Dorsal Body Wall Research Articles

Mirror, a Drosophila homeobox gene in the iroquois complex, is required for sensory organ and alula formation

The Drosophila notum, the dorsal body wall of the thorax, is subdivided genetically into longitudinal domains (Calleja, M., Moreno, E., Pelaz, S. and Morata, G. (1996) Science 274, 252-255). Two homeobox genes clustered in the iroquois complex, araucan and caupolican, regulate proneural genes and are required for development of sensory bristles in the lateral notum (Gomez-Skarmeta, J. L., del Corral, R. D., de la Calle-Mustienes, E., Ferres-Marco, D. and Modolell, J. (1996) Cell 85, 95-105). An iroquois-related homeobox gene, mirror, was recently isolated and is localized close to the iroquois complex region (McNeil, H., Yang, C.-H., Brodsky, M., Ungos, J. and Simon, M. A. (1997) Genes and Development 11, 1073-1082; this study). We show that mirror is required for the formation of the alula and a subset of sensory bristles in the lateral domain of the notum. Genetic analysis suggests that mirror and the other iroquois genes interact to form the alula as well as the sensory organs. Based on similarities between mirror and the iroquois genes in their genetic map positions, expression, protein structure and function, mirror is considered a new member of the iroquois complex and is involved in prepatterning sensory precursor cells in the lateral notum.

Regulation and function of tinman during dorsal mesoderm induction and heart specification in Drosophila.

The homeobox gene tinman plays a key role in the specification of Drosophila heart progenitors and the visceral mesoderm of the midgut, both of which arise at defined positions within dorsal areas of the mesoderm. Here, we show that in addition to the heart and midgut visceral mesoderm, tinman is also required for the specification of all dorsal body wall muscles. Thus it appears that the precursors of the heart, visceral musculature, and dorsal somatic muscles are all specified within the same broad domain of dorsal mesodermal tinman expression. Locally restricted activities of tinman are also observed during its early, general mesodermal expression, where tinman is required for the activation of the homeobox gene buttonless in precursors of the "dorsal median" (DM) glial cells along the ventral midline. These observations, together with others showing only mild effects of ectopic tinman expression on heart development, indicate that tinman function is obligatory, but not sufficient to determine individual tissues within the mesoderm. Therefore, we propose that tinman has a role in integrating positional information that is provided by intersecting domains of additional regulators and signals, which may include Wingless, Sloppy Paired, and Hedgehog in the dorsal mesoderm and EGF-signaling at the ventral midline. Previous studies have shown that Dpp acts as an inductive signal from dorsal ectodermal cells to induce tinman expression in the dorsal mesoderm, which, in turn, is needed for heart and visceral mesoderm formation. In the present report, we show that Thickveins, a type I receptor of Dpp, is essential for the transmission of Dpp signals into the mesoderm. Constitutive activity of Tkv in the entire mesoderm induces ectopic tinman expression in the ventral mesoderm, and this results in the ectopic formation of heart precursors in a defined area of the ventrolateral mesoderm. We further show that Screw, a second BMP2/4-related gene product, Tolloid, a BMP1-related protein, and the zinc finger-containing protein Schnurri, are required to allow full levels of tinman induction during this process. It is likely that some of these functional and regulatory properties of tinman are shared by tinman-related genes from vertebrates that have similarly important roles in embryonic heart development.

The mup-4 locus in Caenorhabditis elegans is essential for hypodermal integrity, organismal morphogenesis and embryonic body wall muscle position.

mup-4 is a member of a set of genes essential for correct embryonic body wall muscle cell positions in Caenorhabditis elegans. The mup-4 phenotype is variably expressed and three discrete arrest phenotypes arise during the phase of embryonic development when the worm elongates from a ball of cells to its worm shape (organismal morphogenesis). Mutants representing two of the phenotypic classes arrest without successful completion of elongation. Mutants of the third phenotypic class arrest after completion of elongation. Mutants that arrest after elongation display profound dorsal and ventral body wall muscle cell position abnormalities and a characteristic kinked body shape (the Mup phenotype) due to the muscle cell position abnormalities. Significantly, genetic mosaic analysis of mup-4 mutants demonstrates that mup-4 gene function is essential in the AB lineage, which generates most of the hypodermis (epidermis), a tissue with which muscle interacts. Consistent with the genetic mosaic data, phenotypic characterizations reveal that mutants have defects in hypodermal integrity and morphology. Our analyses support the conclusion that mup-4 is essential for hypodermal function and that this function is necessary for organismal morphogenesis and for the maintenance of body wall muscle position.

