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At least 19 recordsLinked to original sources

PubMed1998

Expression of the tissue-specific transcription factor Pit-1 in the lancelet, Branchiostoma lanceolatum.

Lancelets, known also as amphioxus, are protochordates that share common archetypal features with vertebrates. Recently, several developmental and molecular biology studies have pointed out homologies between anatomical structures of lancelets and vertebrates. We have studied the head region of the lancelet, Branchiostoma lanceolatum, by means of scanning electron microscopy, immunocytochemistry, and Western blotting techniques, to localize the pituitary-specific transcription factor, Pit-1. Immunoreactive Pit-1 protein has been found in cells of two typical structures of the lancelets, the Kölliker's and Hatschek's pits. Moreover, the frontal eye complex, neurons, and the rostral nerves show Pit-1 immunoreactivity. A band of 33 kilodaltons has been resolved in lancelet extracts by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and after Western blotting, the bands have been probed by a monoclonal antibody to rat Pit-1. Our results demonstrate that Pit-1 is expressed in both neurones and receptosecretory epithelial cells of adult lancelets, and that the cells lining the two pits display ultrastructural and immunocytochemical features typical of chemoreceptosecretory/olfactory- and adenohypophyseal-like structures.

Animals↗

PubMed2001

Early expressed genes showing a dichotomous developing pattern in the lancelet embryo.

Lancelets (amphioxus), although showing the most similar anatomical features to vertebrates, never develop a vertebrate-like head but rather several structures specific to this animal. The lancelet anatomical specificity seems to be traceable to early developmental stages, such as the vertebrate dorsal and anterior-posterior determinations. The BMP and Wnt proteins play important roles in establishing the early basis of the dorsal structures and the head in vertebrates. The early behavior of BMP and Wnt may be also related to the specific body structures of lancelets. The expression patterns of a dpp-related gene, Bbbmp2/4, and two wnt-related genes, Bbwnt7 and Bbwnt8, have been studied in comparison with those of brachyury and Hnf-3beta class genes. The temporal expression patterns of these genes are similar to those of vertebrates; Bbbmp2/4 and Bbwnt8 are first expressed in the invaginating primitive gut and the equatorial region, respectively, at the initial gastrula stage. However, spatial expression pattern of Bbbmp2/4 differs significantly from the vertebrate cognates. It is expressed in the mid-dorsal inner layer of gastrulae and widely in the anterior region, in which vertebrates block BMP signaling. The present study suggests that the lancelet embryo may have two distinct developmental domains from the gastrula stage, the domains of which coincide later with the lateral diverticular and the somitocoelomic regions. The embryonic origin of the anterior-specific structures in lancelets corresponds to the anterior domain where Bbbmp2/4 is continuously expressed.

Animals↗

PubMed1997

The main features of the craniate mitochondrial DNA between the ND1 and the COI genes were established in the common ancestor with the lancelet.

We have cloned the mitochondrial DNA fragment extending from tRNA-Leu to the cytochrome oxidase subunit 1 (COI) genes of Branchiostoma lanceolatum, Myxine glutinosa, Lampetra fluviatilis, and Scyliorhinus caniculus and have determined their respective gene sequences and organization. In all four species, this region contains the ND1 and ND2 genes and the genes coding eight tRNAs, namely, tRNA-Ile, -Gln, -Met, -Trp, -Ala, -Asn, -Cys, and -Tyr. The gene order is the same in the hagfish, lamprey and dogfish. In the lancelet, the location of the tRNA genes is slightly different. The mitochondrial code of Myxine, Lampetra, and Scyliorhinus is identical to that of vertebrates. The code used by the lancelet is the same with the exception of AGA (a stop codon in vertebrates), which codes for glycine in the lancelet. From the comparison of the four maps with already published ones for other species, we propose that the main features of the craniate mtDNA between the ND1 and COI genes were established in the common ancestor to cephalochordates and vertebrates more than 400 MYA. The origin of replication of the light-strand (Ori-L), usually located between the tRNA-Asn and tRNA-Cys genes in vertebrates, was not found in the lancelet, hagfish, or lamprey (Lampetra). In contrast, it was found in the dogfish. Thus the position of Ori-L was established for the first time in the common ancestor to the Chondrichthyes and Osteichthyes and remained present in all later-emerging vertebrates.

