To date, nearly 28 distinct genetic loci of autosomal dominant cerebellar ataxias have been identified, among them 18 disease-causing genes have been cloned. Of these, Machado-Joseph disease (MJD), also named as spinocerebellar ataxia type 3 (SCA3), is perhaps the most common subtype among different races and origins in the world. It is a neurodegenerative disease caused by the expansion of a CAG repeat in the coding region of the MJD1 gene, with obvious clinical and genetic heterogeneity. In this review, authors covered the recent advances in molecular genetic of SCA3/MJD.
Ataxin-3 ; Humans ; Machado-Joseph Disease ; genetics ; Molecular Biology ; Mutation ; Nerve Tissue Proteins ; chemistry ; genetics ; metabolism ; Nuclear Proteins ; chemistry ; genetics ; metabolism ; Repressor Proteins ; chemistry ; genetics ; metabolism

Recently, the gene encoding clathrin assembly protein of lymphoid myeloid leukemia (CALM), which is homologous to the AP180, was cloned from rat brain, and its expression differential to AP180 was reported (Kim and Lee, 1999). This gene product promotes the polymerization of clathrin into clathrin cage and found to be a regulator in membrane trafficking between intracellular compartments in eukaryotic cells (Kim et al., 2000). In this study, we have purified the CALM protein from clathrin-coated vesicles of rat liver using the monoclonal antibody against the recombinant N-terminal region of the CALM. The coated proteins extracted from the coated vesicle fraction was further purified by multi-step procedures involving gel-filtration and ion-exchange chromatography and SDS-PAGE. The purified protein with an apparent molecular weight of 100 kD promoted the assembly of clathrin triskelia into clathrin cage. In this respect the CALM protein bears a functional resemblance to the AP180 that has been previously described.
Animal ; Clathrin/*metabolism ; Clathrin-Coated Vesicles/*chemistry ; Liver/*chemistry ; Nerve Tissue Proteins/*isolation & purification ; Phosphoproteins/*isolation & purification ; Rats

OBJECTIVETo investigate the effects of preweaning enrichment on the expression of activity-regulated cytoskeletal protein (Arc), an immediate early gene, and on the long-term memory in rats.

METHODSForty neonatal Sprague-Dawley rats were randomly assigned to control group (standard environment, n=20) and experimental group (enriched environment, n=20). The experimental group received enriched environment exposure from postnatal day 10 until weaning (2 weeks, 20 minutes per day). The open field and novel object recognition tests were performed at postnatal day 28. Arc expression was detected by Western blotting and immunohistochemistry.

RESULTSThere was no significant difference in the open field test between the two groups. However, in the novel object recognition test, the experimental group rats performed significantly better than the control rats after 1 and 24-hr retention. The preference index in the experimental group after 1-hr (59.61%+/-9.61% vs 50.46%+/-9.34%; P<0.05) and 24-hr retention (62.72%+/-14.12% vs 52.39%+/-9.16%; P<0.05 ) was significantly higher than that in the control group. Arc expression in both areas CA1 and DG of hippocampus in the experimental group increased significantly compared with that in the control group (P<0.01).

CONCLUSIONSPreweaning enrichment can up-regulate the expression of immediate early gene, Arc, in the hippocampus of the rats, and promote their long-term memory.

Animals ; Cytoskeletal Proteins ; analysis ; Female ; Hippocampus ; chemistry ; physiology ; Immunohistochemistry ; Male ; Memory ; Nerve Tissue Proteins ; analysis ; Rats ; Rats, Sprague-Dawley

AIM: To study the chemical constituents and bioactivity of the seeds of Crataegus pinnatifida. METHODS: The chemical constituents were isolated and purified by macroporous adsorptive resin D101, silica gel, and ODS column chromatography, and preparative HPLC. Their structures were elucidated on the basis of spectroscopic methods. In addition, the cytotoxic activities of compounds 1-4 were investigated on OPM2 and RPMI-8226 cells. RESULTS: Four compounds were obtained and their structures were identified as (7S, 8S)-4-[2-hydroxy-2-(4-hydroxy-3-methoxyphenyl)-1-(hydroxymethyl)ethoxy]-3, 5-dimethoxybenzaldehyde (1), (+)-balanophonin (2), erythro-guaiacylglycerol-β-coniferyl aldehyde ether (3), buddlenol A (4). CONCLUSION Compound 1 is a novel norlignan, while compounds 1-4 exhibited marginal inhibition on the proliferation of OPM2 and RPMI-8226 cells.
Cell Line ; Cell Proliferation ; drug effects ; Crataegus ; chemistry ; Humans ; Molecular Structure ; Nerve Tissue Proteins ; chemistry ; isolation & purification ; toxicity ; Plant Extracts ; chemistry ; isolation & purification ; toxicity ; Seeds ; chemistry

Clathrin-coated vesicles (CCVs) are involved in protein and lipid trafficking between intracellular compartments in eukaryotic cells. CCVs are composed of clathrin and assembly proteins. The clathrin assembly protein lymphoid myeloid leukemia (CALM) gene, encodes a homologoue of the neuronal clathrin assembly protein AP180. In this study, we characterized the properties of the CALM expressed in E. coli. The molecular weight of bacterially expressed GST-CALM fusion protein was approximately 105 kD on SDS-PAGE. The CALM protein could promote clathrin triskelia into clathrin cages and could bind the preformed clathrin cage. However, 33 kD N-terminal domain of CALM could not bind pre-assembled clathrin cages, but assemble clathrin triskelia into clathrin cages. The CALM protein was bound to SH3 domain through N-terminal domain1, in vitro. The CALM protein is proteolyzed by caspase 3, caspase 8 and calpain through C-terminal domain.
Animal ; Antibodies, Monoclonal ; Calpain/chemistry ; Caspases/chemistry ; Clathrin-Coated Vesicles/metabolism* ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli/metabolism ; Escherichia coli/genetics ; Female ; Glutathione Transferase/genetics* ; Mice ; Mice, Inbred BALB C ; Nerve Tissue Proteins/metabolism ; Nerve Tissue Proteins/metabolism ; Nerve Tissue Proteins/chemistry* ; Phosphoproteins/metabolism ; Phosphoproteins/genetics ; Phosphoproteins/chemistry* ; Protein Binding ; Rabbits ; Recombinant Fusion Proteins/metabolism ; Recombinant Fusion Proteins/genetics ; Recombinant Fusion Proteins/chemistry* ; src Homology Domains

