Mechanisms of arm-tip regeneration in the sea star,Leptasterias hexactis

Summary Wound healing and regeneration following amputation of arm-tips of the sea star, Leptasterias hexactis, are described using light microscopy, SEM, TEM, and [³H] thymidine autoradiography. The process can be divided into a number of stages. Initially, the wound is closed by contractions of the stump-tip. Re-epithelialization then occurs through migration of epidermal cells from adjacent areas over the wound to form a thin wound epidermis. This is converted into a thicker, permanent covering in concurrence with the onset of cell cycle activity in the wound epidermis and adjacent epidermal regions. Histolysis and phagocytosis of damaged tissues occur beneath the new epidermis and a small connective tissue scar develops at the wound site within which muscle differentiates. At this time, elevated levels of [³H]thymidine incorporation are initiated in the sub-epidermal tissues of the arm-tip. A variety of differentiated cell types enter the cell cycle including cells of the parietal peritoneum, lining of the radial water canal, and the dermis. Cell division is accompanied by the development of a small new arm-tip complete with terminal ossicle, terminal tentacle, and optic cushion. The radial water canal, radial nerve, and perivisceral coelom extend by outgrowth into this newly developing tip. Accelerated growth of the regenerate then occurs in a zone just proximal to the new tip. There is no evidence of a blastema-like mass of rapidly dividing undifferentiated cells at the tip of regenerating arms. Arm-tip regeneration in this sea star may therefore be best described as a morphallactic-like process in which a true blastema is not formed, but in which scattered cell proliferation plays an essential role.     

Morphological and mechanical aspects of fission in Ophiocomella ophiactoides (Echinodermata,Ophiuroida)

Summary During fission the ophiuroid Ophiocomella ophiactoides splits across the disc into two halves each of which regenerates to form a complete individual. This paper describes the gross anatomy of the fission plane and the histology, ultrastructure, and mechanical properties of key structures transected during fission.Rupture of the disc integrument appears not to be facilitated by a pre-determined plane of weakness. Comparison of naturally split and artificially split animals showed that at fission a mechanism operates which restricts breakage to the interradial plane of two jaws. The interradial plane is subtended mainly by collagenous ligaments and by muscles linked to the skeletal components by basal lamina-like tendinous fibres. No fission-related adaptations could be identified in the oesophagus, circumoral nerve ring, or circumoral water vascular canal.On the basis of creep tests on isolated preparations of the disc integrument and jaw-frame and the responses of these preparations to ionic manipulation, it is suggested that their behaviour is dominated by collagenous elements that can undergo actively controlled changes in their mechanical properties. A hypothesis is presented which proposes a role for such a mechanism in the initiation and facilitation of fission.Contribution 318 of the Discovery Bay Marine Laboratory, University of the West Indies     

Metabolic profiling of the resurrection plant Haberlea rhodopensis during desiccation and recovery

Desiccation tolerance is among the most important parameters for crop improvement under changing environments. Resurrection plants are useful models for both theoretical and practical studies. We performed metabolite profiling via gas chromatography coupled with mass spectrometry (GC‐MS) and high‐performance liquid chromatography (HPLC) and analyzed the antioxidant capacity of the endemic resurrection plant Haberlea rhodopensis at desiccation and recovery. More than 100 compounds were evaluated. Stress response included changes in both primary and secondary metabolic pathways. The high amounts of the specific glycoside myconoside and some phenolic acids – e.g. syringic and dihydrocaffeic acid under normal conditions tend to show their importance for the priming of H. rhodopensis to withstand severe desiccation and oxidative stress. The accumulation of sucrose (resulting from starch breakdown), total phenols, β‐aminoisobutyric acid, β‐sitosterol and α‐tocopherol increased up to several times at later stages of desiccation. Extracts of H. rhodopensis showed high antioxidant capacity at stress and normal conditions. Myconoside was with the highest antioxidant properties among tested phenolic compounds. Probably, the evolution of resurrection plants under various local environments has resulted in unique desiccation tolerance with specific metabolic background. In our case, it includes the accumulation of a relatively rare compound (myconoside) that contributes alone and together with other common metabolites. Further systems biology studies on the involvement of carbohydrates, phenolic acids and glycosides in the desiccation tolerance and antioxidant capacity of H. rhodopensis will definitely help in achieving the final goal – improving crop drought tolerance.     

