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Nonintegrin laminin receptors in the nervous system: evidence for lack of a relationship to P40.

Laminins are extracellular matrix proteins that mediate their effects on cells through integrin and nonintegrin receptors. Two receptors of 67 and 110 kD that bind laminin with a high affinity (Kd approximately nM) have been reported in neural cells. Here, we discuss these and other nonintegrin laminin receptors that have been implicated in neural function. In addition, we report studies characterizing a 43 kD protein, (P40), immunologically related to the 67 kD laminin receptor, which may be involved in retinal development. In our studies, polyclonal antisera (anti-P-20-A) to a synthetic peptide derived from the sequence of a cDNA for a putative high-affinity laminin receptor (67 kD) detected a protein of 43 kD in immunoblots of adult rat retinas. Immunohistochemistry with this antiserum showed that the retinal immunoreactivity was predominantly localized in the ganglion cell layer of both adult chicken and rat retinas where it appeared to be intracellular. Retinal ganglion cells were shown to be immunoreactive by retrogradely labeling them from the superior colliculus with a lipophilic dye and subsequently with anti-P-20-A antisera. Consistent with the preferential localization of the P-20-A immunoreactivity in ganglion cells, there was a substantial decrease in the amounts of P40 on Western blots following optic nerve section and resulting retinal ganglion cell death. Screening of a rat (PC12 cell) cDNA library with the anti-P-20-A antiserum further confirmed the specificity of the antiserum for the rat homologue of P40. Rat P40 is 97% identical to the mouse and 87% identical to human P40 at the nucleic acid level and 98% at the protein level. Restriction mapping of the rather abundant positive clones in the library that cross-hybridized with a human cDNA probe for P40 indicated that the full-length cDNA of 1.2 kb was the major and perhaps the only cDNA in the library. In Northern blots of adult rat retina, these clones hybridized to a single 1.2-kb transcript. Electroblots of retinal homogenates probed with radioiodinated laminin demonstrated binding to a broad band at 110 kD, but none at 43 kD. Taken together these findings suggest that P40 may not be a laminin receptor and are in keeping with the hydrophilic composition of the protein, its intracellular localization, as well as other features predicted by its nucleic acid sequence.(ABSTRACT TRUNCATED AT 400 WORDS)

Amino Acid Sequence↗

Differentiation of smooth muscle cells in chicken gizzard. Self-catalytic mechanism for the production of smooth muscle cells.

During development of the chicken gizzard, a thick layer of undifferentiated cells (mesenchymal cells) is constructed, and the cells differentiate into smooth muscle cells or connective tissues. We found that the differentiation of smooth muscle cells occurred first near the outer surface of the gizzard and the differentiated area spread to the inside of the gizzard. Therefore, we assumed that the differentiation of most of the smooth muscle cells in the gizzard is induced by differentiated smooth muscle itself. When undifferentiated cells from gizzard of 7-day-old embryo (Hamburger and Hamilton's stages 26-27) were cultured on a coverglass coated with extract of gizzard that contained differentiated smooth muscle cells, the cells attached to the coverglass and differentiated into smooth muscle cells. On the other hand, extract of gizzard from 7-day-old embryo did not induce the differentiation of smooth muscle cells, though it induced the attachment of cells. We found that activity for the differentiation of smooth muscle cells appeared when differentiated smooth muscle cells appeared in developing gizzard. Gizzard contained higher activity for the differentiation of smooth muscle cells than the other tissues. Transforming growth factor-beta (TGF-beta), which induces the differentiation of vascular smooth muscle cells, did not induce the differentiation of smooth muscle cells in gizzard, though extract of aorta induced the differentiation of smooth muscle cells in gizzard. The results obtained here support evidence that the differentiation of most of the smooth muscle cells in gizzard is induced by a self-catalytic mechanism in which differentiated smooth muscle itself induces the differentiation of smooth muscle cells.

Animals↗

Toxicity and occurrence of Balansia on grasses from toxic fescue pastures.

Balansia epichloe (Weese) and B. henningsiana (Moell.) were isolated from grasses in toxic fescue pastures. B. epichole, cultured in a synthetic medium, was toxic to chicken embryos. Thin-layer chromatography and ultraviolet absorption data indicated that in submerged culture the fungus produced compounds with the indole or ergoline nucleus.

Animals↗

Antibodies to the sulphated, high molecular mass mouse tectorin stain hair bundles and the olfactory mucus layer.

