Summary. In placental (eutherian) mammals, a numberof important events take place within the oviductincluding the pre-fertilisation maturation of gametes(including sperm storage), sperm-egg interactions, eggactivation and early embryonic development. Many ofthese events involve interactions of glycoconjugates;both on the surface of the gametes and with thesecretions of the oviductal epithelium and these havebest been studied in eutherian mammals. In marsupials,however, while the oviduct is known to produce theextracellular egg coat, the mucoid layer, that comes tosurround the zona pellucida, its role in the maturation ofgametes is only now being elucidated, particularly in theoocyte. This review emphasises what is known of thestructure and function of the oviduct and its secretions inmarsupials and briefly compares it with data fromeutherians. In particular, knowledge of oviductalglycoconjugates in the structure of the post-ovulatoryoocyte and its vestments around the time of fertilisationin Australian marsupials is outlined.

Key words: Oviduct, Marsupial, Oviductin,Glycoconjugates, Fertilisation

Introduction

In order for normal fertilisation and early embryonicdevelopment to occur, there must be an appropriateenvironment for these processes to occur. The specificmolecular interactions that take place in vivo betweensperm and eggs and how the microenvironment of theoviduct influences these events are poorly known, evenfor the most extensively studied eutherian species (for

recent reviews see Suarez, 2008; Töpfer-Petersen et al.,2008). Whilst the oviduct provides an optimalenvironment "in terms of temperature, pH, osmoticpressure, nutrients, oxygen tension and other factors"(Buhi et al., 1997; for other reviews see Hunter, 1988,2005; Leese, 1988), certain proteins and factors areactively synthesised and secreted which facilitate theevents leading up to, and following, fertilisation andearly embryonic development (Buhi et al., 1997, 2000;Buhi, 2002). In eutherians many of these events can bestudied in vitro, but in Australian marsupials evenrepeatable in vitro fertilisation has yet to be achieved foreven the most extensively studied species, nor is it clearat this stage whether sperm capacitation occurs. Whilethe effect of oviductal secretions on pre-fertilisationmaturation of sperm may be important (Sidhu et al.,1999; Mate et al., 2000), the possible role of secretedoviductal proteins on the post-ovulatory maturation ofthe oocyte has not been investigated. Using the brushtailpossum (Trichosurus vulpecula) and fat-tailed dunnart(Sminthopsis crassicaudata), as marsupial models(Breed, 1994, 1996; Chapman and Breed, 2004), wehave undertaken a histological and immunocytochemicalstudy on the secretions of the oviduct around the time ofovulation to determine their possible contribution to theovulated oocyte and the postovulatory egg coats(Menkhorst and Selwood, 2008). Here we review therole of the oviduct in gamete maturation of marsupials,and we propose an active involvement of oviductalglycoconjugates in sperm-egg interactions.

General morphology of the oviduct

Eutherians

The mammalian oviduct is classically divided intothree regions: the infundibulum, ampulla and isthmus.The infundibulum, and in particular its finger-like

Review

Glycoconjugates within the oviduct and their functionalsignificance with special reference to marsupialsJamie A. Chapman1, Meng Inn Chuah2 and William G. Breed3

1School of Medicine, University of Tasmania, Hobart, Tasmania, Australia, 2Menzies Research Institute, University of Tasmania,

Hobart, Tasmania, Australia and 3Discipline of Anatomical Sciences, School of Medical Sciences, University of Adelaide, Adelaide,

Australia

Histol Histopathol (2010) 25: 121-132

Offprint requests to: Associate Professor Meng Inn Chuah, MenziesResearch Institute, University of Tasmania, Private Bag 24, Hobart,Tasmania 7001 (Australia). e-mail: Inn.Chuah@utas.edu.au

http://www.hh.um.es

Histology andHistopathology

Cellular and Molecular Biology

processes the fimbriae, is the most distal region andgenerally lies in close proximity to the ovary andfunctions in the capture of ovulated oocytes. Theisthmus is the most proximal part which passes into theutero-tubal junction and is generally narrow. It is the sitefor sperm storage prior to ovulation in many mammalianspecies (for reviews see Suarez, 2001, 2008). Theampulla, which lies between the isthmus andinfundibulum, is generally the widest region of theoviduct, at least in eutherians, and is the site wherefertilisation takes place (Hunter, 1988).

Marsupials

In marsupials, however, some differences appear tobe evident in the gross anatomy of the oviduct comparedwith those of eutherian mammals (Fig. 1), even thoughRodger and Bedford (1982) in their studies of theVirginian opossum Didelphis virginiana did notdifferentiate between regions of the duct into separatemorphological regions, except for the fimbriatedinfundibulum. In dasyurid marsupials, as reflected in thefat-tailed dunnart, it appears that, in contrast toeutherians, the ampulla is the narrower part of theoviduct. Due to its proximity to the ovary, this region hasbeen referred to as the "ovarian segment" (Bedford andBreed, 1994). Each oviductal region displays its ownunique ultrastructural and histological features that relateto its biosynthetic and functional properties (Roberts and

Breed, 1996a; Buhi et al., 1997).

Functional histology of the oviduct

Eutherians

The epithelium of the mammalian oviduct consistsof two main cell types: the ciliated cells (CC) and non-ciliated secretory cells (NCC) (Fig. 2). The number,height and synthetic activity of these cells is dependantupon the circulating levels of steroid hormones, inparticular oestradiol (Hunter, 1988; Leese, 1988). Afterovariectomy the oviductal epithelium changes from anormally active simple (or pseudo-stratified) columnar

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Fig. 1. Gross anatomy of a dissected oviduct and uterine horn of abrushtail possum (Trichosurus vulpecula). Note that the isthmusappears wider than the ampulla. Most of the mesosalpinx has beenremoved. Scale bar: 1 cm.

Fig. 2. Scanning (A) and transmission (B) electron microscopy of theoviductal epithelium of a brushtail possum. Two cell types occur: ciliatedcells (CC) and non-ciliated secretory cells (NCC). Scale bars: A, 5 µm;B, 1 µm

epithelium containing the two cell types, to a single-layered cuboidal epithelium consisting of a cell type thatappear morphologically different from either the CC orNCC (for reviews see Hunter, 1988; Leese, 1988; Buhiet al., 2000).

In the rabbit, probably the most analogous eutherianto compare with marsupials due to the similaraccumulation of a thick mucoid postovulatory egg coat,considerable histological differences occur between thedifferent regions of the oviduct. For example, in therabbit infundibulum NCC are rare, but in the isthmusthey increase in abundance where they reach theirmaximum (Greenwald, 1958; Jansen and Bajpai, 1982).The NCC of the rabbit synthesise and store mucin inresponse to oestrogen, while progesterone causes themucin to be discharged where it comes to surround theovulated oocytes as a mucoid coat (Greenwald, 1958).

Marsupials

Marsupial ovulated oocytes also become surroundedby a mucoid coat that varies in thickness across thevarious species (see Selwood, 2000; Menkhorst andSelwood, 2008). Unlike in rabbits, however, thedeposition of this component of the postovulatory eggcoat around marsupial oocytes occurs very rapidly afterovulation (and entry of oocytes into the oviduct),whereas in the rabbit mucin is not released until 10-12hours after ovulation (Greenwald, 1958). These temporaldifferences may reflect the difference in transit of theeggs through the oviducts, which is very short inmarsupials (between 4-7 h: Menkhorst et al., 2007; up to15-20 h: Rodger and Bedford, 1982), but takes 3-4 daysin rabbits (Harper, 1982). Like eutherians, marsupialoviducts also display morphological and ultrastructuralchanges in response to change in levels of steroidhormones (Arnold and Shorey, 1985; Roberts and Breed,1996a).

