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Genome-wide characterization of MADS-box genes and their roles in axillary bud development in tobacco.

A total of 118 NtMADS-box genes were identified in tobacco, revealing their potential roles in axillary bud development. Preliminary overexpression analysis indicated that NtMADS91 promotes axillary bud development. MADS-box transcription factors are core regulators of plant development, but their functions in axillary bud development in Nicotiana tabacum L. have not been systematically elucidated. In this study, 118 NtMADS-box genes were identified from the tobacco genome. Phylogenetic analysis classified them into type I (comprising the Mα and Mγ subfamilies) and type II (comprising the MIKC* and MIKCC clades). Promoter analysis revealed that cis-acting elements were predominantly associated with light and hormone responses. RNA-seq analysis of axillary buds after topping identified 60 differentially expressed NtMADS-box genes, from which 12 candidate genes with significant expression changes were selected. Tissue-specific qRT-PCR revealed that seven of these genes were preferentially expressed in axillary buds, with members of the SOC1 and SVP subfamilies accounting for the majority. Exogenous application of abscisic acid and the strigolactone analog GR24 significantly suppressed the expression of most candidate genes, including NtMADS91. The preliminary overexpression analysis suggested that NtMADS91 may promote axillary bud growth, increasing both the number and length of axillary buds. This study lays a foundation for future dissection of the regulatory mechanisms of the NtMADS-box gene family in axillary bud development and provides promising candidate genes for research related to tobacco axillary bud development.

Nicotiana

Location and variation in number of taste buds in human fungiform papillae.

Serial sections of 182 fungiform papillae, obtained at autopsy from 22 individuals aged 2 days to 90 years, were examined by light microscopy with regard to location and number of taste buds. The taste buds were always found on the convex, dorsal surface of the papillae but otherwise failed to display any preferential location pattern. A total of 262 taste buds, an average of 1.4 per papilla, were found. However, there was considerable variation in the occurrence of taste buds, both from papilla to papilla and from case to case. While the number of taste buds in a single papilla varied from 0 to 27, 63% of the papillae had no taste buds at all, 26% had 1-3 buds and the remainder 4 or more buds. The mean number of taste buds per papilla varied from 0 to 9 between individuals; no dependence upon sex or age could be demonstrated for this variation. The significance of these anatomical findings with regard to physiological studies on taste involving the fungiform papillae is discussed.

Adolescent

[Demonstration, by means of electron microscopy, of the penetration of somitic cells into the mesoblast of the limb buds of reptile embryos (Anguis fragilis, Lacerta viridis)].

An electron microscopic study of the components of anterior limb buds of the slow-worm (Anguis fragilis) and of the green lizard (Lacerta viridis) (embryos of Anguis whose allantoic bud reach 0,7 to 4 mm of length; embryos of Lacerta 2 to 7 days old) provides data on the cytological characteristics of the components of the limb bud at these early stages. 1. The cells of the distal extremity of the somitic processes extending in the limb bud of Anguis and Lacerta, are elongated cells with ovoid nuclei containing large nucleolus; they possess mitochondria always thin and with dense matrix; they are rich in lipid droplets; they possess cilia; they are devoid of myofilaments; endoplasmic reticulum, free ribosomes and polyribosomes are abundant. Golgi networks display signs of activity. These characteristics are also observed in the cells of the "dermatome" layer of the dermo-myotome; and so, it appears probable that the cells of the "dermatome". Furthermore, in Anguis embryos, the cells of the distal extremities of the somitic processes possess numerous lysosomes and a certain number of cells among them, degenerate early. 2. The somatopleural mesoblastic cells of the limb bud of Anguis and Lacerta embryos keep the characters of the cells of the mesodermic layer of lateral plate from which they originate; they have rounded nuclei, cilia, and their mitochondria are always larger and more transparent to electrons, than the ones of cells of the somitic processes and of cells of the epiblastic apical crest. Golgi networks are well developped, endoplasmic reticulum is abundant, lipid droplets are rare. 3. The processes of somites which extend in the dorsal part of the limb bud of Anguis embryos are cords of cells with thin lumina; at the stage of the allantoic bud of 0,6 to 0,8 mm long, the distal extremity of these processes dislocate in group of cells which afterwards dissociate, releasing individual somitic cells which are integrated among the mesoblastic somatopleural cells. In young lizard embryos (2 to 4 days old) the distal extremity of the somitic processes enlarges into a vesicle from which cells are released and penetrate in the mesoblast of the limb bud. 4. The somitic cells released from the somitic processes of Anguis and Lacerta keep--at least at early stages--the cytological characteristics they displayed when they were still in situ in the somitic processes: grounded on the presence or absence of lipid droplets, on the width and density of the mitochondria, the distinction, at these stages, between the somitic and mesoblastic somatoplerual cells is possible; and it is also possible to observe the integration of the somitic cells into the mesoblast. This study brings the demonstration of the cellular contribution of the somites to the formation of the limb bud in Reptiles. 5...