Animals↗

PubMed2000

Left-right asymmetric expression of BbPtx, a Ptx-related gene, in a lancelet species and the developmental left-sidedness in deuterostomes.

The long-standing question of how asymmetric development or asymmetric body structures in lancelets (amphioxus) are phylogenetically related to the body plan of other animals is still untouched. Three anterior structures, the preoral pit, club-shaped gland and mouth, are remarkable asymmetric features in developing lancelets that all open on the left side of the body. A Ptx-related gene, BbPtx is the first identified transcription factor gene with an asymmetrical expression pattern in lancelets similar to that in vertebrates, and thus it may provide a clue for the above question. Expression of the BbPtx gene is first detected at the dorsal margin of the blastopore in early mid-gastrulae and then becomes restricted to the left anterodorsal wall of the primitive gut and to the developing left somitocoelomic system. Expression continues on the left side in the developing preoral pit, club-shaped gland and mouth as well as in the mesoderm at the caudal end. Unlike D-Ptx1 in Drosophila, BbPtx is not coexpressed with a fork head gene in lancelets; instead the two genes are expressed in a complementary fashion on the left side of the embryo. The expression pattern of BbPtx is not compatible with the calcichordate hypothesis of Jefferies, in which the proposed ancestor of chordates rotated its tail 90 degrees counterclockwise in relation to the head/trunk. The expression of both BbPtx and vertebrate Pitx2 in tissues derived from the coelom implies that the left-right asymmetric development has a common origin between cephalochordates and vertebrates. Considering the development of the coelom in deuterostomes, however, left-right asymmetric development involving Pitx2-related genes is rather likely to be a primitive character shared among deuterostomes.

Amino Acid Sequence↗

PubMed1998

Evidence for the presence of the tissue-specific transcription factor Pit-1 in lancelet larvae.

Recent molecular studies have noted the affinity among cephalochordates and vertebrates. In particular, a cluster of vertebrate-like homeobox genes regulates the development of the lancelet Branchiostoma lanceolatum. A previous study has outlined the expression pattern of the pituitary-specific transcription factor Pit-1 in adult lancelets. Pit-1 belongs to the POU family of transcription factors, which, like homeotic proteins, are members of the helix-turn-helix superfamily of proteins. POU is an acronym for Pit-1, Oct-1 and Oct-2, and Unc-86. In the present work, we investigated the head region of premetamorphic larvae of B. lanceolatum, by means of scanning electron microscopy, wholemount and tissue sections immunocytochemistry, and Western blotting assay, to verify the presence and distribution of Pit-1. Immunoreactive Pit-1 protein was detected in the rostral nerves and in a cluster of photoreceptor cells of the frontal eye. At the same time, an electrophoretic band of 33 kDa was shown from extracts of premetamorphic larvae and recognized by a monoclonal antibody to rat Pit-1. On the basis of the immunocytochemical and electrophoretic results, we can assume that Pit-1 may play a neuromodulatory role in the larval central nervous system. Moreover, the spatial and temporal distribution of Pit-1 protein in larva and adult lancelets agrees only in part with that described in embryonic and adult mice, suggesting different molecular controls of regional identity in the nervous system of cephalochordates and vertebrates.

Age Factors↗

PubMed2001

Establishment of left-right asymmetric innervation in the lancelet oral region.

