Search PubMed⌕ Search

Biomedical subjects

S S Koide

Publications and source records attributed to S S Koide.

At least 73 records · Page 4Linked to original sources
PubMed1994

Immunization with synthetic peptide segments of a sperm protein impair fertility in rats.

The nucleotide sequence of the cDNA encoding a sperm protein (rSMP-B) was determined in a previous study. Two peptide segments corresponding to the extracellular domain of the deduced sperm polypeptide were synthesized as multiple antigen peptide (MAP) and designated as rSMP-229 and rSMP-230. Polyclonal antibodies were raised against the two MAPs. Sera obtained from rabbits immunized with rSMP-230 interacted with human and rabbit sperm membrane proteins with estimated molecular sizes of 72 and 20.1 kD, respectively. Adult female and male rats were immunized with the MAPs and their fertilities determined. Immunization of female rats with rSMP-229 and rSMP-230 induced infertility in 25% and 83% of the treated animals, respectively. All male rats immunized with rSMP-229 remained fertile; whereas animals immunized with rSMP-230 did not mate with normal cycling female rats. Three impotent male rats were found to regain their mating potency 45 days after the last booster injection. These findings demonstrated that immunization with rSMP-230 induced a reversible impotency in male rats. Serum testosterone and LH levels were reduced in rSMP-230-immunized male rats and were elevated in rSMP-229-immunized animals. Histopathological examination of sections of testes from male rats immunized with rSMP-230 showed impairment of spermatogenesis and sloughing of germ cells into the lumen of the seminiferous tubules. The testes of male rats immunized with rSMP-229 showed normal morphology and active spermatogenesis with scattered foci of nodular hyperplasia of Leydig cells in the interstitial areas. In conclusion, immunization with synthetic peptide segments corresponding to different domains of a deduced sperm protein induced infertility in a significant number of female rats and transient impotency in male rats.

Amino Acid Sequence↗
PubMed1993

Regulation of serotonin-induced calcium uptake in Spisula oocytes by tricyclic antidepressants.

The effect of tricyclic antidepressants on 5-hydroxytryptamine (serotonin or 5-HT)-induced maturation and Ca2+ uptake in Spisula oocytes was determined. The drugs at a concentration of 20 microM completely blocked germinal vesicle breakdown and 45Ca2+ uptake by Spisula oocytes. The order of effectiveness of the antidepressants were imipramine > desipramine > clomipramine. The present findings suggest that the action of tricyclic antidepressants is to regulate Ca2+ uptake into 5-HT-responsive cells.

Animals↗
PubMed1993

Potentiating effect of epostane on pregnancy terminating activity of RU 486 in the rat.

The effectiveness of a combination of RU 486, an antiprogestin, and epostane, a 3 beta-hydroxysteroid dehydrogenase inhibitor, for termination of pregnancy in female rats was determined. Epostane administered at doses ranging from 8 to 48 mg/Kg/day on days 7, 8, and 9 of pregnancy did not influence the progress of the pregnancy. RU 486 in doses of 1, 2, and 4 mg/Kg/day terminated pregnancy in 0, 40, and 100% of rats, respectively. The combination of epostane (48 mg/Kg/day) plus RU 486 (1 and 2 mg/Kg/day) terminated pregnancy with complete resorption of fetuses in all treated animals. Unexpectedly, the drug combination that terminated pregnancy used a dose of epostane that was not highly effective in lowering serum progesterone levels when epostane was administered alone. Thus, in the rat, the synergistic action of epostane plus RU 486 may not be dependent upon reduced progesterone secretion plus receptor blockage as anticipated. The present results suggest that the combination of epostane plus RU 486 is effective for pregnancy termination.

Abortion, Induced↗
PubMed1993

Inhibition of progesterone action by a factor in human follicular fluid.

A factor was found in human follicular fluid that blocked progesterone-stimulated net uptake of 45Ca2+ in human sperm and progesterone-induced maturation of Xenopus oocyte. The factor was partially purified by ultrafiltration through PM-10 membrane and gel filtration on Sephadex G-25 column. The active fraction is effective at a concentration of 200 micrograms/ml. The present findings suggest that hFF factor may regulate the metabolism of progesterone sensitive cells.

Animals↗
PubMed1993

Identification of a cAMP-dependent protein kinase in bovine and human follicular fluids.

A soluble protein kinase (PK) was purified from bovine and human follicular fluids (FF) by ultrafiltration through a PM-10 membrane followed by chromatography on heparin-agarose, DEAE-cellulose and cellulose phosphate columns. The PK phosphorylated calf thymus histones and endogenous FF proteins having estimated Mrs of 40, 62, 128 and 180 KD. cAMP enhanced PK activity; whereas protein kinase A (PKA)-inhibitor peptide blocked the activity. The present findings suggest that the enzyme is a cAMP-dependent PK.

