Laura Zanetti

Molecular forms of immunoreactive gonadotropin-releasing hormone in hypothalamus and testis of the frog, Rana esculenta.

The hypothalamus and the testis of the frog, Rana esculenta, contain gonadotropin-releasing hormone (Gn-RH)-like peptides which are recognized by an antiserum raised against mammalian Gn-RH. Two molecular forms which coelute with synthetic chicken II and salmon Gn-RH from reverse-phase HPLC were distinguished in the hypothalamus. A single peak coeluting with synthetic chicken II Gn-RH was present in the testis.

FGF/MAPK/Ets signaling renders pigment cell precursors competent to respond to Wnt signal by directly controlling Ci-Tcf transcription

FGF and Wnt pathways constitute two fundamental signaling cascades, which appear to crosstalk in cooperative or antagonistic fashions in several developmental processes. In vertebrates, both cascades are involved in pigment cell development, but the possible interplay between FGF and Wnt remains to be elucidated. In this study, we have investigated the role of FGF and Wnt signaling in development of the pigment cells in the sensory organs of C. intestinalis. This species possesses the basic features of an ancestral chordate, thus sharing conserved molecular developmental mechanisms with vertebrates. Chemical and targeted perturbation approaches revealed that a FGF signal, spreading in time from early gastrulation to neural tube closure, is responsible for pigment cell precursor induction. This signal is transmitted via the MAPK pathway, which activates the Ci-Ets1/2 transcription factor. Targeted perturbation of Ci-TCF, a downstream factor of the canonical Wnt pathway, indicated its contribution to pigment cell differentiation Furthermore, analyses of the Ci-Tcf regulatory region revealed the involvement of the FGF effector, Ci-Ets1/2, in Ci-Tcf transcriptional regulation in pigment cell precursors. Our results indicate that both FGF and the canonical Wnt pathways are involved in C. intestinalis pigment cell induction and differentiation. Moreover, we present a case of direct transcriptional regulation exerted by the FGF signaling cascade, via the MAPK-ERK-Ets1/2, on the Wnt downstream gene Ci-Tcf. Several examples of FGF/Wnt signaling crosstalk have been described in different developmental processes; however, to our knowledge, FGF-Wnt cross-interaction at the transcriptional level has never been previously reported. These findings further contribute to clarifying the multitude of FGF-Wnt pathway interactions.

Transglutaminase activity is related to CAG repeat length in patients with Huntington’s disease

Abstract Huntington’s disease (HD) is a neurodegenerative disorder associated with CAG repeat expansion. We measured transglutaminase (TGase) activity in lymphocytes from 35 HD patients and from healthy individuals to ascertain whether it was altered in this condition. TGase activity was above maximum control levels in 25% of HD patients; it was correlated with the age of the patient and inversely correlated with the CAG repeat length. These results suggest that: (1) HD could be biochemically heterogeneous, and (2) the length of the CAG repeat expansion/TGase ratio could be important in the manifestation of HD.

α- and β-cephalotoxin: Two paralysing proteins from posterior salivary glands of Octopus vulgaris

Abstract 1. 1. Cephalotoxin has been extracted from the posterior salivary glands of Octopus vulgaris and has been fractionated in five components, two of which (α- and β-cephalotoxin) were further purified on Sephadex G-200 superfine. Their molecular weight, as determined on Biogel P-150, are equal to 91,200 and 33,900 respectively. 2. 2. Cephalotoxins are glycoproteins which contain approximately 50% of carbohydrates. Their acidic nature is indicated by the isoelectric point and by the amino acid analysis. 3. 3. α- and β-Cephalotoxin from O. vulgaris , which have been compared with the active components isolated from other Cephalopods, have a strong paralysing effect upon crabs. This has been used for the bioassay studies.

Isolation and characterization of two genes encoding calitoxins, neurotoxic peptides from Calliactis parasitica (Cnidaria)

Among sea anemone neurotoxins, calitoxin, recently isolated from Calliactis parasitica, is a highly toxic peptide of 46 amino acids (aa), whose sequence differs greatly from that of all sea anemone toxins isolated so far. In this study, two genes (clx-1 and clx-2) coding for two highly homologous calitoxins were isolated and characterized from a C. parasitica genomic library. The clx-1 gene encodes the already known calitoxin sequence, named CLX-I, whereas a single bp substitution in the coding region of clx-2 is responsible for a single Glu6-->Lys replacement in a new peptide named CLX-II. The structural organization of the two genes is very similar: two introns and three exons, whose sequences are highly homologous for clx-1 and clx-2 (95% identity). The open reading frame (ORF) of both clx-1 and clx-2 codes for a precursor peptide of 79 aa, whose N-terminus has the feature of a single peptide, while the C-terminus corresponds to the sequences of mature CLX-I and CLX-II. The finding that a pair of basic aa is located upstream from the sequence of both mature toxins strongly suggests that proteolytic events, at specific cleavage sites, are responsible for the release of neurotoxins from their respective precursor molecules.

