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J L Hedrick

Publications and source records attributed to J L Hedrick.

At least 73 records · Page 4Linked to original sources
PubMed1980

Isolation, physicochemical properties, and macromolecular composition of zona pellucida from porcine oocytes.

Oocytes released en masse from pig ovaries were isolated in large quantities by using sieving techniques. The isolated oocytes were gently homogenized, and the largely intact zona pellucida "ghosts" were purified by using sieving techniques. Sufficient amounts of zonae were recovered to permit, for the first time, adequate characterization of the zona pellucida in chemical, physical, and macromolecular terms. The isolated zonae were greater than 93% pure as determined by chemical, enzymatic, and microscopic criteria. The zonae were completely solubilized by a variety of conditions that do not break covalent bonds. The extent of solubilization was a function of pH, ionic strength, temperature, and the presence of various solubilizing agents such as detergents and urea. Chemically, the zonae were composed predominantly of protein (71%) and carbohydrate (19%). After acid hydrolysis of the zonae, no unusual amounts or types of amino acids were detected. The monosaccharides present after hydrolysis were those typically found in animal glycoproteins (Fuc, Man, Gal, GalNAc, and GlcNAc). Sialic acid in glycosidic linkage and sulfate and phosphate esters were present and were considered to be true constituents of the zona pellucida. Other substances detected, but considered contaminants rather than true constituents, included fatty acids (esterified and free) and uronic acids. The binding by several fluorescein-conjugated plant lectins to the in situ zona pellucida was determined by using light microscopy. The binding of the lectins to the zona pellucida was not uniform, indicating that the carbohydrate moieties of the zona pellucida were asymmetrically distributed. The zona pellucida was composed of at least three macromolecules as indicated by immunodiffusion and sodium dodecyl sulfate gel electrophoresis experiments. Determination of the number of macromolecules composing the zona pellucida was compromised by the aggregation and/or microheterogeneity of its constituent macromolecules. We conclude that the zona pellucida is composed of several glycoprotein macromolecules; interaction of these macromolecules to form supramolecular complexes and the integral zona pellucida is dependent on noncovalent forces.

Amino Acids↗
PubMed1977

Alteration of structure and penetrability of the vitelline envelope after passage of eggs from coelom to oviduct in Xenopus laevis.

The vitelline envelope (VE) that surrounds an egg released from the ovary into the coelom of Xenopus laevis differs markedly, in structure and penetrability, from the VE surrounding an oviposited egg. In a coelomic egg, the filaments that form the VE are arranged in distinct fascicles or bundles. The exterior surface of the VE is irregular in contour and is permeated by channels. In an oviposited egg, the filaments are evenly dispersed and lack a fasciculated arrangement; the exterior surface is smooth and no channels are present. The fascicular arrangement of fibrils in the coelomic VE is maintained only at neutral pH, and is not visibly altered by the cortical reaction. VEs from coelomic eggs retain their fasciculated morphology after isolation from the egg. In an in vitro test system, sperm penetrated VEs isolated from oviposited eggs, but failed to penetrate VEs isolated from coelomic eggs. The structural transformation of the VE from the coelomic type to the oviposited type occurs in the first 1-cm segment of the oviduct, prior to addition of jelly to the egg. Neither intact jelly, solubilized jelly, nor jelly extracts were capable of altering the structural organization of coelomic VEs, suggesting that the structural transformation of the VE is effected by some oviducal factor other than jelly.

Animals↗
PubMed1976

Isolation, physicochemical properties, and the macromolecular composition of the vitelline and fertilization envelopes from Xenopus laevis eggs.

As a step toward defining in molecular terms the sperm-triggered block to polyspermy reaction established by the egg at fertilization, vitelline (VE) and fertilization (FE) envelopes were isolated from eggs of the Sounth African clawed toad Xenopus laevis and some of their physicochemical properties determined. Envelopes were isolated after lysis of the fertilized or unfertilized eggs by sieving techniques; isolated envelopes retained their in situ morphology as determined by electron microscopy. The isolated envelopes had different solubility properties and, in general, VE was more readily dissolved by aqueous solvents than FE, although both could be completely dissolved by detergents or chaotropic agents. Changes in envelope solubility correlated with the progression of the cortical reaction implicating a role for cortical granule material in modifying the solubility properties of the envelope. The VE and FE were composed of protein and carbohydrate with no lipid components detected. As determined by immunodiffusion experiments, the FE contained the same antigens as the VE plus components derived from the cortical granules and the innermost jelly layer, J. The macromolecular composition of the envelopes was determined by sodium dodecyl sulfate gel electrophoresis. The VE contained at least 11 glycoproteins with molecular weights ranging from 125 000 to less than 16 000 with two components (40 000 and 33 000) accounting for almost two-thirds of the total stainable material. The FE contained ten glycoproteins that had the same molecular weights as those in the VE. One glycoprotein component underwent a reduction in molecular weight from 77 000 to 67 500 when the VE was converted to the FE. This molecular weight change was interpreted as the probable result of limited proteolysis. In addition, the FE gel electrophoresis patterns contained macromolecular components derived from the cortical granules and jelly layer, J, consistent with the immunodiffusion experiments. These components were absent when the FE was prepared in the absence of Ca2+, suggesting a role for Ca2+ in binding the VE, cortical granules, and J components together. We concluded that the conversion of the glycoproteinaceous VE to FE at fertilization is caused by interaction of the VE with components from the cortical granules and jelly layer J. These interactions are of both a chemical and physical nature.