Animals↗
PubMed1992

Infertility induced in rats by immunization with synthetic peptide segments of a sperm protein.

Three peptide segments (YAL-198, YAL-201 and YAL-212) corresponding to the extracellular domain of a human sperm protein designated as YWK-II antigen were synthesized as multiple antigen peptide (MAP). Male and female rats were immunized with the YWK-II-MAPs and fertility determined. In a group of 12 female rats immunized with YAL-198, seven animals were infertile and two animals were subfertile. When immunized with YAL-201 and YAL-212, 4 and 2 animals were infertile, respectively. In a group of 15 males immunized with YAL-198, 2 animals were infertile and 6 were subfertile. Two animals immunized with YAL-201 were subfertile. All control male and female rats immunized with bovine serum albumin and adjuvant were fertile. Sera obtained from infertile rats immunized with YAL-198 contained higher titers of antibodies compared to those obtained from fertile animals. The present study shows that immunization with synthetic peptide segments of a sperm protein can effectively reduce fertility.

Amino Acid Sequence↗
PubMed1992

Expression of a gene encoding a rabbit sperm membrane protein in mammalian cells.

A general mammalian expression vector designated pSV2-EP was reconstructed by inserting an oligonucleotide fragment into pSV2-dhfr. This vector allowed insertion of cDNAs with EcoRI cohesive ends. The pSV2-EP contains a simian virus 40 (SV40) early promoter, origin for DNA replication, SV40 poly-A site, splicing site, an initiator ATG downstream from the promoter and an EcoRI site for the insertion of cDNA fragment screened from lambda gt11 expression libraries. A recombinant plasmid (pS-VRS-1) was constructed by inserting RSD-1, a cDNA encoding a rabbit sperm tail protein, into the EcoRI site of the pSV2-EP vector. Chinese hamster ovarian (CHO) dhfr-negative cells were cotransformed with pSV2-dhfr and pSVRS-1 by the calcium phosphate method. In selective culture medium without thymidine and hypoxanthine, several cell lines were obtained containing mRNA and DNA that hybridized with RSD-1. One of these transformed cell lines stained intensely with anti-rSMP-B antibodies, demonstrating that the RSD-1 was expressed in the transformed CHO cells.

Animals↗
PubMed1992

Antifertility effects in rats actively immunized with 80 kDa human semen glycoprotein.

A 80 kDa human sperm antigen has been identified using the serum of an infertile woman having circulating antisperm antibodies. The antigen was then purified to homogeneity by gel permeation chromatography using HPLC (protein PAK-125 column) system and on FPLC (superose-12 column) system. The antigen was found to be a glycoprotein. The antigen was mainly localized in the postacrosomal region of the human sperm, while it was localized in the head region of the rat sperm as demonstrated by immunofluorescent staining. The presence of this antigen was also demonstrated in the human prostate and endometrium and in the rat testis; epididymis and the prostate by immunocytochemical staining. The purified protein upon active immunization in female rats caused infertility in 100 percent animals. While in male rats it caused infertility in 90 percent animals. On morphometric analysis of testicular tissue it was observed that there was no significant change in spermatogonia and spermatocytes, but significant decrease in spermatids and sperm number as well as daily sperm production in the immunized male rats. The epididymal spermatozoa were markedly reduced in number and were largely found to be agglutinated. The results suggest that 80 kDa human sperm antigen appears to be a suitable candidate for immunocontraception both in male and female.

Animals↗
PubMed1992

Isolation and sequencing of the cDNA encoding the 75-kD human sperm protein related to infertility.

Serum was obtained from an infertile woman (IS) inducing head-to-head agglutination of human sperm and was used to screen a human testis lambda gt11 cDNA library. A plaque producing the interacting antigen was located. The recombinant lambda gt11 was isolated and cut with EcoRI releasing a 0.7-kb cDNA. Using the 0.7-kb cDNA as a probe, a larger cDNA of 2.4 kb was isolated and its nucleotide sequence determined. It was composed of 2 427 nucleotides with an open reading frame of 1584 nucleotides encoding 528 amino acid residues. The specific antisperm antibody was isolated from IS by epitope selection, using positive plaques of E. coli Y1090. The epitope-selected antibodies interacted with a 75-kD human sperm protein and with a polypeptide in the form of a beta-galactosidase fusion protein in the recombinant lysate of E. coli Y1089, determined by immunoblot. The fusion protein was purified by affinity chromatography on an anti-beta-galactosidase-Sepharose column. It is proposed that production of anti-75-kD antibodies may be the underlying cause of the infertility.

