The mechanism of anteroposterior cell determination in ascidian palleal buds: A gap of positional values triggers posterior formation

Summary Cell-cell interactions during the development of bud polarity in the polystyelid ascidianPolyandrocarpa misakiensis were studied by juxtaposing normally nondjacent bud pieces from marked stocks. These chimeras have been used to determine the prospective fate of each bud piece with respect to positional disparity. The results showed that the posterior end of a future zooid was always established around the proximal boundary region of the two bud pieces. The anteroposterior axis was skewed from this posterior end toward the bud piece arising from a lower (more anterior) level of the parental zooid, suggesting that the lower level tissue may provide a cue which establishes the site of the future anterior end. The role of positional disparity between bud pieces was further examined by sandwiching a host bud between two grafts of various parental positions. The results showed that an additional positional gap resulted in the formation of an additional posterior structure such as the digestive tract. For example, when the grafts were taken from two or more positions posterior to the host bud, biposterior zooids were induced in 20–30% of the cases. This induction ratio increased to 72.4% if two host buds were used instead of a single host. It is, therefore, concluded that the gap of positional values in ascidian buds triggers the formation of the posterior-most positional value. This cellular response to a positional gap contrasts with the intercalation theory that is predicted by the polar coordinate model for pattern formation.     

Patterning of forelimb bud myogenic precursor cells requires retinoic acid signaling initiated by Raldh2

Limb skeletal muscle is derived from cells of the dermomyotome that detach and migrate into the limb buds to form separate dorsal and ventral myogenic precursor domains. Myogenic precursor cell migration is dependent on limb bud mesenchymal expression of hepatocyte growth factor/scatter factor (Hgf), which encodes a secreted ligand that signals to dermomyotome through the membrane receptor tyrosine kinase Met. Here, we find that correct patterning of Hgf expression in forelimb buds is dependent on retinoic acid (RA) synthesized by retinaldehyde dehydrogenase 2 (Raldh2) expressed proximally. Raldh2(-/-) forelimb buds lack RA and display an anteroproximal shift in expression of Hgf such that its normally separate dorsal and ventral expression domains are joined into a single anterior-proximal domain. Met and MyoD are expressed in this abnormal domain, indicating that myogenic cell migration and differentiation are occurring in the absence of RA, but in an abnormal location. An RA-reporter transgene revealed that RA signaling in the forelimb bud normally exists in a gradient across the proximodistal axis, but uniformly across the anteroposterior axis, with all proximal limb bud cells exhibiting activity. Expression of Bmp4, an inhibitor of Hgf expression, is increased and shifted anteroproximally in Raldh2(-/-) limb buds, thus encroaching into the normal expression domain of Hgf. Our studies suggest that RA signaling provides proximodistal information for limb buds that counterbalances Bmp signaling, which in turn helps mediate proximodistal and anteroposterior patterning of Hgf expression to correctly direct migration of Met-expressing myogenic precursor cells.     

Retinoic Acid can Induce a Secondary Axis in Developing Buds of a Colonial Ascidian, Polyandrocarpa misakiensis

We have examined the effect of retinoic acid (RA) on axial pattern formation during bud development of the ascidian, Polyandrocarpa misakiensis . A bead containing various concentrations of RA was implanted into the distal portion of a bud at a site where morphogenic events do not normally occur. Control buds were implanted with beads containing dimethyl sulfoxide (DMSO), the solvent of RA. No apparent effect was observed in these buds containing beads treated with DMSO. In contrast, beads containing 100 μg/ml of RA could induce ectopic structures in the distal portion of buds in about 30% of the cases. The resulting animals had completely duplicated antero-posterior axes. Histological studies showed that, within two days of RA treatment, atrial epithelial cells situated just beneath the implanted bead became thickened and formed a gut rudiment that resembled the posterior structure of the animal. The effect of RA treatment was dose-dependent. The minimum concentration of RA required to induce a secondary axis was 100 ng/ml. Beads containing 1 mg/ml of RA had a lethal effect on the cells that surrounded the beads. These results are discussed in relation to the role of RA in axis formation and the mechanism by which positional values are specified during normal and aberrant bud development in ascidians.     