Crawling motor patterns induced by pilocarpine in isolated larval nerve cords of Manduca sexta.

1. Larval crawling is a bilaterally symmetrical behavior that involves an anterior moving wave of motor activity in the body wall muscles in conjunction with sequential movements of the abdominal prolegs and thoracic legs. The purpose of this study was to determine whether the larval CNS by itself and without phasic sensory feedback was capable of producing patterned activity associated with crawling. To establish the extent of similarity between the output of the isolated nerve cord and crawling, the motor activity produced in isolated larval nerve cords was compared with the motor activity from freely crawling larvae. 2. When exposed to the muscarinic receptor agonist pilocarpine (1.0 mM), isolated larval nerve cords produced long-lasting rhythmic activity in the motor neurons that supply the thoracic leg, abdominal body wall, and abdominal proleg muscles. The rhythmic activity evoked by pilocarpine was abolished reversibly and completely by bath application of the muscarinic-receptor antagonist atropine (0.01 mM) in conjunction with pilocarpine (1.0 mM), suggesting that the response was mediated by muscarinic-like acetylcholine receptors. 3. Similar to crawling in intact animals, the evoked activity in isolated nerve cords involved bilaterally symmetrical motor activity that progressed from the most posterior abdominal segment to the most anterior thoracic segment. The rhythmic activity in thoracic leg, abdominal proleg, and abdominal body wall motor neurons showed intrasegmental and intersegmental cycle-to-cycle coupling. The average cycle period for rhythmic activity in the isolated nerve cord was approximately 2.5 times slower than the cycle period for crawling in intact larvae, but not more variable. 4. Like crawling in intact animals, in isolated nerve cords, bursting activity in the dorsal body wall motor neurons occurred before activity in ventral/lateral body wall motor neurons within an abdominal segment. The evoked bursting activity recorded from the proleg nerve was superimposed on a high level of tonic activity. 5. In isolated nerve cords, bursts of activity in the thoracic leg levator/extensor motor neurons alternated with bursts of activity in the depressor/flexor motor neurons. The burst duration of the levator/extensor activity was brief and remained relatively steady as cycle period increased. The burst duration of the depressor/ flexor activity occupied the majority of an average cycle and increased as cycle period increased. The phase of both levator/extensor motor nerve activity and depressor/flexor motor nerve activity remained relatively stable over the entire range of cycle periods. The timing and patterning of thoracic leg motor neuron activity in isolated nerve cords quantitatively resembled thoracic leg motor activity in freely crawling larvae. 6. The rhythmic motor activity generated by an isolated larval nerve cord resembled a slower version of normal crawling in intact larvae. Because of the many similarities between activity induced in the isolated nerve cord and the muscle activity and movements of thoracic and abdominal segments during crawling, we concluded that central mechanisms can establish the timing and patterning of the crawling motor pattern and that crawling may reflect the output of a central pattern generating network.

Normal Expression and the Effects of Ectopic Expression of the Drosophila muscle segment homeobox (msh) Gene Suggest a Role in Differentiation and Patterning of Embryonic Muscles

