Reproductive biology of Oreochromis niloticus (Perciformes: Cichlidae) at Emiliano Zapata dam, Morelos, Mexico

The reproductive season of tilapia was studied by monthly samplings at Emiliano Zapata dam, Morelos State, Mexico. From February 1999 through February 2000 a sample of 50 fish was taken from the commercial catch (castnet, 6.5 cm of mesh size). The observed sex ratio was 1:1.29 (females:males) (2=10.26; p<0.05). The tilapia reached maturity at 151.3 mm (females) and 152.0 mm (males) of total length. Rainy (August) and dry (February) seasons were determined as the breeding period. Fecundity variation was better correlated with length (r=0.7473; p<0.002) than with weight (r=0.7395; p<0.002). The fecundity ranged between 243 and 847 oocytes per fish, with egg diameter from 300 to 3 700 µm. Intensive breeding activity in August and February coincide with phytoplankton biomass increase

Se analizó la biología reproductiva de la tilapia en la presa Emiliano Zapata, Morelos, México. Para esto se realizaron muestreos de febrero 1999 a febrero 2000. Se tomaron 50 organismos mensuales de la captura comercial obtenidos con una atarraya de 6.5 cm de luz de malla. La proporción sexual fue de1:1.29 hembras:machos (2=10.26; p<0.05). La madurez sexual se alcanza a los 151.3 mm (hembras) y a los 152.0 mm (machos) de longitud total. Se detectaron dos épocas de reproducción para la especie: durante la estación lluviosa (agosto) y durante la estación de secas (febrero). La fecundidad relativa presentó mayor correlación con la longitud (r=0.7473; p<0.002) que con el peso (r=0.7395; p<0.002). Por otra parte, el intervalo para la fecundidad osciló entre 243 y 847 ovocitos por pez, con diámetros de 300 a 3 700 µm. Asimismo, la actividad reproductiva de la especie en Agosto y Febrero, coincide con el incremento de biomasa fitoplanctónica

Reproductive biology of Oreochromis niloticus (Perciformes: Cichlidae) at Emiliano Zapata dam, Morelos, Mexico.

The reproductive season of tilapia was studied by monthly samplings at Emiliano Zapata dam, Morelos State, Mexico. From February 1999 through February 2000 a sample of 50 fish was taken from the commercial catch (castnet, 6.5 cm of mesh size). The observed sex ratio was 1:1.29 (females:males) (chi(2)=10.26; p<0.05). The tilapia reached maturity at 151.3 mm (females) and 152.0 mm (males) of total length. Rainy (August) and dry (February) seasons were determined as the breeding period. Fecundity variation was better correlated with length (r=0.7473: p<0.002) than with weight (r=0.7395; p<0.002). The fecundity ranged between 243 and 847 oocytes per fish, with egg diameter from 300 to 3 700 microm. Intensive breeding activity in August and February coincide with phytoplankton biomass increase.

Reproductive aspects of Oreochromis niloticus (Perciformes: Cichlidae) at Coatetelco Lake, Morelos, Mexico

Sex ratio, size at maturity, maturity stages, fecundity and egg diameter of Oreochromis niloticus from Coatetelco Lake, Morelos State, Mexico, were studied from January to December 1993. Sex ratio (male:female) was approximately 1:1.02. Length at maturity was 117 mm (males) and 120 mm (females). The fecundity ranged between 104 and 709 eggs, with egg diameter from 1,000 to 3,000 microns. The gonadosomatic and hepatosomatic index indicate that the species breeds during summer and winter.

Reproductive aspects of Oreochromis niloticus (Perciformes: Cichlidae) at Coatetelco Lake, Morelos, Mexico.

Sex ratio, size at maturity, maturity stages, fecundity and egg diameter of Oreochromis niloticus from Coatetelco Lake, Morelos State, Mexico, were studied from January to December 1993. Sex ratio (male:female) was approximately 1:1.02. Length at maturity was 117 mm (males) and 120 mm (females). The fecundity ranged between 104 and 709 eggs, with egg diameter from 1,000 to 3,000 microns. The gonadosomatic and hepatosomatic index indicate that the species breeds during summer and winter.

Prothymosin alpha, a mammalian c-myc-regulated acidic nuclear protein, provokes the decondensation of human chromosomes in vitro.

Prothymosin (ProT alpha) is an acidic nuclear protein, widely distributed in mammalian cells, whose expression is regulated by c-myc and linked to cell proliferation. ProT alpha interacts with histone H1 via its acidic domain, and its overexpression provokes the unfolding of chromatin fibers. Here we show that incubation of human native metaphase chromosomes with ProT alpha induces their extensive unravelling suggesting a function of this protein in chromosome decondensation.

Combined effects of troglitazone and muscle contraction on insulin sensitization in Balb-c mouse muscle.

