminal target stimulus CO(2).

CO(2) is frequently used in an experimental pain model and in imaging studies investigating the central processing of trigeminal nociceptive information because of its specific trigeminal stimulation properties. The aim of the current study was (1) to investigate the sensitivity of the NMP to small increments of CO(2) stimulus concentrations (3% CO(2), v/v) and (2) to characterize the sensory input of CO(2) by determining NMP, detection and pain thresholds and by registering subjective verbal descriptions. Ten subjects participated in the first experimental sessions investigating NMP responses to stimuli of 62, 65, 68% CO(2) (v/v) (stimulus duration: 1000 ms). Our statistical analysis revealed a dose-dependent increase of the NMP amplitudes and areas under the curves (AUCs) demonstrating the high dynamic resolution of the NMP. Ten subjects participated in the second experimental sessions determining thresholds for NMP, detection and pain (stimulus duration: 1000 ms). MANOVA analysis revealed significantly different thresholds for detection, NMP and subjective pain judgements (mean and S.D. as percentage CO(2) (v/v): detection: 20.6+/-9.6, NMP: 42.6+/-12.5, pain: 50.4+/-12.0). We could demonstrate the existence of a prepain range below subjective pain thresholds with activation of trigeminal nociceptive sensors resulting in the generation of NMPs. The detection threshold of 20.6% CO(2) (v/v) was surprisingly low, i.e. 22% CO(2) (v/v) below the NMP threshold. The involvement of newly discovered alpha-gustducin positive trigeminal chemosensory cells in CO(2) detection is hypothesized.

Adult↗

Multiple signalling pathways connect chemoattractant receptors and calcium channels in Dictyostelium.

Dictyostelium mutants expressing aequorin were used to study and compare the roles of heterotrimeric G-proteins and the second messengers IP3 and cGMP in regulating folate- and cAMP receptor-activated [Ca2+]i signals. The calcium responses of vegetative cells to folate were dramatically impaired in Gbeta and Galpha4 null mutants but were restored with altered kinetics and temperature-sensitivity in Gbeta null mutants overexpressing wild type and temperature-sensitive Gbeta isoforms. Folic acid receptors thus mediate changes in [Ca2+]i via a Galpha4betagamma-dependent pathway. Neither folate nor cAMP-induced [Ca2+]i signals were significantly altered in PLC null transformants, but [Ca2+]i changes elicited by both attractants were significantly prolonged in two stmF mutants lacking cGMP-specific phosphodiesterase activity. This confirms an important role of cGMP in regulating receptor-activated Ca2+ uptake and/or extrusion systems. This cGMP-dependent part of the Ca2+ response to cAMP stimuli was developmentally down-regulated and all but disappeared by the time the cells reached full aggregation competence after 8 h of starvation. The results suggest that folate and cAMP receptor-activated [Ca2+]i signals are regulated in a complex manner via multiple signalling pathways, one that is G-protein- and cGMP-dependent (present at the vegetative and early poststarvation stage) and another that is G-protein-independent (dominant in fully aggregation-competent cells at approximately 8 h poststarvation).

Animals↗

Sensitivity and transduction mechanisms of responses to general odorants in turtle vomeronasal system.

(a) The responses of the vomeronasal organ to general odorants in the turtle, Geoclemys reevesii, were measured by recording the accessory olfactory bulbar responses. The threshold concentrations of the vomeronasal responses to various odorants were similar to those in main olfactory bulbar responses, indicating that vomeronasal cells lacking cilia and olfactory cells having many cilia have similar sensitivities to general odorants. (b) The vomeronasal epithelium was perfused with 100 mM NaCl solution and the salt-free solution and the effects of NaCl on the vomeronasal responses to various odorants were examined. There was no essential difference between the concentration-response curves for n-amyl acetate and menthone dissolved in 100 mM NaCl solution and those dissolved in the salt-free solution in the whole concentration range examined. The ratios of the magnitudes of vomeronasal responses in the salt-free solution to those in 100 mM NaCl solution were between 1.01 and 1.10 for seven odorants tested. (c) The magnitudes of responses to the odorants were unchanged by changes in NaCl concentrations. The replacement of Na+ with organic cations such as choline+, Bis-Tris propane2+, and N-acetyl-D-glucosamine+ did not affect the magnitudes of the responses to the odorants. The Na channel blocker amiloride also did not affect the responses. (d) The vomeronasal responses were practically unchanged by changes in CaCl2 concentration. The Ca channel blockers diltiazem and verapamil did not affect the responses. (e) The replacement of Cl- with SO4(2-) did not affect the magnitudes of the vomeronasal responses. (f) The present results suggest that ion transport across the apical membranes of vomeronasal receptor cells does not contribute to the responses to odorants in the turtle.

Amiloride↗

Polar location of the chemoreceptor complex in the Escherichia coli cell.

The eukaryotic cell exhibits compartmentalization of functions to various membrane-bound organelles and to specific domains within each membrane. The spatial distribution of the membrane chemoreceptors and associated cytoplasmic chemotaxis proteins in Escherichia coli were examined as a prototypic functional aggregate in bacterial cells. Bacterial chemotaxis involves a phospho-relay system brought about by ligand association with a membrane receptor, culminating in a switch in the direction of flagellar rotation. The transduction of the chemotaxis signal is initiated by a chemoreceptor-CheW-CheA ternary complex at the inner membrane. These ternary complexes aggregate predominantly at the cell poles. Polar localization of the cytoplasmic CheA and CheW proteins is dependent on membrane-bound chemoreceptor. Chemoreceptors are not confined to the cell poles in strains lacking both CheA and CheW. The chemoreceptor-CheW binary complex is polarly localized in the absence of CheA, whereas the chemoreceptor-CheA binary complex is not confined to the cell poles in strains lacking CheW. The subcellular localization of the chemotaxis proteins may reflect a general mechanism by which the bacterial cell sequesters different regions of the cell for specialized functions.

