27 results on '"Zilli, L."'
Search Results
2. Researchers capture images for practical molt stage determination. Global Aquaculture
- Author
-
ZILLI L, R. SCHIAVONE, G. SCORDELLA, V. ZONNO, VILELLA, Sebastiano, Zilli, L, R., Schiavone, G., Scordella, V., Zonno, and Vilella, Sebastiano
- Published
- 2003
3. Characterization of Na+/H+ antiporter activity in PC-Cl3 thyroid cells
- Author
-
VILELLA, Sebastiano, MARSIGLIANTE, Santo, STORELLI, Carlo, SCHIAVONE R, ZILLI L, Vilella, Sebastiano, Schiavone, R, Zilli, L, Marsigliante, Santo, and Storelli, Carlo
- Abstract
Background/Aims: In thyroid cells, the intracellular pH plays a key role in the control of the iodide uptake, since iodide accumulation is associated with an intracellular acidification. In the present paper we studied the kinetic proprieties of the Na+/H+ antiporter (NHE) and the molecular expression of different NHE isoforms in rat thyroid PC-CI3 cells. In addition the intracellular buffer capacity was also evaluated. Methods: pHi was measured using the pH sensitive fluorescent dye BCECF-AM. Amiloride, 5-(N,N-Dimethyl) hydrochloride was used to inhibit NHE activity. RT-PCR and western blot analyses were used to study the expression of NHE mRNA and protein isoforms. Results: PC-CI3 cells shown a resting pHi, in the absence of CO2/HCO3-, of 6.94 +/- 0.1; after an acid load PC-CI3 cells recovered toward resting pHi value, using a Na-dependent H+ extrusion mechanisms which was amiloride sensitive (K-i = 23 muM). The kinetic parameters were K-(Na)app = 10 +/- 2 mM and V-max = 0.23 +/- 0.02 DeltapH/min x 10(5)cells. NHE1, NHE2 and NHE3 were expressed at the mRNA level as well as at the protein level. Conclusion: PC-CI3 cells express a functional Na/H exchange activity and different isciforms (NHE1, NHE2 and NHE3) are expressed in the plasma membrane.
- Published
- 2003
4. Researcher capture images for practical molt stage determination
- Author
-
ZILLI L, SCORDELLA G, SCHIAVONE R, ZONNO V, VILELLA, Sebastiano, Zilli, L, Scordella, G, Schiavone, R, Zonno, V, and Vilella, Sebastiano
- Published
- 2003
5. Effect of the rearing system on the nitritional physiological state of culturated fishes
- Author
-
ZONNO V, SCHIAVONE R, ACIERNO R, TOMA P, MAFFIA, Michele, ZILLI L, VILELLA, Sebastiano, Zonno, V, Schiavone, R, Acierno, R, Toma, P, Maffia, Michele, Zilli, L, and Vilella, Sebastiano
- Published
- 2003
6. Relationship between molting stages and transporter activities in hepatopancreatic R cells of Penaeus japonicus
- Author
-
ZILLI L, SCHIAVONE R, SCORDELLA G, ZONNO V, VILELLA, Sebastiano, Zilli, L, Schiavone, R, Scordella, G, Zonno, V, and Vilella, Sebastiano
- Published
- 2002
7. Effect of cryopreservation on biochemical parameters and motility of the sea bass (Dicentrarchus labrax) sperm
- Author
-
ZILLI L, SCHIAVONE R, ZONNO V, VILELLA, Sebastiano, Zilli, L, Schiavone, R, Zonno, V, and Vilella, Sebastiano
- Published
- 2002
8. Relationship between molting stages and ATPases activity in hepatopancreatic R cells of Penaeus japonicus
- Author
-
ZILLI L, SCHIAVONE R, ZONNO V, SCORDELLA G, C. STORELLI, VILELLA, Sebastiano, Zilli, L, Schiavone, R, Zonno, V, Scordella, G, C., Storelli, and Vilella, Sebastiano
- Published
- 2002
9. Presence of calcium channels in hepatopancreatic cells of different crustacean species
- Author
-
ZILLI L, T. VERRI, V. ZONNO, G. AHEARN, C. STORELLI, VILELLA, Sebastiano, Zilli, L, Verri, T., Zonno, V., Ahearn, G., Storelli, C., and Vilella, Sebastiano
- Published
- 2000
10. Idendification of calcium channels in B cells of Penaeus japonicus hepatopancreas
- Author
-
ZILLI L, MARSIGLIANTE, Santo, V. ZONNO, T. VERRI, G. AHEARN, C. STORELLI, VILELLA, Sebastiano, Zilli, L, Marsigliante, Santo, Zonno, V., Verri, T., Ahearn, G., Storelli, C., and Vilella, Sebastiano
- Published
- 2000
11. Human chorionic gonadotropin induces spermatogenesis and spermiation in 1-year-old European sea bass (Dicentrarchus labrax): Assessment of sperm quality