A specialised and localised regional epithelium ofthe marsupial oviduct, at least in the dasyurids anddidelphids, lines the mucosal crypts of the isthmus.These small out-pockets are connected to the oviductallumen by narrow openings and are lined by cuboidal, or low-columnar, epithelium of characteristicmorphological appearance (Rodger and Bedford, 1982;Breed et al., 1989; Taggart and Temple-Smith, 1991;Roberts et al., 1994; Taggart, 1994; Taggart et al., 1998).In eutherians, similar isthmic crypts have been describedfor species of the shrews, where there appear to be deepisthmic crypts in the genus Crocidura, while ciliatedampullary crypts appear to exist in the genera Blarina,Cryptotis, Sorex, and Myosorex (Bedford, 1996; Bedfordet al., 1998, 2004). In marsupials, the isthmic crypts,where they occur, act as sperm storage sites with someof the sperm that pass through the uterotubal junctioncoming to reside deep within them from the time ofmating until ovulation which can be up to 15 days post-coitus (Hill and O’Donoghue, 1913; Woolley, 1966;Selwood, 1980).

Oviduct-specific glycoproteins

Biosynthesis of oviduct-specific glycoproteins

Eutherians

The oviduct is a highly synthetically active structurewith many proteins being synthesised and secreted by itsepithelium, while many components that enter the lumenare also derived from the serum as a transudate,including albumin and transferrin (Hunter, 1988, 2005).Proteins that are synthesised by the epithelial cellsinclude oviduct-specific glycoproteins (OGP), whichhave been found in a number of mammalian species andare collectively termed "oviductins" (Bleau and St-Jacques, 1989; Malette et al., 1995). These are a secretedform of mucins (Muc9) (Paquette et al., 1995; Hendrixet al., 2001). Many of the proteins that are secreted havebeen found to be stage- and/or region-specific, and arethus only secreted at a specific time of oestrous cycleand/or in a localised region of the oviduct (for reviewsee Gandolfi, 1995).

The ampulla appears to be the major source of OGPin many species including laboratory mice (Kapur andJohnson, 1986, 1988), rabbits (Hyde and Black, 1986),pigs (Buhi et al., 1989, 1990, 1992), sheep (Murray,1993) and cows (Wegner and Killian, 1992), whereas inthe hamster, the OGP is produced by the NCC along theentire length of the oviduct (Roux and Kan, 1995;Martoglio and Kan, 1996), but predominantly by cells inthe isthmus (Abe and Oikawa, 1990a,b, 1991). Theisthmus is also responsible for the secretion of an OGPin the baboon (Verhage et al., 1990).

The OGP of the pig (Buhi et al., 1996), cow (Sendaiet al., 1994), baboon (Donnelly et al., 1991), human(Arias et al., 1994), macaque (Verhage et al., 1997a),hamster (Suzuki et al., 1995), mouse (Sendai et al.,1995) and rabbit (Yong et al., 2002) have been partially,or completely, sequenced. Comparisons between speciesof the deduced amino acid sequences show a high levelof conservation amongst mammalian OGP. For example,the amino acid sequence of the rhesus macaque OGP is98% homologous to that of the olive baboon and 92%homologous to the human OGP (Verhage et al., 1997b).Less closely related species, however, display onlyslightly less homology with the protein sequence ofporcine OGP exhibiting a significant identity (65-78%)and similarity (78-87%) to OGPs from the cow, sheep,human, mouse and hamster (Buhi et al., 1996). Apartfrom eutherian mammals, OGP have also been found tobe present in the oviducts of amphibians (Lindsay et al.,1999) and chickens (Mann, 2008). This suggests thatthese glycoproteins are also likely to be present inmarsupials. A search of the recently published opossum(Monodelphis domestica) genome on the Ensembldatabase (http://www.ensembl.com) using the mouseOGP gene sequence (termed Ovgp1:EMSMUSG00000074340) as a template found aputative opossum OGP gene homologue

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(EMSMODG00000001338) that shared 57% homologyto the lab mouse sequence, around 60% identity to thehuman, chimpanzee, orangutan, and macaque sequences,and 63% identity to the platypus sequence. The putativeopossum OGP gene is located on chromosome 2 andmay transcribe a precursor peptide of 368 aa of around42kDa with one potential N-linked glycosylation site(position 272).

Apart from divergence in protein sequences ofmammalian OGPs, much interspecific variation in thestructure of these molecules may be due to differences intheir carbohydrate components. The number of potentialN-linked glycosylation sites in OGPs, as deduced fromamino acid sequence data, varies widely between speciesranging from only 1 in the rabbit, cow, baboon andmacaque to 8 in the golden hamster. Lectin-bindingexperiments have demonstrated that oligosaccharides ofthe OGP of the mouse, cow and hamster are WGA-reactive providing evidence of terminal α-D-NeuNAc(sialic acid) and/or non-terminal ß-D-(GlcNAc)2residues (Malette and Bleau, 1993), whereas the cowOGP contains Gal(ß1-3)GalNAc residues, thusindicating that the 1 potential N-linked glycosylation sitededuced from its amino acid sequence is indeed utilised(Wegner and Killian, 1992). Nevertheless, potential N-glycosylation sites do not always reflect the N-linkedglycosylation of the mature protein. As indicated above,the hamster OGP has 8 potential N-linked glycosylationsites but in vitro metabolic labelling studies suggest thepresence of only one or two N-glycan chains of around10kDa (Malette and Bleau, 1993). The mature post-translated hamster OGP, however, appears to bedifferentially glycosylated depending upon the stage of

the oestrous cycle and the utilisation of potential N-linked glycosylation sites may vary (McBride et al.,2004).

Marsupials

Most of the work on oviductal secretions ofmarsupials have been derived from investigations intothe origin and composition of the postovulatory eggcoats (Roberts et al., 1994, 1997; Roberts and Breed,1996a,b; Casey et al., 2002; Menkhorst and Selwood,2008). Histologically, it has been determined that in thebrushtail possum and fat-tailed dunnart, the mucoidmaterial secreted by the epithelium of the oviduct is asulphated and acidic glycoprotein as demonstrated bypositive staining with Alcian blue, pH 1.0 - pH 2.5 andPeriodic Acid-Schiff's Reagent (see Hughes, 1977).Histochemistry using fluorescently conjugated lectinshas shown that there is a similarity in the binding oflectins between the ampullary and isthmic oviductepithelial cells of the possum, with the NCC of boththese regions reacting positively to seven out of the ninelectins used (see Table 1). The greatest intensity ofbinding in the NCC was found to be for ß-Galactose (ß-Gal) and N-acetylglucosamine (GlcNAc) and/or sialicacid indicating that these glycoconjugates are majorconstituents of the oviductal secretions in this species.

