Formation of the acrosome and basal body during spermiogenesis in a marine snail,Nerita picea (Mollusca: Archaeogastropoda)

In Nerita picea the proacrosomal granule is formed basally in the early spermatid from one large cisterna of the Golgi body, with which the other Golgi-derived vesicles fuse. After the proacrosomal granule has attached to the plasma membrane and invaginated to form a cup shape, one cisterna of endoplasmic reticulum inserts into the open end and deposits a granular secretion on the inner surface. Subsequently, the proacrosome migrates along the plasma membrane to the apex of the nucleus, but the Golgi body remains basal, as occurs in other archaeogastropods and also many polychaete annelids. However, the final shape and structure of the acrosome is similar to that of mesogastropods. The annulus attaches the distal centriole to the plasma membrane early in spermiogenesis. The production of the flagellum by the distal centriole not only expands the plasma membrane posteriorly but moves the centriolar complex to the nucleus, causing an invagination of the plasma membrane where it is bound by the annulus. During proacrosome migration, the Golgi body secretes a dense tube around the flagellum, and the mitochondria fuse into two spheres at the base of the nucleus. The nuclear plug that closes off the intranuclear canal until this stage rapidly reorganizes itself into two tubes of material inside the canal. The centrioles continue flagellar production, break away from the annulus, and move deep into the intranuclear canal where they fuse together to form the basal body of the sperm. In the maturing spermatid, the two mitochondria fuse into a single sheath that spirals around the flagellum. The annulus does not migrate posteriorly but remains anterior to the midpiece, which is unusual for a filiform sperm. Spermiogenesis in Nerita picea has features in common with both archaeogastropods and mesogastropods but also has some unique features. These observations lend credence to the idea that the Neritidae are a transitional group between Archaeogastropoda and Mesogastropoda.     

A Method for Combined Gross Skeletal Staining and Feulgen Staining of Embryonic Chick Tissues

This paper describes a combined technique for gross skeletal staining and Feulgen staining of avian embryonic limbs. The gross skeletal stain uses Victoria blue B, and the Feulgen stain is done en bloc before the skeletal stain is applied. The method has been useful in determining the cellular origins of supernumerary structures arising from experiments in which quail wing mesoderm is grafted into chick wing buds.     

Endoderm-secreted factor stimulates growth of embryonal carcinoma stem cells

Summary Stem cells of the embryonal carcinoma cell line called H6 can be induced to differnetiate to endoderm-like cells by retinoic acid (3×10⁻⁶ M). We have detected a diffusible and stable factor which is secreted by H6 endoderm-like cells and stimulates the growth of H6 stem cells. The stimulation by the endoderm-like cells is considereably greater than that by mouse fibroblasts or H6 stem cells themselves. No reciprocal stimulation of endoderm-like cells by stem cells occurs. Part but not all of the stimulation might be due to extracellular matrix proteins or to insulin-like growth factor type 2, each of which also stimulates the growth of H6 stem cells. Insulin causes no such stimulation. This work was supported by research rant no. CA-16754 from the National Cancer Institute to J. W. L. E. L. G. was supported by an American Heart Association Medical Student Research Award. Editor's Statement This paper presents a good example of cooperativity between undifferentiated teratoma stem cells and differentiated parietal endoderm-derived countrparts in terms of growth support. It raises the interesting question of the relationship between factors produced by paprietal and visceral endoderm cells. Gordon H. Sato     

Mathematical modelling of primary production in Green Bay (Lake Michigan,USA), a phosphorus-and light-limited system