- Author
-
Schiavone, R, Zilli, L, Vilella, S, Fauvel, Christian, Schiavone, R, Zilli, L, Vilella, S, and Fauvel, Christian
- Abstract
The aims of the present study were (a) to compare sperm quality (percentage of motile spermatozoa, motility duration, density and fertility after cryopreservation) between precocious and normally maturing male European sea bass Dicentrarchus labrax, (b) to examine the potential of human chorionic gonadotropin (hCG) to increase spermiation in precocious males and (c) to examine the potential of hCG to induce spermatogenesis and spermiation in non-precocious 1-year-old males. One hundred precocious and 100 non-precocious fish were each randomly divided in two groups each: control (precocious saline-treated and non precocious saline-treated) and treated (precocious hCG-treated and non precocious hCG-treated). Treated groups were administered weekly with 1000 IU hCG kg(-1) body weight while control groups were injected with physiological solution. Milt volume produced, sperm concentration, motility duration and fertilising ability were assessed every week in each group. The effect of the hormonal treatment on gonadal development was examined based on the gonadosomatic index and testicular histology. The results demonstrate that sperm produced by precocious fish has characteristics (mean value of motility class, mean maximum motility duration, concentration and fertility after cryopreservation) similar (P > 0.05) to those produced by 2-year-old fish. Human chorionic gonadotropin treatment in precocious fish resulted in a significant increase (P < 0.05) of milt volume, without affecting sperm quality. In non-precocious fish, hCG treatment resulted in greater percentage of spermiation (P < 0.05) compared to non-precocious saline-treated group. At the end of the trial (three weeks), 29 out of 50 non-precocious hCG-treated fish were spermiating and, within these 23 produced > 200 mu l per fish of milt. No differences were observed in terms of sperm concentration, motility class, motility duration and fertilizing capacity due to hCG treatment in either precocious, or non-prec
- Published
- 2006
- Full Text
- View/download PDF
12. Ha-ras-1 restriction fragment length polymorphism and susceptibility to colon adenocarcinoma.
- Author
-
Ceccherini-Nelli, L, De Re, V, Viel, A, Molaro, G, Zilli, L, Clemente, C, and Boiocchi, M
- Published
- 1987
- Full Text
- View/download PDF
13. Identification of two nuclear factor-binding domains on the chicken cardiac actin promoter: implications for regulation of the gene
- Author
-
Quitschke, W W, DePonti-Zilli, L, Lin, Z Y, and Paterson, B M
- Abstract
The cis-acting regions that appear to be involved in negative regulation of the chicken alpha-cardiac actin promoter both in vivo and in vitro have been identified. A nuclear factor(s) binding to the proximal region mapped over the TATA element between nucleotides -50 and -25. In the distal region, binding spanned nucleotides -136 to -112, a region that included a second CArG box (CArG2) 5' to the more familiar CCAAT-box (CArG1) consensus sequence. Nuclear factors binding to these different domains were found in both muscle and nonmuscle preparations but were detectable at considerably lower levels in tissues expressing the alpha-cardiac actin gene. In contrast, concentrations of the beta-actin CCAAT-box binding activity were similar in all extracts tested. The role of these factor-binding domains on the activity of the cardiac actin promoter in vivo and in vitro and the prevalence of the binding factors in nonmuscle extracts are consistent with the idea that these binding domains and their associated factors are involved in the tissue-restricted expression of cardiac actin through both positive and negative regulatory mechanisms. In the absence of negative regulatory factors, these same binding domains act synergistically, via other factors, to activate the cardiac actin promoter during myogenesis.