Allantois

The distribution of alkaline phosphatase activity in normal and cross-species regenerated rat and mouse taste buds.

Alkaline phosphatase (ALK Pase) activity can be detected histochemically in the taste buds of rats but not mice. Since taste buds develop, regenerate and are maintained under the influence(s) of the sensory nerve it was decided to study cross-species regenerated buds of these two animals to determine whether the nerve also regulated ALK Pase development in taste cells. Grafts of rats sensory ganglion and mouse tongue or mouse ganglion and rat tongue were combined in the anterior chamber of the eyes of immunologically-deficient nude mice and the cross-species buds that developed at 35 days were examined histochemically for ALK Pase. The results revealed that the rat nerve did not cause ALK Pase to appear in any buds found in mouse tongue grafts and that mouse nerve could support buds containing ALK Pase in rat tongue tissue. Because the cross-species regenerated buds were histochemically characteristic of those normally found in rat or mouse tongue, there is no evidence that the foreign nerve altered gene expression for ALK Pase in the target organ, and the action of the nerve on gustatory epithelium appears to be that of activation and maintenance.

Alkaline Phosphatase

Distribution of taste buds on fungiform and circumvallate papillae of bovine tongue.

The distribution of taste buds on the fungiform and circumvallate papillae of the cow tongue has been determined. The two tongues studied were from Holstein-Friesian cows four to six years of age; they contained 14,765 and 21,691 taste buds, respectively. The tip of the tongue is well supplied with fungiform papillae, and the posterior portion contains the circumvallate papillae. The midportion of the tongue contains relatively few taste papillae. The fungiform papillae contained 1,580 and 1,838 taste buds on the two tongues, respectively, and the circumvallate papillae were estimated to contain 13,185 and 19,853 taste buds. The highest concentration of taste buds therefore occurs in the circumvallate papillae; these relatively few papillae contain approximately 90% of the taste buds. On a circumvallate papilla, taste buds are found only on the papillary sidewall, with none either on the apical surface of the papilla or on the outer wall of the moat.

Animals

The fine structural effect of sialectomy on the taste bud cells in the rat.

Taste buds in the rat and other mammals share a secretory activity with their transduction function as taste receptor. The present work shows the effect of bilateral removal of the main salivary glands on taste bud cells' components related to secretion in the vallate papilla of the rat. In the sialectomized rats remarkable changes were evidence in the dark and intermediate types of taste bud cells, which are known to be the secretory components. Such changes involve hypertrophy of either the protein synthetizing machinery, the smooth endoplasmic reticulum or the Golgi complex. Lucent and coated vesicles associated to Golgi cisternae increased in number but the amount of dense-core vesicles (secretory vesicles) at the apical cytoplasm of cells decreased. Images of exocytosis of secretory products were observed. The hypertrophy of Golgi complex components was clearly detected with the OsO4 impregnation method for light and electron microscopy. Alteration in the acid phosphatase activity of taste bud cells was not observed in the sialectomized rats. These findings suggest that sialectomy stimulates the entire secretory cycle of dark and intermediate taste bud cells. The light taste bud cells, which are not engaged in secretion, are hardly affected by the treatment. Although taste buds in mammals are neuro-dependent structures, present evidence indicates that they are also sensitive to non-neural influences.