Application of mathematical models in the design and evaluation of lake restoration programmes must include due consideration of three basic concepts of model development; 1) that the model framework is appropriately matched to the intended management use, 2) that selection of the proper degree of model complexity is fundamental to the achievement of model credibility and 3) that field and laboratory studies must be designed and interpreted with the aid of the model to insure development of a comprehensive, integrated tool.These concepts are demonstrated for the case of lake restoration efforts in Green Bay (Lake Michigan, USA). Striking gradients in water quality (transparency, algal standing crop, hypolimnetic oxygen depletion) and trophic state occur along the major axis of the bay in response to phosphorus loaded from the Fox River. A simple model for gross primary production is developed to permit calculation of the relative importance of internal carbon production to the total organic carbon budget of the bay. Primary production varies from high rates over a limited photic depth in the turbid, phosphorus-rich waters of the eutrophic portions of the bay to low rates over an extensive photic depth in the transparent, phosphoruspoor reaches of the oligotrophic regions. Internal production accounts for approximately 90% of the total organic carbon loaded to the system over the summer growing season. Water quality management strategies must address the stimulation of primary production by phosphorus loaded from the Fox River in any attempt to lower the standing crop of nuisance algae, improve water clarity, and reduce rates of hypolimnetic oxygen depletion in Green Bay.     

Specific binding of three neurotoxins with phospholipase A2 activity to synaptosomal membrane preparations from the guinea pig brain

Snake presynaptic toxins such as crotoxin, β-bungarotoxin and taipoxin block neuromuscular transmission through inhibiting the release of acetylcholine by their phospholipase A₂ activities. On the other hand, many other phospholipase A₂s show little neurotoxicity. It is likely that the difference lies in whether high affinity binding to nerve cell membranes exists or not. To test this idea, crotoxin, β-bungarotoxin and taipoxin were first radioactively labeled with Na(¹²⁵I) without loss of their neurotoxicity. Using the radioactive toxins we have found that each of the three showed specific binding to synaptosomal membranes from guinea pig brain. In contrast, we could not detect specific binding of a non-neurotoxic pancreatic phospholipase A₂. Crotoxin and taipoxin, but not β-bungarotoxin, also bound specifically to membrane preparation from other tissues. The binding of each toxin was not greatly affected by the other two toxins. The photoaffinity labeling technique has been used to obtain further information about the components which bind crotoxin. For this purpose, (¹²⁵I) crotoxin was derivatized with N-hydroxysuccinimidyl-4-azidobenzoate. Autoradiographic analysis of the membranes following photoirradiation in the presence of the modified crotoxin revealed that an 85K dalton component was preferentially covalently conjugated with the crotoxin analogue in a specific manner.     

Pseudoterminalisation,terminalisation, and non-chiasmate modes of terminal association

Terminal associations occur commonly between meiotic homologues of the two smallest (S10, S11) chromosomes in the northern race of Cryptobothrus chrysophorus when they are either heterozygous or homozygous for distal supernumerary heterochromatic segments. A detailed examination of the origin and behaviour of these associations provides convincing evidence that they are non-chiasmate in character and so cannot be explained by either pseudoterminalisation or terminalisation. The same is true of the terminal associations involved in the persistent pseudomultiples that develop between non-homologues of Heteropternis obscurella when one or both of these carry distal heterochromatic segments. In both situations the C-bands involved in such terminal associations are entire and are never interrupted by non-banded material. In Cryptobothrus, similar associations can also develop between centromere regions when these are heterozygous or homozygous for proximal supernumerary heterochromatic segments.     

Protein composition of the chromosomal scaffold and interphase nuclear matrix

Residual protein structures were prepared from isolated chromosomes and interphase nuclei of in vitro cultured bovine liver cells and the protein compositions were analysed. Chromosomes with minimal cytoplasmic contamination were obtained by a simple procedure using a pH 8 isolation medium containing Triton X-100 and polyamines, and residual protein-DNA complexes were prepared by extraction with 2 M NaCl. Residual protein structures were also obtained by digesting isolated chromosomes with staphylococcal nuclease. Protein compositions of both structures as obtained by SDS-polyacrylamide gel electrophoresis were essentially the same. Residual protein structures were prepared from isolated nuclei by the same procedures. The major nuclear matrix proteins, i.e., the lamins A, B, and C, were not found in the chromosomes and chromosome scaffolds. On the other hand, the residual chromosome structures contained two major polypeptides of 37 and 83 kilodalton relative molecular weights that were absent from the nuclear matrix preparations. A few polypeptides with the same or very similar electrophoretic mobilities were found in the residual structures of both the nuclei and the chromosomes.