- Published
- 1989
- Full Text
- View/download PDF
14. The β actin promoter
- Author
-
Quitschke, W W, Lin, Z Y, DePonti-Zilli, L, and Paterson, B M
- Abstract
Although β actin mRNA is down-regulated during myogenesis, the β actin promoter confers constitutive expression when joined to heterologous genes transfected into a variety of different cell backgrounds, including differentiated muscle. Normal promoter activity is dependent upon the binding of a ubiquitous factor to the CCAAT-box element. Loss or reduction in factor binding correlates with a major reduction in promoter activity both in vivoand in vitro. The binding domain covers approximately 23 base pairs as determined by DNase footprinting. Methylation of A and G residues in and adjacent to the CCAAT box results in the loss of factor binding. Mutations across the binding domain indicate that the sequence GCCAATCAG within the domain is sufficient as a recognition sequence for factor binding. This binding is not competed by the α cardiac actin CCAAT sequence. Bandshift experiments demonstrate a predominant single band of similar mobility in nuclear extracts from various cells and tissues, with the exception of HeLa cells. The prevalence of the factor and its recognition sequence in a variety of promoters suggests that this factor has a common role in the transcriptional activation of several eukaryotic promoters.
- Published
- 1989
- Full Text
- View/download PDF
15. A 40-base-pair sequence in the 3' end of the beta-actin gene regulates beta-actin mRNA transcription during myogenesis.
- Author
-
DePonti-Zilli, L, Seiler-Tuyns, A, and Paterson, B M
- Abstract
In an earlier report, evidence was presented that the down-regulation of beta-actin mRNA during myogenesis was controlled by a region 3' to the promoter of the gene. In this paper we report the location of this regulatory sequence, determined by deletion analysis and the use of chimeric genes, transfected stably into the mouse myogenic cell line C2C12. The domain responsible for the reduction in beta-actin mRNA levels is at most 40 base pairs long and is located just 5' to the canonical polyadenylylation signal in the gene. Placement of this sequence in the corresponding 3' position both in the alpha-cardiac-actin gene and in the neomycin-resistance gene in pSV2-neo confers the beta-actin mRNA regulatory pattern when these constructs are stably introduced into C2C12 cells. Nuclear run-on experiments indicate that transcriptional control can account for the decrease observed in beta-actin mRNA levels during myogenesis for both the endogenous as well as the transfected beta-actin gene constructs. This 3' transcriptional control sequence is conserved in all of the vertebrate beta-actin genes sequenced and is not similar to any of the 3' processing-adenylylation or termination sequences described previously. This mode of gene regulation may reflect a more general mechanism involved in the process of gene suppression during development.
- Published
- 1988
- Full Text
- View/download PDF
16. Molecular Mechanisms Determining Sperm Motility Initiation in Two Sparids (Sparus aurata and Lithognathus mormyrus)
- Author
-
Loredana Zilli, Sebastiano Vilella, Carlo Storelli, Roberta Schiavone, Zilli, L, Schiavone, R, Storelli, Carlo, and Vilella, Sebastiano
- Subjects
Male ,Proteome ,Motility ,Biology ,sperm ,Dephosphorylation ,Lithognathus mormyrus ,Cyclic AMP ,Animals ,Protein phosphorylation ,sperm motility and transport ,sparid ,Sperm motility ,Molecular mass ,phosphorylation ,Osmolar Concentration ,motility initiation ,Cell Biology ,General Medicine ,Anatomy ,Cyclic AMP-Dependent Protein Kinases ,Spermatozoa ,Sperm ,Sea Bream ,Cell biology ,Reproductive Medicine ,Potassium ,Sperm Motility ,Phosphorylation ,Calcium ,signal transduction - Abstract
Molecular mechanisms involved in sperm motility initiation in two sparids (Sparus aurata and Lithognathus mormyrus) have been studied. Our comparative study demonstrates that osmolality is the key signal in sperm motility activation in both species, whereas K(+) and Ca(2+) do not have any role. The straight-line velocity that resulted, however, was significantly different when measured in sperm activated with non-ionic and/or calcium-free solutions with respect to that measured in seawater-activated sperm. In both species, motility initiation depends on cAMP-dependent protein phosphorylation. The phosphorylation/dephosphorylation patterns that resulted in gilthead and striped sea bream were quite different. In gilthead sea bream, the phosphorylated proteins have molecular weights of 174, 147, 138, 70, and 9-15 kDa, whereas the dephosphorylated proteins have molecular weights of 76, 57, and 33 kDa. In striped sea bream, phosphorylation after sperm motility activation occurred on proteins of 174, 147, 103, 96, 61, 57, and 28 kDa, whereas only one protein of 70 kDa resulted from dephosphorylation. Matrix-assisted laser desorption ionization-time of flight analyses allowed identification of the following proteins: In gilthead sea bream, the 9-15 kDa proteins that were phosphorylated after motility activation include an A-kinase anchor protein (AKAP), an acetyl-coenzyme A synthetase, and a protein phosphatase inhibitor, and in striped sea bream, 103- and 61-kDa proteins that were phosphorylated after motility activation were identified as a phosphatase (myotubularin-related protein 1) and a kinase (DYRK3), respectively.
