Biomedical subjects

S S Koide

Publications and source records attributed to S S Koide.

At least 217 records · Page 12Linked to original sources

PubMedDate not supplied

Immune responses in rats following oral immunization with attenuated Salmonella typhimurium expressing human sperm antigen.

The HSD-I gene codes a human sperm membrane protein (hSMP-1) and has been assigned the accession number U12978. The gene is located on human chromosome 9, region p12-p13. When the 1.7-kb cDNA of HSD-I was digested sequentially with EcoRI, BamHI, and HindIII, a 550-bp cDNA fragment was formed, which codes for the extracellular domain. This fragment was cloned into the asd+ vector pYA3149 to construct pYA3149R. The recombinant plasmid was used to transform an avirulent deltacva, deltacrp, deltaasd vaccine strain of Salmonella typhimurium chi4550. The hSMP-1 component was localized on the surface of the head of mature rat spermatozoa by an immunofluorescence technique using polyclonal anti-hSMP-1 antibodies. Since rat sperm contain hSMP-1, this rodent can be used to assay the immunogenicity of pYA3149R. Female Wistar rats were immunized by oral administration of the recombinant Salmonella. Anti-hSMP-1 antibodies in blood and vaginal washes of immunized animals were determined. Both body fluids contained significant amounts of the antibodies, showing that the recombinant Salmonella is an effective oral immunogen in rats.

Administration, Oral↗

PubMedDate not supplied

Human sperm membrane protein (hSMP-1): a developmental testis-specific component during germ cell differentiation.

Serum was obtained from an infertile woman having antibodies with sperm agglutinating activity. The antibodies interacted with a human sperm membrane protein (hSMP-1) with an estimated Mr of 55 kD. The gene (HSD-1) coding hSMP-1 was isolated from a human testis cDNA expression library and assigned the accession number U12978. The cDNA was conjugated to a prokaryotic expression vector to construct the recombinant vector, pRSET-HSD-I, which was expressed in Escherichia coli. The recombinant hSMP-1 was isolated and used to immunize rabbits to raise polyclonal antibodies. Usingan immunocytochemical technique, hSMP-1 protein was immunolocalized in germ cells of human testis at all stages of spermatogenesis. mRNAs were prepared from 16 different human tissues and analyzed by Northern blot using HSD-1 as probe. A positive reaction was elicited only with testis mRNA. The present findings suggest that the expression of hSMP-1 gene is testis-specific and occurs during the early stages of germ cell differentiation. In a comparative study, the location of the hSMP-I protein in sperm and in germ cells of the seminiferous tubules of rats was determined. The target antigen was immunolocated on the head and tail of rat sperm and in late spermatids and spermatozoa of rat testis. These results suggest that, in the rat, the HSD-1 gene is expressed during spermiogenesis.

Animals↗

PubMedDate not supplied

Evidence for the binding of a human sperm component with diaphanous protein.

The YWK-II component of human sperm membrane is related to the betaA4-amyloid precursor protein (APP) of Alzheimer's disease. A yeast 2-hybrid system was used to screen a mouse testis cDNA expression library for potential ligands capable of interacting with the extracellular domain of the YWK-II component. One of the bound proteins was identified as hDIA1, which has 96% identity with p140mDia. These proteins are members of the formin homology family and participate in cytokinesis and organization of the actin cytoskeleton. By interacting with these diaphanous proteins, the YWK-II component may be involved in germ cell differentiation and in the structural formation of the acrosome.

Amino Acid Sequence↗

PubMedDate not supplied

Partial amino acid sequencing of 80-kDa human sperm antigen (80-kDa HSA).

An 80-kDa human sperm antigen (80-kDa HSA) has been identified as a sperm protein responsible for inducing immunoinfertility. Immunization with the purified protein induced infertility in male and female rats. Immunohistochemical and immunofluorescent studies have demonstrated that the antigen is specific to spermatozoa. The present study describes the partial amino acid sequencing of 80-kDa HSA. The homogeneous protein was electrophoretically transferred onto a PVDF membrane and the excised band of 80-kDa HSA was used to determine the partial N-terminal amino acid sequence. The protein was then subjected to enzymatic digestion with endoproteinase Lys-C and endoproteinase Glu-C. The partial amino acid sequence of the major peptides thus obtained was determined. The digestion with endoproteinase Lys-C generated 4 major peptides, two of which showed partial sequence homology with lactoferrin. Endoproteinase Glu-C digestion produced 3 major peptides. The sequences of the 2 peptides were determined for which no matches were found in the databank. These results confirmed earlier observations that 80-kDa HSA is a sperm-specific protein that is chemically distinct from any other protein involved in normal physiological process. Earlier studies have demonstrated that it is antigenic, efficacious, conserved, and could be a promising candidate for the development of an antifertility vaccine.