Polyclonal antibodies were raised in chickens to the glycosylated forms of the high (H), medium (M) and low (L) molecular mass (MM) mouse tectorins. In the mouse cochlea, all three antibodies stained the tectorial membrane. Antibodies raised to HMM tectorin also stained the hair bundles of both inner and outer hair cells. A number of other mouse tissues were screened with the anti-tectorin antibodies to look for similar or antigenically related molecules. Staining was not observed in any other tissue type with the antibodies directed against the MMM and LMM tectorins. In the nose, the anti-HMM tectorin antibodies stained Bowman's glands and the mucus layer overlying the olfactory epithelium. The surface of the adjacent respiratory epithelium was not stained by these antibodies. HMM tectorin can be specifically radiolabelled by injecting neonatal mice with 35SO4 and undergoes a shift in electrophoretic mobility following treatment with keratanase, an endo-beta-galactosidase from Pseudomonas. However, when centrifuged on shallow CsCl gradients HMM tectorin has a buoyant density similar to that of glycoproteins and does not behave as a typical cartilage type proteoglycan. HMM tectorin does not react with mab 5D4, a monoclonal antibody that recognises keratan sulphate glycosaminoglycan from corneal and skeletal muscle proteoglycan. Unlike antibodies to HMM tectorin, mab 5D4 selectively stains the upper surface of the tectorial membrane, Hensen's stripe and the mucus layer overlying the respiratory epithelium. These studies indicate that the MMM and LMM tectorins may be unique to the cochlea, and that HMM may be a "light' keratan sulphate proteoglycan that is antigenically related to either the mucins or a more specific component of the olfactory mucus layer.

Animals↗

Messenger RNA and protein expression analysis of betaglycan in the pituitary and ovary of the domestic hen.

Betaglycan was originally characterized as the type III receptor for TGFbeta, yet recent research has indicated that betaglycan can serve as an accessory receptor for inhibin. To understand better the action of inhibin in avian follicular development, we have investigated the expression of betaglycan in the pituitary gland and ovary of the hen. In experiments 1 and 2, betaglycan mRNA was detected at 6 kilobases (kb) by Northern blot analysis (n = 5) in chicken pituitary, granulosa, and theca layers and whole ovary. Expression of betaglycan was greatest in the pituitary gland in experiment 1 and greater in the granulosa layer of small yellow follicles (SYF) compared with the granulosa layer of larger follicles. In experiment 2, betaglycan mRNA was more abundantly expressed in the theca layer compared with the granulosa layer for all follicle sizes, although there was no significant difference in betaglycan expression in the theca layer among follicle sizes. In experiment 3, immunohistochemical analysis revealed betaglycan protein in the anterior pituitary as well as in the ovary (n = 4) and SYF (n = 4). Colocalization studies revealed a high abundance of cells within the anterior pituitary expressing both betaglycan and FSH (n = 4). Betaglycan protein was found in the granulosa layer; however, markedly enhanced staining was observed in the theca layer of ovarian follicles. Our results provide evidence for expression of betaglycan mRNA and protein colocalization with FSH in the anterior pituitary, consistent with known inhibin effects. Ovarian localization of betaglycan, particularly in the theca layer, suggests a paracrine role for inhibin in the hen.

Animals↗

[Chicken genome project and its application in genetic research].

Chicken is an important model organism for studying all sorts of biological problems due to its unique biological features. Two strains of chicken, the egg-laying layer and meat-producing broiler, which were generated from selective breeding, come to be ideal models for understanding the molecular mechanisms underlying different phenotypes. The Chicken Genome Project provides a large amount of information for research using chicken as the model. This review summarizes the development of the Chicken Genome Project, as well as the strategies for researching the quantitative traits of different chicken strains using the information of the Chicken Genome. We also give a perspective of the developmental and genetic studies that can be conducted using the chicken as a model organism as well as using its genomic information.

Animals↗

Gene expression of procollagen alpha 1 (I) and alpha 1 (III) in partial-thickness tears of the deep pectoral tendon in chickens.

The purpose of this study was to evaluate the healing process of partial-thickness tears of the deep pectoral tendon in chickens which is considered to be equivalent to the human rotator cuff. The superior side (bursal-side) and the inferior side (joint-side) layers of the deep pectoral tendon of 80 adult chickens were lacerated in the same manner. The mRNA expression levels of alpha 1 (I) and alpha 1 (III) procollagen was at a level 2 weeks after laceration. The mRNA expression level of procollagen alpha 1 (I) was significantly higher than that of procollagen alpha 1 (III) in all experimental periods (p<0.05). The expression levels of procollagen-mRNAs were constitutively higher in bursal-side tears than in joint-side tears throughout the experimental periods (p<0.05), even 52nd weeks after laceration. These observations indicate that the bursal-side layer possesses higher healing ability than the joint-side layer in the chicken's deep pectoral tendon where subacromial impingement is absent, and that the remodeling of partial-thickness tears in chicken models needs more than one year.

