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PubMed1989

Compensatory weight gain in steers recovered from oak bud toxicosis.

Cattle that had recovered from clinical oak bud toxicosis (as assessed by normal BUN and serum creatinine values and good appetite) performed better than did clinically normal herdmates when weight gain and feed efficiency were compared. Both groups had a rate of weight gain (oak bud-exposed cattle, 1.76 kg/steer/d; control cattle, 1.57 kg/steer/d) that exceeded the feedlot mean value (approximately 1.4 kg/d) for cattle (of similar starting weight and diet) that had zeranol implants, and the difference was significant (P less than 0.005) for both groups. The difference in weight gain performance of oak bud-exposed cattle, compared with control cattle, was not highly significant; however, the improved feed efficiency was significantly (P less than 0.05) better than that in controls (2.57 kg of feed/kg of weight gain vs 3 kg of feed/kg of weight gain). On the basis of accepted criteria (increased rate of weight gain and efficiency of feed conversion), the group of oak bud-exposed cattle of this study had compensatory weight gain. Thus, cattle with good evidence of clinical recovery from oak bud toxicosis can be expected to perform in the feedlot at least as well as cattle not exposed to oak bud toxicosis, and may have compensatory weight gain.

Animal Feed↗

PubMed1981

Control of maintenance and anteroposterior skeletal differentiation of the anterior mesenchyme of the chick wing bud by its posterior margin (the ZPA).

If the posterior half of the chick wing bud (stages 17-22) is excised, the anterior half which normally forms humerus (part), radius and digit 2, forms only a single skeletal element, either humerus or humerus fused with reduced radius. Beginning at 18 h after operation, and continuing to 48 h the anterior and distal mesenchyme in such anterior halves becomes necrotic and the AER regresses. By contrast, if the anterior half of the chick wing bud (stages 17-22) is excised, the posterior half develops as in the normal bud, and forms humerus (part), ulna and digits 3, 4 and 5. Such posterior halves develop no more mesenchymal necrosis than the normal contralateral wing buds and the AER remains healthy. Further, if the excision of the posterior part is made in such a way as to leave in place a part of the zone of polarising activity (ZPA), a normal wing with complete skeleton is formed. Thus in order to survive and differentiate, the anterior part of the wing bud needs a factor supplied by the posterior part containing the ZPA. These results support the view that the ZPA plays a role in controlling the anteroposterior differentiation of the normal wing bud.

Animals↗

PubMed1981

A gradation of hyaluronate accumulation along the proximodistal axis of the embryonic chick limb bud.

We are currently investigating the mechanism by which the apical ectodermal ridge (AER) of the embryonic chick limb bud exerts its negative effect on the cytodifferentiation of limb mesenchymal cells directly subjacent to it, and the mechanism by which cytodifferentiation is triggered when the cells leave the influence of the AER. Since there is a gradation of differentiation along the proximodistal axis of the limb bud, we have dissected limb buds into discrete segments along the proximodistal axis, and biochemically examined the accumulation of hyaluronate (HA) and other glycosaminoglycans (GAG) in each segment. The unspecialized subridge region of stage-25 limb buds was separated into distal (segment 1) and proximal (segment 2) regions, and the remaining proximal portion of the limb was separated into four segments (segments 3, 4, 5 and 6) along the proximodistal axis. Stage-24 limb buds were separated into corresponding regions. Since in the proximal regions of the limb (segment 3 through 6), only the cells comprising the central cores of the limb are involved in chondrogenic differentiation, the central core tissue was surgically separated from the peripheral tissue. We have found that HA is by far the predominant GAG accumulated by cells comprising the distal subridge region (representing greater than 50% of the total GAG accumulated during a 90-min labelling period with [3H]glucosamine), and that there is a progressive decline in HA accumulation along the proximodistal axis. The relative and total amount of HA accumulated is highest in the distal subridge region (segment 1), intermediate in the proximal subridge region (segment 2) and lowest in the proximal central core regions of the limb (segments 3 through 6). The striking decrease in HA accumulation in the central core of segment 3 is accompanied by a striking increase in the accumulation of chondroitin sulphate, one of the major constituents of cartilage matrix. In contrast to the central core regions of segments 3 through 6, the relative and total amount of HA accumulated by the peripheral non-chondrogenic regions of these segments remains relatively high, being similar to the accumulation observed in the proximal subridge region. These results indicate that there is a gradation of HA accumulation along the proximodistal axis of both stages-24 and -25 limb buds which correlates with distance of cells from the AER and the state of differentiation of the cells.