Amino Acid Sequence↗

PubMedDate not supplied

Influence of acetazolamide on AQP1 gene expression in testis and on sperm count/motility in epididymis of rats.

Acetazolamide (Ace) is a putative inhibitor of carbonic anhydrase (CA), an enzyme that catalyzes the equilibration of carbon dioxide and carbonic acid and plays a key role in HCO(3)(-) and water reabsorption and acid secretion. Aquaporins (AQPs) are channel-forming membrane glycoproteins that mediate water reabsorption by the renal tubules and other organs of mammals. AQP1 and CAII or CAIV share many common biological properties. Previous studies have shown that AQP1 and CA are located at the same sites in cells of the male reproductive tract. In the present study, Ace at a dose of 40 mg/kg/d x 14, administered per os, suppressed AQP1 gene expression and inhibited CA activity in rat testis. On day 7 of treatment the epididymal sperm motility was significantly reduced, while on day 14 a decrease in sperm count occurred. Ace caused a marked downregulation of AQP1 gene expression; significant suppression occurred on days 7 and 14. Moreover, CA activity was totally blocked throughout the treatment period. The present findings suggest that the reduction of rat sperm motility and count by Ace can be attributed to its capacity to downregulate AQP1 water channel gene expression.

Acetazolamide↗

PubMedDate not supplied

Biphasic effect of GABA on rat sperm acrosome reaction: involvement of GABA(A) and GABA(B) receptors.

The functional relationship between GABA(A) and GABA(B) receptors in regulating acrosome reaction (AR) of rat spermatozoa was demonstrated by studying the differential effects of a GABA(B) agonist and an antagonist on the process. AR rates were determined using the chlortetracycline staining assay. The induction of AR in rat sperm by GABA was found to be a biphasic phenomenon; i.e., AR rates increased with increasing GABA concentrations up to <5 micro M and at higher concentrations of the neurotransmitter (>5 micro M), there was a reductionin the AR rates. This biphasic phenomenon is apparently due to the differential interaction of the neurotransmitter with GABA receptor subtypes in a dose-dependent manner; i.e., GABA(A) receptors (stimulatory) are primarily activated at low concentration of GABA, while GABA(B) receptors (inhibitory) become activated at higher concentrations. This hypothesis is supported by the present findings that treatment with saclofen, a GABA(B) receptor antagonist, did not influence the AR rates effected by GABA at low concentrations; while the AR rates were maintained at the maximum level at higher concentrations of GABA, resulting in the elimination of the biphasic phenomenon. Baclofen, a GABA(B) receptor agonist, blocks the AR activating action of GABA at both low and high concentrations. It would appear that the induction of AR in rat sperm by GABA is regulated by the proportionality of activated GABA(A) and GABA(B) receptors acting as a yin-yang control.

Acrosome↗

PubMedDate not supplied

Eliciting an immune response by plasmid DNA encoding a human sperm protein (HSD-1).

The cDNA encoding a human sperm membrane designated as HSD-1 was isolated from a human testis lambda gt11 cDNA expression library and assigned the accession number U12978 by GenBank. HSD-1 was conjugated to an eukaryotic expression plasmid (pRSV) to construct the recombinant plasmid pRSV-HSD-1. Female mice were inoculated intramuscularly with the plasmid DNA and the expression of HSD-1 was determined. HSD-1 mRNAs were detected in myocytes and endomysial connective tissue cells of the quadriceps muscle by in situ hybridization. Spleen of inoculated animals contained an increased number of cytotoxic T lymphocytes, phagocytes, and plasma cells. Fertility of the treated animals was not affected. Thus, intramuscular inoculation of female mice with the plasmid DNA (pRSV-HSD-1) results in the expression of HSD-1 and may elicit a tissue-mediated immune response.

Animals↗

PubMedDate not supplied

Expression and characterization of an epididymis-specific gene.

