Start Of Gastrulation Research Articles

XIF3, a Xenopus peripherin gene, requires an inductive signal for enhanced expression in anterior neural tissue.

A full-length cDNA clone for the Xenopus intermediate filament gene XIF3 has been isolated. It is very similar in sequence to the rat intermediate filament cDNA clone 73 that is thought to encode the neuronal intermediate filament protein 'peripherin'. By analysing dissected embryos, we show that XIF3 is expressed predominantly in anterior and dorsal structures and most strongly in the brain of the tailbud (stage 26) embryo. In situ hybridization shows XIF3 transcripts to be localized in neural tissue and especially in regions that most probably correspond to the motor neurones of the neural tube and to some cranial nerve ganglia. New XIF3 transcripts are first found at the start of gastrulation at a low level throughout the ectoderm and are not localized to the presumptive neurectoderm. Expression subsequently increases by about 10-fold in neural tissue, and requires an interaction of the mesoderm with overlying ectoderm. Because new transcripts are found predominantly in neural tissue of the head, this response can be used as a marker of anterior neural induction.

The first cleavage plane and the embryonic axis are determined by separate mechanisms in Xenopus laevis: I. Independence in undisturbed embryos

We examined the spatial relationships between the meridian of sperm entry the plane of first cleavage, and the embryonic axis (defined by the neural groove) in eggs of Xenopus laevis. Direct measurement of the angular separations between these embryonic structures in gelatin-embedded eggs confirmed the classical conclusion that the sperm entry point and neural groove tend to form on opposite sides of the egg, and also revealed that the first cleavage plane has a nearly random orientation with respect to the neural groove. We next examined the distortion of the first cleavage plane that results from the normal processes of convergence and extension during gastrulation and neurulation. We permanently marked the first cleavage plane by injecting one blastomere of the two-cell embryo with a fluorescent lineage marker. At the start of gastrulation, the interface between the labeled and unlabeled regions was almost randomly oriented relative to the dorsal blastopore lip, confirming our first set of observations. In embryos with the interface <60° to the plane passing through the midline of the dorsal lip, convergent movements of cells produced a confrontation of labeled and unlabeled cells along much of the dorsal midline. Thus, although the first cleavage plane and the bilateral plane were frequently not congruent, the morphogenetic movements of gastrulation and neurulation brought about an apparent congruence in many half-labeled embryos.

Lineage analysis of transplanted individual cells in embryos of Drosophila melanogaster : IV. Commitment and proliferative capabilities of mesodermal cells.

We describe the results of cell transplantation experiments performed to investigate mesodermal lineages in Drosophila melanogaster, particularly the lineages of the somatic muscles, the visceral muscles and the fat body. Cells to be transplanted were labelled by injecting a mixture of horseradish peroxidase (HRP) and fluorescein-dextran (FITC) in wild-type embryos at the syncytial blastoderm stage. For transplantation cells were removed from the ventral furrow, 8-12 min after the start of gastrulation, and individually transplanted into homotopic or heterotopic locations of unlabelled wild-type hosts of the same age. HRP labelling in the resulting cell clones was demonstrated histochemically in the fully developed embryo; histotypes could be distinguished without ambiguity. Mesodermal cells were already found to be committed to mesodermal fates at the time of transplantation. They developed only into mesodermal derivatives and did not integrate in non-mesodermal organs upon heterotopical transplantation. No evidence was found for commitment to any particular mesodermal organ at the time of transplantation. The majority of somatic muscle clones contributed cells to only one segment. However, clones were not infrequently distributed through two or even three segments. Clones of fat body cells were generally restricted to a small region. However, cells of clones of visceral musculature were widely distributed. With respect to the proliferative abilities of transplanted cells the clones were difficult to interpret due to the syncytial character of the somatic musculature and the fact that the organization of the other organs is poorly understood. Evidence from histological observations of developing normal embryos indicates only three mitoses for mesodermal cells. Clones larger than seven cells were not found when embryos were fixed previous to germ-band shortening; larger clones were found in the fat body and visceral musculature after fixing the embryos at the end of organogenesis. Quantitative considerations suggest that a few mesodermal cells might perform more than three mitoses.

SEM analysis of normal and abnormal gastrulae of lymnaea palustris: [formula omitted][formula omitted] and [formula omitted][formula omitted]. Div. Nat. Sci., New College-USF, Sarasota, Florida, USA

