[Impact of sperm DNA fragmentation on ART outcome]. / La fragmentation de l'ADN du spermatozoïde: impact en Assistance médicale à la procréation.

We have used the sperm chromatin structure assay (SCSA) test in order to determine if correlations can be found between sperm DNA fragmentation and spermogram parameters. Only necrospermia and DNA fragmentation index are strongly correlated (P<0.0001). Neither fertilization rates for ICSI and IVF, nor blastocyst formation rates are impaired by a high DFI. However when the critical DFI>30% is reached, the chances of having ongoing pregnancies after blastocyst transfer are reduced by three. Treatments with antioxidants are of limited efficacy even though we obtained 2 deliveries after DFI treatments with such treatments. New strategies in order to improve the pregnancy rates for these peculiar cases of reduced fertility are discussed.

Sperm DNA fragmentation: threshold value in male fertility.

BACKGROUND: The extent of sperm DNA fragmentation, which can be measured by the TUNEL assay, is one of the determinants of male fertility. However, the clinical application of this test to in-vivo situations is difficult owing to the absence of a statistically validated threshold value. METHODS: The aim of this study was to compare the results of TUNEL assay applied to semen samples from men of proven fertility (n = 47) and patients from an infertile population (n = 66), in order to establish a discriminating threshold value. RESULTS: Infertile patients had a higher mean level of DNA fragmentation than men of proven fertility (40.9 +/- 14.3% versus 13.1 +/- 7.3%, respectively; P < 0.001). The area under the receiver operating characteristics curve was 0.93 for 20% sperm DNA fragmentation. The calculated threshold value for TUNEL assay to distinguish between fertile controls and infertile men was 20%. At this threshold, specificity was 89.4 [95% confidence interval (CI) 83.7-95.1] and sensitivity was 96.9% (95% CI 93.8-100). The positive and negative predictive values of the 20% sperm DNA fragmentation threshold were high: 92.8% (95% CI 87.9-97.5) and 95.5% (95% CI 91.6-99.3), respectively. CONCLUSION: This study demonstrates that sperm DNA fragmentation, as measured by TUNEL assay, is a highly valuable indicator of male fertility.

Longitudinal study of sperm DNA fragmentation as measured by terminal uridine nick end-labelling assay.

BACKGROUND: One major limitation in the use of sperm DNA fragmentation as measured by the TdT (terminal deoxynucleotidyl transferase)-mediated dUTP nick-end labelling (TUNEL) assay is the paucity of solid data on the stability of this parameter. METHODS: The objective of our study was to evaluate variations in the degree of sperm DNA fragmentation, as measured by the TUNEL assay, over a 6 month period. Five donors provided semen samples (total 107) on the average three times per month, and 10 infertility patients provided semen samples every 4 weeks (total 58). RESULTS: The mean percentage of sperm DNA fragmentation for donors was 13.18%, the within-donor standard deviation (SD(W) = 3.79%) was small compared to between-donor (SD(B) = 17.56%). For the group of patients, the mean percentage of sperm DNA fragmentation was 22.44%, with SD(W) of 4.43% within patients and SD(B) of 29.48% between patients. No seasonal rhythm was observed during the study. The intra-class correlation coefficient for all subjects combined was 0.83. Compared to sperm concentration, individual coefficients of variation for sperm DNA fragmentation indicated less variability in four subjects, but were similar in the others. CONCLUSION: This longitudinal study shows that sperm DNA fragmentation is a parameter with good stability (repeatability) over time; it can be taken as a baseline both in healthy fertile men and in patients from infertility couples.

[Sperm DNA integrity as diagnosis and prognosis element of male fertility]. / Intégrité de l'ADN des spermatozoïdes comme élément diagnostique et pronostique de la fertilité masculine.