Myogenesis is a several step process that requires genes involved in specifying mesoderm lineage and genes involved in determining muscle identity, differentiation, and patterning. We report here on the isolation, characterization, and expression pattern of a cDNA clone encoded by the previously uncharacterized Drosophila muscle segment homeobox (msh) gene and its possible role in myogenesis. The amino acid sequence of the msh homeobox domain is highly homologous to the homeodomains of the Drosophila S59 and empty spiracles genes and the Hox 7 and Hox 8 family of vertebrate homeobox genes. In addition, the 5′ end of msh has 52% sequence identity to the 5′ end of the empty spiracles gene and encodes several stretches of amino acids rich in serine, alanine, proline, glutamine, and acidic amino acids, indicating potential domains of regulatory activity. The expression of msh is initially detected at about stage 6 in the dorsal lateral ectoderm of the embryo and later in the developing central (CNS) and peripheral nervous systems. During germ band retraction (stage 12), msh continues to be expressed in cells of the nervous system as well as cells of the somatic mesoderm corresponding mostly to the developing dorsal and lateral somatic body wall muscles. These mesodermal cells, which continue to express msh in daughterless mutant embryos, undergo an increase in cell number in neurogenic mutants. By late stage 14 of embryonic development, msh expression is greatly reduced or absent in most or all mesoderm and muscle but continues in the CNS until hatching. Ectopic expression of msh in the mesoderm results in altered expression of the S59 and nau/Dmyd genes leading to a loss of some muscles and defects in the patterning of others, suggesting that the muscle defects are at the level of recruitment and/or patterning of muscle precursor cells. Thus the similarity of Drosophila msh expression to that of the homologous vertebrate Hox 7 and Hox 8 genes together with the effects of ectopic expression of msh in the mesoderm suggest a role for the msh-like family of genes in mesodermal and muscle differentiation and patterning.

Sprouting and connectivity of embryonic leech heart excitor (HE) motor neurons in the absence of their peripheral target.

The rhythmic pumping of the hearts in the medicinal leech, Hirudo medicinalis, is neurogenic and mediated by a defined circuit involving identified interneurons in a central pattern generator (CPG) and segmentally iterated motor neurons that drive the heart muscle. During early embryogenesis, presumptive heart excitor (HE) motor neurons extend many axon branches into the body wall; they later innervate the heart while retracting the supernumerary peripheral axons, and only much later in development receive synaptic input from the central pattern generator (Jellies, Kopp and Bledsoe (1992) J. Exp. Biol., 170, 71-92.). In this study, HE motor neurons were deprived of an early interaction with the heart by surgical ablation of a circumscribed portion of body wall including the heart primordium. Anatomical and electrophysiological data were obtained using intracellular techniques to examine the hypothesis that peripheral interactions with the developing heart provide instructive cues for the final differentiation of these neurons. Target-deprived HE motor neurons continued to extend multiple axons in ventral, lateral and dorsal body wall throughout late embryonic and into postembryonic stages and they extended anomalous axons within the CNS. This resembles the early embryonic growth of HE motor neurons before heart tube differentiation. Furthermore, HE motor neurons deprived of heart contact exhibited tonic activity similar to the situation during early development before they are contacted by the CPG interneurons. In contrast, sham-operated and contralateral HE motor neurons oscillated normally. These results suggest that heart tube contact is specifically required for at least some aspects of HE development and provide a framework in which to identify cell-cell interactions that are involved in matching neurons and targets to generate behaviorally relevant neural circuits.

Developing neurons use a putative pioneer's peripheral arbor to establish their terminal fields

Pioneer neurons are known to guide later developing neurons during the initial phases of axonal outgrowth. To determine whether they are also important in the formation of terminal fields by the follower cells, we studied the role of a putative leech pioneer neuron, the pressure-sensitive (PD) neuron, in the establishment of other neurons' peripheral arbors. The PD neuron has a major axon that exits from its segmental ganglion to grow along the dorsal-posterior (DP) nerve to the dorsal body wall, where it arborizes extensively mainly in its own segment. It also has two minor axons that project to the two adjacent segments but branch to a lesser degree. We found that the peripheral projections of several later developing neurons, including the AP motor neuron and the TD sensory neuron, followed, with great precision, the major axon and peripheral arbor of the consegmental PD neuron, up to its fourth-order branches. When a PD neuron was ablated before it had grown to the body wall, the AP and TD axons grew normally toward and reached the target area, but then formed terminal arbors that were greatly reduced in size and abnormal in morphology. Further, if the ablation of a PD neuron was accompanied by the induction, in the same segment, of greater outgrowth of the minor axon of a PD neuron from the adjacent segment, the arbors of the same AP neurons grew along these novel PD neuron branches. These results demonstrate that the peripheral arbor of a PD neuron is a both necessary and sufficient template for the formation of normal terminal fields by certain later growing follower neurons.