The largest regional differences in lectin stainingwas with soybean agglutinin (SBA), which is specificfor α-D-GalNAc/α-D-Gal residues. Fluorescence wasfound to occur in the secretory cells of the ampulla butnot elsewhere. This difference in binding was quantifiedusing lectin immunocytochemistry with significantlymore gold labelling with SBA on the secretory granulesof the ampulla than in the isthmus (Fig. 3; Table 2).Greater density of gold labelling of the secretorygranules within the ampulla was also found for Pisumsativum agglutinin (PSA) suggesting differential

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Table 1. Intensity of lectin binding of non-ciliated secretory (NCC) andciliated (CC) epithelial cells from the oviduct of the peri-ovulatorybrushtail possum.

Ampulla Isthmus

Lectin and sugar specificity NCC CC NCC CC

PNA: ßGal-(1-3)-GalNAc ++ +++ ++ +++ECA: ßGal-(1-4)-GlcNAc + ++ + ++Con A: α-D-Man, α-D-Glc + + + +LCA: α-D-Man, α-D-Glc + + + +RCA-I: ß-D-Gal +++ ++ +++ ++WGA: [ß-(1-4)-D-GlcNAc]2, NeuNAc +++ + +++ +SNA: α-NeuNAc-(2-6)-Gal/GalNAc ++ - ++ -SBA: α-D-GalNAc, α-D-Gal ++ - - -LTA: α-L-Fuc - ++ - ++

Fluorescence was qualitatively scored as: -, negative; +, mild; ++,strong; +++, intense. PNA: peanut agglutinin; ECA: Erythrina cristagalliagglutinin; Con A: Concanavalia ensiformis agglutinin; LCA: Lensculinaris agglutinin; RCA-I: Ricinus communis -I agglutinin; WGA: wheatgerm agglutinin; SNA: Sambucus nigra agglutinin; SBA: soybeanagglutinin; LTA: Lotus tetragonolobus agglutinin. Fuc: fucose; Gal:galactose; GalNAc: N-acetylgalactosamine; Glc: glucose; GlcNAc: N-acetylglucosamine; Man: mannose; NeuNAc: N-acetylneuraminic acid(sialic acid). From Wu et al., 1988.

Table 2. Gold labelling density of secretory granules of the NCC in theampulla and isthmus of the brushtail possum of the peri-ovulatoryoviduct.

Lectin and sugar specificity Ampulla Isthmus

PSA: α-D-Man, α-D-Glc 21.8±1.11 9.9±0.59a

RCA-I: ß-D-Gal 51.7±3.12 55.0±3.93WGA: [ß-(1-4)-D-GlcNAc]2, NeuNAc 23.4±2.69 39.6±3.47b

SBA: α-D-GalNAc, α-D-Gal 95.3±8.03 0.7±0.22a

Values are shown as the number of gold particles per secretory granule;mean values ± SEM. PSA: Pisum sativum agglutinin; RCA-I: Ricinuscommunis-I agglutinin; WGA: wheat germ agglutinin; SBA: soybeanagglutinin. a: The labelling densities of secretory granules using theselectins was significantly less in the isthmic NCC than in the ampullaryNCC (p<0.001); b: The labelling densities of secretory granules usingthis lectin was significantly less in the ampullary NCC than in the isthmicNCC (p<0.001); Gal: galactose; GalNAc: N-acetylgalactosamine; Glc:glucose; GlcNAc: N-acetylglucosamine; Man: mannose; NeuNAc: N-acetylneuraminic acid (sialic acid). From Wu et al., 1988.

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Fig. 3. Lectin histochemistry (A, B) and immunocytochemistry (C, D) of the ampullary (A, C) and isthmic (B, D) oviductal epithelium from the brushtailpossum stained with soybean agglutinin (SBA). Note that the apical regions of the secretory cells of the ampullary epithelium (E) are labelled with FITC-SBA (white arrows, A), while the secretory granules (black arrow, C) of the ampullary secretory cells are heavily labelled with gold particles. Thesecretory cells (B) and secretory granules (black arrow, D) of the isthmus remain unstained. Some fluorescence of erythrocytes can be seen within thelamina propria (LP) (A, B); lumen (L). Scale bars: A, B, 45 µm; C, D, 0.5 µm.

localisation of α-D-Mannose residues, whilesignificantly greater labelling of GlcNAc and/or sialicacid was found with wheat germ agglutinin (WGA) inthe isthmus compared to ampullary secretory granules(p<0.05). Only ß-Gal had a similar labelling intensity inthe secretory granules of the ampulla and isthmus. As ineutherians (for review see Abe, 1996), these differencesprobably reflect the regional differences in function ofthe duct epithelium.

Functions of oviduct-specific glycoproteins

Eutherians

Formation of a sperm reservoir within the oviduct isthought to be brought about by binding of spermatozoato the glycoconjugates on the surface of the isthmicepithelial cells (Demott et al., 1995; Lefebvre et al.,1997; Suarez, 1998, 2001, 2002, 2008; Suarez et al.,1998; Green et al., 2001; Talevi and Gualtieri, 2001;Wagner et al., 2002; Hunter, 2008). This binding appearsto reduce the incidence of polyspermy by selectivelyreleasing limited numbers of sperm around the time ofovulation (Hunter, 1973). Binding to the oviductepithelium may also act to maintain sperm viability(Pollard et al., 1991; Smith and Nothnick, 1997), as wellas forming a selective barrier to physiologically'abnormal' sperm (e.g. those sperm that have undergonepremature capacitation and acrosome reaction) (Fazeli etal., 1999). Sperm, in turn, influence the biosyntheticactivities of the oviduct, with the finding that oviductepithelial cells modify their gene and protein syntheticactivity following interaction with sperm (Fazeli et al.,2004; Georgiou et al., 2005).

OGP also facilitate pre-fertilisation maturation ofovulated oocytes. For example, oviductal exposure iscommonly associated with an increase in penetrability ofzonae pellucidae and fertilisability of oocytes (Boatmanet al., 1994; King et al., 1994; Kito and Bavister, 1996),although in the laboratory mouse it does not appear to beessential for in vivo fertilisation (Araki et al., 2003).OGP have been localized to the membranes and/orperivitelline spaces of oocytes, eggs, embryos and theirvestments in a number of species (for reviews see Bleauand St.-Jacques, 1989; Malette et al., 1995; Buhi et al.,2000). This binding of OGP to the gametes and embryosis thought to be facilitated through an OGP-bindingprotein that is similar in structure to non-muscle myosinheavy chain (Kadam et al., 2006).

In eutherians, carbohydrates play an important rolein the events leading up to sperm-egg interactions atfertilisation (for reviews see Benoff, 1997; Shalgi andRaz, 1997; Tulsiani et al., 1997; Dell et al., 1999;Topfer-Petersen, 1999; Topfer-Petersen et al., 2008;Jiménez-Movilla et al., 2009). Once sperm migrate tothe site of fertilisation, initial binding of sperm to thezona pellucida (ZP) is generally thought to take place byway of the ZP glycoconjugates, while the high degree ofhomology of the ZP genes and proteins between

eutherian species means that it is likely theoligosaccharide components of the glycoproteins may beresponsible for a high level of specificity of thisinteraction, if this occurs (Florman and Wassarman,1985; Wassarman, 1988, 1990, 1995; Bleil andWassarman, 1988; Thaler and Cardullo, 1996; Ozgur etal., 1998; Takasaki et al., 1999; Litscher and Wassarman,2007).