- Published
- 2008
17. Evaluation of DNA damage in Dicentrarchus labrax sperm following cryopreservation
- Author
-
Sebastiano Vilella, V. Zonno, Loredana Zilli, Roberta Schiavone, Carlo Storelli, Zilli, L, Schiavone, R, Zonno, V, Storelli, Carlo, and Vilella, Sebastiano
- Subjects
Male ,endocrine system ,Cryoprotectant ,DNA damage ,Aquaculture ,DNA Fragmentation ,DNA laddering ,Biology ,General Biochemistry, Genetics and Molecular Biology ,Cryopreservation ,Andrology ,Animals ,Sea bass ,reproductive and urinary physiology ,urogenital system ,General Medicine ,Anatomy ,Sperm ,Spermatozoa ,Comet assay ,DNA fragmentation ,Bass ,Comet Assay ,General Agricultural and Biological Sciences ,DNA Damage ,Semen Preservation - Abstract
In this paper, DNA laddering analysis and single-cell gel electrophoresis (SCGE) or Comet assay, were used to detect DNA damage in response to a cryopreservation process in sea bass spermatozoa. The results obtained demonstrate that the cryopreservation protocol used to cryopreserve the sea bass sperm cause significantly damage at DNA level. In fact, the degree of DNA damage in frozen-thawed sperm (%DNAT=38.2+/-11.2, MT=498.9+/-166.4, n=3) was different (P
- Published
- 2003
18. D-glucose uptake in isolated cells of lobster hepatopancreatic epithelium
- Author
-
VERRI T, L. ZILLI, A. MANDAL, D. BOSSA, P. K. MANDAL, L. INGROSSO, V. ZONNO, G. AHEARN, C. STORELLI, VILELLA, Sebastiano, Verri, T, Zilli, L, Mandal, A, Bossa, D, Mandal, Pk, Ingrosso, L, Zonno, V, Vilella, Sebastiano, Ahearn, Ga, Storelli, C., L., Zilli, A., Mandal, D., Bossa, P. K., Mandal, L., Ingrosso, V., Zonno, G., Ahearn, and C., Storelli
- Published
- 2000
19. Comparative proteome analysis of cryopreserved flagella and head plasma membrane proteins from sea bream spermatozoa: effect of antifreeze proteins.