Thiazolidinediones, represented by troglitazone, are insulin-sensitizing agents with proven efficacy for the treatment of type 2 diabetes. Exercise is also recommended for patients with type 2 diabetes because it both stimulates glucose uptake directly and it increases insulin sensitivity following exercise. The purpose of this study was to investigate the effects of troglitazone combined with exercise on 2-deoxyglucose (2DG) uptake in both the epitrochlearis and soleus muscle of Balb-c mice. Acute, 1-h treatment with troglitazone (10 or 20 microM), in the presence or absence of insulin, had no effect on 2DG uptake in either muscle. Chronic treatment with troglitazone by feeding enhanced the insulin sensitivity and responsiveness of 2DG uptake primarily in the epitrochlearis. Direct electrical stimulation of in situ muscle was used to model exercise while the contralateral muscle was used as the unexercised control. This model mimicked exercise in that glycogen was depleted, immediate 2DG uptake was enhanced, and there was a post-exercise increase in insulin sensitivity. Troglitazone feeding had no effect on 2DG uptake in the soleus when measured immediately after electrical stimultion. However, 2DG uptake in the unstimulated epitrochlearis from troglitazone-fed mice was elevated when measured immediately after removal such that no additional effects of the electrical stimulation were measured. We found that the insulin-sensitizing effect of troglitazone was not additive to the insulin-sensitizing effect of exercise, which suggests that troglitazone and exercise share similar pathways. A unique finding in this study was the differential response to troglitazone between the epitrochlearis (fast twitch) and the soleus (slow twitch) muscle types. Possible mechanisms are discussed.

Differential expression of thymosins beta(4) and beta(10) during rat cerebellum postnatal development.

The beta-thymosins are a family of actin monomer-sequestering proteins widely distributed among vertebrate classes. The most abundant beta-thymosins in mammalian species are thymosin beta(4) (Tbeta(4)) and thymosin beta(10) (Tbeta(10)), two small peptides (43 amino acids) sharing a high degree of sequence homology. In the present work, we have analyzed the distribution of Tbeta(4) and Tbeta(10) in the developing and adult rat cerebellum using in situ hybridization and immunohistochemistry techniques. Our results show that the temporal and cellular patterns of expression of both beta-thymosins are different. In the young (7 and 18 postnatal days) and adult (1 and 4 months old) rat cerebellum, Tbeta(4) was mainly expressed in the glia (microglia, Golgi epithelial cells and oligodendrocytes), neurons (granule cells and Purkinje cells), and in the capillaries. In 14-month-old rats, the Tbeta(4) immunoreactivity was only detected in some microglia cells. In young and adult animals, most of the Tbeta(10) immunoreactivity was localized in several types of neuronal cells including granule cells, Golgi neurons and Purkinje cells. In old animals, a faint Tbeta(10) signal could be detected in a few Purkinje cells. Our results suggest that each beta-thymosin could play a different function in the control of actin dynamics.

Prothymosin alpha.

Prothymosin alpha (ProT alpha) is a highly acidic protein widely distributed in mammalian cells. Since its discovery in 1984, the biological role of this protein has been controversial. Initially, ProT alpha was considered a thymic factor with a hormonal-like role in the maturation of T-lymphocytes. However, molecular and cellular analyses led to conclude that ProT alpha is a nuclear protein required in proliferation events while failing to show a clear immunological effect. The involvement of ProT alpha in changes in the compaction state of chromatin has been recently elucidated with the demonstration that this protein induces the unfolding of chromatin fibres in a process that seems to be mediated by the interaction of ProT alpha with histone H1. This finding opens up new perspectives in the study of the dynamics of the genetic material in mammalian cells. Furthermore, the relationship between ProT alpha and apoptosis as well as with proliferation makes this protein an attractive target in the search for modulators of cell death and tumour growth.

Expression of the thymosin beta10 gene in normal and kainic acid-treated rat forebrain.

Thymosin beta10 (Tbeta10) is a small actin-sequestering peptide widely distributed in mammalian tissues including nervous system. Here, we analyze the expression of Tbeta10 gene in normal and kainic acid (KA)-treated rat forebrain by in situ hybridization. Our results showed a defined regional pattern of the mRNA encoding for Tbeta10 in both normal and KA-treated rat forebrain. The presence of this transcript in different regions of the rat forebrain, including hippocampus, neocortex and several brain nuclei, provides evidence for the participation of Tbeta10 in the control of the actin dynamics that takes place in neurons. Furthermore, the analysis of the forebrain in KA-treated rats revealed an activation of the Tbeta10 gene linked to gliosis.

Expression of thymosin beta4 messenger RNA in normal and kainate-treated rat forebrain.

