engrailed duplication events during the evolution of barnacles.

Barnacles (Cirripedia) are a subclass of Crustacea. Their peculiar segmentation pattern (few segments, absence of abdominal segments, and, in the parasitic rhizocephalan, loss of segmentation at the adult stage) prompted us to study the engrailed gene, which encodes a homeodomain transcription factor and is expressed in arthropods in the posterior half of each segment. We searched for engrailed genes by PCR in a representative cross section of the Cirripedia. Eight unambiguous engrailed genes were cloned from five species, three genes belonging to the same species (Elminius modestus). This implies two duplication events. Molecular phylogenies were constructed and a cladistic approach was applied to the most informative sites. The results indicate that the older duplication (en.a/en.b) is probably very ancient and concerns the whole cirripedean subclass, whereas the other (en.a1/en.a2) is specific to the Elminius lineage.

Cirripede phylogeny using a novel approach: molecular morphometrics.

We present a new method using nucleic acid secondary structure to assess phylogenetic relationships among species. In this method, which we term "molecular morphometrics," the measurable structural parameters of the molecules (geometrical features, bond energies, base composition, etc.) are used as specific characters to construct a phylogenetic tree. This method relies both on traditional morphological comparison and on molecular sequence comparison. Applied to the phylogenetic analysis of Cirripedia, molecular morphometrics supports the most recent morphological analyses arguing for the monophyly of Cirripedia sensu stricto (Thoracica + Rhizocephala + Acrothoracica). As a proof, a classical multiple alignment was also performed, either using or not using the structural information to realign the sequence segments considered in the molecular morphometrics analysis. These methods yielded the same tree topology as the direct use of structural characters as a phylogenetic signal. By taking into account the secondary structure of nucleic acids, the new method allows investigators to use the regions in which multiple alignments are barely reliable because of a large number of insertions and deletions. It thus appears to be complementary to classical primary sequence analysis in phylogenetic studies.

Barnacle duplicate engrailed genes: divergent expression patterns and evidence for a vestigial abdomen.

Cirripedes (barnacles) are crustaceans that are characterized by a very peculiar body plan, in particular by the lack of an abdomen. To study their body plan, we searched for their engrailed gene. We found two engrailed (en.a/en.b) genes in cirripedes. The two engrailed genes of the rhizocephalan barnacle Sacculina carcini are expressed in the posterior compartment of developing segments and appendages. When the neuroectoderm differentiates into epidermis and neuroderm the expression patterns of en.a and en.b diverge dramatically. en.a expression fades in segment epidermis whereas it is subsequently detected ventrally in reiterated putative neural cells. At the same time, en.b expression increases in the epidermis, which makes it a very good segmentation marker. Five tiny en.b stripes are observed between the sixth thoracic segment and the telson. We interpret these stripes as the molecular definition of vestigial abdominal segments, being the remnant of an ancestral state in keeping with the bodyplan of maxillopod crustaceans. engrailedexpression is the first molecular evidence for a segmented abdomen in barnacles.

Three-partner conversion induced by the P-element transposase in Drosophila melanogaster.

Conversion of one P-derived transposon into another has already been shown to occur with a measurable frequency. However, the mechanism responsible for such replacements has remained controversial. We previously proposed a mechanism involving three partners. We assumed that after excision of the P-element inserted at the target site, the double-strand break was repaired using, first, the homologous P sequences on the sister chromatid, and second, a remote template, the donor P-derived transposon. However, two other mechanisms have been proposed. The first involves two partners only, the broken end and the remote template, while the second involves transposition of the donor into the target P-element, followed by a double recombination event. Here we describe the conversion of a defective P-element using as a remote template an enhancer-trap element that is itself unable to transpose because it lacks 21 bp at its 5' end. This result makes it possible to exclude the possibility that this conversion event occurred after transposition. The new allele was molecularly and genetically characterized. The occurrence of a polymorphism at position 33 of the P-element sequence and of an imperfect copy of the template on the 3' side of the converted transposon confirmed that the sister chromatid was absolutely necessary as a partner for repair. Our results show that targeting of a marked P-element is possible, even when this element is unable to transpose. This provides a means of improving recovery of conversion events by eliminating unwanted transpositions catalyzed by the P transposase.

Molecules and the body plan: the Hox genes of Cirripedes (Crustacea).