Amino Acids↗
PubMed1975

The macromolecular composition of Xenopus laevis egg jelly coat.

The three morphologically and functionally distinct jelly coat layers of Xenopus laevis eggs, J1, J2, and J3, were separated by manual dissection, solubilized with dithiothreitol, and physicochemically analyzed. The chemical composition of the three jelly layers varied from 37 to 48% protein and 63 to 52% carbohydrate. The carbohydrate consisted of hexosamines, galactose, and fucose. Some of the carbohydrate in each of the jelly layers was covalently linked to protein through O-glycosidic bonds as beta elimination of the carbohydrate moiety in the presence of alkali was observed. In agreement with a previous finding, covalently attached sulfate was localized within the innermost jelly coat layer, J1. Cellulose acetate electrophoresis at pH 8.0 resolved a total of nine macromolecular components from the three jelly coat layers differentially staining for protein and carbohydrate: J1 yielded two anodically migrating components; the middle layer J2 yielded two cathodically migrating macromolecular components; the outermost layer J3 contained five species, three anodic and two cathodic. Sodium dodecyl sulfate agarose gel electrophoresis analysis yielded nine unique species, six of which stained coincidently for protein and carbohydrate. Immunoelectrophoresis and Ouchterlony double diffusion analyses using antiserum to total jelly components resolved nine different antigenic species with cross-reactivity between one or two macromolecules in layers J1 and J3. Analytical sedimentation velocity centrifugation revealed eight distinct species all of which exhibited hypersharp schlieren patterns and whose s20,w values were highly concentration dependent. On the basis of these analyses, Xenopus laevis egg jelly layers are composed of at least 8-9 distinct macromolecular species. The majority of these macromolecules are uniquely associated with different jelly coat layers.

Animals↗
PubMed1974

Agglutination of jelly coat and cortical granule components and the block to polyspermy in the amphibian Xenopus laevis.

A block to polyspermy in amphibians is established at fertilization by the conversion of the vitelline envelope to the fertilization envelope. In Xenopus laevis a major ultrastructural change in the envelope at fertilization is the appearance of an electron-dense layer, termed the F layer, between the envelope and the inner-most jelly coat layer, J(1). The F layer is derived, at least in part, from materials released from the cortical granules. Further definition of the origin and chemical nature of the F layer was sought by using isolated cortical granule (CG) exudate and jelly coat layer J(1). In double diffusion experiments, the isolated components interacted in an agglutination reaction producing a band of precipitation. The agglutination involved alpha-galactoside residues and metal ions (Ca(++)). Employing chemically modified jelly, we demonstrated that sulfhydryl-disulfide interchanges were not involved in the agglutination and, with (35)S-labeled jelly, that the agglutinating J(1) component possessed sulfate esters. Both the CG exudate and the J(1) components contained carbohydrate, as evidenced by their lectin reactivity. A number of ionic polymers, both natural and synthetic, were tested as chemical analogs of CG exudate and J(1); none gave an agglutination band. Dissolved jelly coat material from eggs of two different species of frogs agglutinated with CG exudate, while jelly from sea urchin eggs and hyaluronic acid from mammalian eggs did not. Thus, the agglutination reaction was chemically and phylogenetically specific. An electron-dense layer, similar to the F layer, formed on the outer of the vitelline envelope when jellied unfertilized eggs were immersed in CG exudate; such eggs were not fertilizable. We suggest that in Xenopus laevis, and perhaps other organisms as well, an agglutination type of reaction between cortical granule components and egg integuments is a participant in the structural and molecular events establishing a block to polyspermy.

Agglutination↗