Lancelets (amphioxus) exhibit a remarkable asymmetric development in the anterior body region, which is reflected in the peripheral nervous system even at adulthood. Not all of the anterior nerves are involved, but the left third to fifth nerves are clearly asymmetric. To trace the developmental process responsible for asymmetric innervation, the peripheral nerves in the anterior region were studied in pre- and mid-metamorphic larvae, 1-cm-long juveniles, and in adults by using whole-mount immunostaining. The mouth changes in size and location during larval life before moving ventrally and, in conjunction with this change, nerves in the oral region are also modified. The left second nerve initially innervates the oral region, but this connection is secondarily lost. As the mouth expands and shifts posteriorly, the left fifth to ninth nerves join the left third and fourth in the innervation of the oral region. The left third to sixth nerves anastomose with the oral nerve ring, which encircles the mouth on the left side. In the juveniles and adults, there are two nerve plexuses that run parallel to the margin of the oral hood. The innermost of these, the "inner oral-hood nerve plexus", is asymmetrically connected with the left third to fifth nerves on both sides. The other, the "outer oral-hood nerve plexus", is ipsilaterally connected with the third to seventh nerves on both sides. The velar nerve ring is also innervated asymmetrically by the left fourth and fifth nerves. From these observations, we suggest that the oral nerve ring is the precursor of both the inner oral-hood nerve plexus and the velar nerve ring, and that the asymmetric innervation retained in adult lancelets is related to the early anastomosis of the left nerves with the oral nerve ring. We also show that, contrary to the persistent asymmetric innervation, the axonal patterns of the anterior peripheral nervous system in developing lancelets can change.

Animals↗

PubMed1998

Expression of a twist-related gene, Bbtwist, during the development of a lancelet species and its relation to cephalochordate anterior structures.

Mesoderm formation plays a crucial role in the establishment of the chordate body plan. In this regard, lancelet embryos develop structures such as the anteriorly extended notochord and the lateral divertecula in their anterior body. To elucidate the developmental basis of these structures, we examined the expression pattern of a lancelet twist-related gene, Bbtwist, from the late gastrula to larval stages. In late-gastrula embryos, the transcripts of Bbtwist were detected in the presumptive first pair of somites and the middorsal wall of the primitive gut. The expression of Bbtwist was then upregulated in the lateral wall of somites and the notochord. At the late-neurula stage, it was also expressed in the anterior wall of the primitive gut, as well as in the evaginating lateral diverticula. No signal was detected in the left lateral diverticulum when it was separated from the gut, while in the right one, the gene was expressed later during the formation of the head coelom in knife-shaped larvae, and in the anterior part of the notochord in the same larvae. In 36-h larvae, only faint expression was detected in the differentiating notochordal and paraxial mesoderm in the caudal region. These expression patterns suggest that Bbtwist is involved in early differentiation of mesodermal subsets as seen in Drosophila and vertebrates. The expression in the anterior notochord may be related to its anterior expansion. The expression in the anterior wall of the primitive gut and its derivative, the lateral diverticula, suggests that lancelets share the capability to produce a mesodermal population from the tip of the primitive gut with nonchordate deuterostome embryos.

Amino Acid Sequence↗

PubMed1999

The tissue-specific transcription factor Pit-1 is expressed in the spinal cord of the lancelet, Branchiostoma lanceolatum.

The spinal cord of the lancelet Branchiostoma lanceolatum was studied by using a monoclonal antibody to the rat tissue-specific transcription factor, Pit-1. Our previous studies have demonstrated Pit-1 immunoreactivity in different nervous and endocrine structures of the head region of adults and in the rostral central nervous system (CNS) of larval lancelet. Our present results show the presence of Pit-1-like protein in dorso-lateral nerve cells and ependymocytes of the adult spinal cord. Using double immunofluorescence techniques, we have revealed the coexistence of the glial fibrillary acidic protein (GFAP) with Pit-1 in groups of laterally located ependymocytes. The occurrence of GFAP, a specific marker of mammalian astrocytes and radial glia, in some lancelet ependymocytes confirms that glial elements are also present in protochordates. Furthermore, other ependymocytes, located in the roof of the central canal and containing Pit-1-like protein exclusively, could be considered as ependymal tanycytes.

Animals↗

PubMed2004

Glycoconjugate profiles of the lancelet (Branchiostoma lanceolatum) ovary: a lectin histochemical study by laser confocal microscopy.