Amino Acid Sequence↗
PubMed1991

Identification and expression of boar sperm proteins in the reproductive tract that interact with antibodies in serum from an infertile woman.

Serum designated as IS obtained from a young healthy infertile woman induced a head-to-head agglutination of ejaculated boar sperm. The immunoglobulin G (IgG) prepared from IS localized to the acrosomal region of the sperm head obtained from the corpus and cauda epididymis as determined by an indirect immunofluorescent method. The IgG interacted with a boar sperm protein with an estimated molecular weight of 45-kDa, determined by sodium dodecyl sulfate-polyacrylamide gel electrophoretic (SDS-PAGE) immunoblotting technique. However, the IgG did not interact with proteins extracted from sperm obtained from the testis and caput epididymis or from non-gonadal tissues including liver, kidney, spleen, muscle and serum. The IgG interacted with additional proteins of about 75- and 38-kDa present in the corpus and cauda epididymal fluids but not those in the caput epididymal fluid. The staining intensity of the 75-kDa band was reduced and that of the 38-kDa was nullified with ejaculated seminal plasma proteins. The interacting proteins were adsorbed when chromatographed on Concanavalin A Sepharose column, suggesting that they are glycoproteins.

Agglutination Tests↗
PubMed1991

Antifertility effects of human sperm antigen in female rats.

The sperm antigens responsible for inducing infertility were identified by Western blot technique using sera from an infertile woman with circulating antisperm antibodies. The 80 Kda was prepared from human sperm by extraction with 0.05% sodium deoxycholate in 0.01 M Tris-HCl buffer, pH 8.4 and fractionation with ammonium sulphate. The supernatant after 40% saturation ammonium sulphate extraction was separated by gelpermeation chromatography, using HPLC (Protein PAK 125 column) and FPLC (superose 12 column) systems. The homogeneity of the protein was established by SDS-PAGE and its molecular size was estimated to be 80 Kda and its isoelectric point was 4.5. The purified protein upon active immunization in female rats caused infertility in 100 percent animals. The data suggest that 80 Kda human sperm antigen has the potential for use as a contraceptive vaccine.

Animals↗
PubMed1991

Regulation of testicular differentiation and testosterone production in the fetal mouse gonad in vitro.

The objective of the present study was to develop a chemically-defined medium in which early stages of testicular differentiation can be investigated in an organ culture system. Mouse gonadal primordia were explanted before and after initiation of morphological sex differentiation, i.e. 11 and 12 day of gestation (d.g.), respectively. We found that a combination of human albumin fraction, insulin (or IGF-I), and sodium pyruvate promoted testicular organization of gonadal explants of 11 d.g., but not those of 12 d.g. Insulin also increased the production of testosterone from testicular explants of 11 d.g., but not those of 12 d.g. For the younger explants, progesterone was more efficient than pregnenolone as a steroid precursor during the first day of culture, but the maximum effect of pregnenolone was much higher than that of progesterone in later stages. The responsiveness to human chorionic gonadotropin increased gradually along with testicular organization. The addition of either serum or pregnenolone prominently increased the activity of delta 5-3 beta hydroxysteroid dehydrogenase in testicular explants of 11 d.g., but not the number of positive cells as demonstrated by histochemical staining. These results suggest that insulin (or IGF-I) is required during the initial phase of testicular organization, which is reflected by an increase in testosterone production and sensitivity to gonadotropins.

3-Hydroxysteroid Dehydrogenases↗
PubMed1991

A follicular fluid factor inhibiting Xenopus oocyte maturation.

A factor was isolated from bovine follicular fluid (FF) that blocked progesterone-induced maturation of Xenopus oocytes. The factor was purified by ultrafiltration through PM-10 membrane, gel filtration on Sephadex G-25 column, gel permeation chromatography on Superose 12 column and reversed-phase HPLC on Partisil ODS-3 column. The active factor is a peptide with an estimated Mr of 8000 and it effectively blocked progesterone-induced maturation of Xenopus oocytes at a concentration of 100 micrograms/ml. On the other hand, it did not influence the spontaneous maturation of isolated cumulus-enclosed or denuded mouse oocytes. The present findings suggest that the FF factor may be an inhibitor or antagonist of progesterone action.

Animals↗
PubMed1991

Ovarian glycosaminoglycans potentiate angiogenic activity of epidermal growth factor in mice.