Interactions between segmental homologs and between isoneuronal branches guide the formation of sensory terminal fields

Process outgrowth and peripheral field innervation by an identified mechanosensory neuron were examined in the intact embryonic leech. The dorsal pressure-sensitive (PD) neurons of the leech CNS are found as bilateral pairs in every segmental ganglion, and are amenable to study at early ages in intact embryos. Each PD has one major axonal projection that putatively pioneers the nerve to the dorsal body wall and branches extensively in its own segment, and two minor projections that innervate, via neighboring ganglia, smaller areas in adjacent segments. We found that adjacent embryonic PD cells form overlapping terminal fields in the body wall, but that the extent of overlap was governed by inhibitory interactions among these fields. When one PD neuron was ablated, the adjacent PD cell changed its peripheral arborization by (1) its major axon producing more filopodia and extending longer side branches toward the ablated cell and (2) its minor axon producing a large arbor in the operated segment. Interestingly, although growth was biased toward the side of the ablated neuron, reduced outgrowth of the PD cell was found on the side away from the ablation, while the total extent of arborization of the PD cell kept relatively constant. Further, we found that axotomy of the major PD projection resulted in extensive outgrowth of its minor projections. These results suggest that a single PD neuron has a limited capacity for growth, each of its branches growing at the expense of the others, and that inhibitory interactions between neighboring PD neurons influence the extent and direction of that growth.

Oogenesis in a placental viviparous onychophoran

This first ultrastruetural study of oogenesis in a placental viviparous onychophoran describes oocyte differentiation, cell interactions and reveals various unusual cellular features. The viviparous onychophoran Plicatoperipatus jamaicensis has paired ovaries medially located, attached to the dorsal body wall by muscular terminal filaments. The rest of the female reproductive tract consists of paired spermathecae oviduct/uteri (hereafter referred to as uterus). Bulbous spermathecae are joined to the oviducts by ducts. Also continuous with the oviduct lumen are two tubular structures whose lumina open to the hemolymph. The uteri contain a progression of developmental stages from implantation through stalked morulae, blastocysts, larvae and juveniles about to be born. Growing oocytes are characterized by large germinal vesicles showing synaptonemal complexes. Oocytes are surrounded by flattened follicle cells that possess extensive bundles of thick and thin filaments. Mature oocytes contain little or no yolk, but are unique among organisms in accumulating a large central reservoir of stored glycogen. The lack of yolk reflects the placental viviparous nature of the reproductive process. The glycogen reservoir provides a rapidly accessible energy source for early developmental stages. Particularly prominent also are unusually extensive and highly elaborate Golgi complexes in the cortical and peri-nuclear ooplasm. While extensive Golgi complexes have been described in oocytes of a variety of species, the particularly exaggerated size and amount of Golgi in these onychophorans suggests they may provide excellent material for the study of Golgi function. The features of the oocyte and placental viviparity show this is an ideal model to investigate the nature of the placental reproductive process analogous to mammals in an invertebrate and its implications to oogenesis.

Interactions between primordial germ cells play a role in their migration in mouse embryos

Primordial germ cells (PGCs) are the founder cell population of the gametes which form during the sexually mature stage of the life cycle. In the mouse, they arise early in embryogenesis, first becoming visible in the extraembryonic mesoderm, posterior to the primitive streak, at 7.5 days post coitum (d.p.c.). They subsequently become incorporated into the epithelium of the hind gut, from which they emigrate (9.5 d.p.c.) and move first into the dorsal mesentery (10.5 d.p.c.), and then into the genital ridges that lie on the dorsal body wall (11.5 d.p.c.). We have used confocal microscopy to study PGCs stained with an antibody that reacts with a carbohydrate antigen (Stage-Specific Embryonic Antigen-1, SSEA-1) carried on the PGC surface. This allows the study of the whole PGC surface, at different stages of their migration. The appearance of PGCs in tissue sections has given rise to the conventional view that they migrate as individuals, each arriving in turn at the genital ridge. In this paper, we show that PGCs leave the hind gut independently, but then extend long (up to 40 microns) processes, with which they link up to each other to form extensive networks. During the 10.5-11.5 d.p.c. period, these networks of PGCs aggregate into groups of tightly apposed cells in the genital ridges. As this occurs, their processes are lost, and their appearance suggests they are now non-motile. Furthermore, we find that PGCs taken from the dorsal mesentery at 10.5 d.p.c. perform the same sequence of movements in culture.(ABSTRACT TRUNCATED AT 250 WORDS)