Marsupials

In marsupials, the role of the oviduct secretions infacilitating gamete maturation and interaction is notknown, although it is clear that the oviduct is responsiblefor the production of at least one of the postovulatoryegg coats, that of the mucoid layer (for reviews seeHughes, 1977; Breed, 1996; Selwood, 2000; Menkhorstand Selwood, 2008). While Selwod (2000) hassuggested that the early deposition of mucoid mightenhance sperm-ZP binding, others have inferred theopposite with suggestions that the mucoid layer reduceschances of polyspermy due to its relatively rapidacquisition and the observation of sperm trapped withinit as the ovulated oocyte passes along the oviduct(Rodger and Bedford, 1982; Selwood, 1982; Breed,1994).

In the brushtail possum proteins from the oviductepithelium bind to the cell membrane of the sperm, andappear to maintain their viability as well as increasingtheir ability to undergo capacitation, binding andpenetration of the ZP (Sidhu et al., 1998, 1999; Mate etal., 2000). In addition, the oviduct epithelium may giverise to the material that accumulates within theperivitelline space of the ovulated oocyte (Chapman andBreed, 2004). Previously described as a “cortical granuleenvelope” that accumulated following oocyte activation(Dandekar et al., 1995), the ruthenium red stainedmaterial within the perivitelline space (known as theperivitelline space matrix) has been shown to arise, notfrom cortical granule exocytosis, but following transit inthe oviduct (Chapman and Breed, 2004). This issupported by comparing lectin immunocytochemistry ofpreovulatory and postovulatory egg coats of brushtailpossum oocytes (see Fig. 4) where it was found thatlectin gold labelling with three of the four lectins used(those of PSA, SBA and Ricinus communis-I agglutinin[RCA-I]) of the ZP around ovarian oocytes was muchlower than that of ovulated oocytes (Fig. 5). Only WGA,specific for GlcNAc and/or sialic acid, did not show astatistical significant difference in labelling. Theseresults demonstrate that the mucoid coat surrounding theovulated oocytes has a greater density of labelling forGlcNAc and ß-Gal, and less for α-D-GalNAc/α-D-Galand α-D-Mannose. Labelling was also evident on themicrovilli and within the perivitelline space of theovulated, but not ovarian, oocytes.

One possibility for the major increase in labelling ofglycoconjugates within the ZP following ovulation isthat some kind of biochemical change has taken place

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which involves a rearrangement of glycoconjugatescoinciding with the morphological change of the ZPfollowing ovulation (Breed et al., 2002). Alternatively,some kind of oocyte/cumulus-specific modification mayhave occurred that adds to, or exposes, previouslymasked glycoconjugates, or glycoconjugates that hadpreviously been inaccessible to the lectins. Reactivity tolectins has been shown to increase in ovarian ZPfollowing removal of masking agents such as sialic acidwith the use of the enzyme neuraminidase, and O-acetylgroups on sialic acids, through saponification withpotassium hydroxide (Chapman et al., 2000).Neuraminidase is present, and appears to play a role infertility, within the oviducts of non-mammalian speciessuch as toads (De Martinez and Olavarria, 1973; DeMartinez et al., 1975; Vitaioli et al., 1990). It has alsobeen suggested that in the human, the cumulus-coronalcells may secrete a factor that increases the penetrabilityof the ZP (Tesarik et al., 1988), while removal of sialicacid from the surface of sperm and ZP increases sperm-ZP binding (Lassalle and Testart, 1994; Banjeree andChowdhury, 1997; Ozgur et al., 1998). Whether a similarmodification occurs to the ZP following ovulation in thebrushtail possum is unknown.

Another possibility is for an increase inglycoconjugate content of the possum ZP followingovulation as a result of an incorporation of OGP asdemonstrated in a number of eutherian species, includingthe hamster (Kan et al., 1988, 1990), pig (Buhi et al.,1993), sheep (Gandolfi et al., 1991), cow (Staros andKillian, 1998) and baboon (Boice et al., 1990). In thehamster, early investigations found that the oviductcontributed a glycoprotein, termed hamster oviductin 1(Hm OV-1), to the post-ovulatory ZP (Kan et al., 1988,

1989), and that its contribution resulted in the addition ofGalNAc residues, not previously identified in theovarian ZP but localised to the secretory granules of theampulla (Kan et al., 1990). Similarly in the possum, α-D-GalNAc/α-D-Gal residues, which are labelled at verylow levels in the ZP surrounding ovarian oocytes, werefound to significantly increase in labelling followingovulation. These glycoconjugates, similar to those in thehamster, were localised to the secretory granules of theampulla, but not the isthmus, and only appeared to beminor contributors to the mucoid coat. These findingssuggest that α-D-GalNAc/α-D-Gal residues, originating

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Fig. 4. Density of gold labelling of the zona pellucida of the ovarian (Ov)and ovulated (Tubal) oocytes and mucoid coats (Mucoid) of the brushtailpossum following incubation in four different lectins. The labellingdensities are shown as the mean number of gold particles /µm2 ± SEM.Asterisks (*) represent significant differences (p<0.05) in gold labeling ofthe same lectin from the previous stage of development. PSA: Pisumsativum agglutinin; RCA-I: Ricinus communis-I agglutinin; WGA: wheatgerm agglutinin; SBA: soybean agglutinin.

Fig. 5. Zona pellucida of the peri-ovulatory ovarian (A) and ovulated (B)oocyte of the brushtail possum incubated with biotinylated-Pisumsativum (garden pea) lectin (biotin-PSA) and labelled with 10nm gold-conjugated goat anti-biotin antibody. Note sparse gold labelling of thezona pellucida (ZP) surrounding the peri-ovulatory oocyte (Oo) (A) butvery dense labelling of the zona surrounding the ovulated oocyte (B)that may be localised to the zona pellucida filaments (arrow). Labellingis also evident on the microvilli (Mv) of the ovulated oocyte (B); coronaradiata cell (Cr); perivitelline space (PVS). PSA binding is specific for α-D-Mannose. Scale bar: 1 µm.

from the ampullary region of the oviduct and transferredto the ZP after ovulation, may play a role in sperm-zonapellucida interactions in this species.

Likewise, labelling with PSA for α-D-Mannose, alsoshowed similar modifications (Fig. 5). Taking the lectinimmuno-gold cytochemistry results of the oviduct, ZPand mucoid coat together, a hypothetical summary of theeffect of duct epithelium secretory activity on theovulated oocytes is presented (Fig. 6). The recentlyovulated oocyte first enters the oviduct via theinfundibulum and reaches the ampulla where fertilisationtakes place (Selwood, 1982; Bedford and Breed, 1994;Jungnickel et al., 2000). This is also the site for theinitial deposition of the mucoid coat, and any influenceof the oviduct secretions on gamete interaction, eitherpositive or negative, is likely to occur within this region.Within the ampulla, therefore, α-D-GalNAc (and/or α-D-Gal) and α-D-Man appear to have the greatest effecton the post-ovulatory ZP (Fig. 6).

Furthermore, the localisation of lectin labelling tothe material within the perivitelline space and on themicrovilli of the oolemma, in conjunction with thefindings of Chapman and Breed (2004), provide furtherevidence for the incorporation of oviductal glycoproteinsinto these areas. The exact role of these incorporatedglycoproteins is unknown, but, the inability to achievesperm-oolemma binding in vitro following pre-incubation of sperm, but not follicular oocytes, withoviduct-conditioned media suggests that they may play arole in sperm-egg fusion (Mate et al., 2000). This isfurther supported by recent findings thatintracytoplasmic sperm injection can induce fertilisationand early embryonic development (Magarey and Mate,2003; Richings et al., 2004) suggesting perhaps that theinability to achieve successful in vitro fertilisation maybe due to an absence of an oviductal influence at thelevel of the oolemma.

