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Query: UNIPROT:P56851 (epididymal)
 11,273 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The protein composition of epididymal fluid and sperm extracts of rats treated with the nitroimidazole compound ornidazole was investigated by two-dimensional gel electrophoresis. Epididymal luminal fluid from the corpus and cauda regions of male animals rendered infertile by ornidazole treatment contained a prominent protein (contraception-associated protein 1, CAP1) with a molecular mass of approximately 25 kDa and an isoelectric point (pI) of 5.8; it was not found in fluids, but was present in sperm, from fertile vehicle-fed rats. Infrared matrix-assisted laser desorption/ionization mass spectrometry indicated that the molecular mass of CAP1 was 20420+/-120 daltons. Analysis of 17 amino acids demonstrated 49% homology to a diuretic hormone from an insect (Acheta domesticus). Densitometric quantitation of CAP1 on silver-stained gels indicated its presence in greater amounts in cauda than in corpus fluid from treated animals, whereas fluid from the rete testis lacked CAP1. In vitro incubations of tissue from the caput, corpus, and cauda epididymidal regions with [35S]methionine gave no hint that CAP1 was a secretion product of the epididymal epithelium. The absence of CAP1 from luminal fluid obtained from the sperm-depleted corpus epididymidis of efferent duct-ligated ornidazole-fed rats suggested a spermatozoal origin. CAP1 was present in spermatozoa from the caput epididymidis but not from the rete testis in control animals. Less CAP1 was present in detergent extracts of cauda sperm from ornidazole-treated rats than in sperm from control animals, suggesting a contraceptive-related displacement of protein from sperm to fluid. The association of ornidazole- and alpha-chlorohydrin-induced infertility with the presence of CAP1 in epididymal fluid, probably originating from spermatozoa, suggests a critical role for this protein in fertilization.
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PMID:Shedding of a rat epididymal sperm protein associated with infertility induced by ornidazole and alpha-chlorohydrin. 960 61

Congenital absence of the vas deferens (CAV), failed vasoepididymostomy, and all irreparable obstructions were once very frustrating conditions, because these patients have normal spermatogenesis, but were previously untreatable (Silber SJ, Ord T, Balmaceda J et al (1990) New England Journal of Medicine 323: 1788-1792). Microsurgical epididymal sperm aspiration (MESA) together with in vitro fertilization was introduced in 1985 and in 1988 to treat these cases, but only modest success was achieved (Temple-Smith PD, Southwick GJ, Yates CA et al (1985) Journal of In Vitro Fertilization and Embryo Transfer 2: 119-122; Silber SJ, Asch R, Balmaceda J et al (1988) Fertility and Sterility 500: 525-528; Silber SJ, Ord T, Balmaceda J et al (1990) New England Journal of Medicine 323: 1788-1792). It is very difficult to predict in which cases epididymal sperm will fertilize and in which cases it will not. The reason for this problem may be either sperm maturation defects that are poorly defined or senescent and pathological changes caused by the obstruction. Thus, intracytoplasmic sperm injection (ICSI) became necessary to achieve consistently good results with MESA. The only factor in these couples which affected success had nothing to do with the sperm origin or quality but rather was simply the age and ovarian reserve of the wife. Clearly, whether sperm was from the epididymis or the testis, frozen or fresh, or whether the male had CAV, or irreparable obstruction from a variety of other causes, made no meaningful difference. The cystic fibrosis genotype, the sperm morphology and the quality of motility had no impact. Furthermore, the only factor in the wife that mattered was her age. It is crucial to screen for CF in both the husband with CAV and also his wife. If the wife is negative for any of the 36 common CF mutations, we feel that it is quite safe to perform MESA-ICSI on the couple. The chances of a male offspring's having CAV are very remote, and the chances of the child's having cystic fibrosis are probably less than in a normal, unscreened population. However, if the wife turns out to be a CF carrier herself (4% incidence in the general population), the couple can still undergo MESA-ICSI, but pre-implantation embryo diagnosis would then be mandatory. We have published the first case of successful pre-implantation embryo diagnosis in a CAV-MESA case in which both partners were carriers of the delta F508 mutation (Liu J, Lissens W, Silber SJ et al (1994) Journal of the American Medical Association 23: 1858-1860. We require this as a routine approach whenever the female is discovered, on screening, to be a CF carrier.
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PMID:The use of epididymal sperm for the treatment of male infertility. 969 14