Thymosin beta4 is a major actin-sequestering peptide widely distributed in mammalian tissues, including the nervous system. In the present study, we analyse the expression of thymosin beta4 in normal and kainate-treated rat forebrain. In untreated animals, thymosin beta4 messenger RNA is mainly expressed in neurons of the hippocampal formation, neocortex and amygdaloid complex, as well as in oligodendrocytes. Other high-expressing areas are the tanycytic ependyma of the infundibulum, the substantia nigra pars compacta, and the supraoptic and premammillary nuclei. In rats treated with kainate, an excitotoxin that induces synaptic activation in the CA1-CA3 pyramidal neurons of the hippocampus, the levels of thymosin beta4 were clearly increased in the hippocampus and neocortex during the first 2-3 h after injection. In the long term, kainate causes neuronal degeneration in the CA1-CA3 regions of the hippocampus and functionally related structures, provoking a depletion of thymosin beta4 messenger RNA in these areas; however, the levels of this transcript are restored two weeks after kainate injection. Moreover, we have found that, in these degenerating zones, gliosis is accompanied by an elevation of the levels of thymosin beta4 messenger RNA, particularly in the CA1-CA3 region of the hippocampus, the lateral geniculate nucleus and the mammillothalamic tract. The present results demonstrate the existence of relatively high levels of thymosin beta4 messenger RNA in several areas of the rat forebrain, indicating that this peptide plays an important role in the regulation of actin polymerization in these regions of the brain. Moreover, the elevation of this messenger RNA after kainate treatment suggests a function of thymosin beta4 in the production and remodelling of neuronal processes. Finally, our findings provide evidence for a participation of this actin-sequestering molecule in the reactivity of certain types of glial cell that follows kainate lesions.

A non-hypervariable human minisatellite strongly stimulates in vitro intramolecular homologous recombination.

Several features indicate that the low polymorphic human minisatellite MsH42 region could be involved in recombination. It contains different well-known recombination motifs, is able to generate single-stranded loops and is specifically recognized by nuclear proteins. These characteristics led us to investigate the possible recombinogenic activity of the MsH42 region in terms of intramolecular recombination. We constructed two plasmids, one of them carrying two copies of the minisatellite region and the other one containing sequences upstream of this repetitive region. We showed that MsH42 strongly stimulates intramolecular in vitro recombination, approximately 22 times more than the control sequence, solely when the source of biological extract is mouse testes, suggesting that MsH42 could be a hotspot involved in meiotic recombination. Furthermore, there is a direct relationship between the frequency of equal crossovers and the enhancement of recombination. Interestingly, the third repeat of the minisatellite array is always involved in the resolution of unequal crossovers leading to minisatellite shortening. As far as we know, our results provide the first evidence that a non-hypervariable minisatellite can enhance homologous recombination.

Prothymosin alpha is a chromatin-remodelling protein in mammalian cells.

Prothymosin alpha (ProTalpha) is an abundant mammalian acidic nuclear protein whose expression is related to cell proliferation. Here we report that in HL-60 cells overexpressing ProTalpha, the accessibility of micrococcal nuclease to chromatin is strongly increased. In the DNA ladder generated by the nuclease activity, the sizes of the mononucleosome (146 bp, the DNA fragment that is bound to the histone octamer) and its multimers correspond to nucleosomes lacking histone H1. The percentage of histone-H1-depleted chromatin (active chromatin) is also higher in the cells overexpressing ProTalpha. On the basis of these and previous findings, we propose a biological role for ProTalpha in the remodelling of chromatin fibres through its interaction with histone H1.

Overexpression of prothymosin alpha accelerates proliferation and retards differentiation in HL-60 cells.

Prothymosin alpha (ProTalpha) is an acidic nuclear protein the expression of which is related to the proliferation and differentiation processes in mammalian cells. In the present study we have stably transfected HL-60 cells, a biological system that allows the study of both proliferation and differentiation, with recombinant vectors encoding sense and antisense ProTalpha mRNA. In the HL-60 cell clones overexpressing ProTalpha we observed an acceleration in the growth rate, whereas expression of the antisense orientation showed the opposite effect. Moreover, cell-cycle analysis demonstrated that the G1-phase was shortened in the cells expressing the sense construct. Before studying how ProTalpha affects differentiation, we showed that the down-regulation of ProTalpha gene during differentiation occurs in all mammalian cell lines (HL-60, K562, U937, MEL C88, N2A and PC12) analysed. The biological effect evoked by the induction of the ProTalpha sense vector was the retardation of cell differentiation, although expression of the antisense construct showed no effect on differentiation. In conclusion, our findings provide evidence that ProTalpha is directly implicated in cellular proliferation and that the maintenance of high levels of ProTalpha inside HL-60 cells is incompatible with their ability to differentiate.