The presence and the distribution of carbohydrate moieties in ripe lancelet (Branchiostoma lanceolatum) oocytes (mean diameter 130 microm) was studied by lectin histochemistry in combination with enzyme and chemical treatments. Binding sites for eight lectins with specificities towards different glycan moieties were studied on sections of the whole body of mature female lancelets. Only three of the lectins tested reacted positively. Concanavalin-A (ConA)-binding glycoconjugates were localized in the cytoplasm, namely in yolk granules, whereas Artocarpus integrifolia (AIA) and Ricinus communis (RCA) agglutinins bound strongly to extracellular coats of the oocyte identified as the jelly coat and vitelline layer. No other tissues of the lancelet body were found to be positive to any lectin tested, except gut enterocytes which reacted strongly with AIA. Reactivity to ConA was abolished by pretreatment of sections with N-glycosidase F but not by mild alkaline hydrolysis, confirming that the glycoconjugates were of the N-linked type. On the contrary, chemical removal of O-linked chains by mild alkaline hydrolysis abolished AIA and RCA reactivity but had no effect on ConA positivity.

Animals↗

PubMed2002

beta-Catenin in early development of the lancelet embryo indicates specific determination of embryonic polarity.

The lancelet (amphioxus) embryo develops from a miolecithal egg and starts gastrulation when it is approximately 400 cells in size, in a fashion similar to that of some non-chordate deuterostomes. Throughout this type of gastrulation, the embryo develops characteristics such as the notochord and hollow nerve cord that commonly appear in chordates. beta-Catenin is an important factor in initiating body patterning. The behavior and developmental pattern of this protein in early lancelet development was examined in this study. Cytoplasmic beta-catenin was localized to the animal pole after fertilization and then was incorporated asymmetrically into the blastomeres during the first cleavage. Asymmetric distribution was observed at least until the 32-cell stage. The first nuclear localization was at the 64-cell stage, and involved all of the cells. At the initial gastrula stage, however, concentrated beta-catenin was found on the dorsal side. LiCl treatment affected the asymmetric pattern of beta-catenin during the first cleavage. LiCl also changed distribution of nuclear beta-catenin at the initial gastrula stage: distribution extended to cells on the animal side. Apparently associated with this change, expression domains of goosecoid, lhx3 and otx also changed to a radially symmetric pattern centered at the animal pole. However, LiCl-treated embryos were able to establish embryonic polarity. The present study suggests that in the lancelet embryo, polarity determination is independent of dorsal morphogenesis.

Animals↗

PubMed1999

Distribution and localization of immunoreactive FMRFamide-like peptides in the lancelet.

Immunofluorescence was used to study the distribution of FMRFamide (Phe-Met-Arg-Phe-NH2) in premetamorphic larvae and adults of the lancelet, Branchiostoma lanceolatum. In the larvae, FMR-Famide-containing presumably neuronal perikarya and fibers were limited to the anterior third of the dorsal nerve cord. Throughout this region, most of the immunoreactive perikarya and fibers were located ventrolaterally and ventrally within the nerve cord; in addition, in the caudal part of the cerebral vesicle, some of the immunofluorescent cells projected cytoplasmic extensions across the slot-like neural canal. In adult lancelets, immunofluorescence was detected in cells of the Hatschek's pit (a probable homologue of the anterior hypophysis of vertebrates); however, no immunofluorescence was detected in the larval preoral pit, which is the ontogenetic precursor of Hatschek's pit. Moreover, the FMR-Famide-containing elements do not show immunoreactivity to other peptides of the FaRPs family such as pancreatic polypeptide (PP). The results suggest that FMRF-amide may be involved in neuroendocrine functions of lancelets.