Animals

Location of taste buds in intact taste papillae by a selective staining method.

Taste buds were found to stain strongly and selectively in intact papillae with highly acidic dyes such as ponceau S. In intact tongues the taste buds in the fungiform, circumvallate and foliate papillae of the cynomolgus monkey and in the fungiform papillae of the rat as well as the taste discs in the fungiform papillae of the frog could be visualized. This method enables a rapid location and counting of taste buds in taste papillae without preparing histological sections. In cynomolgus tongue material fixed in formalin, the dyes penetrate into the buds. In fresh tongues only the taste pore region of the buds stains, which suggests that in vivo taste buds are impenetrable underneath the pore.

Animals

The effect of temperature on the turnover of taste bud cells in catfish.

Renewal of taste bud cells on the barbels of channel catfish was studied. Groups of catfish, held in and acclimitized to 14 degrees C, 18 degrees C, 22 degrees C and 30 degrees C dechlorinated tap water were injected with [3H]thymidine (3.0 muCi/g body weight intraperitoneally). Barbels were sampled at various times after injection and prepared for light microscope autoradiography. Results show that epithelial cells surrounding the taste buds divide and some of their daughter cells migrate into the taste buds. The time at which 50% of the labelled cells have degenerated is taken as the average turnover time or average life span of the taste bud cells. The average life span as well as the time spent inside the taste buds is highly temperature-dependent. At 14 degrees C, 18 degrees C, 22 degrees C and 30 degrees C the average life span is on the order of 40, 30, 15 and 12 days respectively. Further studies indicate that both light and dark staining cells of the taste bud were labelled.

Animals

The occurrence of taste buds in the palate of human adults as evidenced by light microscopy.

There is some uncertainty in the literature as to the existence of taste buds in the palate of the human adult. In those histologic studies in which the ages of the individuals have been reported, taste buds have not been found in the palates of adults, but have been found in fetuses or newborn. However, clinical studies have demonstrated taste perception in the palate of the human adult. Thus, the aim of the present study was to attempt to find taste buds in the human palate in subjects of different ages. In serial sections of selected areas of the palatal mucosa from autopsy material from individuals 0--80 years of age no taste buds could be demonstrated. However, in four of seven subjects aged 25--44 years, one or two taste buds were found in biopsies from areas of the soft palate where taste perception had been demonstrated clinically just prior to excision. Thus the present study indicates that scattered taste buds exist also in the soft palate of human adults.

Adolescent

Biochemical studies of taste sensation. III. Preparation of a suspension of bovine taste bud cells and their labeling with a fluorescent probe.

A method to prepare suspensions of taste bud cells is described. Bovine circumvallate papillae, which contain most of the taste buds in this animal, are incubated in collagenase-containing medium and the epidermal sidewall tissue is then dissected from the inner gelatinous dermis. The sidewall tissue, which contains the taste buds, is gently homogenized by manual operation of an all-glass homogenizer with a loose-fitting pestle. The suspended material is separated on a discontinous Ficoll gradient (2%, 8%, 10%, 12% w/w). The material banding at the 8-2% interface is greatly enriched in spindle-shaped cells that are morphologically similar to taste bud cells as they appear in situ. These cells are not seen when the procedure is done with tissues devoid of taste buds, namely the upper surface of the circumvallate papilla or epithelium from the intermolar eminence. Fluorescence analysis indicates that the hydrophobic probe, 8-anilino-1-naphthalenesulfonate (ANS), binds to relatively nonpolar sites in the suspension. It is postulated that the probe is adsorbing onto the surface membrane of the cell. These preparations may be useful in studying specificity and transduction in taste sensation.

