• The androgen receptor mRNA is up-regulated by testosterone in both the Harderian gland and thumb pad of the frog, Rana esculenta.

    Publication Date: 01/12/1994, on The Journal of steroid biochemistry and molecular biology
    by Varriale B, Serino I

    Alpha 32P-labelled cDNA probe from plasmid containing rat androgen receptor (rAR) has been tested in hybridization experiments using RNAs from the Harderian gland and thumb pad of the edible frog, Rana esculenta. Northern blot analysis has shown a high degree of homology between the rAR cDNA and the frog androgen receptor mRNA (fAR mRNA); this has been supported by both the hybridization conditions (high stringency) and the molecular size of fAR mRNA which is quite similar to those described in mammals (9.4 kb). The role of androgens has been further investigated with respect to the kinetics of expression of fAR mRNA in in vivo experiments. In both the Harderian gland and thumb pad, testosterone has increased the levels of fAR mRNA as compared with the untreated groups. The use of cyproterone acetate (CPA) in combination with testosterone has resulted in a loss of the increase in fAR mRNA as compared to testosterone-treated groups, while CPA alone has resembled the control group. In primary cultures of frog Harderian gland and thumb pad cells, the steady-state levels of fAR mRNA have been increased in the cells exposed to testosterone as compared to those not exposed. These findings confirm that, in these androgen target tissues, testosterone exerts an up-regulation on its own receptors, increasing the accumulation of fAR mRNA in the same way as oestrogens up-regulate the expression of their own receptors in Xenopus liver and oviduct cells.

  • Nitric oxide and long-term habituation to novelty in the rat.

    Publication Date: 17/11/1994, on Annals of the New York Academy of Sciences
    by Papa M, Pellicano MP, Sadile AG

    The role of nitric oxide in learning and memory processes has been tested in the albino rat by a histochemical and a behavioral study, following behavioral habituation to spatial novelty. Histochemically, the neural consequences of behavioral testing were mapped in the brain by staining for NADPH-d, known to be a NOS, whereas behaviorally the formation of LTH has been interfered with by posttrial NOS-inhibition. In the histochemical study, adult male Sprague-Dawley rats were tested in a Làt-maze and sacrificed at different time intervals thereafter. Handled unexposed rats served as controls. The brains were perfused with aldheide and processed for NADPH-d staining. In unexposed control rats the basal expression of NADPH-d was low and scattered. It pertained to few cells in the neostriatum, cerebral cortex, and CA1 hippocampal regions. In contrast, rats that had been exposed for the first time to the maze (spatial novelty) showed NADPH-d activity in the dorsal hippocampus (granule cells, few hilar neurons, and some CA1 pyramidal cells), the caudate-putamen complex, the cerebellum, and in all layers of somatosensory cortex. The positivity was not due to activity per se, since immediately after exposure it was not different from baseline. In contrast, it was present by 2 h and decreased significantly 24 h later. In addition, a strong neuronal discharge induced by the convulsant pentylentetrazol did not induce NADPH-d 2 h afterwards. The staining was prevented by pretreatment with the NMDA receptor antagonist CPP (5 mg/kg) or with the NOS inhibitor L-NOARG (10 mg/kg). In the behavioral study, rats were given an intraperitoneal injection of 1-10 mg/kg (L-NOARG) or vehicle immediately following exposure to a Làt-maze. The highest dose used (10 mg/kg) disrupted habituation of the vertical component only, known to be mainly of emotional meaning. Conversely, both doses disrupted emotional habituation based on defecation scores. The data indicate that the formation of LTH to novelty triggers a cascade of neurochemical events also involving NOS neurons. Further, the widespread induction of NADPH-d by exposure to novelty suggests that spatial and emotional information processing activate neural networks across different organizational levels of the CNS.

  • [Ambulatory monitoring of arterial pressure in the diagnosis and therapeutic evaluation of dysautonomic diseases: observations in a case of Shy-Drager syndrome].

    Publication Date: 01/11/1994, on Cardiologia (Rome, Italy)
    by Irace L, Lus G, Spadaro P, Ducceschi V, De Marco N, Sarubbi B, Iacono A, Cotrufo R

    Shy-Drager syndrome is a very rare disease affecting the autonomic nervous system. Usefulness of beta-adrenergic chronic therapy has already been focused in these cases. Ambulatory blood pressure monitoring may help to study the cardiocirculatory adaptation in Shy-Drager syndrome patients.