OBJECTIVETo investigate the distribution, changes and a possible role for retinal dopamine transporter (DAT) in experimental myopia in chickens.

METHODSTwo-day-old chickens were divided into four groups. Chicken eyes were fitted with lenses of -10D,-20D and translucent goggles unilaterally. Normal eyes were used as controls. After 3 wk, all chickens were given an intramuscular injection of (125)I-beta-CIT 2beta-carbomethoxy-3beta-(4-iodophenyl)tropane and sacrificed two hours post injection. Retinal pigment epithelium (RPE) and the neural retina were obtained together or RPE was dissected out from the neural retina. Radioactive DAT from each specimen was assayed by gamma-counter.

RESULTSRetinal DAT was detected in RPE specimens rather than in the neural retina in all eyes. Radioactive DAT in myopic eyes was higher, compared with control eyes.

CONCLUSIONSRetinal DAT is mainly located in the RPE and may be involved in the formation of lens induced myopia (LIM) and form deprivation myopia (FDM). These methods may provide a new approach for further studying the role of the dopamine system in experimental myopia.

Animals ; Chickens ; Dopamine Plasma Membrane Transport Proteins ; Eye ; growth & development ; Membrane Glycoproteins ; Membrane Transport Proteins ; analysis ; physiology ; Myopia ; metabolism ; Nerve Tissue Proteins ; Retina ; chemistry

Antisperm antibodies can lead to immunological infertility. Further research on the target antigens of antisperm antibodies may help to discover the causal relationship of antisperm antibodies to infertility. This paper summarizes the structure and function of the six target antigens of antisperm antibodies found recently, so as to discover the causal relationship of the antibodies to infertility and provide a basis for screening a vaccine for immunological contraception.
Amyloid beta-Protein Precursor ; chemistry ; immunology ; Antibodies ; immunology ; Antigen-Antibody Reactions ; Cytoskeletal Proteins ; Glycoproteins ; chemistry ; immunology ; Humans ; Infertility, Male ; etiology ; Male ; Nerve Tissue Proteins ; chemistry ; immunology ; Nuclear Pore Complex Proteins ; Nuclear Proteins ; Spermatozoa ; immunology ; Zyxin

We present a congenital plexiform cellular tumor with high mitotic activities arising in the right thigh of a 3-days-old infant. This subcutaneous tumor measured 6.5 x 4.5 cm in diameter with multinodular, whitish, elastic cut surface. Microscopically, the tumor was composed of fascicles of closely packed uniform spindle cells with frequent nuclear palisadings. The most remarkable finding was frequent mitoses(4-5/10 HPF). The tumor cells were strongly positive for S-100 protein, myelin basic protein and vimentin, and weakly positive for Leu 7. On electron microscopic examination, the spindle cells were found to be surrounded by continuous basal lamina and had interlocking long cytoplasmic processes. Although the prognosis of this tumor is difficult to predict, aggressive behavior such as recurrences may be suggested, but it is less likely to be metastatic. Further accumulation of similar unusual cases may be helpful in evaluation of its biologic behavior.
Case Report ; Human ; Infant, Newborn ; Male ; Neoplasm Proteins/analysis ; Nerve Sheath Tumors/chemistry/*congenital/pathology/surgery ; Soft Tissue Neoplasms/chemistry/*congenital/pathology/surgery ; Thigh ; Tumor Markers, Biological/analysis

Animals ; Globins ; analysis ; chemistry ; genetics ; physiology ; Humans ; Hypoxia-Ischemia, Brain ; metabolism ; prevention & control ; Nerve Tissue Proteins ; analysis ; chemistry ; genetics ; physiology ; Signal Transduction

Dishevelled proteins are multifunctional and highly conserved. These proteins are also required for the specification of cell fate and polarity by secreted Wnt proteins. To investigate the molecular mechanism of Dishevelled in mediating Wnt signal transduction, a mouse 11.5dpc embryo library was screened by yeast-two-hybrid system to find mouse Dishevelled2 DEP domain and C-terminal interacting proteins. 15 possitive clones were identified from 4.1 x 10(6) transformants. The DNA sequences of the positive AD/library plasmids were determined. The BLAST results revealed that one of the positive clones contained N-terminus cDNA fragments (amino acids 6-122) of Gli3 protein. The interaction between Dv12 and Gli3 detected by yeast two-hybrid system suggests that Gli3 might play a role in some biological processes with Dishevelled.
Adaptor Proteins, Signal Transducing ; chemistry ; genetics ; physiology ; Animals ; Dishevelled Proteins ; Gene Library ; Kruppel-Like Transcription Factors ; physiology ; Mice ; Nerve Tissue Proteins ; physiology ; Phosphoproteins ; chemistry ; genetics ; physiology ; Plasmids ; Signal Transduction ; Two-Hybrid System Techniques ; Wnt Proteins ; physiology ; Zinc Finger Protein Gli3