Differential expression and localization of brain-type and mitochondrial creatine kinase isoenzymes during development of the chicken retina: Mi-CK as a marker for differentiation of photoreceptor cells

The expression and the cellular- as well as subcellular-distribution of brain-type B-CK and mitochondrial Mi-CK during development of the chicken retina was studied by immunoblotting, immunofluorescence and immunogold methods. B-CK expression and accumulation in retina was high from early stages of embryonic development on, decreased slightly around hatching and remained high again during adulthood. At early stages of development (days 2-5), B-CK was more or less evenly distributed over the entire retina with the exception of ganglion cells, which were stained more strongly for B-CK than other retinal precursor cells. Then, at around day 10, the beginning of stratified immunostaining by anti-B-CK antibody was noted concomitant with progressing differentiation. Finally, a dramatic increase in staining of the differentiating photoreceptor cells was seen before hatching (day 18) with weaker staining of other cell types. At hatching, as in the adult state, most of the B-CK was localized within rods and cones. Thus, during retinal development marked changes in the immunostaining pattern for B-CK were evident. By contrast, Mi-CK expression was low during development in ovo and rose just before hatching with a predominant accumulation of this isoenzyme within the ellipsoid portion of the inner photoreceptor cell segments. Mi-CK accumulation in the retina coincided with functional maturation of photoreceptors and therefore represents a good marker for terminal differentiation of these cells. B-CK, present from early stages of retina development, seems to be relevant for the energetics of retinal cell proliferation, migration and differentiation, whereas the simultaneous expression of both B- and Mi-CK around the time of hatching indicates a coordinated function of the two CK isoforms as constituents of a PCr-circuit involved in the energetics of vision, which, in autophagous birds, has to be operational at this point in time.     

Classification of types of ontogenetic reproduction

The object of this paper is to present an original classification of ontogenetic reproduction. The main general criterion used is the degree and type of phylogenetic differentiation. In relation to this criterion, criteria are given for the classification of the fundamental types of ontogenetic reproduction and for the classification of the types of ontogenetic generation cycles. Between the fundamental types of ontogenetic reproduction and the types of ontogenetic generation cycles there is a hierarchical relationship which shows that the former are components of the latter. Between the well-defined types of ontogenetic reproduction there exist many intermediate types.     

Influence of sarmesin on the cardiac and vascular actions of angiotensin II

Summary The angiotensin II (ANG II) receptor blocker properties of sarmesin and its influence on the homotropic cooperativity of ANG II receptors were studied in two experimental models: isolated rabbit aorta and isolated rabbit atria. The results show that: (i) sarmesin is a specific competitive antagonist of ANG II receptors, with high affinity (pA₂=8.93 in the isolated aorta and 8.66 in the isolated atria); and (ii) the slope of the concentration-response curves to ANG II and the Hill coefficient increased in the presence of sarmesin, the latter suggesting an enhancement of the positive homotropic cooperativity of ANG II receptors. These results may be explained overall by the reciprocal negative modulation of receptor affinity between sarmesin and ANG II, due, possibly, to opposite effects on the binding of G-proteins to ANG II receptors.     

Genetic variability in the European minnow,Phoxinus phoxinus (L.)

An electrophoretic study of genetic variation at 11 loci was performedfor a population of European minnows, Phoxinus phoxinus (L.). Ten loci, EST-1 ^*, EST-2 ^* EST-3 ^*,GPD-1 ^*,GPD-2 ^*,GPI-1 ^*,GPI-2 ^*,MPI ^*,6PGD ^* and PGM ^* were polymorphic. IDH ^*wasmonomorphic. The mean number of heterozygotic loci over all 176 fish was 3.05 ± 0.104(SE). Observed mean heterozygosity was 0.28±0.058(SE) and expected mean heterozygosity was 0.27±0.054(SE). EST-2 ^*, EST-3 ^* andPGM ^* were not in Hardy-Weinberg equilibrium. Length,condition, parasite numbers or male breeding characters, i.e. red colorationand tubercles, were not influenced by single enzyme loci.     