Karyotypic differences and evolutionary tendencies of some species from the subgenusObliquodesmus Mlad. of genusScenedesmus Meyen (Chlorophyta, Chlorococcales)

Karyotype structures ofScenedesmus acuminatus (Lagerch.) Chod. andScenedesmus pectinatus Meyen are compared. The karyotype ofS. acuminatus (n = 5) is described for the first time. It reveals four large metacentric and one large submetacentric chromosomes (4M + 1SM). The established karyotype differences have been helpful in clarifying the taxonomic position of these two species. The cytological analyses of other related clonal cultures suggest an evolutionary transition fromS. pectinatus towardsS. regularis throughS. pectinatus f.regularis, which correlates with the morphological data about their variability. These results are discussed from the cytogenetic, morphological and evolutionary point of view. On the basis of the karyotypic analysis, it was confirmed that from a taxonomic point of viewS. pectinatus, S. acuminatus andS. regularis are separate biological species.     

L-citrulline supplementation reverses the impaired airway relaxation in neonatal rats exposed to hyperoxia

Background

Hyperoxia is shown to impair airway relaxation via limiting L-arginine bioavailability to nitric oxide synthase (NOS) and reducing NO production as a consequence. L-arginine can also be synthesized by L-citrulline recycling. The role of L-citrulline supplementation was investigated in the reversing of hyperoxia-induced impaired relaxation of rat tracheal smooth muscle (TSM).

Methods

Electrical field stimulation (EFS, 2–20 V)-induced relaxation was measured under in vitro conditions in preconstricted tracheal preparations obtained from 12 day old rat pups exposed to room air or hyperoxia (>95% oxygen) for 7 days supplemented with L-citrulline or saline (in vitro or in vivo). The role of the L-citrulline/L-arginine cycle under basal conditions was studied by incubation of preparations in the presence of argininosuccinate synthase (ASS) inhibitor [α-methyl-D, L-aspartate, 1 mM] or argininosuccinate lyase inhibitor (ASL) succinate (1 mM) and/or NOS inhibitor [N^ω-nitro-L-arginine methyl ester; 100 μM] with respect to the presence or absence of L-citrulline (2 mM).

Results

Hyperoxia impaired the EFS-induced relaxation of TSM as compared to room air control (p < 0.001; 0.5 ± 0.1% at 2 V to 50.6 ± 5.7% at 20 V in hyperoxic group: 0.7 ± 0.2 at 2 V to 80.0 ± 5.6% at 20 V in room air group). Inhibition of ASS or ASL, and L-citrulline supplementation did not affect relaxation responses under basal conditions. However, inhibition of NOS significantly reduced relaxation responses (p < 0.001), which were restored to control level by L-citrulline. L-citrulline supplementation in vivo and in vitro also reversed the hyperoxia-impaired relaxation. The differences were significant (p <0.001; 0.8 ± 0.3% at 2 V to 47.1 ± 4.1% at 20 V without L-citrulline; 0.9 ± 0.3% at 2 V to 68.2 ± 4.8% at 20 V with L-citrulline). Inhibition of ASS or ASL prevented this effect of L-citrulline.

Conclusion

The results indicate the presence of an L-citrulline/L-arginine cycle in the airways of rat pups. L-citrulline recycling does not play a major role under basal conditions in airways, but it has an important role under conditions of substrate limitations to NOS as a source of L-arginine, and L-citrulline supplementation reverses the impaired relaxation of airways under hyperoxic conditions.     