A truncated cDNA coding a rabbit epididymal protein (BE-20) was identified in a previous study. In the present study the full-length cDNA was isolated by the method of rapid amplification of cDNA ends. The BE-20 cDNA consisted of 585 bp with a poly(A) tail of 26 residues and an open reading frame composed of 369 bp encoding a deduced polypeptide containing 123 amino acid residues with a calculated molecular mass of 13 kDa. The N-terminus-contained a leucine-rich segment. BE-20 cDNA has about 76.8% homology with the HE4 gene of human epididymis. Northern blot analysis of mRNAs prepared from 17 different human tissues was performed using as probe a 0.5-kb DNA fragment corresponding to a segment of BE-20 cDNA. Positive reaction was elicited only with epididymal mRNA. A DNA fragment corresponding to a section of the open reading frame of BE-20 cDNA was cloned in Escherichia coli under the control of the T7 promoter. The cellular content of the expressed recombinant protein comprised about 55% of the total protein. The chromatographically purified bacterial product migrated as a single band with an estimated M(r) of 15 kDa on analysis by SDS-PAGE. In conclusion, BE-20 cDNA is expressed only in the epididymis. It is structurally related to the four-disulfide core family of extracellular proteinase inhibitors and may be involved in sperm maturation.

Amino Acid Sequence↗

PubMedDate not supplied

Molecular cloning and characterization of a novel testis-specific nucleoporin-related gene.

A 20-kDa sperm membrane protein cDNA, designated as RSD-1, was isolated by epitope selection from a rat testis lambda gtll expression library. RSD-1 was used as a probe to screen a human testis lambda ZAPII cDNA expression library. A cDNA designated as BS-63 was isolated and found to consist of 1933 bp with an open reading frame of 1824 bp and assigned the accession number U64675 by GenBank. The deduced polypeptide consisted of 608 amino acid residues containing XFXFG or FG motifs that are characteristic of nuclear pore complex (NPC) proteins and act as potential binding sites for Ran. The N-terminal region has high homology with RanBP2/Nup358, a nucleoporin component, showing that BS-63 is a member of the NPC family. Northern blot analysis of mRNAs prepared from various human tissues shows that BS-63 is transcribed in two forms: 6.0 and 8.5 kb. The 8.5-kb transcript was present in low amounts in several somatic tissues, whereas the 6.0-kb transcript is expressed only in testis. In situ hybridization analysis of human testis sections showed that BS-63 mRNA is expressed only in germ cells at all stages of spermatogenesis. Sertoli cells did not transcribe the gene.

Amino Acid Sequence↗

PubMedDate not supplied

Identification of the sperm antigen interacting with antibodies in serum from an infertile woman.

Serum (IS) obtained from an infertile woman induced head-to-head agglutination of human sperm. The immunoglobulin G (IgG) fraction of the IS was prepared by ammonium sulfate fractionation and DEAE cellulose chromatography. The IgG localized to the post-acrosomal region of the sperm, determined by indirect immunofluorescence and interacted with a human sperm protein with an estimated Mr of 80 kDa, determined by immunoblotting. The identity of the interacting sperm antigen was verified by isolating the 80 kDa sperm protein by affinity chromatography. The present results suggest that the anti-80 kDa antibodies may be responsible for the infertility.

Antibodies↗

PubMedDate not supplied

Immunoglobulin-Fc binding factor in human male reproductive organs.

Human seminal plasma, testis, seminal vesicle, epididymis, and prostate contain a component with an estimated Mr of 20 kD that binds human immunoglobulin-Fc. The factor did not bind goat-IgG-Fc, immunoglobulins of human, rat, mouse, goat, horse, or rabbit sera and did not interact with antibodies raised against Fc gamma receptors. The present findings show that this Fc binding factor in seminal plasma is a secretory product of the testis and accessory sex organs. It binds human Ig-Fc but does not meet the criteria of an Fc receptor. Additional IgG-Fc binding proteins with estimated Mr of 90, 88, and 86 kD were detected in the prostate, testis, and seminal vesicle, respectively.

Genitalia, Male↗

PubMedDate not supplied

Binding of a specific subclass of immunoglobulins by a human seminal plasma component.

Human seminal plasma contains a component that binds immunoglobulins (Ig). The Ig binding factor was purified by ammonium sulphate precipitation, preparative isoelectrofocusing and gel filtration chromatography and found to bind strongly human IgGl and mouse IgM. This seminal plasma component may possess immunosuppressive activity and may modulate the activities of the immunosurveillance system of the reproductive tract.

Animals↗

PubMedDate not supplied

Monoclonal antibody inducing human sperm agglutination.

A mouse fused splenic cell line producing antibody that induces agglutination of human sperm was cultivated. The antibody was localized to the midpiece and tail of human spermatozoa by an indirect immunofluorescence method. The interacting antigen was identified in a sperm membrane protein preparation by SDS gel/protein blot immunofluorescence method. The molecular weight of the antigen was in the vicinity of 84 Kdaltons.

Animals↗

PubMedDate not supplied

Hyaluronic acid-like substance from mouse ovaries with angiogenic activity.