- Author
-
Zilli L, Beirão J, Schiavone R, Herraez MP, Gnoni A, and Vilella S
- Subjects
- Animals, Cell Membrane drug effects, Cell Membrane metabolism, Cell Survival drug effects, Cryoprotective Agents pharmacology, Electrophoresis, Gel, Two-Dimensional, Fertilization drug effects, Fish Proteins metabolism, Male, Mass Spectrometry, Sperm Head drug effects, Sperm Motility drug effects, Antifreeze Proteins pharmacology, Cryopreservation, Flagella metabolism, Membrane Proteins metabolism, Proteome metabolism, Proteomics methods, Sea Bream metabolism, Sperm Head metabolism
- Abstract
Cryopreservation induces injuries to fish spermatozoa that in turn affect sperm quality in terms of fertilization ability, motility, DNA and protein integrity and larval survival. To reduce the loss of sperm quality due to freezing-thawing, it is necessary to improve these procedures. In the present study we investigated the ability of two antifreeze proteins (AFPI and AFPIII) to reduce the loss of quality of sea bream spermatozoa due to cryopreservation. To do so, we compared viability, motility, straight-line velocity and curvilinear velocity of fresh and (AFPs)-cryopreserved spermatozoa. AFPIII addition to cryopreservation medium improved viability, motility and straight-line velocity with respect to DMSO or DMSO plus AFPI. To clarify the molecular mechanism(s) underlying these findings, the protein profile of two different cryopreserved sperm domains, flagella and head plasma membranes, was analysed. The protein profiles differed between fresh and frozen-thawed semen and results of the image analysis demonstrated that, after cryopreservation, out of 270 proteins 12 were decreased and 7 were increased in isolated flagella, and out of 150 proteins 6 showed a significant decrease and 4 showed a significant increase in head membranes. Mass spectrometry analysis identified 6 proteins (4 from isolated flagella and 2 present both in flagella and head plasma membranes) within the protein spots affected by the freezing-thawing procedure. 3 out of 4 proteins from isolated flagella were involved in the sperm bioenergetic system. Our results indicate that the ability of AFPIII to protect sea bream sperm quality can be, at least in part, ascribed to reducing changes in the sperm protein profile occurring during the freezing-thawing procedure. Our results clearly demonstrated that AFPIII addition to cryopreservation medium improved the protection against freezing respect to DMSO or DMSO plus AFPI. In addition we propose specific proteins of spermatozoa as markers related to the procedures of fish sperm cryopreservation.
- Published
- 2014
- Full Text
- View/download PDF
20. Improving sperm cryopreservation with antifreeze proteins: effect on gilthead seabream (Sparus aurata) plasma membrane lipids.
- Author
-
Beirão J, Zilli L, Vilella S, Cabrita E, Schiavone R, and Herráez MP
- Subjects
- Animals, Cell Membrane metabolism, Cell Survival drug effects, Dimethyl Sulfoxide pharmacology, Fatty Acids metabolism, Fatty Acids, Unsaturated metabolism, Male, Sperm Head drug effects, Sperm Head ultrastructure, Sperm Tail drug effects, Sperm Tail ultrastructure, Antifreeze Proteins pharmacology, Cell Membrane drug effects, Cryopreservation methods, Cryoprotective Agents pharmacology, Membrane Lipids metabolism, Perciformes metabolism, Semen Preservation methods
- Abstract
Changes in the plasma membrane lipid composition have been related to a decrease in sperm quality during cryopreservation. Antifreeze proteins (AFPs) have been tested in different species because of their ability to depress the freezing point and their potential interaction with membranes, but controversial effects were reported. In the present study we analyzed separately the lipid composition of two sperm membrane domains, head plasma membrane (HM) and flagellar membrane (FM), after cryopreservation with an extender containing 5% dimethyl sulfoxide (DMSO) either alone or with AFPI or AFPIII (1 μg/ml). We used sperm from a teleost, Sparus aurata, because the lack of acrosome avoids changes of lipid profiles due to capacitation process or acrosomal losses during freezing/thawing. Comparing with the control (cryopreservation with 5% DMSO alone), the addition of AFPIII increased the velocity, linearity of movement, and percentage of viable cells. In addition, freezing with DMSO alone increased the phosphatidyl-serine content as well as the saturated fatty acids and decreased the unsaturated ones (mainly polyunsaturated) both in HM and FM. These changes in the lipid components were highly avoided with the addition of AFPIII. HM had a higher amount of saturated fatty acids than FM and was more affected by cryopreservation without AFPs. The percentage of viable cells was positively correlated with the amount of unsaturated fatty acids in the HM, whereas the motility parameters were positively correlated with both FM and HM amount of unsaturated fatty acids. AFPs, especially AFPIII, seem to have interacted with unsaturated fatty acids, stabilizing the plasma membrane organization during cryopreservation and contributing to improve sperm quality after thawing.
- Published
- 2012
- Full Text
- View/download PDF
21. Evidence for the involvement of aquaporins in sperm motility activation of the teleost gilthead sea bream (Sparus aurata).