Animals↗

Genetic diversity of Chinese native chicken breeds based on protein polymorphism, randomly amplified polymorphic DNA, and microsatellite polymorphism.

Genetic diversity of Chinese native chicken breeds was investigated using protein polymorphism, randomly amplified polymorphic DNA (RAPD), and microsatellite polymorphism. Imported broiler and layer breeds were also included in the analysis. The results from protein polymorphism did not show distinct differences between Chinese native chicken and imported broilers; however, there were small significant differences between these two types of chickens. The results from RAPD indicated that gene diversity within a population was large in Chinese native chickens, intermediate in broilers, and low in layers and that there were small differences between Chinese native chickens and both broilers and layers. A great difference between broilers and layers was observed. Microsatellite polymorphism data showed that genetic diversity was high in the Chinese native chickens and low in layers and that there was a close relationship between Chinese native chickens and broiler but a remote relationship between Chinese native chickens and layers. The wide genetic diversity of Chinese native breeds can meet different requirements of breeding for chicken quality in China.

Alkaline Phosphatase↗

Surface-sensitive polarized Raman spectroscopy of biological tissue.

In a two-layer diffusing medium, polarized light directly backscattering off the superficial layer will partially retain its sense of polarization, whereas deeper-probing light will be increasingly depolarized by diffusion. This effect has been studied in both elastic scattering and fluorescence contexts. We apply this method to Raman scattering in two two-layer models with a highly diffusing lower layer of glucose powder and an upper layer of either clear plastic or chicken skin. We employ detection of orthogonal polarization states to generate a Raman spectrum of only the superficial layer by combining the orthogonal signals.

Animals↗

A long term observation of antibody status to chicken anaemia virus in individual chickens of breeder flocks.

The antibody status to chicken anaemia virus (CAV) in four layer breeder flocks was evaluated. Sera were periodically collected from the same 17 to 20 individual chickens of each flock ranging in age from 10 to 63 weeks old. The neutralising and fluorescence antibody were detectable in individual chickens during the observation periods ranging from 13 to 44 weeks. A high prevalence of both neutralising and fluorescence antibodies was observed; however, the prevalence of fluorescence antibody in older chickens was lower than that of neutralising antibody. The geometric mean (GM) of neutralising antibody titres, after all the chickens examined had seroconverted in flocks 1, 2 and 4, ranged from 373.2 to 2940.6. In flock 1, the GM titre at 63 weeks old was significantly lower than that at 37 and 52 weeks old. In flock 4, the GM titre at 48 weeks old was significantly lower than that at 24 and 35 weeks old. In flock 2, the GM titre at more than 31 weeks old significantly increased compared with that at 25 weeks old; this tendency was not seen in the GM of the fluorescence antibody titres. The results indicate that immunity to CAV can last a long time in naturally infected individual chickens.

Animals↗

Third-generation synchrotron x-ray diffraction of 6-microm crystal of raite, approximately Na3Mn3Ti0.25Si8O20(OH)2.10H2O, opens up new chemistry and physics of low-temperature minerals.

The crystal structure of raite was solved and refined from data collected at Beamline Insertion Device 13 at the European Synchrotron Radiation Facility, using a 3 x 3 x 65 microm single crystal. The refined lattice constants of the monoclinic unit cell are a = 15.1(1) A; b = 17.6(1) A; c = 5.290(4) A; beta = 100.5(2) degrees; space group C2/m. The structure, including all reflections, refined to a final R = 0.07. Raite occurs in hyperalkaline rocks from the Kola peninsula, Russia. The structure consists of alternating layers of a hexagonal chicken-wire pattern of 6-membered SiO4 rings. Tetrahedral apices of a chain of Si six-rings, parallel to the c-axis, alternate in pointing up and down. Two six-ring Si layers are connected by edge-sharing octahedral bands of Na+ and Mn3+ also parallel to c. The band consists of the alternation of finite Mn-Mn and Na-Mn-Na chains. As a consequence of the misfit between octahedral and tetrahedral elements, regions of the Si-O layers are arched and form one-dimensional channels bounded by 12 Si tetrahedra and 2 Na octahedra. The channels along the short c-axis in raite are filled by isolated Na(OH,H2O)6 octahedra. The distorted octahedrally coordinated Ti4+ also resides in the channel and provides the weak linkage of these isolated Na octahedra and the mixed octahedral tetrahedral framework. Raite is structurally related to intersilite, palygorskite, sepiolite, and amphibole.

Journal Article↗