Concluding remarks

The present brief review provides some evidencethat secretions of the oviduct affect gamete maturationand interaction in Australian marsupials. The secretionsof the oviduct not only appear to be necessary forsuccessful prefertilisation maturation of spermatozoaand formation of the mucoid coat around the ovulatedoocyte, but also for the composition of the post-ovulatory ZP to facilitate sperm binding. In addition,material also appears to accumulate within theperivitelline space. The regional variation ofglycosylation of the oviductal secretions in marsupialssuggests that there is indeed a complex interactionbetween the oviduct and gametes. While it has beenpreviously postulated that some variation may exist forthe secretion of the mucoid layer between the ampullaand isthmus (Roberts and Breed, 1996a), such variationin terms of the specific glycosylation patterns of theoviductal glycoproteins and their possible contribution tothe oocyte and the post-ovulatory ZP in marsupials issuggested by these observations.

References

Abe H. (1996). The mammalian oviductal epithelium: regional variationsin cytological and functional aspects of the oviductal secretory cells.Histol. Histopathol. 11, 743-768.

Abe H. and Oikawa T. (1990a). Study of the differentiation of secretorycells in the golden hamster oviductal epithelium by use of amonoclonal antibody. J. Exp. Zool. 254, 97-106.

Abe H. and Oikawa T. (1990b). Ultrastructural evidence for anassociation between an oviductal glycoprotein and the zonapellucida of the golden hamster egg. J. Exp. Zool. 256, 210-221.

Abe H. and Oikawa T. (1991). Immunocytochemical localization of anoviductal zona pellucida glycoprotein in the oviductal epithelium ofthe golden hamster. Anat. Rec. 229, 305-314.

128

Oviducts in marsupials

Fig. 6. Diagrammatic representation of thepossible glycoconjugate contribution by theampullary and isthmic epithelium of the oviductto the post-ovulatory oocyte of the brushtailpossum. Note: The larger the arrows, thegreater the contribution.

Araki Y., Nohara M., Yoshida-Komiya H., Kuramochi T., Ito M., HoshiH., Shinkai Y. and Sendai Y. (2003). Effect of a null mutation of theoviduct-specific glycoprotein gene on mouse fertilization. Biochem.J. 374, 551-557.

Arias E.B., Verhage H.G. and Jaffe R.C. (1994). Complementarydeoxyribonucleic acid cloning and molecular characterization of anestrogen-dependent human oviductal glycoprotein. Biol. Reprod. 51,685-694.

Arnold R. and Shorey C.D. (1985). Structure of the oviductal epitheliumof the brush-tailed possum (Trichosurus vulpecula). J. Reprod. Fert.73, 9-19.

Banerjee M. and Chowdhury M. (1997). Localization of a 25kDa humansperm surface protein: its role in in-vitro human sperm capacitation.Mol. Hum. Reprod. 3, 109-114.

Bedford J.M. (1996). What marsupial gametes disclose about gametefunction in eutherian mammals. Reprod. Fertil. Dev. 8, 569-580.

Bedford J.M. and Breed W.G. (1994). Regulated storage andsubsequent transformation of spermatozoa in the fallopian tubes ofan Australian marsupial, Sminthopsis crassicaudata. Biol. Reprod.50, 845-854.

Bedford J.M., Bernard R.T.F. and Baxter R.M. (1998). The 'hybrid'character of the gametes and reproductive tracts of the Africanshrew, Myosorex varius, supports its classif ication in theCrocidosoricinae. J. Reprod. Fert. 112, 165-173.

Bedford J.M., Mock O.B. and Goodman S.M. (2004). Novelties ofconception in insectivorous mammals (Lipotyphla), particularlyshrews. Biol. Rev. 79, 891-909.

Benoff S. (1997). Carbohydrates and fertilization: an overview. Mol.Hum. Reprod. 3, 599-637.

Bleau G. and St-Jacques S. (1989). Transfer of oviductal proteins to thezona pellucida. In: The mammalian egg coat: Structure and function.Dietl J. (ed) Springer-Verlag. Berlin. pp 99-110.

Bleil J.D. and Wassarman P.M. (1988). Galactose at the nonreducingterminus of O-linked oligosaccharides of mouse egg zona pellucidaglycoprotein ZP3 is essential for the glycoprotein's sperm receptoractivity. Proc. Natl. Acad. Sci. USA 85, 6778-6782.

Boatman D.E., Felson S.E. and Kimura J. (1994). Changes inmorphology, sperm penetration and fertilization of ovulated hamstereggs induced by oviductal exposure. Hum. Reprod. 9, 519-526.

Boice M.L., McCarthy T.J., Mavrogianis P.A., Fazleabas A.T. andVerhage H.G. (1990). Localization of oviductal glycoproteins withinthe zona pellucida and perivitelline space of ovulated ova and earlyembryos in baboons (Papio anubis). Biol. Reprod. 43, 340-346.

Breed W.G. (1994). How does sperm meet egg? - In a marsupial.Reprod. Fertil. Dev. 6, 485-506.

Breed W.G. (1996). Egg maturation and fertilization in marsupials.Reprod. Fertil. Dev. 8, 617-643.

Breed W.G., Leigh C.M. and Bennett J.H. (1989). Sperm morphologyand storage in the female reproductive tract of the fat-tailed dunnart,Sminthopsis crassicaudata (Marsupialia: Dasyuridae). Gamete Res.23, 61-75.

Breed W.G., Hope R.M., Wiebkin O.W., Spargo S. and Chapman, J.A.(2002). Structural organisation and evolution of the marsupial zonapellucida. Reproduction 123, 13-21.

Buhi W.C. (2002). Characterization and biological roles of oviduct-specific, oestrogen-dependent glycoprotein. Reproduction 123, 355-362.

Buhi W.C., Alvarez I.M. and Kouba A.J. (1997). Oviductal regulation offertilization and early embryonic development. J. Reprod. Fert.

Suppl. 52, 285-300.Buhi W.C., Alvarez I.M. and Kouba A.J. (2000). Secreted proteins of the

oviduct. Cells Tissues Organs 166, 165-179.Buhi W.C., Vallet J.L. and Bazer F.W. (1989). De novo synthesis and

release of polypeptides from cyclic and early pregnant porcineoviductal tissue in explant culture. J. Exp. Zool. 252, 79-88.

Buhi W.C., Alvarez I.M., Sudhipong V. and Dones-Smith M.M. (1990).Identification and characterization of de novo-synthesized porcineoviductal secretory proteins. Biol. Reprod. 43, 929-938.

Buhi W.C., Ashworth C.J., Bazer F.W. and Alvarez I.M. (1992). In vitrosynthesis of oviductal secretory proteins by estrogen-treatedovariectomized gilts. J. Exp. Zool. 262, 426-435.

Buhi W.C., Alvarez I.M., Choi I., Cleaver B.D. and Simmen F.A. (1996).Molecular cloning and characterization of an estrogen-dependentporcine oviductal secretory glycoprotein. Biol. Reprod. 55, 1305-1314.