SETDB1 regulates microtubule dynamics

ObjectivesSETDB1 is a methyltransferase responsible for the methylation of histone H3‐lysine‐9, which is mainly related to heterochromatin formation. SETDB1 is overexpressed in various cancer types and is associated with an aggressive phenotype. In agreement with its activity, it mainly exhibits a nuclear localization; however, in several cell types a cytoplasmic localization was reported. Here we looked for cytoplasmic functions of SETDB1.MethodsSETDB1 association with microtubules was detected by immunofluorescence and co‐sedimentation. Microtubule dynamics were analysed during recovery from nocodazole treatment and by tracking microtubule plus‐ends in live cells. Live cell imaging was used to study mitotic kinetics and protein–protein interaction was identified by co‐immunoprecipitation.ResultsSETDB1 co‐sedimented with microtubules and partially colocalized with microtubules. SETDB1 partial silencing led to faster polymerization and reduced rate of catastrophe events of microtubules in parallel to reduced proliferation rate and slower mitotic kinetics. Interestingly, over‐expression of either wild‐type or catalytic dead SETDB1 altered microtubule polymerization rate to the same extent, suggesting that SETDB1 may affect microtubule dynamics by a methylation‐independent mechanism. Moreover, SETDB1 co‐immunoprecipitated with HDAC6 and tubulin acetylation levels were increased upon silencing of SETDB1.ConclusionsTaken together, our study suggests a model in which SETDB1 affects microtubule dynamics by interacting with both microtubules and HDAC6 to enhance tubulin deacetylation. Overall, our results suggest a novel cytoplasmic role for SETDB1 in the regulation of microtubule dynamics.

SETDB1 association with microtubules inhibits microtubule polymerization and enhances their instability. SETDB1 may affect the microtubules by interacting with HDAC6 to enhance HDAC6 tubulin deacetylation activity.     

Anti-phospholipase C antibodies inhibit the lectin-induced proliferation of human lymphocytes

A novel approach was used to assess the role of phosphoinositide hydrolysis in the mitogenic action of phytohemagglutinin (PHA) or concanavalin A (ConA). The treatment of human peripheral blood leukocytes (PBL) with monospecific antibodies against phospholipase C (PLC) produced a dose-dependent inhibition (up to 100%) of PHA (10 g/ml) or ConA (25 g/ml) proliferative effects. Thus, the activation of membrane-bound PLC is asine-qua-non condition for lectin-induced proliferation of T lymphocytes. The key-role of PLC versus protein kinase C (PKC) is stressed by the fact that the inhibition of PKC with Hidaka's compound H-7 (40 M) produced only a partial blockade (about 25%) of lectin mitogenic effect.To whom correspondence should be addressed.     

An enlarged European Union challenges priority settings in conservation

Biodiversity is declining worldwide under increasing human pressure. Since the location of and the threats are unevenly distributed and the resources available for conservation are limited, prioritization is essential to reduce the losses. Most conservation efforts until now proved to be ineffective in stopping the present worldwide decline of threatened species. We focus on the European Union (EU) after the repeated enlargements in the last decade, from 15 to 27 countries, by considering the present conservation priorities that have shifted towards a continental scale approach. The situation in the EU indicates that despite the differences in wealth across countries, there are no significant differences in the number and surface of protected areas between them, so re-evaluating conservation priorities at a continental scale and a reallocation of funds is required. A major limitation in priority settings for conservation is data availability. We recommend including in the decision process data provided by phylogeographic studies. This will prevent the decline of populations and species with evolutionary potential from centres of speciation and climate refugia. Recent EU members from central and eastern Europe still retain high biodiversity with a rather good conservation status. A large number of areas with high evolutionary potential identified by phylogeographic studies are located there and should be considered priorities within the context of global changes, as a proactive approach. We recommend a periodic re-evaluation of the status of species and habitats based on current research results, harmonization between the priority species listed in the conventions, directives and Red Lists at both EU and national levels.