- Author
-
Zilli L, Schiavone R, Chauvigné F, Cerdà J, Storelli C, and Vilella S
- Subjects
- Animals, Blotting, Western, Dose-Response Relationship, Drug, Image Processing, Computer-Assisted, Immunohistochemistry, Male, Mercuric Chloride pharmacology, RNA, Messenger metabolism, Reverse Transcriptase Polymerase Chain Reaction, Spermatozoa drug effects, Video Recording, Aquaglyceroporins metabolism, Aquaporin 1 metabolism, Sea Bream physiology, Sperm Motility physiology, Spermatozoa metabolism
- Abstract
The expression of aquaporins in the spermatozoa of the marine teleost gilthead sea bream (Sparus aurata) and their involvement in the motility activation process were investigated. Sperm motility was activated by a hyperosmotic shock, but it was completely inhibited by 10 microM HgCl(2), such inhibition being partially recovered by beta-mercaptoethanol (ME). Conventional RT-PCR using primers specific for S. aurata aquaglyceroporin (glp) and aquaporin 1a (aqp1a) demonstrated the presence of both mRNAs in spermatozoa. Heterologous expression in Xenopus laevis oocytes showed that 10 and 100 microM HgCl(2) equally inhibited water and solute transport through S. aurata aquaporin 1a and S. aurata aquaglyceroporin, but treatment with ME only recovered aquaporin 1a-mediated water permeability. Western blot analysis using isoform-specific antisera on protein extracts from spermatozoa revealed bands that corresponded to the predicted molecular mass of S. aurata aquaglyceroporin (31 kDa) and S. aurata aquaporin 1a (28 kDa). The antisera also demonstrated that both aquaporins were localized in the head and flagellum of the spermatozoa. However, the immunoreaction at the plasma membrane of the spermatozoa head was more intense after the hyperosmotic activation, suggesting the translocation of both aquaporin 1a and aquaglyceroporin into the plasma membrane after the osmotic shock. This study therefore provides the first direct demonstration for the presence of aquaporins in fish sperm. The different sensitivities of S. aurata aquaporin 1a and S. aurata aquaglyceroporin to ME may explain the failure of this reducing agent to fully recover the mercurial inhibition of sperm motility, suggesting that these aquaporins may play different physiological roles during the activation and maintenance of sperm motility in sea bream.
- Published
- 2009
- Full Text
- View/download PDF
22. Molecular mechanisms determining sperm motility initiation in two sparids (Sparus aurata and Lithognathus mormyrus).
- Author
-
Zilli L, Schiavone R, Storelli C, and Vilella S
- Subjects
- Animals, Calcium pharmacology, Cyclic AMP physiology, Cyclic AMP-Dependent Protein Kinases metabolism, Cyclic AMP-Dependent Protein Kinases physiology, Male, Osmolar Concentration, Potassium pharmacology, Proteome analysis, Sperm Motility drug effects, Spermatozoa chemistry, Spermatozoa metabolism, Sea Bream metabolism, Sea Bream physiology, Sperm Motility physiology
- Abstract
Molecular mechanisms involved in sperm motility initiation in two sparids (Sparus aurata and Lithognathus mormyrus) have been studied. Our comparative study demonstrates that osmolality is the key signal in sperm motility activation in both species, whereas K(+) and Ca(2+) do not have any role. The straight-line velocity that resulted, however, was significantly different when measured in sperm activated with non-ionic and/or calcium-free solutions with respect to that measured in seawater-activated sperm. In both species, motility initiation depends on cAMP-dependent protein phosphorylation. The phosphorylation/dephosphorylation patterns that resulted in gilthead and striped sea bream were quite different. In gilthead sea bream, the phosphorylated proteins have molecular weights of 174, 147, 138, 70, and 9-15 kDa, whereas the dephosphorylated proteins have molecular weights of 76, 57, and 33 kDa. In striped sea bream, phosphorylation after sperm motility activation occurred on proteins of 174, 147, 103, 96, 61, 57, and 28 kDa, whereas only one protein of 70 kDa resulted from dephosphorylation. Matrix-assisted laser desorption ionization-time of flight analyses allowed identification of the following proteins: In gilthead sea bream, the 9-15 kDa proteins that were phosphorylated after motility activation include an A-kinase anchor protein (AKAP), an acetyl-coenzyme A synthetase, and a protein phosphatase inhibitor, and in striped sea bream, 103- and 61-kDa proteins that were phosphorylated after motility activation were identified as a phosphatase (myotubularin-related protein 1) and a kinase (DYRK3), respectively.