A variety of sialic acids contained in the rat epididymis during post-natal development were examined by means of lectin and carbohydrate histochemistry. Epididymides from male Sprague-Dawley rats on post-natal days 14, 21, 30, 39, 49, 56 and 70 were fixed in Bouin's fluid and embedded routinely in paraffin wax. Hydrated sections were subjected either to the lectin methods using biotinylated Sambucus sieboldiana lectin or Maackia amurensis lectin or to the selective periodate oxidation-phenylhydrazine-thiocarbohydrazide-silver protein-physical development technique with or without saponification. The results revealed that sialic acids appeared in the epididymal epithelium at day 14, followed by particular distribution patterns corresponding to cell differentiation during days 21-39. High-level O-acetylation of sialic acids was observed in the principal cells of the initial segment and proximal caput after day 39. These results suggest that sialic acids with different linkages and O-acetylation become adult in distribution at the 'differentiation' period under the influence of androgen, before spermatozoa reach the epididymal lumen. Such carbohydrates may be correlated, at least in part, with sperm-binding sialoproteins, which increase dramatically during the window between days 21 and 39.
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PMID:Histochemical studies on sialic acids in the epididymis during post-natal development of the rat. 987 Jul 67

Sialoprotein "anti-agglutinin," previously shown to inhibit sperm head-to-head agglutination, is found in both boar epididymal and seminal plasma. The present report characterizes anti-agglutinin by mass spectrometry, by N-terminal amino acid sequence analysis, and by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) and Western blotting techniques to assess phosphate content of the molecule. Anti-agglutinin had the SDS-PAGE mobility of approximately 25 kDa. By electrospray ionization-mass spectrometry, however, mass spectra of anti-agglutinin were characterized by two major peaks (19,379-19,382 Da and 19,395-19,397 Da) and several minor peaks. Mass spectrometry of tryptic peptide fragments of deglycosylated anti-agglutinin and amino acid sequence analysis revealed that the protein has a unique peptide-mass fingerprinting of fragments (12,668 Da, 5,209 Da, 1,226 Da, and 1,168 Da) and a novel N-terminal amino acid sequence (KTDDY AISGA KEEEF YDYME ELYAV), respectively. Additionally Western blot techniques, using commercially available monoclonal antibodies, were used to detect presence of phosphothreonine and phosphoserine substituents, but two different monoclonal antibodies did not detect phosphotyrosine. Moreover, treatment with two different alkaline phosphotases converted the molecule, as assessed by SDS-PAGE and detection by silver stain, from the parent form of about 25 kDa to forms of approximately 19 kDa (similar to that assigned by mass spectrometry) and/or 15 kDa. Original antiserum generated toward, and reacting with native anti-agglutinin, reacted only with 19 kDa form. These results are consistent with the conclusion that the native anti-agglutinin may be a novel protein that is phosphorylated at serine and/or threonine residues.
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PMID:Biochemical characterization of sialoprotein "anti-agglutinin" purified from boar epididymal and seminal plasma. 1060 79

Rat outer dense fibres were isolated from cauda epididymal spermatozoa using mechanical and chemical dissection methods. Sperm tail isolation procedures were monitored by phase-contrast microscopy and the purity of the outer dense fibres was verified by electron microscopy. SDS-PAGE of isolated outer dense fibres revealed at least nine Coomassie brilliant blue stained bands, and 12 silver staining bands. The most abundant proteins were a large band between 26.5 and 32.5 kDa, and 84 kDa, 21.5 kDa and 15.5 kDa bands. The amino acid composition of the total rat outer dense fibres and seven isolated proteins showed similar compositions, being abundant in aspartic and glutamic acid, serine, glycine and leucine. However, the content of cysteine and proline was highly variable among the isolated proteins. Immunofluorescence microscopy demonstrated that a polyclonal antiserum to isolated rat outer dense fibres showed positive staining localized to the mid-piece of rat and rabbit spermatozoa. However, there was crossreactivity in the principal piece as well as the mid-piece of the human spermatozoa. The antiserum also showed crossreactivity in the perforatorium of rat sperm heads and the acrosome and equatorial segment of rabbit sperm heads. These data indicate that it is technically possible to isolate proteins from the outer dense fibres that will enable further studies of the amino acid sequences of sperm tail proteins.
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PMID:Isolation and characterization of rat sperm tail outer dense fibres and comparison with rabbit and human spermatozoa using a polyclonal antiserum. 1061 60