Recent progress in reproductive biology has improved comprehension physiology of the spermatozoa and on the fertilization mechanisms. This new knowledge has carried out the elaboration of tests on male fertility based on sperm genomic integrity. This review presents some of these techniques and brings a reflexion element on the application and use of sperm DNA integrity in the investigation of male fertility. The single cell gel electrophoresis (COMET assay), Sperm Chromatin Structure Assay (SCSA), In Situ Nick Translation (NT: Nick Translation) and Terminal Uridine Nick-End Labelling (TUNEL assay) are actually the most currently used techniques for the measure of sperm DNA integrity in research clinic. From a certain point of view, TUNEL assay, SCSA, COMET assay and NT assay are complementary. The TUNEL and COMET can measure single and double strand breaks of DNA, the SCSA can detect the abnormalities in the chromatin compaction and the NT assay can detect the single strand breaks of DNA. The exact origin of sperm DNA fragmentation is not established yet. However, several mechanisms have been proposed: defect in the chromatin compaction during spermiogenesis; reactive oxygen species production by immature spermatozoa; apoptosis during spermatogenesis. It becomes important to consider the possible consequences of the oocyte fertilization by a spermatozoon having a high degree of DNA fragmentation. The use in routine of some of these tests must however pass by a standardization of the inter laboratory protocols and obviously, by the establishment of both in vivo and in vitro discriminating threshold values in order for these tests to present a good predictive value for pregnancy outcome.

Innocuity of tetradecyl-dimethyl-benzyl ammonium fluoride on the DNA of human spermatozoa.

Tetradecyl-dimethyl-benzyl-ammonium fluoride (TDBAF) is a powerful new spermicide. In cases of failure of local contraception, there is a theoretical risk that a spermatozoon exposed to a sublethal concentration of spermicide might suffer DNA strand breaks, with transmission of damaged DNA to the oocyte. The present study aims to ascertain the innocuity of TDBAF in this regard. Spermatozoa from 32 fertile men were exposed to a sublethal concentration of TDBAF. The degree of DNA fragmentation was measured by modified terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin end labeling (TUNEL) assay, pulsed field gel electrophoresis of purified sperm DNA and radioenzymatic labeling. With the TUNEL assay, a statistically significant increase in sperm labeling was observed after TDBAF treatment compared to the controls. However, pulsed field gel electrophoresis and enzymatic labeling of purified DNA showed no evidence of augmented DNA fragmentation in TDBAF-treated sperm. Under electron microscopy, TDBAF caused sperm chromatin decondensation. In the TUNEL assay, this phenomenon would allow easier access of terminal transferase to its DNA substrate, explaining the apparent increase in DNA fragmentation in the presence of TDBAF. In conclusion, the use of TDBAF as a spermicide does not place progeny at heightened risk of abnormalities should pregnancy occur. In regard to the TUNEL assay, we suggest that pretreatment of spermatozoa with TDBAF promotes a more accurate measurement of sperm DNA fragmentation.

Organization of a gene coding for an oviduct-specific glycoprotein (oviductin) in the hamster.

Hamster oviductin, a high molecular weight glycoprotein secreted by the oviducts, is believed to participate in fertilization and protection of the tubal epithelium. Expression of the oviductin gene is confined strictly to nonciliated secretory cells of the oviduct and is regulated by hormones. The objective of this study was to characterize the genomic organization and to identify potential regulatory elements implicated in the control of transcription of the oviductin gene. Polymerase chain reaction was performed on hamster genomic DNA, yielding 2.2 kb of the 5' flanking region as well as 13.6 kb of genomic sequence comprising the entire coding sequence of the oviductin gene distributed in 11 exons. Sequencing of the 5' flanking region revealed, among other elements, an almost perfect estrogen-responsive element (GGTCACTGTGACT), an atypical TATA box (TATTAA), and a perfect inverted Sp1 site located between the transcription start site and the atypical TATA box. Primer extension analyses indicated that the hamster oviductin transcript possesses an unusually short 5' untranslated region of only 14 nucleotides. The distinct organization of the hamster oviductin gene in the vicinity of the transcription start site provides an interesting ground for further functional studies.

Lack of association between smoking and DNA fragmentation in the spermatozoa of normal men.

Male factor infertility patients can have anomalies in their sperm nuclei, displaying high levels of loosely packaged chromatin and damaged DNA. The primary objectives of this study were to compare the extent of DNA fragmentation in the spermatozoa of healthy light and heavy smokers versus non-smokers, and to investigate its correlation with concentrations of the smoking markers cotinine and cadmium. A secondary objective was to compare the concentrations of blood cadmium and serum cotinine with corresponding concentrations in seminal plasma. Ninety-seven healthy male volunteers were divided into three groups: non-smokers, light and heavy smokers. There was no difference between the three groups with respect to age, number of ejaculations per week, serum testosterone concentration, and parameters of semen analysis. The percentages of DNA fragmentation in spermatozoa were not statistically different in the heavy smokers (12.11%), light smokers (11.66%) and non-smokers (20.41%). Serum and seminal plasma concentrations of cotinine were significantly higher in heavy smokers compared with the other groups (P < 0.0001). Median values for blood cadmium concentration were higher in heavy smokers (4.50 microg/l) than in light smokers (0.20 microg/l) and non-smokers (0.20 microg/l) (P < 0.001). Cadmium concentration in seminal plasma was significantly higher in heavy smokers (0.20 microg/l) than in light smokers (0.10 microg/l) and non-smokers (0. 10 microg/l) (P < 0.05). In summary, our results indicate no association between smoking and DNA fragmentation in the spermatozoa of healthy men.