ATP-Binding Cassette Transporters↗

Light, fluorescence and electron microscopic studies of rabbit subclavian glomera.

The subclavian glomera (aortic bodies) of young New Zealand white rabbits were studied with the light, fluorescence, and electron microscopes. Two cell types were identified: type I, granule-containing (chief) cells, and type II, agranular (sustentacular) cells. The type I cells possessed large nuclei, the normal complement of cytoplasmic organelles and numerous electron-opaque cytoplasmic granules. The type II cells were agranular with attenuated cytoplasmic processes which partially or completely ensheathed the type I cells. The glomera were well vascularized. Capillary endothelial cells contained numerous pinocytotic vesicles, but few fenestrae. Two profiles of nerve terminals were observed. One, apposing the type I cells, contained numerous electron-lucent vesicles, several dense-cored vesicles, mitochondria and possessed membrane specializations resembling those usually observed in synaptic zones. The other profile contained abundant mitochondria and a few electron-lucent and dense-cored vesicles. Structural specializations were not observed on the apposed membranes of these terminals or adjacent to type II cells. Fluorescence histochemistry revealed an intense yellow-green fluorescence in the glomera, which indicated the presence of biogenic amines, possibly primary catecholamines or an indolamine. The electron-opaque granules observed in the type I cells were believed to be the storage sites for these amines. The subclavian glomera were found to be morphologically similar to the carotid body which is a known chemoreceptor.

Animals↗

Tetrahydropyrimidine derivatives inhibit binding of a Tat-like, arginine-containing peptide, to HIV TAR RNA in vitro.

The ability of a small molecule, 2-methyl,4-carboxy,5-hydroxy-3,4,5,6-tetrahydropyrimidine (THP(A)), which accumulates intracellularly in various streptomyces, to inhibit the interaction of Tat peptide (R52) with TAR RNA is presented. Using gel-shift assay, we found that the inhibition constant Ki of THP(A) is 50-100 nM, which is in the range of the binding constants of Tat peptide and protein. THP(A) is approximately 10(6) times more tightly bound than the free L-arginine. The high binding affinity may be attributed to the special delocalized positive charge on the NCN group and the hydroxyl group at the 5 position of this molecule. A model for THP(A)-TAR interaction, analogous to the arginine guanidinum group-TAR interaction, is presented. The relatively high uptake of THP(A) by mammalian cells warrants in vivo Tat/TAR inhibition studies.

Bacterial Proteins↗

Interactions between hypoxia and hypercapnic acidosis on calcium signaling in carotid body type I cells.

The effects of hypercapnic acidosis and hypoxia on intracellular Ca(2+) concentration ([Ca(2+)](i)) were determined with Indo 1 in enzymatically isolated single type I cells from neonatal rat carotid bodies. Type I cells responded to graded hypoxic stimuli with graded [Ca(2+)](i) rises. The percentage of cells responding was also dependent on the severity of the hypoxic stimulus. Raising CO(2) from 5 to 10 or 20% elicited a significant increase in [Ca(2+)](i) in the same cells as those that responded to hypoxia. Thus both stimuli can be sensed by each individual cell. When combinations of hypoxic and acidic stimuli were given simultaneously, the responses were invariably greater than the response to either stimulus given alone. Indeed, in most cases, the response to hypercapnia was slightly potentiated by hypoxia. These data provide the first evidence that the classic synergy between hypoxic and hypercapnic stimuli observed in the intact carotid body may, in part, be an inherent property of the type I cell.

Acidosis↗

Expression of GDNF and GFR alpha 1 in mouse taste bud cells.

GDNF (glial cell line-derived neurotrophic factor) affects the survival and maintenance of central and peripheral neurons. Using an immunocytochemical method, we examined whether the taste bud cells in the circumvallate papillae of normal mice expressed GDNF and its GFR alpha 1 receptor. Using double immunostaining for either of them and NCAM, PGP 9.5, or alpha-gustducin, we additionally sought to determine what type of taste bud cells expressed GDNF or GFR alpha 1, because NCAM is reported to be expressed in type-III cells, PGP 9.5, in type-III and some type-II cells, and alpha-gustducin, in some type-II cells. Normal taste bud cells expressed both GDNF and GFR alpha 1. The percentage of GDNF-immunoreactive cells among all taste bud cells was 31.63%, and that of GFR alpha 1-immunoreactive cells, 83.21%. Confocal laser scanning microscopic observations after double immunostaining showed that almost none of the GDNF-immunoreactive cells in the taste buds were reactive with anti-NCAM or anti-PGP 9.5 antibody, but could be stained with anti-alpha-gustducin antibody. On the other hand, almost all anti-PGP 9.5- or anti-alpha-gustducin-immunoreactive cells were positive for GFR alpha 1. Thus, GDNF-immunoreactive cells did not include type-III cells, but type-II cells, which are alpha-gustducin-immunoreactive; on the other hand, GFR alpha 1-immunoreactive cells included type-II and -III cells, and perhaps type-I cells. We conclude that GDNF in the type-II cells may exert trophic actions on type-I, -II, and -III taste bud cells by binding to their GFR alpha 1 receptors.

Animals↗