Description of Neoechinorhynchus carinatus n. sp. (Acanthocephala: Neoechinorhynchidae) from the Sharpfin Chubsucker, Erimyzon tenuis, of Louisiana and Mississippi

Neoechinorhynchis carinatus n. sp. is described from Erimyzon tenuis (Agassiz) from southern Louisiana and Mississippi. Neoechinorhynchus carinatus is distinguished from other Neoechinorhynchus species, with unequal lemnisci, of North American freshwater fishes by having a dorsal body wall that is usually 2-3× thicker than the ventral body wall and small proboscis hooks with lateral hooks of the anterior circle being the largest and hooks of the middle and posterior circles about equal in length

Development of the Deformed protein pattern in the embryo of the honeybee Apis mellifera L. (Hymenoptera).

We have raised antiserum against part of the Deformed (Dfd) protein of the honeybee and describe here the expression pattern of the Dfd protein during honeybee embryogenesis. Dfd protein is first stained in the prospective gnathal region of the cellular blastoderm. This circumferential band corresponds to the distribution of Dfd mRNA described earlier, and to the blastodermal Dfd expression pattern in Drosophila. Using an antibody against the engrailed (en) protein of Drosophila, we found that at the beginning of gastrulation Dfd expression in the honeybee, as in Drosophila, is restricted to the future intercalary, mandibular and maxillary segments. During gastrulation, the mesodermal nuclei loose the Dfd label gradually from anterior to posterior, and in the ectoderm the most posterior ventral cells loose Dfd while retaining en staining; thus, in contrast to what has been described for Drosophila, the posterior Dfd expression border seems to move forward ventrally to the parasegmental boundary within the maxillary segment. In the late germ band, the lateral tips of the Dfd-expressing band are connected across the dorsal side by a row of amnion cells with strongly staining large nuclei. After dorsal closure, a narrow stripe of Dfd-staining dorsal cells behind the neck region may indicate that the maxillary segment contributes to the dorsal body wall posterior to the head capsule. Thus, apart from some minor deviations, the Dfd expression pattern in the honeybee strongly resembles that in Drosophila prior to head involution. This is compatible with the assumption that head involution (which is a special adaption in higher dipterans) ensues after a rather conserved course of early head development in which Dfd appears to play a basic role.

Comments on the morphology of the pre-oral spine in Argulus (Crustacea: Branchiura).

A histological study of the pre-oral spine and associated structures in Argulus japonicus reveals that two exceptionally long and slender muscles casuse retraction of the spine into the spinal sheath. Originating from the body wall mid-dorsally the two muscles proceed through the nerve ring and the blood-filled lumen of the spinal sheath before being inserted on a bolster of tissue constituting the base of the spine. Movement of the spinal sheath is affected by two muscles originating from a common apodeme in the dorsal body wall, just posterior to the nauplius eye, and inserted on the lateral walls of the base of the spinal sheath. Two ducts, each leading from a glandular complex in the lateral parts of the carapace, proceed rostrally through the haemocoelic spaces. After passing the tritocerebrum laterally they accompany the retractor muscles of the spine on their course to the base of the spine. Passing through the bolster of tissue at the base of the spine the two ducts run all the way up to the tip of the spine where they open separately. The glandula praeboscoidalis is not associated with the pre-oral spine since its duct runs posteriorly in the wall of the proboscis. The labial spines are associated with giant glandular cells located beneath the optic tracts. © 1992 Wiley-Liss, Inc.

Epithelio--mesenchymal interactions are critical for Quox 7 expression and membrane bone differentiation in the neural crest derived mandibular mesenchyme.