Short-term oxidative status of lens and aqueous humor after excimer laser photorefractive keratectomy.

PURPOSE: Oxidation in the anterior ocular segment is associated with cataractogenesis and possible complications to corneal endothelium. We investigated whether oxidation occurred in the rabbit anterior ocular segment shortly after excimer laser photorefractive keratectomy (PRK). METHODS: Rabbits were treated unilaterally with PRK, the other eye serving as a control. Aqueous humor sampled shortly after treatment was assayed spectrophotometrically for hydrogen peroxide using ferrous oxidation in xylenol in the presence (group 1; n=10), or absence of oxygen (group 2; n=8). Oxidized glutathione and malondialdehyde levels were measured in lenses by spectrophotometry and HPLC. RESULTS: Hydrogen peroxide concentration of aqueous humor was not different between treated (77 +/- 36 microM) and control eyes (88 +/- 34 microM) in the oxygen group or the nonoxygenated group (treated eyes: 6.7 +/- 5.4 microM and control eyes: 5.5 +/- 4.7 microM). Peroxide levels did not correspond to endogenous H2O2 but presumably reflected action on ascorbic acid. There was no difference in the percent of oxidized glutathione between experimental and control eyes. Malondialdehyde could not be detected in the lens of treated or control eyes, despite good sensitivity in recovery assays. CONCLUSION: Based on these assays, there is no evidence that PRK oxidizes the aqueous humor or the lens of treated rabbits within 10 minutes of treatment.

Mammalian chitinase-like proteins.

Mammals express genes coding for proteins that show significant similarity to chitinases of family 18 glycosyl hydrolases. These chitinase-like proteins have no chitinase activity due to changes in critical residues in the putative active center. One of these is oviductin, a high molecular weight glycoprotein most likely involved in fertilization and protection of the tubal epithelium owing to its mucin character. Another is YKL-40 (HCgp39) produced in association with tissue remodeling. Such proteins could have a general function in morphogenesis.

Measurement of hydrogen peroxide in biological samples containing high levels of ascorbic acid.

The physiological concentration of hydrogen peroxide in the aqueous humor was reported to range between 25 and 60 microM, and conditions leading to elevated levels could have important damaging effects such as cataract formation. However, the high concentration of ascorbic acid in aqueous humor, which is 20 times that of plasma, was recently shown to interfere in the dichlorophenol-indophenol assay for hydrogen peroxide. The actual concentration of hydrogen peroxide in this fluid has become a controversial issue. In the present study, we used the method of ferrous oxidation of xylenol orange (FOX1 assay) performed in a nitrogen atmosphere to accurately measure low levels of hydrogen peroxide, even in the presence of ascorbic acid at concentrations normally found in aqueous humor. Contrary to values reported in the literature, we observed that the concentration of hydrogen peroxide in the rabbit aqueous humor is less than 5 microM, which is the detection limit of the method.

Allelic polymorphism and chromosomal localization of the human oviductin gene (MUC9).

OBJECTIVE: To determine if the gene coding for human oviductin (the estrogen-dependent, oviduct-specific glycoprotein with an affinity for the zona pellucida) shows length polymorphism in the region of tandem repeats. To determine the frequencies of the length alleles in health and disease. DESIGN: Descriptive fundamental and clinical studies. SETTING: Fertility clinic and research center, university teaching hospital. PATIENT(S): Fertile women, women with a history of ectopic pregnancy or tubal disease, and women with stage I or II endometriosis. INTERVENTION(S): Blood samples were drawn for DNA analysis. MAIN OUTCOME MEASURE(S): Length and sequence of the region of tandem repeats. RESULT(S): Four different length alleles of the human oviductin gene were identified. Their relative frequencies in pathologic cases were not statistically significant compared with those found in normal fertile women. The human genome contains a single copy of the oviductin gene located on chromosome 1p13. CONCLUSION(S): The human oviductin gene codes for a glycoprotein that shares the characteristics of epithelial mucins. Because eight epithelial mucin genes have been identified so far, we therefore propose to name this gene MUC9. The biologic function of the protein is likely to include protection of the early embryo and of the fallopian tube itself.