Herbivory modifies conifer phenology: induced amelioration by a specialist folivore

Herbivory by Zeiraphera canadensis Mut. & Free. (Lepidoptera: Tortricidae), an early season folivore of white spruce [ Picea glauca(Moench) Voss], has been associated with a shift in the timing of bud burst by its host during the subsequent year. We tested the hypothesis that a herbivory-induced shift in the phenology of bud development improves the window for colonisation of white spruce buds by Z. canadensis. Feeding on cortical tissue of elongating shoots caused the destruction of apical buds and an interruption of apical dominance in the year following herbivory. White spruce compensated for damage with the activation of dormant buds; mainly at proximal positions along shoots. As a result, half of all active buds on previously damaged branches were located immediately adjacent egg sites (i.e. previous year's bud scales), whereas <10% of active buds on intact shoots were situated there. More than 40% of newly emerged larvae colonised the basal buds of damaged shoots versus just 10% for intact shoots. Previous herbivory also influenced the initiation of bud burst. All buds flushed 2 days earlier on damaged shoots and date of bud burst was inversely correlated to bud density, indicating that short damaged shoots with large numbers of buds were stronger sinks for nutrients required for bud development. Egg hatch was best synchronized with early bursting buds on damaged branches. As a consequence, 89% of first-instar larvae successfully colonised buds on damaged branches while only 55% were successful on undamaged branches. Improved survival of larvae in the year following herbivory was a direct result of the evolved response by white spruce to the interruption of apical dominance. The pattern of herbivory by Z. canadensis may have evolved as a strategy to enhance the quality of white spruce for their offspring.     

Homeobox genes and gut development

The gut of vertebrates exhibits a common anteroposterior regional differentiation. The role of homeobox genes in establishing this pattern is inferred by their sites of expression. It is suggested that the primary source of positional information is in the endoderm, which subsequently establishes a 'dialogue' with the surrounding visceral layer of the lateral plate mesoderm. This results in the anatomical and physiological specialization of the adult gut.     

Pattern of cell proliferation during budding in the colonial ascidian Diplosoma listerianum

Many invertebrates reproduce asexually by budding, but morphogenesis and the role of cell proliferation in this diverse and nonconserved regeneration-like process are generally poorly understood and particularly little investigated in didemnid ascidians. We here analyzed cell proliferation patterns and telomerase activity during budding in the colonial didemnid ascidian Diplosoma listerianum, with special focus on the thoracic bud where a new brain develops de novo. To help define developmental stages of the thoracic bud, the distribution of acetylated tubulin was also examined. We found extensive cell proliferation in both the thoracic and abdominal buds of D. listerianum as well as higher telomerase activity in bud tissue compared to adult tissues. In the parent adult, proliferation was found in various tissues, but was especially intense in the adult esophagus and epicardial structures that protrude into the proliferating and developing buds, confirming these tissues as the primary source of the cells that form the buds. The neural complex in the thoracic bud forms from a hollow tube that appears to separate into the neural gland and the cerebral ganglion. Whereas most of the bud undergoes proliferation, including the hollow tube and the neural gland, the cerebral ganglion shows little or no proliferation. Pulse-chase labeling experiments indicate that the ganglion, as well as the myocardium, in adult zooids are instead composed of postmitotic cells.     