Anilino Naphthalenesulfonates

Histochemical observations on the taste buds of the marmosets (Callithrix jacchus and Callithrix penicillata).

The epithelial cells in the taste buds of C. jacchus and C. penicillata show a moderate amount of ribonucleic acid an a concentration of a PAS-positive diastase-resistant material at their apical part. These cells are devoid of UDPG-GT, phosphorylases, G-6-PA, alanyl aminopeptidase, leucine aminopeptidase, cholinesterase and MAO; they present a weak reaction of F-1, 6-P Ald, LDH, SDH, MDH, cytochrome oxidase, beta-OHBDH, nonspecific esterase and acid phosphatase and a stronger reaction to ADH, NADPH2-TR, ATPases, alpha-GPDH, alkaline phosphatase, 5-nucleotidase and GDH. Although some enzymes (alkaline phosphatase, 5-nucleotidase and ATPases) have an almost uniform reactivity by the several taste buds, the other ones react with a lesser intensity in the smaller uniform reactivity by the several taste buds, the other ones react with a lesser intensity in the smaller taste buds of the fungiform papillae. As a rule the apical part of the cells shows a stronger enzymatic reactivity. The taste buds of the marmosets are penetrated by acetylcholinesterase positive nerve fibers whereas the autonomic ganglia in the connective tissue contain both-acetyl and butyrylcholinesterase.

Adenosine Triphosphatases

[Uptake of (14C) leucine and protein synthesis in potato tuber buds and effect of abscisic acid on these processes].

The uptake of [14C] leucine and its incorporation into proteins of dormant and growing potato tuber buds were studied. It was found that the label uptake was increased at the beginning of the growth period, whereas the dynamics of this process were not changed in comparison with the dormant buds samples. The rate of [14C] leucine incorporation into proteins was increased in the growing buds; this increase was not, however, due to the increase in the uptake of the labelled precursor and was probably caused by activation of the protein synthesis. In contrast, the activation of protein synthesis was accompanied by changses is the dynamic incorporation of [14C] leucine into the protein at the end of dormancy. The effect of abscisic acid (10(-7) M) on the protein synthesis was not connected with its action of the uptake of labelled precursor and depended on the physiological state of buds and incubation time. A possible mechanism of regulatory effect of abscisic acid on protein synthesis in potato tuber buds is discussed.

Abscisic Acid

Postembryonic changes in the optic primordia and optic bud in the flesh fly Sarcophaga ruficornis fabr. (Diptera: Sarcophagidae).

Differentiation of the optic lobe anlagen begin in the brain of second instar. Each is an elongated disc of cortical cells placed on the dorsolateral border of each protocerebrum. In the late second instar the disc elongates and its two ends bend inwards which gradually separate from the central region, thus giving three imaginal discs. The protocerebral neuropile extends into these discs and medulla interna and externa are formed. The rudiments of compound eyes (cephalic complex) appear in the early laid larva. These are attached with the brain and pharyngeal wall separately. The posterior portion of cephalic complex (optic bud), after establishing a nervous association with the central optic lobe anlage (lamina ganglionaris), forms the compound eye. Ech optic bud is attached to the brain by a non-nervous stalk. The epiblast cells of the optic bud do not migrate into the brain and the lamina is formed by the proliferation of the central imaginal disc. The reorientation of the optic lobe anlagen starts in the late third instar and the medulla interna divides into two unequal lobes. In 2 day pupa the nerve fibres from the lamina travel into the optic stalk and the optic nerve is formed. The epiblast cells of the optic bud differentiate to form a peripheral epithelial layer which becomes pigmented and gets apposed to the lateral boundary of the brain. The central epiblast cells of the optic bud form several ommatidia. The optic nerve degenerates gradually and various components of the compound eye are formed by the epiblast cells. Chiasm internum is present but chiasm externum is absent.

Animals

Epithelial differentiation and taste buds in the soft palate of the monkey, Macaca irus.