  • Protein synthesis in nerve endings from squid brain: modulation by calcium ions.

    Publication Date: 01/10/1994, on The Biological bulletin
    by Benech J, Crispino M, Chun JT, Kaplan BB, Giuditta A
    DOI: 10.1086/BBLv187n2p269

  • Conversion of enkephalin and dermorphin into delta-selective opioid antagonists by single-residue substitution.

    Publication Date: 15/08/1994, on European journal of biochemistry
    by Tancredi T, Salvadori S, Amodeo P, Picone D, Lazarus LH, Bryant SD, Guerrini R, Marzola G, Temussi PA

    The properties of di- and tri-peptides containing 1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid (Tic) in second position suggest that the message domain of opioid peptides can be composed of only two residues [Temussi, P. A., Salvadori, S., Amodeo, P., Guerrini, R., Tomatis, R., Lazarus, L. H., Picone, D. & Tancredi, T. (1994) Biochem. Biophys. Res. Commun. 198, 933-939]. As a crucial test of the possibility that the Tyr-Tic segment be a message domain in longer peptide sequences, we have inserted it in the sequences of two typical opioid peptides: [Leu]enkephalin, a non-selective agonist, and dermorphin, a selective mu agonist. Here we report the synthesis and biological activity of [L-Tic2]enkephalin, [L-Tic2]dermorphin, [L-Tic2]dermorphin carboxylic acid and [D-Tic2]dermorphin: all [L-Tic2]peptides were converted from agonists to delta-selective antagonists. The NMR conformational study in a dimethylsulfoxide/water cryoprotective mixture at low temperature shows diagnostic side-chain--side-chain NOEs in the spectra of all [L-Tic2]peptides and hints that the 90 degrees arrangement of the the two aromatic rings found in the cis-Tyr-L-Tic moiety, typical of N-methyl naltrindole and other delta-selective opiate antagonists, is responsible for the antagonist activity of all these peptides.

  • Kinesin mRNA is present in the squid giant axon.

    Publication Date: 01/07/1994, on Journal of neurochemistry
    by Gioio AE, Chun JT, Crispino M, Capano CP, Giuditta A, Kaplan BB

    Recently, we reported the construction of a cDNA library encoding a heterogeneous population of polyadenylated mRNAs present in the squid giant axon. The nucleic acid sequencing of several randomly selected clones led to the identification of cDNAs encoding beta-actin and beta-tubulin, two relatively abundant axonal mRNA species. To continue characterization of this unique mRNA population, the axonal cDNA library was screened with a cDNA probe encoding the carboxy terminus of the squid kinesin heavy chain. The sequencing of several positive clones unambiguously identified axonal kinesin cDNA clones. The axonal localization of kinesin mRNA was subsequently verified by in situ hybridization histochemistry. In addition, the presence of kinesin RNA sequences in the axoplasmic polyribosome fraction was demonstrated using PCR methodology. In contrast to these findings, mRNA encoding the squid sodium channel was not detected in axoplasmic RNA, although these sequences were relatively abundant in the giant fiber lobe. Taken together, these findings demonstrate that kinesin mRNA is a component of a select group of mRNAs present in the squid giant axon, and suggest that kinesin may be synthesized locally in this model invertebrate motor neuron.

  • Visuospatial imagery in Alzheimer disease.

    Publication Date: 01/06/1994, on Perceptual and motor skills
    by Grossi D, Becker JT, Trojano L
    DOI: 10.2466/pms.1994.78.3.867

    The purpose of the present study was to investigate the imagery ability of 8 "probable" Alzheimer disease patients using a standard task sensitive to the disruption of the imagery system. These patients were selected based on their good performance on a prerequisite clock-setting task, and with 8 matched control subjects were then required to compare the angles made by the hands on clock faces using only imagined stimuli. There were no significant differences between patients and controls in performance on the clock-imagery task. These data are consistent with models of information processing which postulate the relationships between visual imagery and a short-term memory store and include a central executive system for allocation of cognitive resources.

  • Effect of testing procedure on Corsi's block-tapping task in normal subjects and Alzheimer-type dementia.