Buhi W.C., O'Brien B., Alvarez I.M, Erdos G. and Dubois D. (1993).Immunogold localization of porcine oviductal secretory proteinswithin the zona pellucida, perivitelline space, and plasma membraneof oviductal and uterine oocytes and early embryos. Biol. Reprod.48, 1274-1283.

Casey N.P., Martinus R. and Selwood L. (2002). Outer egg coats of themarsupial conceptus: secretion and protein composition. Mol.Reprod. Dev. 62, 181-194.

Chapman J.A. and Breed W.G. (2004). The egg coats of the brushtailpossum Trichosurus vulpecula following ovulation and artificialactivation, with particular reference to the origin of the perivitellinespace matrix. In: The biology of australian Possums and Gliders.Goldingay R.L. and Jackson S.M. (eds). Surrey Beatty and Sons.Chipping Norton. pp 426-433.

Chapman J.A., Wiebkin O.W. and Breed W.G. (2000). Interspecificvariation of zona pellucida glycoconjugates in several species ofmarsupial. J. Reprod. Fertil. 119, 111-120.

Dandekar P., Mate K.E. and Talbot P. (1995). Perivitelline space ofmarsupial oocytes: extracellular matrix of the unfertilized oocyte andformation of a cortical granule envelope following the corticalreaction. Mol. Reprod. Dev. 41, 368-373.

Dell A., Morris H.R., Easton R.L., Patankar M. and Clark G.F. (1999).The glycobiology of gametes and fertilisation. Biochim. Biophys.Acta 1473, 196-205.

De Martinez N.R. and Olavarria J.M. (1973). Metabolism of sialic acid intoad oviduct. Biochim. Biophys. Acta 320, 301-310.

De Martinez N.R., Mendez B.A. and Olavarria J.M. (1975).Topographical distribution of sialic acids and related enzymes intoad oviduct. Comp. Biochem. Phys. B 50, 603-607.

Demott R.P., Lefebvre R. and Suarez S.S. (1995). Carbohydratesmediate the adherence of hamster sperm to oviductal epithelium.Biol. Reprod. 52, 1395-1403.

Donnelly K.M., Fazleabas A.T., Verhage H.G., Mavrogianis P.A. andJaffe R.C. (1991). Cloning of a recombinant complementary DNA toa baboon (Papio anubis) estradiol-dependent oviduct-specificglycoprotein. Mol. Endocrinol. 5, 356-364.

Fazeli A., Duncan A.E., Watson P.F. and Holt W.V. (1999). Sperm-oviduct interaction: induction of capacitation and preferential bindingof uncapacitated spermatozoa to oviductal epithelial cells in porcinespecies. Biol. Reprod. 60, 879-886.

Fazeli A., Affara N.A., Hubank M. and Holt W.V. (2004). Sperm-inducedmodification of the oviductal gene expression profile after naturalinsemination in mice. Biol. Reprod. 71, 60-65.

129

Oviducts in marsupials

Florman H.M. and Wassarman P.M. (1985). O-linked oligosaccharidesof mouse egg ZP3 account for its sperm receptor activity. Cell 41,313-324.

Gandolfi F. (1995). Functions of proteins secreted by oviduct epithelialcells. Microsc. Res. Techniq. 32, 1-12.

Gandolfi F., Modina S., Brevini T.A.L., Galli C., Moor R.M. and Lauria A.(1991). Oviduct ampullary epithelium contributes a glycoprotein tothe zona pellucida, perivitelline space and blastomeres membraneof sheep embryos. Eur. J. Basic Appl. Histochem. 35, 383-392.

Georgiou A.S., Sostaric E., Wong C.H., Snijders A.P.L., Wright P.C.,Moore H.D. and Fazeli A. (2005). Gametes alter the oviductalsecretory proteome. Mol. Cell. Proteomics 4, 1785-1796.

Green C.E., Bredl J., Holt W.V., Watson P.F. and Fazeli A. (2001).Carbohydrate mediation of boar sperm binding to oviductal epithelialcells in vitro. Reproduction 122, 305-315.

Greenwald G.S. (1958). Endocrine regulation of the secretion of mucinin the tubal epithelium of the rabbit. Anat. Rec. 130, 477-495.

Harper M.J.K. (1982). Sperm and egg transport. In: Reproduction inmammals. I. Germ cells and fertilization. Austin C.R. and Short R.V.(eds). Cambridge University Press. Cambridge. pp 102-127.

Hendrix E., Hewetson A., Mansharamani M. and Chilton B.S. (2001).Oviductin (Muc9) is expressed in rabbit endocervix. Endocrinology142, 2151-2154.

Hill J.P. and O’Donoghue C.H. (1913). The reproductive cycle in themarsupial Dasyurus viverrinus. Quart. J. Microsc. Sci. 59, 133-174.

Hughes R.L. (1977). Egg membranes and ovarian function duringpregnancy in monotremes and marsupials. In: Reproduction andevolution: Proceedings of the 4th Symposium on ComparativeBiology of Reproduction. Calaby J.H. and Tyndale-Biscoe C.H.(eds). Australian Academy of Science. Canberra. pp 281-291.

Hunter R.H.F. (1973). Polyspermic fertilization in pigs after tubaldeposition of excessive numbers of spermatozoa. J. Exp. Zool. 183,57-64.

Hunter R.H.F. (1988). The fallopian tubes: Their role in fertility andinfertility. Springer-Verlag. Berlin.

Hunter R.H.F. (2005). The Fallopian tubes in domestic mammals: howvital is their physiological activity? Reprod. Nutr. Dev. 45, 281-290.

Hunter R.H.F. (2008). Sperm release from oviduct epithelial binding iscontrolled hormonally by peri-ovulatory Graafian follicles. Mol.Reprod. Dev. 75, 167-174.

Hyde B.A. and Black D.L. (1986). Synthesis and secretion of sulphatedglycoproteins by rabbit oviduct explants in vitro. J. Reprod. Fert. 78,83-91.

Jansen R.P.S. and Bajpai V.K. (1982). Oviduct acid mucusglycoproteins in the estrous rabbit: ultrastructure and histochemistry.Biol. Reprod. 26, 155-168.

Jiménez-Movilla M., Martínez-Alonso E., Castells M.T., Izquierdo-RicoM., Saavedra M.D., Gutiérrez-Gallego R., Fayrer-Hosken R.,Ballesta J. and Avilés M. (2009). Cytochemical and biochemicalevidences for a complex tridimensional structure of the hamsterzona pellucida. Histol. Histopathol. 24, 599-609.

Jungnickel M.K., Molinia F.C., Harman A.J. and Rodger J.C. (2000).Sperm transport in the female reproductive tract of the brushtailpossum, Trichosurus vulpecula, following superovulation andartificial insemination. Anim. Reprod. Sci. 59, 213-228.

Kadam K.M., D’Souza S.J.D., Bandivdekar A.H. and Natraj U. (2006).Identification and characterization of oviductal glycoprotein-bindingprotein partner on gametes: epitopic similarity to non-muscle myosinIIA, MYH9. Mol. Hum. Reprod. 12, 275-282.

Kan F.W.K., St-Jacques S. and Bleau G. (1988). Immunoelectronmicroscopic localization of an oviductal antigen in hamster zonapellucida by use of a monoclonal antibody. J. Histochem. Cytochem.36, 1441-1447.

