Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:3.2.1.23 (beta-galactosidase)
14,648 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

This study reports on the binding of beta-galactosidase obtained from different organs of the rat urogenital tract to membranes of these organs. Homologous and cross binding saturation assays indicated that: (1) high-affinity sites that recognize fructose-6-phosphate derivates (FPR) are present in spermatozoa from the rete testis, epididymal membranes and testes, although the latter may reflect binding to testicular spermatozoa; (2) the membranes of the other organs studied do not have FPR; (3) the FPR of the epididymis does not recognize enzymes purified from other organs of the reproductive tract. These results suggest that the FPR-binding system belongs to a peculiar transport route that permits maturing spermatozoa to acquire hydrolytic enzymes secreted by the epididymal epithelium. In the epididymis and seminal vesicles more than 50% of the enzymatic activity of beta-galactosidase was recovered in cytosol, suggesting that the enzyme is located mainly in the secretory fluid of these organs.
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PMID:Organ-specific binding system for beta-galactosidase in the male reproductive tract. 856 94

Prolonged treatment with tamoxifen induces changes in the male reproductive tract in rats. In this study changes in the protein content of the rat epididymal fluid as a consequence of prolonged treatment with tamoxifen are reported. Among five lysosomal enzymes measured in the epididymal fluid, alpha-mannosidase (alpha-MAN) significantly diminished, but other enzymes did not. Electrophoretic analysis of fluids showed that proteins of estimated molecular weight 25, 60, 80-85 and 180 kDa decreased in the treated rats. We also detected an increase in the binding of beta-galactosidase (beta-GAL) to caudal spermatozoa in treated rats. These changes may be related in part to the loss of fertilizing capacity of spermatozoa after tamoxifen treatment.
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PMID:Changes in the content of rat epididymal fluid induced by prolonged treatment with tamoxifen. 983 49

Spermatogonial stem cells can be transplanted from a fertile donor mouse to the testis of an infertile recipient where they establish spermatogenesis and produce spermatozoa. In the present study we investigated whether treatment of recipient mice with the gonadotropin-releasing hormone (GnRH) agonist leuprolide acetate could alter the efficiency of colonization by donor spermatogonial stem cells in the recipient testis. Six recipient mice were treated with busulfan to destroy endogenous spermatogenesis followed by injection of leuprolide acetate to three of the mice. Testis cells from mice carrying the ZFlacZ transgene, which produces beta-galactosidase in spermatids, were used as donor cells for transplantation to allow for identification of donor spermatogenesis in the recipient testis by staining for enzyme activity. The extent of donor cell colonization was compared between leuprolide treated recipients and untreated control mice 3 months after transplantation. Efficiency of colonization by donor cells was markedly enhanced in recipient mice treated with the GnRH agonist leuprolide acetate, which makes the technique of spermatogonial transplantation applicable to a wide range of experimental situations. The present study also indicates that this technique can be used as a biological assay system to investigate factors controlling the establishment and progression of spermatogenesis.
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PMID:Leuprolide, a gonadotropin-releasing hormone agonist, enhances colonization after spermatogonial transplantation into mouse testes. 983 81

Coinjection of unfertilized mouse oocytes with sperm heads and exogenous DNA encoding either a green fluorescent protein (GFP) or beta-galactosidase reporter produced 64 to 94 percent transgene-expressing embryos, reflecting DNA-sperm head association before coinjection. Nonselective transfer to surrogate mothers of embryos in the GFP series generated about 20 percent offspring expressing the integrated transgene. These data indicate that exogenous DNA can reproducibly be delivered into an oocyte by microinjected spermatozoa and suggest an adaptable method of transgenesis.
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PMID:Mammalian transgenesis by intracytoplasmic sperm injection. 1036 36