In the present study we report the identification of a novel epididymis-specific secretory glycoprotein, E-3, which is a sperm-associated isoantigen containing defensin- and lectin-like motifs. E-3 was detected in rat epididymal fluid and in sperm extracts by two-dimensional (2-D) Western blotting using rat hyperimmune sera raised against rat sperm. The immunoreactive spot of approximately 28 kDa with an isoelectric point (pI) of 3.5 was cored from silver-stained gels. Microsequencing by tandem mass spectrometry and database searches revealed several peptides to be novel sequences. Degenerate deoxyinosine-containing primers corresponding to the novel peptides were used in rapid amplification of cDNA ends and polymerase chain reaction to clone E-3 from a rat epididymal cDNA library. A 449-base pair nucleotide sequence was subsequently obtained consisting of a complete open reading frame (ORF) of 111 amino acids, which showed similarity to the defensin and lectin families. The first 21 amino acids constituted a putative signal peptide, suggesting that E-3 is a secretory protein. Mature E-3 protein corresponding to amino acids 22-111 was expressed in E. coli, and chickens were immunized with recombinant E-3 (rE-3). The resulting anti-rE-3 antisera recognized the recombinant immunogen as well as a "native" protein of 28 kDa, pI 2.5-3.5 in both epididymal fluid and in sperm extracts on 2-D Western blots. Northern hybridization indicated that E-3 mRNA was present in the epididymis but not in testis or other tissues, and that E-3 mRNA was predominantly expressed in the corpus and cauda of the epididymis, but not in the initial segment or caput. Similarly, Western blots detected the E-3 protein only in the epididymal fluid and sperm from the corpus and caudal regions. Finally, indirect immunofluorescence localized E-3 on the entire tail, and with less intensity on the head of the sperm. These observations indicate that E-3 is a secreted epididymal protein that becomes associated with the sperm as it transits through the corpus and cauda. The presence of a defensin-like motif suggests that E-3 may play a role in protecting the sperm from microbial infections in the epididymis and in the female reproductive tract.
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PMID:Cloning and characterization of a novel sperm-associated isoantigen (E-3) with defensin- and lectin-like motifs expressed in rat epididymis. 1249 25

The goals of the present study were to determine the availability of progesterone (P4) receptor (P4r) in mouse sperm during maturation and capacitation and to make the first steps toward a characterization of P4r. It has been proposed that P4 is able to induce an acrosomal reaction (AR) by using a membrane P4r. This induction was verified in sperm isolated from the cauda epididymis (fully mature) when incubated in specific conditions that capacitate sperm. First, we set up the conditions in our laboratory to induce an AR in mature and capacitated sperm triggered by P4 that was detected by a chlortetracycline (CTC) assay. Then, we examined sperm isolated from the caput epididymis (immature) incubated under conditions that support cauda sperm capacitation and found that the AR could not be detected. Moreover, P4 was unable to induce the AR when it was applied to sperm isolated from either region and incubated under conditions that did not support capacitation. These results can be explained by changes in P4r availability. A suitable marker for P4r is the gold (Au)-P4 complex. This marker shows a binding capacity that can be visualized directly by electron microscopy (EM) and indirectly by silver-enhanced methods with light microscopy. The Au-P4 complex was localized in capacitated cauda sperm at the dorsal edge of the head. Using these techniques, we observed a significant decrease in both noncapacitated cauda sperm and caput sperm (whether incubated in capacitating media or not). Genomic P4r could be responsible for the signal detected, but antibodies against the P4 nuclear receptor did not recognize any sites in the sperm by immunostaining methodology. Instead, a 44-kd protein band was detected in the sperm by a ligand blot assay. In conclusion, P4 promotes the AR in capacitated cauda sperm but is unable to do so in noncapacitated or immature sperm because the availability of P4r increases during epididymal transit and after capacitation. The P4r responsible for this behavior is different from a classical nuclear receptor-on the basis of the immunostaining results-and is probably a protein close to 44 kd-on the basis of the ligand assay results.
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PMID:Progesterone receptor availability in mouse spermatozoa during epididymal transit and capacitation: ligand blot detection of progesterone-binding protein. 1282 1