Among arthropods, Cirripedia (barnacles) are remarkable in that they completely lack abdominal segments. This feature prompted us to study the Hox genes of three cirripede species, representing a wide array of the diversity of these organisms, a segmented sessile barnacle, Elminius modestus (Thoracica), the parasite of a crab, Sacculina carcini (Rhizocephala), and the burrowing barnacle Trypetesa lampas (Acrothoracica). Using PCR amplification of genomic DNA and cDNA and library probing, we have found seven clear cirripedian homologues of the eight homeotic Hox genes known in insects, including labial and proboscipedia homologues, that were not previously reported in crustaceans. In addition we have isolated a divergent Antp-like gene, named Diva, that we homologize to the ftz gene of insects. The homeotic gene abdominalA (abdA) was not retrieved from any of these three cirripede species. By contrast, we have found all eight homeotic homologue genes, including abdA, in Ulophysema oeresundense, a crustacean possessing a well-developed abdomen, belonging to the Ascothoracica, generally thought to be the sister group of Cirripedia. Since we have found in barnacles homeobox-containing genes that are more divergent from the Antennapedia type than the typical abdA, we believe that a bona fide abdA gene would not have escaped our search. Hence, the abdA gene has been lost or is profoundly derived in sequence during the evolution leading to the cirripedian lineage. If confirmed, the lack of abdA would represent the first case in which the loss of a homeotic gene is correlated with a change in body plan during the evolution of metazoans.

The caudal gene of the barnacle Sacculina carcini is not expressed in its vestigial abdomen.

We report the characterization of a caudal gene from the rhizocephalan cirripede Sacculina carcini and its embryonic and larval expression patterns. Cirripedes are maxillopodan crustaceans that are devoid of any complete abdominal segment at the adult stage. We currently explore the genetic basis of this peculiar body plan. In a previous study we have shown that they probably lack the abdominalA gene, while possessing the other Hox genes shared by arthropods. However, at least a part of the genetic program might be conserved, since the engrailed.a and engrailed.b genes are expressed in a posterior region that we interpret as a relic of an ancestral abdomen. Here we show first that the Sacculina caudal gene is expressed early in embryogenesis, which makes it the earliest genetic marker evidenced in the development of Sacculina and of any other crustacean species. It is expressed later in the embryo in the caudal papilla, a posterior proliferating zone of cells. During the larval stages, the caudal gene is first expressed in the whole thoracic region; then its expression regresses to the posterior end of the larva. Surprisingly, it is never expressed in the vestigial abdomen. This lack of expression of the Sacculina caudal gene in a posterior region, at odds with what is known in all other studied metazoan species, might be correlated with the defective development of the abdomen.

The use of the Magnetip double-J ureteral stent in urological practice.

The Magnetip* double J type ureteral stent has been used in a wide variety of clinical urological settings. We reviewed the use of the stent in 50 patients. In 45 patients stents were placed in conjunction with extracorporeal shock wave lithotripsy, stone manipulation, obstruction due to pregnancy or malignancy, pyeloplasty and ureteroneocystostomy. In 83 per cent of the attempts the stent was placed successfully. Retrieval with the Magnetriever* was accomplished in 86 per cent of the cases (100 per cent in female and 76 per cent in male patients). Details of stent use are described.

Cloning and expression of the engrailed.a gene of the barnacle Sacculina carcini.

Cirripedia (barnacles) constitute a crustacean monophyletic taxon which is very well defined by several synapomorphies. In particular, all cirripedes are composed of six thoracic segments, but are devoid of any complete abdominal segment. This body plan is preserved in the adult in non-parasitic groups, while the parasitic rhizocephalan cirripedes completely lose arthropodian segmentation at the adult stage. These traits make them a particularly favourable model for studying the formation and maintenance of segmental identity. For the above reasons, it seemed worthwhile to look at the segmentation gene engrailed in a cirripede. A complete engrailed.a cDNA was isolated from larvae of the rhizocephalan cirripede Sacculina carcini. Its expression was monitored during larval development by use of the monoclonal antibody MAb4D9 directed against the Drosophila homologous proteins. The Sacculina engrailed.a gene is expressed during the second and third larval stages in stripes within a posterior area corresponding to the presumptive trunk segments. Surprisingly, these stripes appear in a posterior to anterior sequence. Six engrailed.a stripes characterize the thoracic segments of the cirripedean ground plan.