Revisiting the fate of buds: size and position drive bud mortality and bursting in two coexisting Mediterranean Quercus species with contrasting leaf habit

Understanding the relationships between bud size and position and bud fate through time is crucial for identifying and subsequently modeling the mechanisms underlying tree architecture. However, there is a lack of information on how bud size drives crown architectural patterns in coexisting tree species. We studied bud demography in two coexisting Mediterranean oak species with contrasting leaf habit (Quercus ilex, evergreen; Q. faginea, deciduous). The main objective was to analyse the effect of bud size on the fate of buds with different positions along the shoot (apical, leaf axillary and scale-cataphyll axillary buds). The number, length and position of all buds and stems were recorded in marked branches during 4 years. Study species presented different strategies in bud production and lifespan. The evergreen species showed greater mortality rate than the deciduous one, which produced larger buds. Bud size and position were highly related since apical buds where longer than axillary ones and bud length declined basipetally along the stem. Apical buds had also higher chances of bursting than axillary ones. Within positions, longer buds presented a higher probability of bursting than shorter ones, although no absolute size threshold was found below which bud bursting was impaired. In Q. ilex, four-year-old buds were still viable and able to burst, whereas in Q. faginea practically all buds burst in their first year or died soon after. Such different bud longevities may indicate contrasting strategies in primary growth between both species. Q. ilex is able to accumulate viable buds for several ages, whereas Q. faginea seems to rely on the production of large current-year buds with high bursting probability under favourable environmental conditions.     

The Morphogenesis of Regeneration Buds in Hordeum bulbosum L.--A Perennial Grass

The tillering phase in Hordeum bulbosum L. is terminated whenthe newly-formed axillary buds no longer emerge as tillers,but differentiate into dormant regeneration buds. The patternof development of the axillary buds differs during the tilleringphase and the post-tillering phase. During the former, accumulationof leaf primordia corresponds to the age of the bud, i.e., leafnumber per bud increases basipetally along the shoot. Duringthe post-tillering phase, leaf number per bud decreases basipetallyfrom the base of the future bulb internode. This transitionis brought about by an acceleration in the rate of accumulationof leaf primordia which is more sustained in the buds situatedcloser to the base of the bulb internode. These positional differencesin the morphogenesis of the regeneration buds are reflectedin their physiological responses during the relaxation of dormancyand activation of the buds.     

Specification of pharyngeal endoderm is dependent on early signals from axial mesoderm.

The development of taste buds is an autonomous property of the pharyngeal endoderm, and this inherent capacity is acquired by the time gastrulation is complete. These results are surprising, given the general view that taste bud development is nerve dependent, and occurs at the end of embryogenesis. The pharyngeal endoderm sits at the dorsal lip of the blastopore at the onset of gastrulation, and because this taste bud-bearing endoderm is specified to make taste buds by the end of gastrulation, signals that this tissue encounters during gastrulation might be responsible for its specification. To test this idea, tissue contacts during gastrulation were manipulated systematically in axolotl embryos, and the subsequent ability of the pharyngeal endoderm to generate taste buds was assessed. Disruption of both putative planar and vertical signals from neurectoderm failed to prevent the differentiation of taste buds in endoderm. However, manipulations of contact between presumptive pharyngeal endoderm and axial mesoderm during gastrulation indicate that signals from axial mesoderm (the notochord and prechordal mesoderm) specify the pharyngeal endoderm, conferring upon the endoderm the ability to autonomously differentiate taste buds. These findings further emphasize that despite the late differentiation of taste buds, the tissue-intrinsic mechanisms that generate these chemoreceptive organs are set in motion very early in embryonic development.     

Basipetal Gradient of Axillary Bud Inhibition Along a Rose (Rosa hybridaL.) Stem: Growth Potential of Primary Buds and their Two Most Basal Secondary Buds as Affected by Position and Age