Animals↗

PubMed1993

Neural crest cell migration into the limb bud of avian embryos.

The colonization of limb buds by neural crest cells was studied in quail-chick chimeras and in chick embryos using HNK-1 and DiI staining and the LD-DOPA reaction. Two populations of neural crest cells were found to colonize the limb bud. They migrate successively and use different routes of migration. The first population migrates within the limb bud subectodermally at stages before the limb is innervated. In the wing bud the migration route is localized postaxially and in the leg bud preaxially. Two cell types were identified differentiating from this first population: melanoblasts and Merkel cells. The second population of crest cells invades the limb bud at a later stage. These cells follow the routes of ingrowing nerves and migrate along a dorsal and a ventral path which correspond to the position of nerves for extensor and flexor muscles. Crest cells were found here also in the absence of nerves. Schwann cells and terminal glial cells develop from this second population of neural crest cells.

Animals↗

PubMed1994

Sorting out of limb bud cells in monolayer culture.

To examine differences in the surface properties of limb bud cells, we mixed cells from the progress zone at different stages or from different positions along proximodistal axis of chick and quail wing limb buds. To identify the origin of cells, a chick-specific antibody was used in a mixed culture of chick and quail cells, or cells from one of the stages were labeled with a fluorescent dye, PKH-26. Within 18 hours in mixed culture, cells segregated from each other and formed patches of various sizes. The process zone cells at early developmental stages mixed homogeneously with the cells from proximal region of old limb buds and the progress zone cells at late stages mixed homogeneously with the cells from distal region of the old limb buds. These results suggest that surface properties of cells in progress zone change during limb bud development and vary along the proximodistal and anteroposterior axes of the limb bud and that these differences in surface property may correspond to the positional values for limb pattern formation.

Animals↗

PubMed1994

[Congenital hand malformation--local disturbance in the limb bud and longitudinal deficiency: an experimental study on the pathogenesis].

Experimental congenital malformation produced in fetal chickens to investigate any correlation between a local disturbance in the limb buds and longitudinal deficiency, and to elucidate the pathogenesis of radial and ulnar ray deficiencies. Fertilized White Leghorn eggs were incubated at 38.0 degrees C and windowed. The right wing bud of the embryos was locally cauterized using a bipolar microcoagulator under a microscope. Cauterization was carried out from 2.5 days to 5.5 days of development, at 12-hour intervals. Each wing bud was divided into 6 areas: axial (distal and proximal), preaxial (distal and proximal) and postaxial (distal and proximal). After cauterization the eggs were sealed with micropore tape and further incubated until the 21st-23rd day. The chick wings were then removed and investigated macroscopically and roentgenographically. The wing buds had been examined microscopically after cauterization on the 3rd day until ray deficiencies were produced on the 5th day. Radial ray deficiencies were produced by cauterization in the distal preaxial area, and ulnar ray deficiencies by cauterization in the distal postaxial area. These results indicated a peculiarity of the longitudinal axis. However, the critical periods of the radial and the ulnar ray deficiencies were the same; from immediately after the formation of the wing bud to before the formation of the digital plate. The patterns of the combined defects in the digits and of other associated deformities were quite similar to those found in human malformations. Microscopically necrosis in the mesoderm and in the overlying ectoderm of the pre and postaxial areas of the tip of a wing bud caused defects in the cartilaginous primordium in the same ray.

Animals↗

PubMed1996

Mouse molar morphogenesis revisited by three dimensional reconstruction. I. Analysis of initial stages of the first upper molar development revealed two transient buds.