Fate of hamster oviductin in the oviduct and uterus during early gestation.

Oviductins are a family of glycoproteins which are synthesized and secreted by oviductal secretory cells and which, upon their secretion in the lumen of the oviduct, become associated with postovulatory oocytes and developing embryos. Recently, we showed that hamster oviductin is maximally secreted in the oviduct at the time of ovulation and is later associated with a certain population of uterine epithelial cells, where it is subsequently endocytosed and degraded. In light of these results, this study was conducted to follow the fate of hamster oviductin in the oviduct and uterus during early gestation. Using a monoclonal antibody against hamster oviductin, immunofluorescence and immunogold labeling revealed that during early gestation, immunoreactivity to oviductin in the uterus gradually diminished to an almost total disappearance at time of implantation. However, the strong labeling intensity remained unchanged in the oviduct. Biochemical analyses demonstrated that a degradation of oviductin occurs in the uterus, and a loss of immunoreactivity was also observed as gestation progressed, so that by the time of implantation, immunoreactivity to oviductin was barely detectable. The decrease of oviductin along the uterine epithelium at the time of blastocyst attachment and its final disappearance at implantation suggest that this glycoprotein could be a potential modulator of uterine receptivity.

The COI mitochondrial gene encodes a minor histocompatibility antigen presented by H2-M3.

We found that (LP x C57BL/6)F1 mice could raise a CTL response against parental C57BL/6 cells. These CTLs recognized a maternally transmitted, H2-M3wt-restricted, minor histocompatibility Ag (MiHA) that is widely distributed among many strains of mice and encoded by the COI mitochondrial gene. The wild-type MiHA is the COI N-terminal hexapeptide. Sequencing the 5' end of the COI gene in LP and C57BL/6 mice showed that the LP allele arose by a T-->C transition in the third codon, which caused substitution of threonine for isoleucine. Molecular characterization of this MiHA and the demonstration that it is presented exclusively by H2-M3: 1) support the concept that differential expression of MiHA in MHC-identical animals is caused by polymorphism of the MiHA gene proper; 2) expand our knowledge of the repertoire of self-peptides naturally presented by H2-M3 and show that this MHC class I molecule can present short endogenous peptide ligands; and 3) suggest that mitochondrial DNA mutations that modify the repertoire of H2-M3-associated mitochondrial peptides are representative of mitochondrial DNA mutations in general.

Selenium-vitamin E supplementation in infertile men. Effects on semen parameters and micronutrient levels and distribution.

In order to verify the hypothesis that selenium (Se) and vitamin E (Vit E) could improve male fertility, nine oligoasthenoteratozoospermic men were supplemented for a period of 6 mo with Se and Vit E. Compared to the baseline period (presupplementation) of 4 mo, statistically significant increases were observed for Se and Vit E levels, sperm motility, percent live, and percent normal spermatozoa. These improvements are likely to be "supplementation-dependent," since all of the parameters returned to baseline values during the posttreatment period. None of the couples reported a pregnancy during the study. The HPLC analysis conducted on the serum of one of the patients showed the existence of at least six different Se-containing peaks, whose Se content was affected by supplementation. The mechanism(s) involved in these improvements of semen parameters is presently under investigation.

Allelic polymorphism in the hamster oviductin gene is due to a variable number of mucin-like tandem repeats.

Oviductins are high-molecular-weight glycoproteins specifically secreted by the oviduct. These proteins bind to the zona pellucida of the ovulated oocyte and remain associated with the embryo during its transit in the oviduct. They may be involved in fertilization and early embryonic development. In order to explore their putative biological function, the cDNA sequence corresponding to oviductin in the golden hamster was determined. We found that the deduced amino acid sequence of this heavily O-glycosylated protein presents characteristics typical of mucins, including serine- or threonine-rich tandem repeats. Analysis of several cDNA clones and of genomic DNA revealed the presence of a single copy gene with two frequent alleles differing in the number of repeats. Comparison with oviductin sequences from other mammals indicates a high degree of conservation amongst species, except for the repeat region which shows divergence, notably in the number of repeats. Based on its biochemical and genetic properties, hamster oviductin can now be classified as a secretory mucin. This concept provides a new insight in the elucidation of its biological role: oviductin could possibly provide the oviduct and the oocyte with a protective coating ensuring normal tubal function and embryonic development.