InRosa hybridaL. cv. Ruidriko ‘Vivaldi’®, theeffect of position on growth and development potentials of axillarybuds was investigated by single internode cuttings excised alongthe floral stem and its bearing shoot. The experiments werecarried out in both glasshouses and in a phytotron. The studyfirstly concerned the development of the primary shoot fromthe onset of bud growth until anthesis. The primary shoot wasthen bent horizontally to promote the growth of the two mostproximal secondary buds, the collateral buds, already differentiatedinside the primary bud. They gave rise to basal shoots. In thebasipetal direction, the axillary buds along the floral stemexhibited both an increase in the lag time before bud growthand a decrease in bud growth percentage, demonstrating the existenceof a physiological basipetal gradient of inhibition intrinsicto the buds or due to short range correlations. The same basipetalgradient of inhibition was observed along the floral stem andits bearing shoot, demonstrating that the age of the bud wasnot a major factor in determining the rate of bud growth. Afterbending the primary shoot, the percentage of collateral budgrowth was also affected by the cutting position. The more proximalthe cutting, the lower the sprouting ability of collateral buds.The growth potential of these buds appeared to be already determinedinside the main bud before cutting excision.Copyright 1998 Annalsof Botany Company Axillary bud; basal shoot; cutting; development; endodormancy; growth; paradormancy; position; primary shoot;Rosa hybridaL.; rose; secondary bud; topophysis.     

Interaction of endodormancy and paradormancy in raspberry (Rubus idaeus L.)

Bud break in protected Northern European raspberry crops is often poor and uneven with many of the sub-apical buds remaining in a dormant state. In order to improve bud break and therefore yields, the mechanism controlling bud dormancy must be determined. Canes of the biennial cultivar ‘Glen Moy’ were forced as isolated single nodes, trisections, or as intact canes after different lengths of cold storage chill unit (CU) accumulation in order to determine whether the buds were in an endodormant or paradormant state. The results showed that buds on the lower parts of the intact canes remained in a dormant state long after buds from higher up the intact cane and also the single nodes from all parts of the cane had emerged from the deepest phase of endodormancy. This would imply that these buds were being held in a paradormant state until large amounts of chilling units (>1000 CU) had been accumulated. The trisected cane portions revealed almost no significant differences in bud break levels throughout the experiment when compared with the single nodes. This suggests that removal of the apical part of the cane would be effective in improving bud break by reducing the paradormant condition. A period of secondary dormancy was also observed in the intact canes which may also exacerbate the poor bud break observed in protected crops. This was not seen in the single nodes or the trisected canes which indicates that treatments which reduce paradormancy may also minimise the risk of secondary dormancy. By identifying the phase of bud dormancy which causes poor bud break, attention can now be focused on methods which overcome paradormancy in protected crops. Such methods might include tipping (removal of the cane apex), horizontal training methods, more efficient chilling methods, and chemical treatments.     

Morphogenèse de l'appareil génital ectodermique femelle d'Eurygaster austriaca (Schrank, 1776) (Heteroptera : Scutelleridae)

Female ectodermal genital apparatus of Eurygaster austriaca (Heteroptera : Scuttelleridae) originates from 3 primary buds, which are situated on 3 different regions and from one secondary bud. The external genitalia arise from 8–9th and 9–10th buds. The common oviduct with its fingershaped tubules, results from a median invagination of the 7–8th intersegmental area. The spermatheca is a dorso-apical invagination of the 8–9th bud. The vagina and its annexes originate from fused 8–9th and 9–10th buds and then, they have a bisegmental origin. The genital orifice, or secondary gonopore, also has the same origin.     

Expression of a homeobox gene product in normal and mutant zebrafish embryos: evolution of the tetrapod body plan

An antibody was used to detect antigens in zebrafish that appear to be homologous to the frog homeodomain-containing protein XlHbox 1. These antigens show a restricted expression in the anteroposterior axis and an anteroposterior gradient in the pectoral fin bud, consistent with the distribution of XlHbox 1 protein in frog and mouse embryos. In the somitic mesoderm, a sharp anterior limit of expression coincides exactly with the boundary between somites 4 and 5, and the protein level fades out posteriorly. A similar, graded expression of the antigen is seen within the series of Rohon-Beard sensory neurons of the CNS. We also immunostained the mutant spt-1 ('spadetail'), in which the trunk mesoderm is greatly depleted and disorganized in the region of XlHbox 1 expression. The defects stem from misdirected cell movements during gastrulation, but nervertheless, newly recruited cells that partially refill the trunk mesoderm express the antigen within the normal span of the anteroposterior axis. This finding suggests that the mutation does not delete positional information required for activation of the XlHbox 1 gene.     