Early stages of tooth development in the maxillary cheek region in the mouse were investigated by combined analysis of histological sections, computer assisted 3D reconstructions and morphometry. In ED 12.5 embryos, 3D reconstructions revealed an accessory epithelial bud (R1) and a large bud (R2), which appeared as a single bud-shaped epithelium in frontal sections. This developmentally most advanced dental epithelium in the mouse embryonic maxilla until ED 13.5, generally considered as the bud of the first molar, regressed during later development. Meanwhile the bud and cap of the first upper molar originated more posteriorly, from ED 13.5. The regression of R1 and R2 was associated with epithelial apoptosis. Apoptotic cells and bodies were apparent on sections in the R1 epithelium from ED 12.5. The R2 epithelium maintained the large bud-shaped appearance on sections, representing the largest part of the dental epithelium in the maxillary cheek region until ED 14.0; apoptoses were detected there as late as from ED 13.5. During regression, the R2 rudiment was transformed into the medial and lateral epithelial ridges, posteriorly in continuity with the arising cap of the first molar. The reduced R1 epithelium seemed to contribute to the medial ridge. These results should be taken into consideration in the interpretation of early odontogenesis in the upper jaw in the mouse. The interesting problem of the identification of tooth homology of the rudiments should be elucidated by further comparative morphological and paleontological investigations.

Animals↗

PubMed1998

The morphogenesis of mouse vallate gustatory epithelium and taste buds requires BDNF-dependent taste neurons

The developmental absence of brain-derived neurotrophic factor (BDNF) in null mutant mice caused three interrelated defects in the vallate gustatory papilla: sparse innervation, a reduction in the area of the gustatory epithelium, and fewer taste buds. On postnatal day 7, the stunted vallate papilla of bdnf null mutant mice was 30% narrower, the trench walls 35% reduced in area, and the taste buds 75% less abundant compared with wild-type controls. Quantitative assessment of innervation density was carried out to determine if the small trench walls and shortage of taste buds could be secondary consequences of the depletion of gustatory neurons. The diminished gustatory innervation was linearly associated with a reduced trench wall area (r=+0.94) and fewer taste buds (r=+0.96). Residual taste buds were smaller than normal and were innervated by a few surviving taste neurons. We conclude that BDNF-dependent taste neurons contribute to the morphogenesis of lingual gustatory epithelia and are necessary for both prenatal and postnatal mammalian taste bud formation. The gustatory system provides a conspicuous example of impaired sense organ morphogenesis that is secondary to sensory neuron depletion by neurotrophin gene null mutation.

Journal Article↗

PubMed1998

Expressed sequence tags of radish flower buds and characterization of a CONSTANS LIKE 1 gene.

Expressed sequence tag (EST) analysis was conducted for young flower buds of radish plants. Among a total of 66 ESTs examined, 40 showed a significant similarity to previously identified genes. Twenty-eight ESTs were similar to proteins identified in other plants, 11 were similar to eukaryotic proteins other than plants, and one was similar to a prokaryotic protein. Four clones were selected for further studies. EST clone 81, which showed a homology to germin-like proteins was expressed more abundantly in leaves and roots as compared to flower buds. Clone 105 was highly homologous to the translation inhibitor protein and was expressed in all three organs, but the expression level was higher in flower buds and roots. Another EST clone, 133, which shared a significant similarity with the Ran-binding protein, hybridized to two different size transcripts that were detectable only in flower buds. Clone 39 was a homolog of CONSTANS, which is a gene involved in controlling the flowering time in Arabidopsis. The cDNA clone of EST clone 39 containing the entire open reading frame was obtained and designated as RsCOL1 (Raphanus sativus CONSTANS LIKE 1). It was 1049 bp long and contained an open reading frame of 307 amino acid residues (calculated molecular mass = 33.1 kDa). The RsCOL1 protein contained two putative zinc finger motifs in the amino terminal region which were 59% identical to the corresponding region of the Arabidopsis CO protein. The radish protein also contained a predicted nuclear localization domain in the carboxyl terminal region which was 87% identical to the corresponding region of CO. DNA blot analysis revealed that the radish genome contained several genes similar to RsCOL1. RNA blot analysis showed that RsCOL1 was strongly expressed in flower buds at the early bolting stage, and the expression level declined as the flower bud matured. The transcript was also detectable in leaves and roots. In mature flowers, the RsCOL1 transcript was present primarily in carpels.

Amino Acid Sequence↗

PubMed1996

Differential expression of brain-derived neurotrophic factor and neurotrophin 3 mRNA in lingual papillae and taste buds indicates roles in gustatory and somatosensory innervation.