Epidermal growth factor (EGF) has been shown to induce capillary proliferation. In the course of our investigation on the interaction of ovarian components with EGF, we observed that partially purified glycosaminoglycans (GAGs) isolated from mouse ovaries enhanced the angiogenic activity of EGF when applied simultaneously to the lateral wall of the sheath of musculus rectus abdominis. Mouse EGF from submandibular glands embedded in Elvax 40 implanting on the musculus rectus abdominis induced neovascularization in a dose-dependent manner. When 0.5 micrograms ovarian GAGs was embedded in the implant with a low amount of EGF that induced only slight neovascularization (0.5 or 1 microgram/implant), the angiogenic activity of the growth factor was markedly enhanced. The active GAG component was isolated by chromatography on Dowex 1-x2. The fraction eluted with 0.5 M NaCl possessed the greatest activity to potentiate the angiogenic activity of EGF. When the reaction mixture of GAGs and EGF was treated with 1% cetylpyridinium chloride, the angiogenic activity was identified with the supernatant. On the other hand, after incubating EGF with 0.5 M NaCl fraction, the angiogenic activity of EGF was identified with the precipitate (GAG fraction) of the cetylpyridinium chloride-treated reaction mixtures. These findings show that ovarian GAGs potentiate the angiogenic activity of EGF by interacting or complexing with EGF.

Abdominal Muscles↗
PubMed1991

Differential effects of epidermal growth factor on the differentiation of type A spermatogonia in adult mouse cryptorchid testes in vitro.

The effect of epidermal growth factor (EGF) on testicular germ cell differentiation was investigated. Testicular fragments from surgically prepared cryptorchid testes of adult mice were cultured for 9 days in serum-free media containing various concentrations of EGF. Histological sections of testis were examined under a light microscope and each type of germ cell and mitotic cell in the seminiferous tubules was counted per 1000 Sertoli cells. EGF at concentrations ranging from 100 to 200 ng/ml induced differentiation of type A spermatogonia. The observed maximal stimulatory activity of EGF at a concentration of 100 ng/ml was 30% of the positive control cultures treated with calf serum. EGF at concentrations ranging from 1 to 100 ng/ml significantly inhibited the mitotic activity of FSH, FSH plus retinol, or FSH plus fetuin on type A spermatogonia and their differentiation. The number of type A spermatogonia in testes cultured with FSH, FSH plus retinol, or FSH plus fetuin decreased when EGF was added. On the other hand, EGF stimulated the differentiation of type A spermatogonia induced with fetuin but did not influence retinol-induced differentiation. It is proposed that EGF inhibits testicular germ cell differentiation by blocking the proliferation of type A spermatogonia stimulated by FSH.

Animals↗
PubMed1991

Properties of an immunoglobulin binding factor from human seminal plasma.

A soluble factor (IBF) in human seminal plasma that binds serum immunoglobulins (Ig) of various species was purified to homogeneity by ammonium sulfate precipitation, preparative isoelectrofocusing, and gel filtration chromatography. The purified IBF interacted weakly with Fc and F(ab')2 fragments and not with Fab. It interacted with anti-Leu 11b and polyclonal anti-Fc gamma RIII antibodies, but not with other anti-Fc gamma R antibodies (32.2, IV.3 and 3G8). IBF is probably a non-glycosylated protein with isoelectric point ranging from 5.1 to 5.8. The estimated Mr of the purified native IBF is 27 kD, determined by sodium dodecyl sulfate polyacrylamide electrophoresis (SDS-PAGE) under non-reducing condition. In its native form, IBF did not bind Ig or interact with anti-Fc gamma R antibodies. Following SDS-PAGE under reducing condition, IBF migrated as a single protein with an estimated Mr of 16 kD and interacted with Ig of various species and with anti-Leu 11b antibodies. When carboxymethylated, however, IBF did not bind IgG. The present results suggest that free sulfhydryl groups of IBF is required for Ig binding.

Antibodies↗
PubMed1991

Identification of pyruvate carboxylase in 3T3-L1 cells by transblot and its correlation with differentiation into adipocytes.

The 3T3-L1 preadipocytes treated with insulin, dexamethasone and 3-methyl-1-isobutylxanthine (IBMX) two days before reaching monolayer undergo differentiation into adipocytes. Cell lysates were prepared from these cells under various conditions and analyzed by SDS-PAGE and transblot. Peroxidase-conjugated avidin used to detect endogenous proteins interacted strongly with a protein with an estimated molecular weight of 120 kDa, corresponding to pyruvate carboxylase, in the differentiated 3T3-L1 cells. On the other hand, this protein was not detected in undifferentiated 3T3-L1 cells.

1-Methyl-3-isobutylxanthine↗