Glycosaminoglycans prepared from extracts of non-luteal mouse ovaries (JCL-ICR strain) were assayed for neovascularization by implanting Elvax films, impregnated with test samples, on the lateral wall of the sheath of m. rectus abdominis in adult mice of the same strain. Neovascularization occurred in a dose-dependent manner. When purified by chromatography on Dowex 1-x2 and DEAE Sephadex columns, fractions eluted with 0.5 M NaCl showed strong neovascularizing activity. On further purification by high performance liquid chromatography using TSK gel DEAE 2 SW column, the fraction with a retention time nearly coincident with that of hyaluronic acid possessed high neovascularizing activity. The activity of this fraction was markedly reduced when treated with streptococcal hyaluronidase. The present results suggest that glycosaminoglycans, especially a hyaluronic acid-like substance, are involved in ovarian neovascularization.

Animals↗

PubMedDate not supplied

Identification of IgG and Fc-binding proteins in human seminal plasma and sperm.

Human seminal plasma contains two novel soluble proteins capable of binding IgG and Fc, but not Fab. The IgG- and Fc-binding proteins were identified by immunoblotting using IgG of various species, Fc and Fab fragments. Their estimated molecular sizes are 16-kD and 20-kD. The monoclonal antibody (MAb), Leu 11b, raised against Fc gamma RIII interacts with the 16-kD protein, whereas other mAbs (32.2, IV.3, and 3G8) raised against FcR did not. The 16-kD protein is capable of binding IgG of several species (human, mouse, rabbits, and goat), whereas the 20-kD protein interacted only with human IgG-Fc fragment. The 16-kD IgG and the 20-kD Fc-binding proteins were found in the PBS extract of human sperm. Human seminal plasma/sperm contain an IgG- and a Fc-binding protein with estimated Mr of 16 and 20 kD, respectively.

Antibodies, Monoclonal↗

PubMedDate not supplied

Gene expression of human sperm component related to A4 amyloid precursor protein.

Monoclonal antibodies were raised against a specific human sperm protein and designated as the YWK-II mAb. The partial cDNA encoding the protein was isolated from a rat testis lambda gt11 expression library and the amino acid sequence of the protein was deduced. The cytoplasmic-transmembrane domains of the deduced protein had high homology with the A4 amyloid precursor protein of Alzheimer's disease. To evaluate the stage of spermatogenesis when the gene was expressed, single-stranded 35S-labeled RNA probes were prepared from the cDNA. By an in situ hybridization technique the mRNA for the antigen was detected in germ cells at all stages of spermatogenesis. The finding that the gene is expressed in spermatogonia suggests possible involvement in the initiation of germ cell differentiation or in the detachment of spermatogonia from the basement membrane.

Amyloid beta-Peptides↗

PubMedDate not supplied

Purification and characterization of a boar sperm protein interacting with immunoglobulins from an infertile woman.

Immunoglobulin G (IgG) fraction was prepared from a serum obtained from an infertile woman (IS) that induced sperm agglutination of human and boar sperm. The antisperm antibodies interacted with a 50-kD boar sperm protein, determined by immunoblot. The 50-kD protein was extracted with deoxycholate (DOC) and purified by affinity chromatography on concanavalin A column, ion exchange chromatography on CM52 column, affinity chromatography on IS IgG-protein G-agarose, and by preparative electrophoresis with electroelution. The purified 50-kD protein migrated as a single homogeneous band when analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis and by Western blot. The production of antibodies against specific sperm proteins may be a cause of immunological infertility.

Animals↗

PubMedDate not supplied

Expression of a gene encoding a specific human sperm protein in Chinese hamster ovarian cells.

The cDNA (RSD-2) encoding a human sperm protein (YWK-II) was isolated from a rat testis lambda gt11 expression library. Its nucleotide sequence was determined. The deduced polypeptide showed high identity with the transmembrane-cytoplasmic domains of A4 amyloid precursor protein of Alzheimer's disease. The RSD-2 was inserted into the EcoRI site of the pSV2-EP vector to construct the pSVRS-2 vector. Chinese hamster ovarian (CHO-dhfr) cells were cotransformed with pSV2-neo and pSVRS-2. mRNA and chromosomal DNA prepared from the transformed cells interacted with [32P]RSD-2 as probe by dot hybridization. The production of the YWK-II protein was determined by staining with the YWK-II mAb by an indirect immunofluorescence technique. There was marked staining of the cytoplasm. The RSD-2 cDNA encoding the YWK-II sperm was expressed in the transformed CHO cells. The pSV2-EP vector and the CHO cell expression system can be utilized to produce sperm proteins for antifertility studies.

Amyloid beta-Protein Precursor↗