Although many studies have demonstrated the dependency of taste bud function and/or survival on intact innervation, relatively few have dealt with the development of taste bud innervation. Using in situ hybridization histochemistry, we show that brain-derived neurotrophic factor (BDNF) and neurotrophin 3 (NT3) mRNA are expressed in a specific pattern in the taste buds, tongue papillae, and lingual epithelium during development and that expression persists into adulthood. BDNF mRNA is expressed in a fraction of the taste cells of the developing and adult taste buds in rats, showing different labeling intensities among the labeled cells. NT3 and mRNA seems to be located in areas other than those where BDNF mRNA is expressed, mainly in the superior epithelial surfaces of circumvallate papillae, the outer surface epithelium of foliate papilae, the superior surface and the lateral epithelium of the fungiform papillae, and the epithelium of the filiform papillae. NT3 mRNA labeling is also observed among muscle and connective tissue of the tongue. The morphological appearance, expression of NT3 mRNA, and ramification of nerve fibers in defined epithelial structures in the posterior wall of the anterior filiform papillae suggest the existence of a mechanosensory apparatus in these papillae. Nerve growth factor and neurotrophin 4 probes did not give rise to selective labeling in tongue, although their presence cannot be totally excluded. Based on present and prior studies, we suggest that BDNF is needed during initiation and for maintenance of gustatory innervation of taste buds and gustatory papillae and that NT3 is mainly needed for somatosensory innervation of the tongue.

Animals↗

PubMed1997

Light and dark cells of rat vallate taste buds are morphologically distinct cell types.

Cells of mammalian taste buds have been classified into morphological types based on ultrastructural criteria, but investigators have disagreed as to whether these are distinct cell types or the extremes of a continuum. To address this issue, we examined taste buds from rat vallate papillae that had been sectioned transversely, rather than longitudinally, to their longest axis. In these transverse sections, dark (Type I) and light (Type II) cells were easily distinguished by their relative electron density, shape and topological relationships. Cells with electron-lucent cytoplasm (light cells) were circular or oval in outline, while those with electron-dense cytoplasm (dark cells) had an irregular outline with sheetlike cytoplasmic projections that separated adjacent light cells. A hierarchical cluster analysis of 314 cells across five morphological parameters (cell shape and area, and nuclear ellipticity, electron density and invagination) revealed two distinct groups of cells, which largely corresponded to the dark and light cells identified visually. These cells were not continuously distributed within a principal components factor solution. Differences in the means for dark and light cells were highly significant for each morphological parameter, but within either cell type, changes in one parameter correlated little with changes in any other. These analyses all failed to reveal cells with a consistent set of intermediate characteristics, suggesting that dark and light cells of rat vallate taste buds are distinct cell types rather than extremes of a continuum. Sections of taste buds were stained with antibodies to several carbohydrates, then observed by indirect immunofluorescence. Optical sections taken with a confocal laser-scanning microscope showed that the Lewis antigen was present only on spindle-shaped cells with circular or oval outlines and lacking transverse projections; these characteristic shapes matched those of light cells seen by electron microscopy. The H blood group antigen and the 2B8 epitope appeared at most cell-cell interfaces in the bud and are present on dark cells and possibly on some light cells. These findings relate molecular markers to morphological phenotypes and should facilitate future studies of taste cell turnover, development and regeneration.

Animals↗

PubMed1996

Relation of TGF-beta 2 to inhibition of limb bud chondrogenesis by retinoid in rats.

A newly synthesized retinoid, Am-80 (4-[5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthalenyl] carbamoxyl) benzoic acid, induced limb reduction defects in rats in vivo. The present study was designed to investigate the mechanisms of its teratogenic action and also to examine whether transforming growth factor-beta (TGF-beta) is involved in limb reduction defects by using a rat limb bud cell culture system. Am-80 inhibited DNA synthesis and chondrogenesis of limb bud cells from day 12 embryos at concentrations of 1-4 ng/ml in a concentration-dependent manner. Quantities of TGF-beta 1 and 2 in culture medium from limb bud cells treated with Am-80 were evaluated using sandwich ELISAs. Although TGF-beta 1 was not detected in conditioned medium with or without Am-80, TGF-beta 2 increased in a concentration-dependent manner at drug concentrations that inhibited DNA synthesis and chondrogenesis. In addition, when TGF-beta 2 was exogenously added to limb bud cell culture medium, the synthesis of glycosaminoglycan (GAGs) was reduced even without Am-80 treatment. Neutralization of TGF-beta 2 with antibody prevented the inhibition of GAGs synthesis induced by Am-80. These results suggest that TGF-betas, especially beta 2, are involved in the inhibition of chondrogenesis caused by Am-80 in limb bud cells.