- Published
- 2008
- Full Text
- View/download PDF
23. Analysis of calcium concentration fluctuations in hepatopancreatic R cells of Marsupenaeus japonicus during the molting cycle.
- Author
-
Zilli L, Schiavone R, Storelli C, and Vilella S
- Subjects
- Adenosine Triphosphatases metabolism, Animals, Cytoplasm metabolism, Fura-2, Mitochondria metabolism, Penaeidae metabolism, Spectrometry, Fluorescence, Calcium metabolism, Hepatopancreas metabolism, Molting physiology, Penaeidae physiology
- Abstract
In this study we examined the fluctuations of the intracellular calcium concentration in isolated hepatopancreatic R cells during the four molting stages of the prawn Marsupenaeus japonicus. In addition, we used the Fura-2-AM fluorescence technique to investigate the release of calcium from mitochondria and ATP-sensitive calcium stores (endoplasmic reticulum (ER), Golgi, and nucleus) into cytoplasm during the molting cycle. Results demonstrate that both the cytosolic free calcium concentration and the total cell calcium (free, bound to calcium-binding proteins, and stored in amorphous form) in the R cells strictly depend upon the molting cycle. Interestingly, the total cell calcium was higher (approximately 10 mmol l(-1)) in postmolt than in premolt (approximately 1 mmol l(-1)) and intermolt (approximately 0.3 mmol l(-1)). The calcium released from mitochondria was higher during premolt than during postmolt and intermolt, but the amount of calcium released from ATP-sensitive calcium stores was similar during all four stages. All together, our results suggest that the mitochondria-ATP-sensitive calcium stores system does not play a key role in calcium storage during the molting cycle but that it is involved in transcellular calcium flux. We hypothesize that lysosome or membrane-clad concretion vacuoles could represent the main site of calcium storage in hepatopancreatic R cells.
- Published
- 2007
- Full Text
- View/download PDF
24. Effect of cryopreservation on sea bass sperm proteins.
- Author
-
Zilli L, Schiavone R, Zonno V, Rossano R, Storelli C, and Vilella S
- Subjects
- Animals, Electrophoresis, Gel, Two-Dimensional, Fish Proteins isolation & purification, Male, Peptide Mapping, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Xenopus Proteins isolation & purification, Zebrafish Proteins isolation & purification, Bass metabolism, Cryopreservation, Fish Proteins metabolism, Semen Preservation, Spermatozoa metabolism
- Abstract
In the present study we used two-dimensional polyacrylamide gel electrophoresis (2-DE) and matrix-associated laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry to verify whether the protein expression of sea bass sperm was affected by the cryopreservation procedure. The protein profiles differed between fresh and frozen-thawed semen as revealed by visual inspection and by image analysis software. We identified 163 spots in fresh sperm; among these, 13 were significantly decreased and 8 were absent in two-dimensional gel obtained with cryopreserved sperm. Five of these spots were analyzed with MALDI-TOF, but only three showed a significant match in the databases used in bio-informatics analysis (PeptIdent, Mascot, and MS-Fit). In particular, spot 5 showed homology with a novel protein of zebrafish (similar to SKB1 of human and mouse), spot 13 showed homology with amphibian G1/S-specific cyclin E2, and spot 20 showed homology with the hypothetical protein DKFZp566A1524 of Brachidanio rerio. The present work shows that the use of the cryopreservation procedure causes the degradation of sperm proteins and among these, two could be at least partially responsible for the observed decrease in sperm motility duration and the lower hatching rate of eggs fertilized with cryopreserved sperm.