Changes in the pattern of protein synthesis were analyzed during the in vitro development of the micromere-primary mesenchyme cell line of the sea urchin embryo. Micromeres were isolated and cultured from 16-cell stage embryos, and primary mesenchyme cells were isolated and cultured from early gastrulae. Both cell isolates developed normally in culture with about the same timing as their in situ counterparts in control embryos. Newly synthesized proteins were labeled with [3H]valine at several stages of development and were analyzed by two-dimensional polyacrylamide gel electrophoresis and fluorgraphy. The electrophoretic pattern of labeled proteins changed dramatically during development. More than half of the analyzed proteins underwent qualitative or quantitative changes in their relative rates of valine incorporation and these changes were highly specific to this cell line. Almost all of the changes were initiated prior to gastrulation and many prior to hatching. The highest frequency of changes in the micromere pattern of protein synthesis occurred between hatching and the start of gastrulation. This peak of activity coincided with the normal time of ingression of the primary mesenchyme and preceded the differentiation of spicules by more than 30 hr. Most of the observed changes were characterized as either decreases in the synthesis of proteins that showed maximum incorporation at the 16-cell stage or increases in the synthesis of proteins that showed maxima in the fully differentiated cells. Very few proteins exhibited transient synthetic maxima at intermediate stages. Thus, the program of protein synthesis associated with the development of micromeres consists largely of a switch in emphasis from early to late proteins, with the primary time of switching being between hatching and the onset of gastrulation.

Temporal and spatial regulation of fibronectin in early Xenopus development

Pattern formation and temporal control of gene expression in Xenopus development were investigated using fibronectin as a biochemical marker. We determined the spatial localization of fibronectin in the embryo by immunofluorescence and the temporal program of its expression by biosynthesis studies and Western blotting techniques. At the start of gastrulation, fibronectin is localized on the roof of the blastoccel which serves as the surface upon which mesodermal cells will migrate. However, since we find fibronectin secreted by all parts of the embryo, localization is probably achieved through spatially localized receptors that bind secreted fibronectin. Fibronectin levels and fibronectin synthesis rates increase following the midblastula stage. This increase is independent of transcription and therefore involves activation of maternal RNA for fibronectin. Since this message mobilization also occurs in activated but unfertilized eggs, this event must be regulated separately from the midblastula transition.

The program of protein synthesis during the development of the micromere-primary mesenchyme cell line in the sea urchin embryo

Changes in the pattern of protein synthesis were analyzed during the in vitro development of the micromere-primary mesenchyme cell line of the sea urchin embryo. Micromeres were isolated and cultured from 16-cell stage embryos, and primary mesenchyme cells were isolated and cultured from early gastrulae. Both cell isolates developed normally in culture with about the same timing as their in situ counterparts in control embryos. Newly synthesized proteins were labeled with [ 3H]valine at several stages of development and were analyzed by two-dimensional polyacrylamide gel electrophoresis and fluorgraphy. The electrophoretic pattern of labeled proteins changed dramatically during development. More than half of the analyzed proteins underwent qualitative or quantitative changes in their relative rates of valine incorporation and these changes were highly specific to this cell line. Almost all of the changes were initiated prior to gastrulation and many prior to hatching. The highest frequency of changes in the micromere pattern of protein synthesis occurred between hatching and the start of gastrulation. This peak of activity coincided with the normal time of ingression of the primary mesenchyme and preceded the differentiation of spicules by more than 30 hr. Most of the observed changes were characterized as either decreases in the synthesis of proteins that showed maximum incorporation at the 16-cell stage or increases in the synthesis of proteins that showed maxima in the fully differentiated cells. Very few proteins exhibited transient synthetic maxima at intermediate stages. Thus, the program of protein synthesis associated with the development of micromeres consists largely of a switch in emphasis from early to late proteins, with the primary time of switching being between hatching and the onset of gastrulation.

The fine structure of Comanthus japonica (Echinodermata: Crinoidea) from zygote through early gastrula

The fine structure of the early embryo of Comanthus has been described by scanning and transmission electron microscopy at approximately 20-min intervals from zygote (20 min) through early gastrula (260 min). In normally developing (and presumably monospermic) embryos, some non-fertilizing sperm were invariably trapped in the perivitelline space; this suggests that there is an effective block to polyspermy at the level of the plasma membrane. No trace of a hyaline layer is encountered in the pervitelline space. At first cleavage, which begins unilaterally at the animal pole, the contractile ring filaments are rather thick (50–150 Å) in comparison to those known for other marine invertebrates. From first cleavage through early gastrula, the lateral surfaces of the blastomeres are broadly adherent, and there is an intercellular material, presumably an adhesive, in the intercellular space. The blastocoel first appears during the four-cell stage. From the eight-cell stage through the start of gastrulation, only one opening, the vegetal pore, connects the blastocoel with the perivitelline space. Gastrulation begins at the 50–100-cell stage, while the vegetal pore is still open, and a clearly defined blastula stage is bypassed. Gastrulation is by a novel process, which I have called holoblastic involution. At gastrulation the eight most vegetal blastomeres, which encircle the vegetal pore, shoot out erect, unbranched filopodia for many microns through the blastocoel. The filopodia adhere to the blastocoelic surfaces of the animal blastomeres and contract, pulling the vegetal blastomeres into the blastocoel. The migrated vegetal blastomeres adhere to one another, forming the entoderm in the vegetal region of the embryo; the remaining blastomeres become the ectoderm. Soon after the completion of cell migration, the entodermal blastomeres appear to cast off their contractile microappendages and adhesive membranes into the blastocoel.