In higher vertebrates, branchial arch mesenchyme (ectomesenchyme) is derived from the cephalic neural crest. The ectomesenchyme of the mandibular arch yields the Meckel's cartilage and several membrane bones. We previously reported the isolation of a quail homeobox gene, Quox 7. In common with its mouse counterpart Hox 7, Quox 7 is highly expressed in the medioventral part of the mandibular arch and later in the precursor cells of the membrane bones. Since bone differentiation from ectomesenchyme is strictly dependent upon a signal provided by the mandibular epithelium, we decided to see whether the regulation of Quox 7 gene activity might be correlated with epithelio--mesenchymal interactions. Quox 7 expression was studied in E3 mandibular ectomesenchyme cultured in vitro or grafted on the chick chorioallantoic membrane either alone or recombined with the homotopic and heterotopic epithelia. We found that Quox 7 mRNA was undetectable after 48 h in cultures of mesenchyme alone while it remained abundant in non-cartilaginous tissue of the mandibular arch ectomesenchyme recombined with its own epithelium. The signal provided by the mandibular epithelium for Quox 7 expression can also arise from various heterotopic epithelia, e.g. of dorsal or ventral body wall and of limb bud. Thus the effect of the epithelium on Quox 7 expression in mesenchymal cells strictly parallels that on bone formation. These results strongly suggest that the epithelio-mesenchymal interactions have an essential role on the regulation of Quox 7 gene, the product of which seems to be, in turn, necessary for the execution of the skeletal developmental program in the facial area.

Morphological and histochemical comparison of the cells elicited by ectopic bone implants and tibial osteoclasts

Pellets of mineralized and demineralized bone and a composite mixture of mineralized and demineralized, devitalized bone particles were implanted subcutaneously on the dorsal body wall of young adult rats. Two weeks post-implantation, the pellets were removed and processed for histochemical and morphological analyses. Rat proximal tibia was also processed for evaluation. The levels of tartrate-resistant acid phosphatase (TRAP) activity in the multinucleated giant cells (MNGCs) from each of the three implants and from osteoclasts were assessed using an image analyzer. The osteoclasts from the proximal tibia and the majority of MNGCs from the demineralized implants demonstrated high levels of TRAP activity. MNGCs from the mineralized implants showed either a low level or absence of TRAP activity. Most MNGCs from the composite implants exhibited a low level of TRAP activity; however, there was a population of cells that demonstrated a high level of reaction product, similar to that seen in the tibia and demineralized implant. Morphologically, osteoclasts from the proximal tibia and from the osteogenic demineralized implant exhibited ruffled borders. A small population of MNGCs from the composite implant also revealed osteoclastic features. In summary, MNGCs from the mineralized implant did not exhibit a level of TRAP reaction product or morphology similar to osteoclasts, while the majority of cells from the demineralized implant and a subpopulation of the MNGCs elicited by the composite implant did demonstrate TRAP expression and morphology similar to osteoclasts. The expression of osteoclastic characteristics in cells at an ectopic site may be dependent on accessory signals from the skeletal microenvironment; such signals appear to be absent from or incomplete in the mineralized implants but appear to be present when demineralized bone particles are implanted.

Torsion in the asteroid ray.

Although torsion (axial twisting) is rare in living structures, some starfish routinely use ray torsion in the righting process. This study examined the torsional properties of the ray and dorsal body wall of Echinaster spinulosus (Echinodermata: Asteroida: Spinulosidae). The body wall of the ray is composed of a reticular skeleton composed of CaCO3 ossicles sandwiched between collagenous layers in which the fibers are orthogonally oriented. G, the shear modulus of the dorsal body wall, determined from a torsion pendulum, was 95.3 MPa, and damping was extremely pronounced (Δ = 1.36). The torque-rotation curve was J-shaped, and G determined from this data was 176 MPa. Torsional stress relaxation was rapid, with the relaxation spectrum skewed to the left and τmax = 2.3 for anaesthetized tissue and 2.7 for unanaesthetized tissue. Analysis of ray geometry showed that the ambulacral groove reduced torsional stiffness by about two orders of magnitude compared to an ungrooved ray. Ray taper reduced the stiffness of the tip relative to the root. The shear stress distribution determined for the ray cross-section showed stress concentrations around the ambulacral arch, the outer body wall surface, and the inner body wall surface. The ossicles are situated to minimize torsional stiffness of the ray.