After in vitro incubation of rat epididymal fat pads with radioactive palmitic acid, the distribution of the label in the different lipid classes and in different triglycerides was determined by silica gel and silver nitrate-silica gel thin-layer chromatography (TLC).The radioactivity of the diglycerides was approximately half of the triglycerides. This ratio did not change with alteration in the time of incubation. It remained unaltered even after a subsequent 10-min incubation in a nonradioactive medium.When the fat pads were incubated, first with(14)C-, then with(3)H-labeled palmitic acid, the(3)H/(14)C ratio was slightly lower in diglycerides than in triglycerides.The fully saturated molecules contained 38% of the radioactivity of triglycerides. Addition of oleic acid or norepinephrine to the labeled palmitic acid-containing medium decreased this value. Subsequent incubation with these compounds did not alter the distribution of radioactivity.
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PMID:The distribution of labeled palmitic acid into the diglycerides and triglycerides of rat adipose tissue. 1780 55

Toll-like receptor-4 (Tlr-4), a key pattern recognition receptor involved in innate immune response, is activated by saturated fatty acids (SFAs). To investigate the involvement of this receptor in obesity caused by consumption of diets high in fat, we utilized male Tlr-4-deficient 10ScN mice and 10J controls. Mice were fed either low fat (low-fat control (LFC)), high unsaturated fat (high-fat control (HFC)), or high saturated fat + palmitate (HFP) diets ad libitum for 16 weeks. Relative to the LFC diet, the HFC diet resulted in greater epididymal fat pad weights and adipocyte hypertrophy in both Tlr-4-deficient and normal mice. However, the 10ScN mice were completely protected against the obesigenic effects of the HFP diet. Moreover, macrophage infiltration and monocyte chemotactic protein-1 (MCP-1) transcript abundance were lower in adipose tissue of 10ScN mice fed the HFP diet, and the hyperinsulinemic response was negated. Tlr-4-deficient mice also had markedly lower circulating concentrations of MCP-1 and much less nuclear factor-kappaB (NFkappaB) protein in nuclear extracts prepared from adipose tissue, irrespective of diet. In contrast, Tlr-4 deficiency did not attenuate the induction of tumor necrosis factor-alpha (TNF-alpha) or interleukin-6 (IL-6) expression in adipose tissue. These data indicate that Tlr-4 deficiency selectively protects against the obesigenic effects of SFA and alters obesity-related inflammatory responses in adipose tissue.
Obesity (Silver Spring) 2008 Jun
PMID:Tlr-4 deficiency selectively protects against obesity induced by diets high in saturated fat. 1842 Dec 79

The acyl-CoA dehydrogenases (ACADs), which catalyze the rate-limiting step in the mitochondrial beta-oxidation spiral, were investigated in white adipose tissue (WAT) of C57Bl/6 mice treated with 10 mg/kg/day rosiglitazone. Rosiglitazone was also administered to PPAR-alpha knockout mice. ACAD abundance and activity were determined using western blotting and an ACAD enzyme activity assay. Rosiglitazone increased ACAD activity in both epididymal and inguinal WAT but not in brown adipose tissue, liver, or muscle. Given the known function of PPAR-alpha in regulating the expression of ACAD genes in liver, it was hypothesized that PPAR-alpha may be involved in upregulating the ACADs during rosiglitazone-mediated adipose tissue remodeling. However, the effect of rosiglitazone on adipose tissue ACAD activity was the same in wild-type and PPAR-alpha knockout mice. In conclusion, rosiglitazone increases expression and activity of ACAD enzymes in WAT independently of PPAR-alpha.
Obesity (Silver Spring) 2009 Jan
PMID:The regulation of acyl-CoA dehydrogenases in adipose tissue by rosiglitazone. 1894 67


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