Animals↗

PubMed2001

Characterisation of hoxa gene expression in the chick limb bud in response to FGF.

We tested a diffusion gradient model for setting up overlapping domains of Hoxa gene expression in the chick limb bud. The model is based on morphogen production at the limb bud tip where the apical ridge is located and assumes that cells respond to a series of concentration thresholds. Consistent with the model, Hoxa13 gene expression rapidly switches off when the ridge is removed from stage 21/22 buds, while Hoxa11 and Hoxa10 expression is stable; Hoxa13 expression can be initiated and maintained in absence of the ridge by FGF soaked beads; the Hoxa13 domain first expands quickly and then slows up and the size is related to the dose of FGF4. Contrary to the model, addition of FGF4 to early limb buds does not activate Hoxa13 prematurely nor extend the Hoxa13 expression domain proximally. Therefore FGF4 signalling is necessary but not sufficient for Hoxa gene expression in the limb bud.

Animals↗

PubMed1990

Analysis of taste bud innervation based on glycoconjugate and peptide neuronal markers.

Primary gustatory neurons and their peripheral and central processes were evaluated histochemically in the geniculate and petrosal cranial nerve ganglia, lingual fungiform taste buds, and the nucleus of the solitary tract (NST) using 1) the plant lectin Griffonia simplicifolia I-B4, which binds specifically to D-galactose residues and selectively labels primarily nonpeptide-containing peripheral somatosensory neurons, and 2) calcitonin gene-related peptide immunoreactivity (CGRP-IR), which labels most peptidergic somatosensory neurons. Lectin reactivity was expressed by the vast majority of geniculate and petrosal ganglion cells, while CGRP-IR labeled very few cells. Peripherally, gustatory intragemmal axons penetrating fungiform taste buds were labeled only by the lectin and were depleted following chorda tympani transection. However, both lectin-labeled and CGRP-IR subpopulations of somatosensory perigemmal axons surrounding the taste buds were observed and were eliminated by section of the lingual nerve. The differing brainstem projection patterns of lectin-reactive vs. CGRP-IR central axons reflected their distinct ganglionic origins and the differential distributions of lectin reactivity and CGRP-IR among taste buds. Central lectin-reactive terminals were found throughout the entire rostrocaudal extent of the NST, including its rostral lateral "gustatory" zone; the extensive lectin-reactive visceral afferent projection can be presumed to have originated mainly from the large proportion of lectin-labeled neurons in the nodose ganglion. The lectin also prominently and selectively labeled the area postrema. CGRP-IR central terminals, however, was relatively sparse and restricted primarily to the caudal and medial "visceral" divisions of the NST. The results are discussed with respect to the possible functional implications of cell surface glycoconjugate expression by gustatory axons innervating taste bud receptor cells of the tongue.

Animals↗

PubMed1991

HVEM serial-section analysis of rabbit foliate taste buds: I. Type III cells and their synapses.

Serially sectioned rabbit foliate taste buds were examined with high voltage electron microscopy (HVEM) and computer-assisted, three-dimensional reconstruction. This report focuses on the ultrastructure of the type III cells and their synapses with sensory nerve fibers. Type III cells have previously been proposed to be the primary gustatory receptor cells in taste buds of rabbits and other mammals. Within rabbit foliate taste buds, type III cells constitute a well-defined, easily recognizable class and are the only taste bud cells observed to form synapses with intragemmal nerve fibers. Among 18 type III cells reconstructed from serial sections, 11 formed from 1 to 6 synapses each with nerve fibers; 7 reconstructed type III cells formed no synapses. Examples of both convergence and divergence of synaptic input from type III cells onto nerve fibers were observed. The sizes of the active zones of the synapses and numbers of vesicles associated with the presynaptic membrane specializations were highly variable. Dense-cored vesicles 80-140 nm in diameter were often found among the 40-60 nm clear vesicles clustered at presynaptic sites. At some synapses, these large dense-cored vesicles appeared to be the predominant vesicle type. This observation suggests that there may be functionally different types of synapses in taste buds, distinguished by the prevalence of either clear or dense-cored vesicles. Previous investigations have indicated that the dense-cored vesicles in type III cells may be storage sites for biogenic amines.