Size variations in the mucin-type domain of hamster oviductin: identification of the polypeptide precursors and characterization of their biosynthetic maturation.

The recent sequencing of a cDNA clone for hamster oviductin revealed that this zona pellucida-associated glycoprotein is a particularly intriguing chimeric molecule because it encloses regions of significant similarity with chitinase-related proteins as well as a carboxyterminal mucin-type domain. This domain contains contiguous Ser/Thr-rich repeated stretches of 15 amino acids; similar units are also found in the deduced sequence of human oviductin. Such structural domains constitute a central feature of mucins. We amplified this region from 16 hamster oviductin cDNA clones and identified three length variants. In order to elucidate the biosynthetic maturation of the glycoprotein, a high-titer antiserum against synthetic peptides derived from internal sequences of hamster oviductin was produced and used in pulse-chase experiments. Two major and one minor polypeptide precursors were identified from tunicamycin-treated cell lysates and in vitro translated products from oviductal poly(A)+ RNA. Their apparent molecular masses correlate with the predicted lengths of the three size variants identified by polymerase chain reaction (PCR) amplification. Using glycosylation and transport inhibitors, we sought to dissect the posttranslational sequential steps leading to the final maturation of hamster oviductin and proposed a compartmental model for its biosynthesis. The polypeptide precursors are rapidly converted in the endoplasmic reticulum into an N- and O-glycosylated premature form of 80-90 kDa (time < 20 min), which is further O-glycosylated and sulfated in the trans-Golgi network, giving rise to the secreted species of 160-350 kDa. The polymorphism in the heavily O-glycosylated region of hamster oviductin is predicted to increase the heterogeneity of the glycoprotein. Such changes may alter the putative biological function of the different variants mediated by their mucin-type domain, such as protection of the embryo and/or adhesion-related phenomena.

Hormonal control of the biosynthesis of hamster oviductin.

In several mammalian species, the epithelial secretory cells of the oviduct synthesize and secrete specific glycoproteins that become associated with the zona pellucida of the ovulated oocyte. These glycoproteins are collectively designated as oviductins. A monoclonal antibody directed against hamster oviductin was used to study the ontogeny of this glycoprotein. Indirect immunofluorescence experiments performed on sections of hamster oviduct revealed that the glycoprotein begins to be secreted in 10-day-old females and that all of the oviductal secretory cells showed fluorescent staining by day 14. The intensity of the immunofluorescence reaction reached a maximum in the 28-day-old females. The oviducts of the 7-day-old hamster incorporated [35S]methionine in vitro into several proteins; however, the production and secretion of detectable amounts of radiolabeled oviductin only began at 14 days of age and reached a maximum at day 28 of age. It appears that the ontogeny of oviductin parallels the hormone dependent changes leading to sexual maturation and that its maximum secretion is already established at the time of the first ovulatory cycle. These results are substantiated by the fact that the production of oviductin is induced in estradiol-treated, but not progesterone or non-treated prepubertal animals, as determined by indirect immunofluorescence experiments.

Oviductins possess chitinase- and mucin-like domains: a lead in the search for the biological function of these oviduct-specific ZP-associating glycoproteins.

Over the last 10 years considerable progress has been made in the immunological and biochemical characterization of oviduct-specific glycoproteins. It is now well established that a subclass of these secretory products, designated as oviductins, associate with the zona pellucida of the ovulated oocyte and with the early embryo. Recent reports on the cloning of cDNAs of oviductins from various species, including that of golden hamster (Mesocricetus auratus) oviductin by our laboratory, allowed us to compare their deduced amino acid sequences with those of other proteins. Optimal alignment analysis showed that oviductins contain regions of significant similarity with catalytically inactive mammalian members of the bacterial and microfilarial chitinase protein family. Most importantly, a close examination of the hamster and human deduced amino acid sequences revealed that both glycoproteins possess contiguous Ser/Thr rich repeated units, clustered in their carboxy-terminal portions. These mucin-type motifs are similar in the hamster and human glycoprotein, although hamster oviductin contains more of these complete units. This striking feature might indicate that these molecules play a similar role to mucin-type glycoproteins, e.g., in protecting the oocyte and early embryo against attacks from their environment. We propose a model whereby oviductins are targeted to the oocyte via the interaction of their chitinase-like domains with specific oligosaccharide moieties of the zona pellucida. Once localized to this structure, oviductin molecules would act as a protective shield around the oocyte and early embryo by virtue of their densely glycosylated mucin-type domains.