Molecular characterization of a new human minisatellite that is able to form single-stranded loops in vitro and is recognized by nuclear proteins.

We report the isolation of a new low polymorphic GC-rich human minisatellite locus (MsH42) that contains different recombination motifs and is homologous to sequences involved in immunoglobulin class-switching. Furthermore, we show that MsH42 undergoes slipped-strand mispairing during PCR indicating its ability to generate single-stranded loops. Specific DNA-protein complexes were detected in band-shifting experiments using nuclear extracts from mouse testes and human NC-37 cells. The possible implications of this minisatellite in recombination events is discussed.

A new EcoRI family of satellite DNA in lampreys.

Satellite DNA sequences have been studied in several groups of organisms. However, until now this type of sequence has not been characterized in cyclostomata, an evolutionarily important class of vertebrates. In the present work, we report the molecular characterization of a new family of satellite DNA in lampreys (Petromyzon marinus). Digestion of lamprey DNA with EcoRI identified a series of very abundant AT-rich (60% A+T) repeating units, with short stretches of AT, that are multimers of 370 bp. Southern blot analysis and comparison with the satellite DNA sequences deposited in the databases indicate that this new family of satellite DNA is exclusive to lampreys. The distribution of this EcoRI satellite DNA on lamprey chromosomes was analyzed by in situ hybridization. The evolutionary origin of this satellite is briefly discussed.

Thymosin beta10 mRNA expression during early postimplantation mouse development.

The beta-thymosins are a family of monomeric actin sequestering peptides that regulate actin dynamics within the cells. During embryogenesis the control of actin polymerization is essential in processes such as cell migration, angiogenesis and neurogenesis. Here we report that the levels of thymosin beta10 (Tbeta10) mRNA strongly increase during early postimplantation mouse embryogenesis as well as during in vitro P19 cell differentiation, indicating that this peptide plays an important role in early development. Moreover, analysis of the spatial distribution of Tbeta10 mRNA in 9.5-12.5 days postcoitum mouse embryos showed a remarkable presence of this transcript in mesenchymal structures as well as in the mantle layer of spinal cord. Interestingly, we observed differences in the distribution of the mRNAs encoding Tbeta10 and Tbeta4, another member of the beta-thymosin family, suggesting different roles for these peptides during mouse embryogenesis.

High levels of mouse thymosin beta4 mRNA in differentiating P19 embryonic cells and during development of cardiovascular tissues.

The self assembly of actin and the large number of actin-binding proteins are important in the establishment of cell shape and function during embryogenesis. Thymosin beta4 (Tbeta4) is a small acidic peptide that participates in the regulation of actin polymerization in mammalian cells. In the present work, we report the presence of the mRNA encoding for Tbeta4 in mouse embryonic stem cells and its induction in P1 9 embryonal cells stimulated to differentiate into ectodermal-like (neurons and glia) or mesodermal-like cells (cardiac and skeletal muscle). The induction of Tbeta4, mRNA in P19 cells was confirmed by in situ hybridization analysis of early mouse postimplantation embryos. Noteworthy, we observed an important hybridization signal in several areas of the embryo specially in blood vessels and in heart tissues, suggesting a role for this peptide in angiogenesis. In conclusion, the results presented here demonstrate the expression of Tbeta4 gene during early embryogenesis which immediately suggests an important role for this peptide in developmental processes requiring actin-based functions such as the formation of cardiovascular system.

The thymosin beta 4 gene is strongly activated in neural tissues during early postimplantation mouse development.

We studied the temporal and spatial distribution of the mRNA encoding for thymosin beta 4 (T beta 4), a small acidic actin-sequestering peptide, during the early postimplantation mouse development. Analysis of total embryo RNA demonstrated a strong activation of T beta 4 gene after gastrulation and coincident with neurulation. In situ hybridization showed that T beta 4 mRNA was strongly expressed in the central nervous system and peripheral ganglia, paralleling the gradient of neuronal differentiation. An intense signal was also observed in intraventricular macrophages and blood vessels. The role of T beta 4 in mammalian neuroembryogenesis is discussed.

Prominent expression of the actin-sequestering peptide Fx gene in the hippocampal region of rat brain.

We have analyzed by in situ hybridization the distribution of Fx (an actin-sequestering peptide) mRNA in the brain of young and old rats. The strongest Fx mRNA-specific hybridization signal was located in the hippocampo-entorhinal cortex; this mRNA was also found in the piriform cortex, amygdala, lateral septum and neocortex. Northern blot analysis confirmed the prominent expression of the Fx transcript in the hippocampus and showed a notably lower amount of this mRNA in the hippocampus of old rats suggesting an influence of aging on the expression of this gene. The possible implication of Fx in synaptic plasticity and in long-term potentiation is discussed.