The mouse polydactylous mutation,luxate (lx), causes anterior shift of the anteroposterior border in the developing hindlimb bud

Pattern formation along the anterior-posterior axis of the vertebrate limb is established upon activation of Sonic Hedgehog (SHH) in the zone of polarizing activity (ZPA). Since many mouse mutants with preaxial polydactyly show ectopic expression of Shh at the anterior margin of the limb buds, it has been thought to be a primary defect caused by these mutations. We show here that the mouse mutation luxate (lx) exhibits dose-dependent reduction in the size of the Fgf8 expression domain in the ectoderm from the initial stage of limb development. This aberration was independent of Fgf10 expression in the limb mesenchyme. Shh was induced in the mesenchyme underlying the posterior end of the Fgf8 expression domain, indicating an anterior shift of Shh expression in lx hindlimb buds. Prior to the ectopic induction of Shh, the expression domains of genes downstream from Shh, namely dHAND, Gli1, Ptc and Gre, which are normally expressed in posterior mesenchyme of limb buds, expanded anteriorly on the lx hindlimb buds. Conversely, the expression domains of anterior mesenchymal markers such as Gli3and Alx4 decreased in size. Thus, ectopic Shh is not a primary defect of the lx mutation. Rather, our results indicate that the lx mutation affects the positioning of the anteroposterior border in developing hindlimb buds.     

Fgf signaling controls pharyngeal taste bud formation through miR-200 and Delta-Notch activity

Taste buds, the taste sensory organs, are conserved in vertebrates and composed of distinct cell types, including taste receptor, basal/presynaptic and support cells. Here, we characterize zebrafish taste bud development and show that compromised Fgf signaling in the larva results in taste bud reduction and disorganization. We determine that Fgf activity is required within pharyngeal endoderm for formation of Calb2b(+) cells and reveal miR-200 and Delta-Notch signaling as key factors in this process. miR-200 knock down shows that miR-200 activity is required for taste bud formation and in particular for Calb2b(+) cell formation. Compromised delta activity in mib(-/-) dramatically reduces the number of Calb2b(+) cells and increases the number of 5HT(+) cells. Conversely, larvae with increased Notch activity and ascl1a(-/-) mutants are devoid of 5HT(+) cells, but have maintained and increased Calb2b(+) cells, respectively. These results show that Delta-Notch signaling is required for intact taste bud organ formation. Consistent with this, Notch activity restores Calb2b(+) cell formation in pharyngeal endoderm with compromised Fgf signaling, but fails to restore the formation of these cells after miR-200 knock down. Altogether, this study provides genetic evidence that supports a novel model where Fgf regulates Delta-Notch signaling, and subsequently miR-200 activity, in order to promote taste bud cell type differentiation.     

Determination of polarity and bilateral asymmetry in palleal and vascular buds of the ascidian Botryllus schlosseri.

Reversal of the bilateral asymmetry of the zooids was induced in a series of colonies of Botryllus schlosseri. Palleal buds from colonies with normal or reversed bilateral asymmetry were isolated in the early stages from the parental zooids and cultured in the vascularized tunic of the same colony or of another colony with opposite asymmetry. Vascular budding was induced in colonies with either type of asymmetry.The bud polarity was shown to depend on the vascularization; the test vessel entering the isolated palleal bud always causes the entrance point to become the posterior end of the developing zooid. On the contrary, the bilateral asymmetric type is predetermined in the bud primordium; the isolated palleal buds develop the type of asymmetry of their parents, even when grafted in the test of a colony with opposite asymmetry. Since the same was also true of the vascular buds, it is concluded that the information for the kind of bilateral asymmetry to be developed is conveyed by the epidermal envelope of the bud. The epidermis of the parental zooids influences the palleal buds, whereas the wall of the test vessels, epidermal extrusions of the zooids, influences the vascular buds.