Kan F.W.K., St-Jacques S. and Bleau G. (1989). Immunocytochemicalevidence for the transfer of an oviductal antigen to the zonapellucida of hamster ova after ovulation. Biol. Reprod. 40, 585-598.

Kan F.W.K., Roux E., St-Jacques S. and Bleau G. (1990).Demonstration by lectin-gold cytochemistry of transfer ofglycoconjugates of oviductal origin to the zona pellucida of oocytesafter ovulation in hamsters. Anat. Rec. 226, 37-47.

Kapur R.P. and Johnson L.V. (1986). Selective sequestration of anoviductal fluid glycoprotein in the perivitelline space of mouseoocytes and embryos. J. Exp. Zool. 238, 249-260.

Kapur R.P. and Johnson L.V. (1988). Ultrastructural evidence thatspecialized regions of the murine oviduct contribute a glycoprotein tothe extracellular matrix of mouse oocytes. Anat. Rec. 221, 720-729.

King R.S., Anderson S.H. and Killian G.J. (1994). Effect of bovineoviductal estrus-associated protein on the ability of sperm tocapacitate and fertilize oocytes. J. Androl. 15, 468-478.

Kito S. and Bavister B.D. (1996). Kinetics of sperm penetration andfertilization in vitro in hamster follicular and oviductal ova. J. Exp.Zool. 274, 373-383.

Lassalle B. and Testart J. (1994). Human zona pellucida recognitionassociated with removal of sialic acid from human sperm surface. J.Reprod. Fert. 101, 703-711.

Leese H.J. (1988). The formation and function of oviduct fluid. J.Reprod. Fert. 82, 843-856.

Lefebvre R., Lo M.C. and Suarez S.S. (1997). Bovine sperm binding tooviductal epithelium involves fucose recognition. Biol. Reprod. 56,1198-1204.

Lindsay L.L., Wieduwilt M.J. and Hedrick J.L. (1999). Oviductin, theXenopus laevis oviductal protease that processes egg envelopeglycoprotein gp43, increases sperm binding to envelopes, and istranslated as part of an unusual mosaic protein composed of twoprotease and several CUB domains. Biol. Reprod. 60, 989-995.

Litscher E.S. and Wassarman P.M. (2007). Egg extracellular coatproteins: from fish to mammals. Histol. Histopathol. 22, 337-347.

Magarey G.M. and Mate K.E. (2003). Ferti l ization fol lowingintracytoplasmic sperm injection of in vivo and in vitro maturedoocytes from an Australian marsupial, the tammar wallaby(Macropus eugenii). Zygote 11, 339-346.

Malette B. and Bleau G. (1993). Biochemical characterization ofhamster oviductin as a sulphated zona pellucida-bindingglycoprotein. Biochem. J. 295, 437-445.

Malette B., Paquette Y., Merlen Y. and Bleau G. (1995). Oviductinspossess chitinase- and mucin-like domains: a lead in the search forthe biological function of these oviduct-specific ZP-associatingglycoproteins. Mol. Reprod. Dev. 41, 384-397.

Mann K. (2008). Proteomic analysis of the chicken egg vitellinemembrane. Proteomics 8, 2322-2332.

Martoglio A-M. and Kan F.W.K. (1996). Immunohistochemicallocalization of oviductin in the endometrial lining of the goldenhamster (Mesocricetus auratus) during the estrous cycle and earlygestation. Histochem. J. 28, 449-459.

Mate K.E., Sidhu K.S., Molinia F.C., Glazier A.M. and Rodger J.C.(2000). Sperm binding and penetration of the zona pellucida in vitrobut not sperm-egg fusion in an Australian marsupial, the brushtailpossum (Trichosurus vulpecula). Zygote 8, 189-196.

130

Oviducts in marsupials

McBride D.S., Boisvert C., Bleau G. and Kan F.W.K. (2004). Evidencefor the regulation of glycosylation of golden hamster (Mesocricetusauratus) oviductin during the estrous cycle. Biol. Reprod. 70, 198-203.

Menkhorst E. and Selwood L. (2008). Vertebrate extracellularpreovulatory and postovulatory egg coats. Biol. Reprod. 79, 790-797.

Menkhorst E., Ezard N. and Selwood L. (2007). Induction of ovulationand natural oestrous cycling in the stripe-faced dunnart, Sminthopsismacroura. Reproduction 133, 495-502.

Murray M.K. (1993). An estrogen-dependent glycoprotein is synthesizedand released from the oviduct in a temporal- and region-specificmanner during early pregnancy in the ewe. Biol. Reprod. 48, 446-453.

Ozgur K., Patankar M.S., Oehninger S. and Clark G.F. (1998). Directevidence for the involvement of carbohydrate sequences in humansperm-zona pellucida binding. Mol. Hum. Reprod. 4, 318-324.

Paquette Y., Merlen Y., Malette B. and Bleau G. (1995). Allelicpolymorphism in the hamster oviductin gene is due to a variablenumber of mucin-like tandem repeats. Mol. Reprod. Dev. 42, 388-396.

Pollard J.W., Plante C., King W.A., Hansen P.J., Betteridge K.J. andSuarez S.S. (1991). Fertilizing capacity of bovine sperm may bemaintained by binding to oviductal epithelial cells. Biol. Reprod. 44,102-107.

Richings N.M., Shaw G., Temple-Smith P.D. and Renfree M.B. (2004).Intra-cytoplasmic sperm injection in a marsupial. Reproduction 128,595-605.

Roberts C.T. and Breed W.G. (1996a). Variation in ultrastructure ofmucoid coat and shell membrane secretion of a dasyurid marsupial.Reprod. Fert. Dev. 8, 645-648.

Roberts C.T. and Breed W.G. (1996b). The marsupial shell membrane:an ultrastructural and immunogold localization study. Cell TissueRes. 284, 99-110.

Roberts C.T., Breed W.G. and Mayrhofer G. (1994). Origin of the oocyteshell membrane of a dasyurid marsupial: an immunohistochemicalstudy. J. Exp. Zool. 270, 321-331.

Roberts C.T., Selwood L., Leigh C.M. and Breed W.G. (1997).Antiserum to the egg coats of the fat-tailed dunnart (Marsupialia,Dasyuridae) cross-reacts with the egg coats of other marsupial andeutherian species. J. Exp. Zool. 278, 133-139.

Rodger J.C. and Bedford J.M. (1982). Induction of oestrus, recovery ofgametes, and the timing of fertilization events in the opossum,Didelphis virginiana. J. Reprod. Fert. 64, 159-169.

Roux E. and Kan F.W.K. (1995). Stage-specific immunolabeling foroviductin in the secretory granules of the oviductal epithelium of thegolden hamster during the estrous cycle. Anat. Rec. 241, 369-376.

Selwood L. (1980). A timetable of embryonic development of thedasyurid marsupial Antechinus stuartii (Macleay). Aust. J. Zool. 28,649-668.

Selwood L. (1982). A review of maturation and fertilization in marsupialswith special reference to the dasyurid: Antechinus stuartii. In:Carnivorous marsupials. Archer M. (ed). Royal Zoological Society ofNew South Wales. Sydney. pp 65-76.

Selwood L. (2000). Marsupial egg and embryo coats. Cells TissuesOrgans 166, 208-219.

Sendai Y., Abe H., Kikuchi M., Satoh T. and Hoshi H. (1994).Purification and molecular cloning of bovine oviduct-specificglycoprotein. Biol. Reprod. 50, 927-934.