Animals↗

PubMed2004

Ventral tail bud mesenchyme is a signaling center for tail paraxial mesoderm induction.

A large body of evidence from several systems indicates that formation of the vertebrate tail is morphogenetically continuous with gastrulation, including neural inducing activity in descendants of the gastrula organizer. However, the signaling centers and molecular events regulating tail mesoderm induction and its organized elongation remain poorly defined. In mammals, the ventral ectoderm ridge (VER) is essential to maintain ongoing formation of paraxial mesoderm and somitogenesis in cultures of intact tail. Avian tail buds contain a similar VER structure. Here, we report that the chick ventral tail bud operates as a signaling center for paraxial mesoderm induction. By using "organizer" style grafting assays to early host embryos, we found that ventral tail bud was able to induce elongated paraxial mesodermal extensions and that the ventral tail bud mesenchyme underlying the VER is both necessary and sufficient for the induction in this assay system. Our observations combined with those of others suggest that interplay between several different signaling centers in the amniote tail bud regulates the coordinate induction and elongation of axial and paraxial structures in the developing tail.

Animals↗

PubMed1993

Electron microscopic immunocytochemistry of glutamate-containing nerve fibers in the taste bud of mudpuppy (Necturus maculosus).

The presence of glutamate immunoreactivity (glu-IR) in the nerve fibers of the mudpuppy taste bud was investigated by electron microscopy. Pre-embedding staining with avidin-biotin-peroxidase complex (ABC) and post-embedding staining with 5 nm colloid gold conjugates were used separately to identify immuno-stained structures. We have found the following: 1) the majority of the nerve fibers innervating the mudpuppy taste bud are unmyelinated; 2) about 85% of nerve fibers located at the base of the taste bud and about 60% of the nerve fibers located between the taste cells show glu-IR by pre-embedding staining; 3) there is a preferential staining of the glu-IR in the nerve fibers of the mudpuppy taste bud; and 4) the distribution of the colloidal gold particles in the nerve fibers is 1.5 to 2 times denser than that of the staining in the connective tissue background or cellular profiles of taste cells. From the distribution and pattern of the nerve fibers obtained in the thick and thin sections, we conclude that the mudpuppy taste bud is innervated by glutamate-containing unmyelinated nerve fibers.

Animals↗

PubMed2005

Chorda tympani nerve transection at different developmental ages produces differential effects on taste bud volume and papillae morphology in the rat.

Chorda tympani nerve transection (CTX) results in morphological changes to fungiform papillae and associated taste buds. When transection occurs during neonatal development in the rat, the effects on fungiform taste bud and papillae structure are markedly more severe than observed following a comparable surgery in the adult rat. The present study examined the potential "sensitive period" for morphological modifications to tongue epithelium following CTX. Rats received unilateral transection at 65, 30, 25, 20, 15, 10, or 5 days of age. With each descending age at the time of transection, the effects on the structural integrity of fungiform papillae were more severe. Significant losses in total number of taste buds and filiform-like papillae were observed when transection occurred 5-30 days of age. Significant reduction in the number of taste pores was indicated at every age of transection. Another group of rats received chorda tympani transection at 10, 25, or 65 days of age to determine if the time course of taste bud degeneration differed depending on the age of the rat at the time of transection. Taste bud volumes differed significantly from intact sides of the tongue at 2, 8, and 50 days post-transection after CTX at 65 days of age. Volume measurements did not differ 2 days post-transection after CTX at 10 or 25 days of age, but were significantly reduced at the other time points. Findings demonstrate a transitional period throughout development wherein fungiform papillae are highly dependent upon the chorda tympani for maintenance of morphological integrity.

Age Factors↗