The sea urchin (Sphaerechinus granularis) codes different H2B histones to assemble sperm and embryo chromatin.

Histones from sperm and gastrula nuclei of the sea urchin Sphaerechinus granularis were analyzed by three different electrophoretic methods, stained with specific color reactions and compared with one another. The histones of the two cell types showed identical properties in none of the three analytical methods employed but the greatest differences involved the H2B type of histones. H2B from embryo chromatin had properties similar to those of the corresponding molecule from calf thymus. In the sperm chromatin no such molecule was observed but two additional ones were found that differed from embryo H2B in size, charge and specific colour reactions. The differences were such that different genes may be postulated for the synthesis of the H2B histones found in sperm and in embryo.     

Single-stranded regions in DNA isolated from different developmental stages of the sea urchin

Long-term labeled sea urchin embryo (Strongylocentrotus purpuratus) DNAs were examined for size of recovered pieces, single-strandedness, and length of continuous double-stranded regions. Sizing on neutral sucrose gradients indicates that morula stage DNA sediments predominantly at 31 S, blastula stage DNA at 27 S, and gastrula stage DNA as a broad range of sizes of greater than 29 S. Treatment of [3H]thymidine-labeled DNA with Aspergillus oryzae S1 nuclease removes 19% of the 3H from morula stage DNA, 4% of the 3H from blastula stage DNA, and less than 0.1% of the 3H from gastrula stage DNA. Sedimentation of S1 nuclease treated [3H]DNAs on alkaline sucrose gradients indicates that in native morula stage DNA there is a nick or gap in one strand approximately every 9700 base pairs, in native blastula stage DNA about every 3300 base pairs, and very few nicks or gaps in native gastrula stage DNA.     

A kinetic analysis of the changes in membrane permeability induced by fertilization in the egg of the sea urchin Sphaerechinus granularis

Fertilization-induced modifications in egg membrane permeability to labeled adenine and l-leucine substrates have been studied by the initial rate-pulse technique, using the sea urchin Sphaerechinus granularis. Kinetic parameters have been determined for unfertilized eggs as well as for fertilized eggs at various developmental stages. Samples were usually processed according to a centrifuge-wash technique, since Millipore filtration was found to be inadequate in the case of leucine.Several arguments are presented which support the view that the uptake of adenine occurs through a process of simple physical diffusion across the cell membrane. It has been demonstrated, on the other hand, that the transport of l-leucine involves a carrier-mediated exchange-diffusion process.Kinetic data point to the existence of major fluctuations in general egg membrane permeability which appear to be largely independent of the mitotic cycle and of the mechanisms involved in the transport of the substrates studied. A unitary hypothesis is presented to account for these fertilization-induced changes in membrane permeability and which does not involve the synthesis or recruitment of previously inactive carriers.     

Ultrastructural,fluorescence microscopic and microfluorimetric study of the innervation of the axial complex in the sea urchin,Sphaerechinus granularis (Lam.)

Summary In juxtaposition with the contractile epithelia of the axial complex of the sea urchin, Sphaerechinus granularis, several types of nerve fibers with different vesicle populations were determined. Nerve terminals, filled with clear vesicles and dense core vesicles, form synaptoid neuromuscular junctions. Close to the somatocoelic epithelia of the axial and terminal sinus septa, numerous axon profiles form a nerve plexus. Among the epithelial cells covering the plexus, two types of nerve cells can be distinguished which presumably produce neurosecretory and aminergic granules, respectively. Monoamine fluorescence (formaldehyde-induced fluorescence, Falck-Hillarp technique) was analyzed microspectrofluorimetrically. The emission spectrum of the fluorophores occurring in the present material shows a maximum at 475 nm and is characteristic of catecholamines; the excitation maximum at 380 nm after formaldehyde treatment is typical of catecholamines at low pH only. Since the peak ratio (370:320 nm) does not change after HCl-vapor treatment, the fluorophores are likely to be indicative of dopamine.     