The mouse monoclonal antibody (mAb) A36 produced by us and shown to induce extensive, "tangled" sperm agglutination was used to isolate cDNAs encoding its cognate antigen. Three overlapping cDNA clones specifically recognized by the mAb were isolated from a human testis cDNA expression library in lambdagt11. Sequencing of these cDNAs yielded the complete nucleotide sequence of a 3-kilobase cDNA that encodes the mAb-related polypeptide, designated sperm antigen-36 (SA-36), composed of 558 deduced amino acids. SA-36 cDNA contained a 5' untranslated region of 234 nucleotides (nt), an open reading frame of 1674 nt, and a 3' untranslated region of 1138 nt. SA-36 cDNA displayed > 99% homology to glucose phosphate isomerase (GPI)/neuroleukin (NLK) mRNA. This surprising homology was confirmed in Western blots demonstrating that mAb A36 reacted specifically with GPI obtained from rabbit muscle and from baker's yeast. Moreover, polyclonal, monospecific antibodies produced against beta-galactosidase/SA-36-3 fusion protein stained human spermatozoa and caused intensive agglutination of these cells in a manner similar to that with the mAb. Taken together, the data presented here demonstrated that mAb A36 cognate sperm surface antigen, encoded by SA-36 cDNA, is a GPI/NLK-like protein involved in sperm agglutination.
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PMID:Cloning of a glucose phosphate isomerase/neuroleukin-like sperm antigen involved in sperm agglutination. 1072 72

In birds, the ovum is surrounded by a glycoprotein coat known as the inner perivitelline layer (IPVL), which is analogous to the mammalian zona pellucida and, as such, is the site of initial sperm binding and induction of acrosomal exocytosis (the acrosome reaction). In this study, we demonstrate that oligosaccharides isolated from chicken-IPVL glycoproteins are capable of inducing the acrosome reaction in chicken spermatozoa. Preparations containing only O-linked glycans were unable to induce the acrosome reaction whereas N-linked oligosaccharides released from the IPVL by PNGaseF treatment could induce the acrosome reaction. Addition of galactose to terminal N-acetyglucosamine residues suppressed the acrosome reaction-inducing capacity of the oligosaccharide preparation; however, this capacity could be restored by co-incubation with beta-galactosidase. This evidence suggests that the acrosome reaction-inducing factor is probably an N-linked oligosaccharide with terminal N-acetyl-glucosamine residues.
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PMID:Induction of acrosomal exocytosis in chicken spermatozoa by inner perivitelline-derived N-linked glycans. 1107 59

To date, two different zona binding assays have been described in the literature. Both assays, however, require a large quantity of human zonae which vary immensely in quality. Furthermore, an inverted microscope with micromanipulation equipment is necessary, which makes both assays relatively complicated and time-consuming, and requires skilled staff. Therefore, we developed a new, highly sensitive zona binding assay using a bioluminescence-enhanced system which employs a pool of solubilized zona pellucida and is easier for routine use. In the detection system, light emission by the luciferin-luciferase system is measured. Because of the limited availability of human zonae pellucidae, this new assay was first developed in the porcine system. The new bioluminogenic substrate D-luciferin-O-beta-galactopyranoside (Lu-Gal) was synthesized, purified and characterized. Synthesis of Lu-Gal resulted in purity better than 99.998%. Analytical data and spectra were appropriate. In terms of the kinetic data, Lu-Gal is a highly sensitive and specific substrate for beta-galactosidase. Using the given chemical conditions, nonlabelled zonae bound competitively to boar spermatozoa, which resulted in a high sensitivity and specificity. By the addition of 10 nonlabelled zonae, the binding of labelled zonae was almost completely inhibited. Corresponding results were obtained when the bioluminescent system was compared with the hemizona assay. On the other hand, spermatozoa of other species (bull, hamster and man) showed only low binding to the porcine zonae or none at all. Competitive displacement was not observed, indicating the inter-species specificity of the assay.
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PMID:Development of a new, highly sensitive zona pellucida binding assay using a bioluminescence-enhanced detection system. 1147 33

The possibility of inadvertent exposure of gonadal tissue to gene therapy vectors has raised safety concerns about germline infection. We show here that the receptor for coxsackie B viruses and adenoviruses 2 and 5 (CXADR) is expressed in mouse germ cells, suggesting the possibility that these viruses could infect germ cells. To directly assess the risk of germline infection in vivo, we injected an adenovirus carrying the germ-cell-specific protamine promoter fused to the bacterial lacZ reporter gene into the left ventricular cavity of mice and then monitored expression of the reporter gene in germ cells. To differentiate between infection of stem cells and differentiating spermatogenic cells, we analyzed expression of the reporter cassette at different times after viral delivery. Under all conditions tested, mice did not express the Escherichia coli beta-galactosidase protein in developing spermatids or in mature epididymal spermatozoa. Primary germ cells cultured in vitro were also refractory to adenoviral infection. Our data suggest that the chance of vertical germline transmission and insertional mutagenesis is highly unlikely following intracoronary adenoviral delivery.
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PMID:Absence of germline infection in male mice following intraventricular injection of adenovirus. 1173 45