Animals↗

PubMed1997

Larval locomotion of the lancelet

The ontogeny of locomotion in the Florida lancelet (Branchiostoma floridae) is described for the early developmental stages through to metamorphosis. Recently hatched larvae swam at speeds up to 1 mm s-1 using their epidermal cilia; this speed decreased to approximately 0.2 mm s-1 by 60 h after fertilization. Changes in cilia-powered fluid flow could be related to changes in the distribution and density of the epidermal cilia during development. Cilia-powered hovering was the dominant behaviour until metamorphosis. The amount of energy expended by ciliating larvae ranged from 10(-9) to 10(-11) W depending upon the age of the larvae and the model used for estimating the power output. The majority of the energy expended was in the ciliary sublayer next to the body. The first muscular movements were seen in larvae 16 h old. These simple flexions increased in complexity during the first 72 h until a complete undulatory (approximately sinusoidal) wave was propagated down the body in the adult manner. The frequency of undulatory beating increased to approximately 10 Hz during the first 48 h, and the larval head showed a large degree of yaw. Lancelet larvae were also capable of high-speed undulations 510 times faster than regular swimming motions. In contrast to ciliating larvae, the energy expended during undulation was at least an order of magnitude greater (10(-8) to 10(-6) W) and radiated beyond the ciliary sublayer.

Journal Article↗

PubMed1998

Early development of the peripheral nervous system in a lancelet species.

The developmental pattern of the lancelet (amphioxus) peripheral nervous system from embryos to larvae has been studied by using wholemount immunostaining and transmission electron microscopy. The peripheral nerves first appeared on the anterior dorsal surface of the medulla at the middle neurula stage, when the anterior nerve cord was just closing. A single axon with a large growth cone was the progenitor of each nerve. The nerve roots adopted an asymmetric arrangement soon after. The first nerve, likely a pair of pure sensory nerves, sprouted from the anterior tip of the nerve cord. This nerve may be comparable topographically to the preoptic nerve (the posterior branch of the terminal nerve) in lungfishes. However, the neuron that first extends its axon was located in the medulla, as in the other posterior nerves. One of the extramedullary primary sensory neurons, the corpuscles of de Quatrefages, appeared in larvae with the mouth and two anterior gill pores. Their axons were seemingly fasciculated with the efferent axon of the first nerve. The second nerve, the most complex one to appear during embryonic and early larval development, innervated the preoral pit and the buccal region. The third and fourth nerves on the left side also innervated the buccal region. The larval innervation patterns in the anterior region differed from the adult organization, suggesting a segmental rearrangement of the nerve supply during development. There was no evidence to dichotomize the peripheral nerves into cranial and spinal nerves, as exist in vertebrates. These characteristics of the peripheral nervous system in the lancelet indicate that this animal has a rather derived or primitive developmental system of peripheral nerves, making the analysis of homology with vertebrates difficult.

Animals↗

PubMed1981

Monoamines in the brain of the lancelet, Branchiostoma lanceolatum. A fluorescence-histochemical and electron-microscopical investigation.

Three types of monoamine-containing neurones and fibres can be discriminated in the brain of the lancelet. Two types of elongated cerebrospinal fluid-contacting neurons, located in the ventral and the dorsolateral part of the brain, exhibit formaldehyde-induced catecholamine fluorescence. These neurones contain dense-core vesicles 75-100 nm in diameter. Their apical portion possesses cilia, displaying a 9 X 2 + 2 arrangement of their internal tubuli, and projecting into the ventricle. Basal processes from the ventrally situated perikarya abut upon the meninx and may discharge their catecholamines into the circulatory system. Fibres exhibiting catecholamine fluorescence originate from the dorsolaterally situated perikarya and run ventrocaudally to the neuropil, where they form numerous swellings of the bouton en passant type. A third type of perikarya in the posterior part of the brain displays specific green fluorescence. Further, neurones characterized by a specific yellow fluorescence are present in the anterior part of the brain and the anterior part of the neural tube. The rapid photodecomposition of the latter fluorophore indicates that these cells contain an indolamine.

Animals↗

PubMed1995

A histochemical study of the distribution of lectin binding sites in the developing oocytes of the lancelet Branchiostoma belcheri.