The effect of feeding or starvation on resource allocation to body components during the reproductive cycle of the sea urchin Sphaerechinus granularis (Lamarck)

To determine the effects of feeding or starvation on resource allocation to body components during the reproductive cycle of Sphaerechinus granularis, sea urchins were placed in laboratory tanks and either fed ad libitum or starved during two different periods of their biological cycle, i.e. the mature stage and the recovery stage. The urchin growth was monitored over the whole experimental period, the gonad, gut, lantern indices and organic matter levels of different organs were determined at the end of the experiment. During the mature stage sea urchins in good nutritional conditions did not increase in size, but allotted energy to gonad production and stored reserves in body wall. Limiting food stopped the gonadal growth without complete regression. During the recovery period food allowed somatic growth, i.e. test growth and the storage of reserves in gonad somatic cells. This somatic production did not occur under food-limited conditions and the resources allotted for survival and maintenance were taken from different body components.     

Preparation and characterization of histone H1 from the sperm of the sea-urchin Sphaerechinus granularis.

The separation and purification of histone H1 from the sperm of the sea-urchin Sphaerechinus granularis is described. Physical studies were used to compare this histone H1 molecule with H1 histones from other species. C.d. and 270 MHz n.m.r. spectroscopy indicate that, despite significant compositional differences from other sea-urchin sperm H1 histones, their secondary and tertiary structures are very similar. A large difference in helicity was, however, found between S. granularis histone H1 and calf thymus histone H1, and their n.m.r. and fluorescence spectra also differ considerably. It is concluded that secondary structure and tertiary structure have not been conserved in the evolution of the H1 histone family.     

Mitochondrial division in non nucleated sea urchin eggs

An electron microscopic analysis of non nucleated and parthenogenetically activated sea urchin eggs shows figures that can be interpreted as dividing mitochondria. This is in agreement with our previous observations showing that the egg nucleus negatively controls mitochondrial DNA synthesis.     

Effects of Bonellin on fertilization and cleavage of the eggs of the sea urchin,sphaerechinus granularis

We have studied the structural changes occurring in the eggs of the sea urchin Sphaerechinus granularis treated with bonellin, the green pigment of the sea worm Bonellia viridis, which is responsible for the masculinization of the larva of this animal. The two major targets of bonellin appear to be the cortical structures and the nuclear membrane, while the mitochondria do not appear to be affected. As a result of bonellin treatment, cleavage is prevented while nuclear divisions proceed. The possibility is discussed that the alteration of the cortical structures may interfere with the assembly of the surface microfilaments and hence with the formation of the cleavage furrows.     

Different aggregation properties of sea urchin embryonic cells at different developmental stages. II. Stage specific response to fibronectin and collagen

Fibronectin and collagen were added to cells dissociated from embryos at the blastula and at the 16 cell stages. Both molecules stimulate aggregation of cells dissociated from blastula but they do not affect aggregation of blastomeres dissociated from the 16 cell stage. The stage-specific response to fibronectin and collagen appears to be related to the onset of new functional role(s) of the two molecules at the blastula stage.     

Nutrient allocation in the gonads of the sea urchin Arbacia dufresnii in different stages of gonadal development

This study reports nutrient allocation in different stages of gonadal development for two populations of the sea urchin Arbacia dufresnii off the Patagonian coast of Argentina (Nuevo Gulf and San Jorge Gulf). The biochemical composition of gonads was used to assess nutrient allocation by measuring ash, soluble protein, lipid and trichloroacetic acid-soluble carbohydrate concentrations, and absolute contents over a 24-month period. Reproductive output in terms of energy was calculated for females. Results were correlated with histological stage of the gonads. Soluble proteins were the main component for the Nuevo Gulf population while unmeasured organic material (i.e. insoluble proteins and nucleic acids, especially in testes) was prevalent in gonads from San Jorge Gulf. Soluble protein and lipid concentrations followed the gonadal cycle, while carbohydrate concentration was almost negligible, especially in the Nuevo Gulf population. The different patterns in the gonadal cycle in the two populations were reflected in the biochemical composition of gonads. Concentrations and contents of the biochemical components and reproductive output were higher in the population from San Jorge Gulf owing to the larger size of gonads and gametes. These findings contribute to the better understanding of the plasticity of the reproductive biology of A. dufresnii in different environments.