Search PubMed⌕ Search

Biomedical subjects

J Serratosa

Publications and source records attributed to J Serratosa.

48 records · Page 3Linked to original sources

Nuclear growth and chromatin relaxation-condensation cycle in hepatocytes during the proliferative activation of rat liver.

In order to quantify the changes in nucleolar and nuclear volumes and in chromatin condensation produced during proliferative activation we have carried out morphometric studies on hepatocyte nuclei during rat liver regeneration using electron microscopy. To minimize the artefactual effects produced by fixation on subcellular structures we have fixed the livers by perfusion with glutaraldehyde. The mean values for the nucleolar and nuclear volumes were progressively increased until 28 h after 66% partial hepatectomy. The maximum values raised for the nuclei and nucleoli at this time were 3 and 4.28 times, respectively, those of controls. Later, nuclear and nucleolar volumes progressively declined. Two waves of diminution in nuclear electron-dense material were produced after hepatectomy. The first occurred between 0 and 12 h, with minimum values 1.34 times lower than those from control animals at 8 h. The second occurred between 12 and 28 h, with minimum values 2.56 times lower than those from control rats at 24 h. These two waves in chromatin relaxation correlate very well with the transcriptional changes described by other authors during the pre-replicative, replicative and mitotic phases of liver regeneration.

Animals↗

Calcium transport from blood into the bile in normal and regenerating rat liver.

We have studied calcium movement from blood into the bile by injecting 45Ca2+ intravenously and measuring the radioactivity appearing in the bile. 45Ca2+ started to appear in the bile at 3 min and maximum values were observed at 5 min after its administration. The amount of calcium secreted into the bile was proportional to the blood calcium concentration indicating that the main pathway involved in calcium movement behaved as a non-saturable system. We have also studied the 45Ca2+ circulation from blood into the bile in rats subjected to a partial hepatectomy. Thereafter, the calcium transported into the bile per gram of liver increased by about 50 per cent. Since bile flow behaved in a similar way, the biliar calcium concentration remained unmodified after hepatectomy. Determination of the activities of the Ca2+ transporting systems in isolated plasma membrane fractions from regenerating livers showed no modification in these activities suggesting that the elevation in calcium movement observed after hepatectomy is not due to an increase in the circulation of Ca2+ through the transhepatocyte pathway, an observation compatible with the absence of saturation in the transport.

Animals↗

Calmodulin may decrease cell surface sialic acid and be involved in the expression of fibronectin during liver regeneration.

The decrease of sialic acid in plasma membrane glycoproteins and the expression of cell surface fibronectin were studied during the pre-replicative phase of liver regeneration. The aim of this study was to correlate these cell-surface events to the intracellular surge of calmodulin observed a few hours after partial hepatectomy. The fact that calmodulin decreased the specific activity of UDP-N-acetyl-D-glucosamine 2'-epimerase, a key regulatory enzyme in the biosynthesis of glycoprotein sialic acids, and that trifluoperazine prevented the desialylation indicates that the membrane desialylation is a calmodulin-dependent process. On the other hand, Western blotting using anti-rat fibronectin antibody in trifluoperazine-treated animals suggests that calmodulin may also be involved in the surface expression of fibronectin in regenerating hepatocytes.

Animals↗

Changes in sinusoidal plasma membrane enzyme activities during the pre-replicative phase of liver regeneration.

Changes in a range of plasma membrane enzyme activities during the early period of liver regeneration are thought to be related to the initiation of DNA synthesis and the triggering of cellular activation. The sinusoidal plasma membrane was isolated from control and partially hepatectomized animals at various intervals during the pre-replicative phase. The specific activities of 5'-nucleotidase, (Na+ + K+)-ATPase, Ca2+-ATPase, Mg2+-ATPase showed that after partial hepatectomy changes in the enzyme activities at the sinusoidal plasma membrane region occur. These changes are probably related to the remodeling of the cell-surface that occurs before the division of hepatocytes.

5'-Nucleotidase↗

[Changes in sialic acid content of the plasma membrane in hepatocellular proliferation].

The content of sialic acid bound to the sinusoidal region of plasma membrane during the prereplicative phase after the intravenous injection of a solution containing triiodothyronine, amino acids, glucagon and heparin (T.A.G.H. solution) has been measured. The results obtained show that an important decrease in sialic acid content is produced as it occurs in the hepatic cells of hepatectomized animals. In order to know if sialidase activity is involved in the decrease of sialic acid content during liver regeneration, the activity of sinusoidal plasma membrane sialidases during the prereplicative phase after the partial hepatectomy has been studied. No modifications of sialidase activity were detected during this period of time indicating that this decrease in sialic acid content has to be produced by other mechanisms such as diminution in the synthesis of precursor molecules. On the other hand due to the importance of Ca2+-calmodulin complexes in the activation of the hepatic cell proliferation the possible implication of this complex on the loss of sialic acid, observing the effect of trifluoperazine (inhibitor of Ca2+-calmodulin complexes) during the prereplicative phase of liver regeneration has been studied. The results show a delay in the decrease of the amount of sugar studied from 10 to 12 hours compared to the results obtained with the hepatectomized rats that have not received trifluoperazine.