Genital ridges exert long-range effects on mouse primordial germ cell numbers and direction of migration in culture

The functional gametes of all vertebrates first arise in the early embryo as a migratory population of cells, the primordial germ cells (PGCs). These migrate to, and colonise, the genital ridges (GR) during the early organogenesis period, giving rise to the complete differentiating gonad. PGCs first become visible by alkaline phosphatase staining in the root of the developing allantois at 8.5 days post coitum (dpc). At 9.5 dpc they are found in the wall of the hind-gut and, during the following three days, they migrate along the hind-gut mesentery to the dorsal body wall, and then to the genital ridges. By 12.5 dpc, the great majority of PGCs have colonised the genital ridges. During this period the number of PGCs increases from less than 100 to approximately 4000. In a previous paper (Donovan et al. 1986), we showed that 10.5 dpc PGCs can be explanted from the hind-gut mesentery, and will spread and migrate on feeder cell layers. We showed also that the intrinsic ability of PGCs to spread and migrate changes as they colonise the genital ridges. In this paper, we examine extrinsic factors that control PGC behaviour in vitro. Using PGCs taken from 8.5 dpc embryos, at the beginning of their migratory phase, we show that culture medium conditioned by 10.5 dpc genital ridges causes an increase in the number of PGCs in these cultures. We also show that PGCs migrate towards 10.5 dpc genital ridges in preference to other explanted organs. These experiments show that genital ridges exert long-range effects on the migrating population of PGCs.(ABSTRACT TRUNCATED AT 250 WORDS)

Gross and Histological Morphology of the Swimbladder and Lack of Accessory Respiratory Structures in Lepidogalaxias salamandroides, an Aestivating Fish from Western Australia

The swimbladder morphology of Lepidogalaxias salamandroides was investigated. This small (maximum size is 67 mm SL) freshwater fish is endemic to southwestern Australia. It apparently survives droughts by aestivating in the bottom mud. The physostomous swimbladder is poorly developed, consisting of a thin serous membrane that extends from the dorsal body wall to the gut. It does not have the tissue normally associated with a teleost swimbladder. Instead, it is composed of a double-layered membrane that consists of squamous epithelium with a thin basement layer of collagen fibers. It appears more like dorsal mesentery, and almost certainly cannot be used in aerial respiration. Comparisons were made with Australian galaxiids and an American umbrid fish all of which possess a well-developed physostomous swimbladder. The skin of L. salamandroides contains numerous goblet (mucous) cells, and the opercular chamber lacks adaptations for air breathing. We suggest four hypotheses whereby L. salamandroides can survive the desiccation of its habitat without air breathing.

Structure and mechanics of starfish body wall.

The structure of the dorsal body wall of the starfish Echinaster spinulosus was studied using polarized light microscopy of frozen tissues, scanning electron microscopy and histology. The collagen fibres of the body wall form a three-dimensional orthogonal web. Voids in the web contain ossicles and papulae. The orthogonal web delivers dimensional stability but allows shear necessary for ray torsion. The ossicles and fibres interact to load the fibres in tension and the ossicles in compression. Strain rates of the dorsal body wall were measured on live animals during typical movements. Uniaxial tension tests of the body wall yielded Young's moduli of 267 MPa (longitudinal), 249 MPa (transverse) and 353 MPa (bias); curves were essentially linear. The body wall was approximately linearly viscoelastic and showed hysteresis at 0.01 Hz. Stress relaxation over five decades of time (in seconds) yielded relaxation spectra with peaks in relaxation time at 2.96-3.35, depending on test direction. Stress relaxation caused the connective tissue to soften. The surface of fractured stress-relaxed tissue revealed wispy, dissociated fibril tufts, whereas unrelaxed fractures produced blunt-ended fibre bundles. Neural control was necessary for body wall integrity.

Timing of postvitellogenic ovarian changes in the ridgeback prawn Sicyonia ingentis (Penaeidae) determined by ovarian biopsy

The relative timing of postvitellogenic oocyte changes in the ridgeback prawn ( Sicyonia ingentis) was studied using a simple ovarian biopsy technique. Prawn ovaries were biopsied by puncturing the dorsal body wall and ovary with a needle attached to a syringe containing crustacean Ringers. Oocytes were removed and staged microscopically. The technique had no significant effect on survival, number of spawns, time of spawning or egg quality compared with unsampled controls. The timing of the cortical specialization (CS) stage, the germinal vesicle breakdown (GVBD) stage and ovulation were studied using repeated ovarian biopsies. The mean (± SD) time from the initiation of the cortical specialization stage to spawning was 97 ± 53 h and 84 ± 22 h in two groups of animals subjected to repeated ovarian biopsies. Ovulation required less than a few hours. It was concluded that the frequency of cortical specialization stage oocytes was an accurate index of spawning activity.