    Publication Date: 01/06/1994, on Perceptual and motor skills
    by Trojano L, Chiacchio L, De Luca G, Fragassi NA, Grossi D
    DOI: 10.2466/pms.1994.78.3.859

    Corsi's block-tapping task was given to 30 normal subjects and 38 Alzheimer-type demented patients following two different procedures. The first is the most widely standardized (scoring criterion: 3 correct reproductions out of 5 sequences), while the second is more lenient since it does not require subjects to replicate a certain performance three times. Demented patients' scores were lower than those of controls in both conditions, and scores on the two tasks were significantly correlated for patients and controls. However, the requirement of replicating the visuospatial memory performances was more detrimental for demented patients than for controls so the two procedures cannot be considered equivalent.

  • Target cells modulate dopamine transporter gene expression during brain development.

    Publication Date: 09/05/1994, on Neuroreport
    by Perrone-Capano C, Tino A, di Porzio U

    We have analysed the expression of the dopamine transporter (DAT) gene and compared it with that of tyrosine hydroxylase, neuronal GABA transporter and synaptic vesicle monoamine transporter genes during pre- and post-natal development of rat mesencephalic dopaminergic (DA) neurones. Our results show that DAT transcripts are not detectable until embryonic day (E) 15, whilst those of the other genes analysed are already present at E12. In vitro, the level of DAT gene transcription in mesencephalic E13 DA neurones is increased in coculture with target striatal cells. Thus striatal targets cells regulate, at the transcriptional level, a key step of dopaminergic neurotransmission during DA neurone development.

  • Docosahexaenoic acid and signaling pathways in rabbit colon.

    Publication Date: 01/04/1994, on Molecular pharmacology
    by Calderaro V, Parrillo C, Balestrieri ML, Giovane A, Filippelli A, Rossi F

    The effects of one of the main components of fish oil, docosahexaenoic acid (DHA), on prostaglandin (PG) and Ca2+ signaling pathways were examined in intact mucosa and freshly isolated crypt cells of rabbit descending colon. Preincubation of serosal mucosa for 20 min with 1 microM DHA fully suppressed the short-circuit and transepithelial conductance increase induced by serosal addition of 10 microM arachidonic acid (AA). DHA at 1 microM also prevented the Cl- secretion promoted by 10 microM AA, as estimated by unidirectional 36Cl flux measurements (net flux = 0.68 +/- 0.30 versus -1.91 +/- 0.20 microEq/hr/cm2, four experiments, p < 0.001), whereas it did not affect the electrophysiological and ion flux responses to PGE2. Addition of 1 microM DHA to the serosal side of the mucosa also inhibited the PG cascade activation elicited by AA (PG synthesis and second messenger cAMP increase). In vitro assays of colonic cyclooxygenase activity showed that 1 microM DHA inhibited (with a 20-min lag) cyclooxygenase activity to the same extent as 5 microM indomethacin (approximately 82% and 80%, respectively). DHA also affected the Ca2+ signaling pathway; in isolated crypt cells, the cytosolic free Ca2+ concentration ([Ca2+]i) dropped by 49 +/- 7.6% (mean +/- standard error, six experiments) after incubation with 1 microM DHA. The sustained phase of the [Ca2+]i response to 500 nM concentrations of the intracellular Ca(2+)-ATPase inhibitor thapsigargin was also inhibited within 150 sec upon 1 microM DHA addition (141 +/- 5.8 versus 243 +/- 8.2 nM [Ca2+]i mean +/- standard error, eight experiments, p < 0.01). The [Ca2+]i-lowering effect of DHA, which was not achieved by incubation with other free fatty acids, was not prevented by removal of Na+ from the incubation medium (-46 +/- 4.3% versus -47 +/- 3.8%, mean +/- standard error, four experiments), nor it was mediated by cAMP-, protein kinase C-, or calmodulin-dependent mechanisms. The incubation of highly purified basolateral membranes of crypt cells with 1 microM DHA for 1 min produced a 5-fold increase (IC50 = 0.25 microM) in the plasma membrane Ca(2+)-ATPase activity (34.3 +/- 2.73 versus 6.02 +/- 0.50 nmol/mg of protein/min, mean +/- standard error, four experiments, p < 0.0001), thus indicating that the DHA effects on the Ca2+ pathway were mediated mainly by an increase in plasma membrane Ca2+ pump activity. These findings suggest that DHA is a powerful modulator of the cellular response to activation of PG and Ca2+ signaling pathways.