Animals↗

Quantitative and qualitative changes of plasma membrane glycoproteins in the early period of liver regeneration.

Alterations of cell surface glycoconjugates have been observed in many developing systems and may be important in the physiological control of growth and differentiation. Liver regeneration after partial hepatectomy is a suitable model in which to study the regulatory mechanisms of cell proliferation in vivo. We have isolated the sinusoidal plasma membrane of hepatocytes at different times after partial hepatectomy. The sialic acid content and the SDS-polyacrylamide gel electrophoresis pattern of glycoproteins were determined. A decrease of periodic acid-Schiff-profiles, a change in the binding capacities of 125I-concanavalin A, a reduction of the sialic acid content and the appearance and disappearance of specific components have been observed during the pre-replicative phase of liver regeneration. These findings during this early period are consistent with the active involvement of the plasma membrane glycoproteins in the transition of cells to the proliferative state.

Animals↗

Low-resolution autoradiography in rat brain after administering labelled CDP-choline.

14C-Methyl labelled cytidine diphosphate choline (CDP-choline, citicoline, Somazina) at a dose of 10 microCi has been administered orally to rats. Half of animals were sacrificed at 5 h, and other half at 24 h after administration. Autoradiography preparations of brain were isolated according to a relevant method for optical microscopy examination. Extra- and intracellular distribution of the drug after different unlabelling times has been studied. The intracellular radioactivity was checked 24 h after administration.

Animals↗

High-resolution autoradiography in mouse brain 24 h after radiolabelled CDP-choline administration.

A group of mice was given 10 microCi/animal of cytidine diphosphate choline (CDP-choline, citicoline, Somazina) (methyl 14C). Animals were sacrificed 24 h after administration and their brains isolated for electron microscopy and autoradiography. The main purpose of the present study was to examine the intracellular distribution of the drug and its possible specificity site in specific organelles and cellular structures. While a large quantity of radioactivity appeared in cellular membranes, no selectivity was in its distribution thus being detected in several cellular organelles and free hyaloplasm.

Animals↗

High-resolution autoradiography in mouse brain and cerebellum 10 days after radiolabelled CDP-choline administration.

A dose of 5 microCi of cytidine diphosphate choline (CDP-choline, citicoline, Somazina) (methyl 14C) was administered to mice. Animals were sacrificed 10 days after administration and their brain and cerebellum isolated for autoradiography and electron microscopy. Sections were prepared in such a manner that a general topographic assignment of radioactivity distribution to different layers and brain structures could be made. Regarding the intracellular distribution of the different cellular types observed, Purkinje's cells exhibited in a density of focus emission higher than the others.

Animals↗

Effect of lindane on the myelination process in the rat.

After lindane administration at several doses, brain myelin fractions of litters of male and female Wistar rats show a significant diminution of CNP (2',3'-cyclic nucleotide 3'-phosphodiesterase) activity. Furthermore, the immunohistochemical study of brains by means of a MBP (myelin basic protein) specific antibody reveals myelin deficits in some brain regions after lindane treatment. This loss of myelin protein is dose dependent. The deficit in myelin cannot be attributed to undernourishment of lindane-administered rats. This work shows the vulnerability of the developing central nervous system (CNS) to lindane and the correlation between a decrease in the CNPase activity and a deficit of MBP during the period of study of these animals.

2',3'-Cyclic-Nucleotide Phosphodiesterases↗

New nuclear functions for calmodulin.

The data reported here summarize a series of results which reveal new functions for nuclear calmodulin (CaM). The addition of CaM inhibitors to cultures of proliferating NRK cells blocked the activity of the cyclin-dependent protein kinases 4 (cdk4) and 2 (cdk2), which are enzymes implicated in the progression of G1 and in the onset of DNA replication, respectively. CaM modulates the activity of cdk4 by regulating the nuclear location of both cdk4 and cyclin D, its associated regulatory subunit. By using CaM-affinity chromatography, we have recently identified two new nuclear CaM-binding proteins: (i) the protein La/SSB, which is an autoantigen implicated in several autoimmune diseases such as lupus erythematosus and Sjögren's syndrome (since La/SSB participates in the process of transcription mediated by RNA polymerase III, CaM could be involved in the regulation of this process); and (ii) the protein SAP145, a member of the spliceosome-associated proteins (SAPs) which is a subunit of the splicing factor SF3(b). This finding suggests the involvement of CaM in pre-mRNA splicing. Finally, a screening for new CaM-binding proteins in the fission yeast performed by using the phage display analysis, revealed that several nucleolar-ribosomal proteins associate to CaM, suggesting that CaM modulates ribosomal assembly and/or function.

Animals↗