A combination of light, transmission and scanning electron microscopy was employed to demonstrate the occurrence, arrangement and structure of taste buds in the oral mucosa of the soft palate of monkeys (Macaca irus). Taste buds are found in aggregates confined to 0.15 to 0.3 mm wide, round islands of keratinizing epithelium embedded in the normally non-keratinizing integument. Topography, configuration and structure of these epithelial islands and their taste buds are described, and the question of a developmental and functional interrelationship between epithelial differentiation and properties, and taste bud function is discussed.

Animals

Cryo-electron tomography reveals coupled flavivirus replication, budding and maturation.

Flaviviruses replicate their genomes in replication organelles (ROs) formed as bud-like invaginations on the endoplasmic reticulum (ER) membrane, which also functions as the site for virion assembly. While this localization is well established, it is not known to what extent viral membrane remodeling, genome replication, virion assembly, and maturation are coordinated. Here, we imaged tick-borne flavivirus replication in human cells using cryo-electron tomography. We find that the RO membrane bud is shaped by a combination of a curvature-establishing coat and the pressure from intraluminal template RNA. A protein complex at the RO base extends to an adjacent membrane, where immature virions bud. Naturally occurring furin site variants determine whether virions mature in the immediate vicinity of ROs. We further visualize replication in mouse brain tissue by cryo-electron tomography. Taken together, these findings reveal a close spatial coupling of flavivirus genome replication, budding, and maturation.

Journal Article

The reversible effect of colchicine on the taste bud cells of the central circumvallate papilla in the rat.

It is believed that differentiation and maintenance of taste buds in vertebrates is dependent on the trophic function of their sensory nerve supply. In the present work, colchicine was injected into the circumvallate papilla of the rat. This produced a reversible blockade of neuroplasmic transport and disappearance of taste buds. Colchicine inhibited the further differentiation of bud cells, but apparently did not change the life cycle of the cells present already at the time of injection. It is speculated that the neurotrophic factors in this particular cell system are effective to induce cell differentiation only.

Acetylcholinesterase

Fine structure of taste buds in the rat.

The taste buds of rat circumvallate papillae contain three distinct types of cells. The type I (dark) cell is characterized by the presence of dense round granules, which are precursor to the dense substance of the taste pore. The granules are discharged into the pore by exocytosis. The type II (light) cell is filled with numerous vesicles and smooth-surfaced endoplasmic reticulum. The type III cell contains in its basal cytoplasm characteristic dark-cored vesicles and masses of clear vesicles, and makes synapse-like contacts with nerve fibers. The fine structure of foliate buds corresponds to that of circumvallate papillae, while fungiform buds differ in their apical regions. In the latter the pore is filled with vesicles alone, and the type I cell contains rod-shaped granules of moderate density. When polysaccharides were examined by means of the periodic acid-silver methenamine and the periodic acid-thiocarbohydrazide methods, slightly positive reactions were found on the dense granules of the type I cell and the dense substance in the pore, whereas the membranes of the pore vesicles, apical cytoplasmic processes and cytoplasmic vesicles in type II cells showed intense reactions.

Animals

The fine structural localization of adenosine triphosphatase activity on the taste bud in the fungiform papillae of the rat.

The localization of Mg++ activated adenosine triphosphatase (ATPase) activity in rat fungiform taste buds was demonstrated by electron microscopic histochemistry. Reaction product was found on the cell membrane of the taste cells, but not within their cytoplasm. Especially strong reaction occurred in the taste pore. Besides, enzymes were observed in axon-Schwann cell and axon-type I cell interspace, but not in the contacts of the nerve fiber and type II cell. The taste bud could well be distinguished from the surrounding epithelium by the existence of the reaction product. ATPase activity was absent at the synaptic portion where type III cells faced nerve fibers. No reaction product was seen along the basal lamina. In fungiform buds, the nerve fibers were frequently shown to make contact with each other, and these sites were devoid of deposits for ATPase.

Adenosine Triphosphatases