Polyamines and cell proliferation in the sea star Pycnopodia helianthoides

1. Diamines (putrescine and cadaverine) and polyamines (spermidine and spermine) were extracted from tissues of the sea star Pycnopodia helianthoides, separated and quantitated using reversed-phase high-performance liquid chromatography (RP-HPLC). Simultaneous measurements of levels of protein and DNA and rates of incorporation of 14C-thymidine were carried out. 2. The most abundant polyamine in tissues was spermidine (0.3873-2.5282 nmol/mg tissue) followed by spermine (0.103-1.5517 nmol/mg tissue), putrescine (0.2096-0.5322 nmol/mg tissue) and cadaverine (0.022-0.6064 nmol/mg tissue). 3. An unknown molecule with derivatization and elution behaviour similar to that of polyamine standards was detected in all tissues. 4. Protein levels ranged from 20.47 mg/g tissue in the body wall to 48.44 mg/g tissue in the pyloric caecum. 5. DNA levels were lowest in the ovary (0.25 mg/g tissue) and highest in the testis (5.62 mg/g tissue). 6. Incorporation of 14C-thymidine was highest in the testis. Testicular tissue had the highest spermidine/spermine ratio (5.4). A significant correlation between the spermidine/spermine ratio and 14C-thymidine incorporation (expressed either as DPM/g tissue or DPM/mg protein) suggests that polyamines are implicated in the regulation of cell proliferation in the sea star P. helianthoides.     

Evidence for the influence of vitamin C on age- and heat exposure-dependent deterioration of biochemical function in rat's liver and kidney

Molecular mechanisms responsible for age-dependent deterioration of biochemical functions have not been completely revealed as yet. We studied the role of ascorbic acid food supplementation in young and aged acute heat-exposed rats. The duration of heat exposure (40±0.5 °C) for heat-exposed Wistar rats, at the age of 35 days and 22–24 months, was approximately 2 h. In the aged heat-unexposed animals cholesterol and triglycerides were considerably high, whereas tissues ascorbic acid, glutathione and methylglyoxal were significantly low. Administration of vitamin C reverted these age-associated differences to the status comparable to young rats. The role of vitamin C supplementation was almost the same in young heat-exposed animals. In this direction in young rats suppression of LTC₄ synthesis is evident during acute heat exposure as a result of vitamin C treatment. The importance of vitamin C treatment for young heat-exposed rats is in the protection of apoptosis, if it is determined across the LTC₄ changes. In contrary, in old heat-exposed rats, vitamin C does not suppress the apoptotic processes. The results suggest that oxidative and apoptotic processes in the liver and the kidney as a result of the acute heat exposure is presumably subject of ascorbic acid deficiency.     

Karyotypic analysis of two algae species Scenedesmus incrassatulus Bohl and Scenedesmus antennatus Bréb (Chlorophyta, Chlorococcales)

The karyotypes (number, morphology and size of the chromosomes) of two algae species of Scenedesmus genus, S. incrassatulus and S. antennatus, were studied. The karyotype of S. incrassatulus (n=4) was asymmetric, characterized by two large metacentric, one large submetacentric and one small metacentric chromosomes. The karyotype assembly of S. antennatus (n=6) reveals two metacentrics and four submetacentrics. This karyotype was symmetric. The general chromosomal formulae of both species, as well as the total average metaphase length of their haploid set are presented. The results of chromosomal studies of other related species are compared and discussed. Data from the karyotypic analysis showed that S. incrassatulus, S. antennatus and S. obliquus are separate biological species from taxonomical point of view.     

L-type Ca2+ channels’ involvement in IFN-γ-induced signaling in rat ventricular cardiomyocytes

Journal of Physiology and Biochemistry - The purpose of this study was to examine the effects of interferon-γ (IFN-γ) on calcium movement in rat ventricular myocytes. L-type Ca2+ currents...