Structure of two biologically active sesquiterpenoid amino-quinones from the marine spongeDysidea avara

The structures of 2 minor constituents of the marine spongeDysidea avara, which induce developmental aberrations in sea-urchin eggs, are described. The structures were confirmed also by a simple synthesis from avarol (1).

Posidonia ecosystem—V. Phenolic compounds from marine phanerogames, Cymodocea Nodosa and Posidonia Oceanica

Abstract 1. 1. A preliminary study of the phenolic compounds in two marine phanerogames, Cymodocea nodosa and Posidonia oceanica , has been undertaken. 2. 2. Several phenols have been isolated and identified by their chemical and physical properties.

Gonadotropin-releasing hormone in elasmobranch (electric ray, Torpedo marmorata) brain and plasma: Chromatographic and immunological evidence for chicken GnRH II and novel molecular forms

Gonadotropin-releasing hormone (GnRH) peptides in the brain, testis and plasma of an electric ray (Torpedo marmorata) were investigated by gel filtration chromatography, reverse phase high performance liquid chromatography and radioimmunoassay with region-specific antisera. In the brain, two major forms of GnRH were demonstrated. One form had identical chromatographic and immunological properties to chicken GnRH II, and the second, novel, molecular form had structural features in common with mammalian, chicken II and salmon GnRHs. A minor, early-eluting immunoreactive peak, possibly also a novel GnRH, was also evident. Immunoreactive GnRH was not detected in the testis. In the plasma, a single major early-eluting immunoreactive peak was demonstrated. This peak, identical to the minor peak observed in the brain, is likely to represent a novel form of GnRH which has immunological properties in common with mammalian, chicken II and salmon GnRHs. Immunoreactive GnRH was not detected in the plasma of species from other vertebrate classes, including rabbit, chicken, monitor lizard, clawed toad, frog, cichlid fish and lamprey. The finding of chicken GnRH II in a species of Chondrichthyes adds further support to our hypothesis that this widespread structural variant may represent an early-evolved and conserved form of GnRH. The presence of a GnRH molecular form in the plasma of the electric ray suggests that GnRH may reach target organs (pituitary and gonads) via the general circulation in some species of Chondrichthyes.

A new transglutaminase‐like from the ascidian Ciona intestinalis 1

A cDNA clone encoding a transglutaminase (TGase) was isolated from a cDNA library prepared from the larval stage of Ciona intestinalis. The cDNA sequence has an open reading frame encoding a protein of 696 amino acids and is about 36% identical to 11 other TGase sequences. In addition, the critical residues thought to form the catalytic center are conserved. The Ciona TGase (CiTGase) has an extension of 39 amino acids in the NH2‐terminal region similar to that reported for keratinocyte TGases. A phylogenetic analysis among other types of TGases demonstrated that CiTGase represents a new type of the enzyme.

Fibroblast growth factor signalling controls nervous system patterning and pigment cell formation in Ciona intestinalis

During the development of the central nervous system (CNS), combinations of transcription factors and signalling molecules orchestrate patterning, specification and differentiation of neural cell types. In vertebrates, three types of melanin-containing pigment cells, exert a variety of functional roles including visual perception. Here we analysed the mechanisms underlying pigment cell specification within the CNS of a simple chordate, the ascidian Ciona intestinalis. Ciona tadpole larvae exhibit a basic chordate body plan characterized by a small number of neural cells. We employed lineage-specific transcription profiling to characterize the expression of genes downstream of fibroblast growth factor signalling, which govern pigment cell formation. We demonstrate that FGF signalling sequentially imposes a pigment cell identity at the expense of anterior neural fates. We identify FGF-dependent and pigment cell-specific factors, including the small GTPase, Rab32/38 and demonstrated its requirement for the pigmentation of larval sensory organs.