The distribution of carbohydrate moieties in lancelet (Branchiostoma belcheri) oocytes has been studied at different stages of development, using a peroxidase-labeled lectin incubation technique, the PAS-reaction and Alcian Blue staining. Binding sites of 5 lectins, indicating the presence of different sugar moieties (Wheat germ agglutinin (WGA) for N-acetylglucosamine, Concanavalin A (Con A) for glucose/mannose, Helix pomatia agglutinin (HPA) for N-acetyl-D-galactosamine, Ricinus communis agglutinin (RCA-I) for galactose and Ulex europaeus agglutinin (UEA-I) for fucose), were identified and were shown to undergo considerable variation during oocyte development. In the previtellogenic stage, HPA, RCA-I and UEA-I were not identified on the oocyte surface, but WGA and Con A gave strongly positive reactions at this site. In the cytoplasm, 4 lectins (Con A, HPA, RCA-I and UEA-I) gave a weak or moderate reaction, and Con A was also observed in the perinuclear region. In vitellogenic oocytes, these 4 lectins were found to also bind to the nuclear envelope, karyoplasm and nucleolus, and, with the exception of Con A, could also be found in the nuclei of more mature stages. The cytoplasmic yolk granules and Golgi vesicles of the vitellogenic oocyte, were moderately positive for Con A, HPA, RCA-I and UEA-I, but HPA, RCA-I and UEA-I were only weakly bound at the oocyte surface. In mature oocytes, all 5 lectins bound moderately or strongly to yolk granules and cell surface. HPA, RCA-I and UEA-I bound moderately or strongly to various nuclear compartments.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed2001

Immunocytochemical localization of serotonin in embryos, larvae and adults of the lancelet, Branchiostoma floridae.

Serotonin (5-hydroxytryptamine) is a biogenic amine distributed throughout the metazoans and has an old evolutionary history. It is involved as a developmental signal in the early morphogenesis of both invertebrates and vertebrates, whereas in adults it acts mainly as a neurotransmitter and gastrointestinal hormone. In vertebrates, serotonin regulates the morphogenesis of the central nervous system and the specification of serotonergic as well as dopaminergic neurons. The present study uses, as an experimental model, an invertebrate chordate, the lancelet Branchiostomafloridae, characterized by its remarkable homologies with vertebrates that allows the 'bauplan' of the probable ancestor of vertebrates to be outlined. In particular, the involvement of serotonin as a developmental signal in embryos and larvae, as well as a neurotransmitter and gastrointestinal hormone in adult specimens of Branchiostoma floridae, gives further support to a common origin of cephalocordates and vertebrates.

Animals↗

PubMed1997

A unique cDNA coding for subunits 8 and 6 of mitochondrial adenosine triphosphatase of the lancelet Branchiostoma lanceolatum, an ancestor of vertebrates.

A cDNA encoding subunits 8 and 6 of mitochondrial ATPase of the cephalochordate lancelet (Branchiostoma lanceolatum), a direct ancestor of vertebrates, has been cloned and sequenced. A unique transcript encodes the 8 and 6 subunits. In the course of isolation of the 5' end of the ATPase 8 subunit gene, we also determined the sequence of the 5' adjacent tRNA-Lys gene. The anticodon used by mitochondrial tRNA-Lys is TTT.

Adenosine Triphosphatases↗

PubMed1982

Twitch activation in Ca2+ -free solutions in the myotomes of the lancelet (Branchiostoma lanceolatum).

The question of whether a Ca2+ influx is necessary to activate twitches in the very thin myotome cells of the lancelet was reinvestigated. Though twitches were blocked in EGTA containing Ca2+- free bathing solutions, they reappeared under certain conditions: a) when the Mg2+ level was lowered, b) when small amounts of caffeine were added, and c) when Cl- in the bathing solution was partly replaced by SCN-. In Ca2+ -containing solutions the changes a) to c) increased the twitch height. The results suggest that a Ca2+ release from the intracellular stores is involved in twitch activation and that external Ca2+ modifies the coupling between excitation and Ca2+ release rather than initiating contractile activation.

Action Potentials↗