Phosphatidylcholine-binding proteins of bovine seminal plasma modulate capacitation of spermatozoa by heparin.

Bovine seminal plasma (BSP) contains four similar acidic proteins, previously designated as BSP-A1, BSP-A2, BSP-A3, and BSP-30-kDa. These proteins are secreted by the seminal vesicles and coat the surface of the spermatozoa after ejaculation. The binding site of BSP proteins on the sperm surface has been identified as choline phospholipids on the plasma membrane. This study was undertaken to determine whether BSP proteins modulate capacitation of bovine spermatozoa induced by heparin. Bovine epididymal spermatozoa were washed and incubated in buffer containing BSP proteins and then washed and incubated with heparin. The percentage of capacitated spermatozoa was determined under the microscope after the acrosome reaction has been initiated with the addition of lysophosphatidylcholine. The results demonstrated that epididymal sperm undergo the acrosome reaction only in the presence of BSP proteins. This effect was concentration-dependent and reached a maximum level of a 3-5-fold increase at 20-40 micrograms/ml BSP protein concentrations. In contrast, ribonuclease (purified from bovine seminal fluid) or seminal fluid proteins depleted of BSP proteins (by sequential absorption of BSP proteins on gelatin-Agarose and DEAE-Sephadex columns) showed no significant potentiating activity. The purified BSP proteins were more active than crude alcohol precipitates of bovine seminal plasma. These results indicate that BSP proteins are regulatory factors of capacitation.

Biochemical characterization of hamster oviductin as a sulphated zona pellucida-binding glycoprotein.

Oviductins are a family of glycoproteins, synthesized and released by oviductal secretory cells, which bind to the zona pellucida of the oocyte after ovulation. Hamster oviductin migrates as diffuse species of 160-350 kDa during SDS/PAGE under reducing as well as non-reducing conditions. In this report, we describe the one-step purification of hamster oviductin using either immuno- or lectin-affinity chromatography. Probing with specific lectins showed that the glycoprotein contains terminal alpha-D-GalNAc, and either terminal alpha-D-NeuAc or non-terminal beta-D-(GlcNAc)2 residues, but fails to react with concanavalin A and Ulex Europeus A-1 lectins which are specific for branched alpha-D-mannose and alpha-L-fucose moieties respectively. Intraovarian oocytes do not contain this glycoprotein and we demonstrate here that the immunoaffinity-purified oviductin readily binds to their zonae pellucidae in vitro, thus mimicking the in vivo phenomenon. Two major immunologically related forms of hamster oviductin (named alpha and beta) were characterized using one- and two-dimensional gel electrophoresis. The alpha-form (160-210 kDa) has an acidic pI of 3.5-4.5 and the beta-form (approx. 210-350 kDa) is localized at the cathodic site in the isoelectric focusing dimension; in between these two major forms lies a smear of minor-charge isomers. Peptide mapping of both major forms with papain and Staphylococcus aureus V8 protease yielded fragments of identical size. Moreover, the two forms share the same N-terminal sequence which display no significant homology with other reported proteins. Treatment with trifluoromethanesulphonic acid showed that a protein with the size and pI of the alpha-form can be generated from the beta-form. Both the alpha- and beta-forms are sulphated on O-linked oligosaccharide side chains but are not phosphorylated. Collectively, these results suggest that the hamster oviductin polymorphism observed in two-dimensional PAGE is a consequence of different glycosylation patterns and not the polypeptide chain itself. Hamster oviductin is mostly O-glycosylated and contains a few N-linked oligosaccharide side chains (approx. 10 kDa). We propose that hamster oviductin is a mucin-type glycoprotein which might act as a protective secretion influencing the first steps of the reproductive process necessary for the normal triggering of fertilization and early embryonic development.