- Published
- 2005
- Full Text
- View/download PDF
25. Adenosine triphosphate concentration and beta-D-glucuronidase activity as indicators of sea bass semen quality.
- Author
-
Zilli L, Schiavone R, Zonno V, Storelli C, and Vilella S
- Subjects
- Animals, Biomarkers metabolism, Cryopreservation, Female, Fertilization physiology, Male, Semen Preservation, Adenosine Triphosphate metabolism, Bass metabolism, Glucuronidase metabolism, Semen metabolism
- Abstract
The most common parameters used to evaluate sperm quality are motility rate and duration and fertilization ability. In this study, chemical and biochemical parameters of sea bass (Dicentrarchus labrax) sperm were investigated to find an alternative method for evaluating sperm fertilization ability before and after cryopreservation. The biochemical and chemical analyses were performed with fresh and frozen-thawed sperm and seminal plasma. To cryopreserve sperm, 250-microl straws were used. Fertilization ability was evaluated by inseminating eggs (obtained from hormonally stimulated females) with fresh and cryopreserved sperm. The results revealed a linear relationship (P < 0.05) between semen fertilization capacity and some seminal plasma (beta-D-glucuronidase activity, potassium concentration) and sperm (ATP concentration, aspartate aminotransferase activity) parameters. Variations in semen fertilization rate could be best described by two multiple regression models: one including the sperm parameters and another including the seminal plasma parameters. For practical application, the use of simple regression models is of value. Fertilization rate in both fresh and cryopreserved sperm was reliably predicted by determining the ATP concentration or the beta-D-glucuronidase activity or both.
- Published
- 2004
- Full Text
- View/download PDF
26. Isolation and characterization of the CRIPTO autosomal gene and its X-linked related sequence.
- Author
-
Dono R, Montuori N, Rocchi M, De Ponti-Zilli L, Ciccodicola A, and Persico MG
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, Blotting, Southern, Cell Line, Chromosome Mapping, GPI-Linked Proteins, Humans, Intercellular Signaling Peptides and Proteins, Mice, Molecular Sequence Data, Multigene Family genetics, Mutation genetics, Tumor Cells, Cultured, Chromosomes, Human, Pair 3, Epidermal Growth Factor, Growth Substances genetics, Membrane Glycoproteins, Neoplasm Proteins genetics, X Chromosome
- Abstract
We have previously reported on the identification of a cDNA clone encoding a novel human growth factor, named "CRIPTO," that is abundantly expressed in undifferentiated human NTERA-2 clone D1 (NT2/D1) and mouse (F9) teratocarcinoma cells. We now report the organization and nucleotide sequence of two related genomic sequences. One (CR-1) corresponds to the structural gene encoding the human CRIPTO protein expressed in the undifferentiated human teratocarcinoma cells, and the other (CR-3) corresponds to a complete copy of the mRNA containing seven base substitutions in the coding region representing both silent and replacement substitutions. The 440 bp 5' to the CAP site of CR-1 are preserved in CR-3. CR-1 maps to chromosome 3, and CR-3 maps to Xq21-q22. Southern blot analysis reveals that multiple CRIPTO-related DNA sequences are present in the human as well as in the mouse genome.
- Published
- 1991
27. The beta actin promoter. High levels of transcription depend upon a CCAAT binding factor.
- Author
-
Quitschke WW, Lin ZY, DePonti-Zilli L, and Paterson BM
- Subjects
- Actins metabolism, Animals, Base Composition, Base Sequence, Binding, Competitive, Chick Embryo, Chromosome Deletion, DNA-Binding Proteins analysis, Molecular Sequence Data, Nuclear Proteins metabolism, Regulatory Sequences, Nucleic Acid, Transcription Factors physiology, Actins genetics, DNA-Binding Proteins physiology, Promoter Regions, Genetic, Transcription, Genetic
- Abstract
Although beta actin mRNA is down-regulated during myogenesis, the beta actin promoter confers constitutive expression when joined to heterologous genes transfected into a variety of different cell backgrounds, including differentiated muscle. Normal promoter activity is dependent upon the binding of a ubiquitous factor to the CCAAT-box element. Loss or reduction in factor binding correlates with a major reduction in promoter activity both in vivo and in vitro. The binding domain covers approximately 23 base pairs as determined by DNase footprinting. Methylation of A and G residues in and adjacent to the CCAAT box results in the loss of factor binding. Mutations across the binding domain indicate that the sequence GCCAATCAG within the domain is sufficient as a recognition sequence for factor binding. This binding is not competed by the alpha cardiac actin CCAAT sequence. Bandshift experiments demonstrate a predominant single band of similar mobility in nuclear extracts from various cells and tissues, with the exception of HeLa cells. The prevalence of the factor and its recognition sequence in a variety of promoters suggests that this factor has a common role in the transcriptional activation of several eukaryotic promoters.
- Published
- 1989
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.