The introduction of mammalian artificial chromosomes (ACs) into zygotes represents an alternative, more predictive technology for the production of recombinant proteins in transgenic animals. The aim of these experiments was to examine the effects of artificial chromosome microinjection into bovine pronuclei on embryo development and reporter gene expression. Bovine oocytes aspirated from 2-5 mm size follicles were matured in vitro for 22 hr. Mature oocytes were fertilized in vitro with frozen- thawed bull spermatozoa. Artificial chromosome carrying either beta-galactosidase (Lac-Z) gene or green fluorescence protein (GFP) gene were isolated by flow cytometry. A single chromosome was microinjected into one of the two pronuclei of bovine zygotes. Sham injected zygotes served as controls. Injected zygotes were cultured in G 1.2 medium for 7 days. Hatched blastocysts were cultured on blocked STO cell feeder layer for attachment and outgrowth of ICM and trophectoderm cells. The results showed a high zygote survival rate following LacZ-ACs microinjection (74%). However, the blastocyst development rate after 7 days of culture was significantly lower than that of sham injected zygotes (7.5 vs. 22%). Embryonic cells positive for Lac-Z gene were detected by PCR in three of nine outgrowth colonies. In addition, GFP gene expression was observed in 15 out of 85 (18%) embryos at the arrested 2-cell stage to blastocyst stage. Six blastocysts successfully outgrew, three outgrowths were GFP positive for up to 3 weeks in culture. We conclude that the methodology for artificial chromosome delivery into bovine zygotes could lead to viable blastocyst development, and reporter gene expression could be sustained during pre-implantation development.
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PMID:Expression of a reporter gene after microinjection of mammalian artificial chromosomes into pronuclei of bovine zygotes. 1174 53

The activities of acid beta-glucuronidase, alpha-mannosidase, alpha-glucosidase, alpha-galactosidase, beta-galactosidase and beta-N-acetylglucosaminidase were analysed in seminal plasma and spermatozoa from 26 infertile men with varicocele and from 36 men of normal fertility. Semen samples from ten men with non-obstructive azoospermia were used as control specimens that contained the other components of semen. Spermatozoa were solubilized by both physical (homogenization) and chemical (Triton-X100) methods to obtain the soluble and non-soluble fractions. The activities of several glycosidases measured both in seminal plasma and spermatozoa were directly correlated with the numbers of spermatozoa and sperm motility, confirming previous studies. As some infertile patients with varicocele have normal semen parameters, whereas others have low numbers of spermatozoa and low sperm motility, the varicocele patients were prospectively divided into two groups: one (n = 15) with normal spermiograms and the other (n = 11) with abnormal spermiograms. The activities (expressed in mU ml(-1)) of alpha-mannosidase, beta-galactosidase and beta-N-acetylglucosaminidase in seminal plasma of normozoospermic infertile patients with varicocele were significantly higher than those of fertile controls, but not when expressed in U per 10(8) spermatozoa. The activities of beta-glucuronidase, alpha-mannosidase, beta-galactosidase and beta-N-acetylglucosaminidase in seminal plasma when expressed in U per 10(8) spermatozoa in varicocele patients with abnormal spermiograms were significantly higher than in those of men of normal fertility. The activity of alpha-mannosidase in the soluble fraction of sperm homogenates, expressed as U per 10(8) spermatozoa, was significantly higher in infertile patients with varicocele and abnormal spermiograms than in controls. In the non-soluble fraction of spermatozoa from infertile patients with varicocele, there was an increase in the expression of beta-galactosidase and beta-N-acetylglucosaminidase activities compared with the fraction of spermatozoa from fertile subjects. In summary, infertile patients with varicocele displayed an overexpression of acid alpha-mannosidase, beta-galactosidase and beta-N-acetylglucosaminidase activities in seminal plasma and spermatozoa that may be associated with functional defects in spermatozoa as these glycosidases play an important role in mammalian fertilization.
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PMID:Abnormal expression of acid glycosidases in seminal plasma and spermatozoa from infertile men with varicocele. 1188 18


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