Sendai Y., Komiya H., Suzuki K., Onuma T., Kikuchi M., Hoshi H. andAraki Y. (1995). Molecular cloning and characterization of a mouseoviduct-specific glycoprotein. Biol. Reprod. 53, 285-294.

Shalgi R. and Raz T. (1997). The role of carbohydrate residues inmammalian fertilization. Histol. Histopathol. 12, 813-822.

Sidhu K.S., Mate K.E. and Rodger J.C. (1998). Sperm-oviduct epithelialcell monolayer co-culture: an in vitro model of sperm-female tractinteractions in a marsupial, the tammar wallaby (Macropus eugenii).J. Reprod. Fert. 114, 55-61.

Sidhu K.S., Mate K.E., Molinia F.C., Glazier A.M. and Rodger J.C.(1999). Secretory proteins from the female reproductive tract of thebrushtail possum (Trichosurus vulpecula): binding to sperm andeffects on sperm survival in vitro. Reprod. Fert. Develop. 11, 329-336.

Smith T.T. and Nothnick W.B. (1997). Role of direct contact betweenspermatozoa and oviductal epithelial cells in maintaining rabbitsperm viability. Biol. Reprod. 56, 83-89.

Staros A.L. and Killian G.J. (1998). In vitro association of six oviductalfluid proteins with the bovine zona pellucida. J. Reprod. Fert. 112,131-137.

Suarez S.S. (1998). The oviductal sperm reservoir in mammals:mechanisms of formation. Biol. Reprod. 58, 1105-1107.

Suarez S.S. (2001). Carbohydrate-mediated formation of the oviductalsperm reservoir in mammals. Cells Tissues Organs 168, 105-112.

Suarez S.S. (2002). Formation of a reservoir of sperm in the oviduct.Reprod. Domest. Anim. 37, 140-143.

Suarez S.S. (2008). Regulation of sperm storage and movement in themammalian oviduct. Int. J. Dev. Biol. 52, 455-462.

Suarez S.S., Revah I., Lo M. and Kolle S. (1998). Bull sperm binding tooviductal epithelium is mediated by a Ca2+-dependent lectin onsperm that recognizes Lewis-a trisaccharide. Biol. Reprod. 59, 39-44.

Suzuki K., Sendai Y., Onuma T., Hoshi H., Hiroi M. and Araki Y. (1995).Molecular characterization of a hamster oviduct-specif icglycoprotein. Biol. Reprod. 53, 345-354.

Taggart D.A. (1994). A comparison of sperm and embryo transport inthe female reproductive tract of marsupial and eutherian mammals.Reprod. Fert. Dev. 6, 451-472.

Taggart D.A. and Temple-Smith P.D. (1991). Transport and storage ofspermatozoa in the female reproductive tract of the brown marsupialmouse, Antechinus stuartii (Dasyuridae). J. Reprod. Fert. 93, 97-110.

Taggart D.A., Breed W.G., Temple-Smith P.D., Purvis A. and ShimminG. (1998). Reproduction, mating strategies and sperm competition inmarsupials and monotremes. In: Sperm competition and sexualselection. Birkhead T.R. and Moller A.P. (eds). Academic Press. SanDiego. pp 623-666.

Takasaki S., Mori E. and Mori T. (1999.) Structures of sugar chainsincluded in mammalian zona pellucida glycoproteins and theirpotential roles in sperm-egg interaction. Biochim. Biophys. Acta1473, 206-215.

Talevi R. and Gualtieri R. (2001). Sulfated glycoconjugates are powerfulmodulators of bovine sperm adhesion and release from the oviductalepithelium in vitro. Biol. Reprod. 64, 491-498.

Tesarik J., Pilka L. and Travnik P. (1988). Zona pellucida resistance tosperm penetration before the completion of human oocytematuration. J. Reprod. Fert. 83, 487-495.

Thaler C.D. and Cardullo R.A. (1996). Defining oligosaccharidespecificity for initial sperm-zona pellucida adhesion in the mouse.

131

Oviducts in marsupials

Mol. Reprod. Dev. 45, 535-546.Töpfer-Petersen E. (1999). Carbohydrate-based interactions on the

route of a spermatozoon to fertilization. Hum. Reprod. Update 5,314-329.

Töpfer-Petersen E., Ekhlasi-Hundrieser M. and Tsolova M. (2008).Glycobiology of fertilization in the pig. Int. J. Dev. Biol. 52, 717-736.

Tulsiani D.R.P., Yoshida-Komiya H. and Araki Y. (1997). Mammalianfertilization: a carbohydrate-mediated event. Biol. Reprod. 57, 487-494.

Verhage H.G., Mavrogianis P.A., Boice M.L., Li W. and Fazleabas A.T.(1990). Oviduct epithelium of the baboon: hormonal control and theimmuno-gold localization of oviduct-specific glycoproteins. Am. J.Anat. 187, 81-90.

Verhage H.G., Fazleabas A.T., Mavrogianis P.A., O'Day-Bowman M.B.,Donnelly K.M., Arias E.B. and Jaffe R.C. (1997a). The baboonoviduct: characteristics of an oestradiol-dependent oviduct-specificglycoprotein. Hum. Reprod. Update 3, 541-552.

Verhage H.G., Mavrogianis P.A., Boomsma R.A., Schmidt A., BrennerR.M., Slayden O.V. and Jaffe R.C. (1997b). Immunologic andmolecular characterization of an estrogen-dependent glycoprotein inthe rhesus (Macaca mulatta) oviduct. Biol. Reprod. 57, 525-531.

Vitaioli L., Ricci R., Bellini L., Baldoni E., Antuzzi D. and Bolognani L.(1990). Sialic acid and neuraminidase activity in the frog oviduct:comparative biochemical investigation in the different tracts duringthe reproductive cycle. Comp. Biochem. Phys. B 95, 35-38.

Wagner A., Ekhlasi-Hundrieser M., Hettel C., Petrunkina A., WaberskiD., Nimtz M. and Topfer-Petersen E. (2002). Carbohydrate-basedinteractions of oviductal sperm reservoir formation - studies in thepig. Mol. Reprod. Dev. 61, 249-257.

Wassarman P.M. (1988). Zona pellucida glycoproteins. Annu. Rev.Biochem. 57, 415-442.

Wassarman P.M. (1990). Profile of a mammalian sperm receptor.Development 108, 1-17.

Wassarman P.M. (1995). Towards molecular mechanisms for gameteadhesion and fusion during mammalian fertilization. Curr. Opin. CellBiol. 7, 658-664.

Wegner C.C. and Killian G.J. (1992). Origin of oestrus-associatedglycoproteins in bovine oviductal fluid. J. Reprod. Fert. 95, 841-854.

Woolley P. (1966). Reproduction in Antechinus spp. and other dasyuridmarsupials. Symp. Zool. Soc. Lond. 15, 281-294.

Wu A.M., Sugii S. and Herp A. (1988). A guide for carbohydratespecificities of lectins. In: The molecular immunology of complexcarbohydrates. Wu A. and Adams L.G. (eds). Plenum Press. NewYork. pp 819-847.

Yong P., Zheng G.U., Luo J.P., Wang J.R. and Tso J.K. (2002).Antibodies against the C-terminal peptide of rabbit oviductin inhibitmouse early embryo development to pass 2-cell stage. Cell Res. 12,69-78.

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