  • Novel 3-beta-methoxysteroids from the Senegalese sponge Microscleroderma spirophora.

    Publication Date: 01/03/1994, on Steroids
    by Costantino V, Fattorusso E, Mangoni A, Aknin M, Gaydou EM

    The Senegalese sponge Microscleroderma spirophora has been found to produce exclusively very unusual 3 beta-methoxysteroids in place of the common 3 beta-hydroxysteroids. Six different methoxysteroids (three of which are novel compounds) have been isolated and identified by spectroscopic means (MS, IR, 1H and 13C NMR), and their taxonomic significance discussed. A new method for the determination of the configuration at C-24 in saturated 24-ethyl side chains is also proposed.

  • A critical review of mental imagery defects.

    Publication Date: 01/03/1994, on Brain and cognition
    by Trojano L, Grossi D
    DOI: 10.1006/brcg.1994.1012

    A critical, clinical review of single-case reports and group studies on visual mental imagery deficits is offered. Neuropsychological findings demonstrate that mental imagery relies upon dissociable processes which are localized in left-hemisphere posterior areas. Imagery defects thus may often be associated with visual recognition and naming impairments. On the other hand, the right hemisphere seems to play an analogous role in imagery and perception. Current theoretical models of imagery do not appear fully capable of accounting for available clinical data.

  • Selective opioid dipeptides.

    Publication Date: 15/02/1994, on Biochemical and biophysical research communications
    by Temussi PA, Salvadori S, Amodeo P, Bianchi C, Guerrini R, Tomatis R, Lazarus LH, Picone D, Tancredi T
    DOI: 10.1006/bbrc.1994.1133

    The surprising change of selectivity induced by the change of chirality in peptides containing the tetrahydro-3-isoquinoline carboxylic acid (Tic) in second position, interpreted as a conformational preference induced on the Tyr-Xaa-Phe domain, can instead be attributed to the Tyr-Tic message domain. The relative spatial disposition of the aromatic ring of delta-selective non peptidic opiates is compatible with a message domain, in opioid peptides, of only two residues. This hypothesis was tested through the synthesis of Tyr-L-Tic-NH2, Tyr-D-Tic-NH2, Tyr-L-Tic-Ala-NH2, Tyr-L-Tic-Ala-OH and Tyr-D-Tic-Ala-NH2. Peptides containing Tyr-L-Tic- behave as very selective delta antagonists and those containing Tyr-DTic- as non selective agonists. This is the first case of opioid peptides containing a two-residue message domain and of opioid dipeptides with substantial opioid activity.

  • c-fos spontaneous expression during wakefulness is reversed during sleep in neuronal subsets of the rat cortex.

    Publication Date: 01/01/1994, on Journal of physiology, Paris
    by Grassi-Zucconi G, Giuditta A, Mandile P, Chen S, Vescia S, Bentivoglio M

  • Neurofilament proteins are synthesized in nerve endings from squid brain.

    Publication Date: 01/09/1993, on Journal of neurochemistry
    by Crispino M, Capano CP, Kaplan BB, Giuditta A

    It is generally believed that the proteins of the nerve endings are synthesized on perikaryal polysomes and are eventually delivered to the presynaptic domain by axoplasmic flow. At variance with this view, we have reported previously that a synaptosomal fraction from squid brain actively synthesizes proteins whose electrophoretic profile differs substantially from that of the proteins made in nerve cell bodies, axons, or glial cells, i.e., by the possible contaminants of the synaptosomal fraction. Using western analyses and immunoabsorption methods, we report now that (a) the translation products of the squid synaptosomal fraction include neurofilament (NF) proteins and (b) the electrophoretic pattern of the synaptosomal newly synthesized NF proteins is drastically different from that of the NF proteins synthesized by nerve cell bodies. The latter results exclude the possibility that NF proteins synthesized by the synaptosomal fraction originate in fragments of nerve cell bodies possibly contaminating the synaptosomal fraction. They rather indicate that in squid brain, nerve terminals synthesize NF proteins.