Following passage through the oviduct, the coelomic envelope of Discoglossus pictus (amphibia) acquires fertilizability upon reorganization, conversion of gp 42 to gp 40, extensive glycosylation, and formation of a specific layer.
This paper describes the morphological and biochemical changes in Discoglossus pictus coelomic oocyte envelope (CE) following passage through the oviduct. As in other anurans, in this species, the transformation of the envelope into vitelline envelope (VE) leads to the acquisition of fertilizability and involves the cleavage of a glycoprotein. In addition, several features, typical of Discoglossus pictus, were observed. A new layer, VE-D, forms underneath the VE region facing the site of sperm entrance, the dimple. In the VE, arrowhead-like bundles of fibrils are perpendicularly oriented toward the dimple. Ultrastructural observations and staining with UEA-I suggested that VE-D might have a role in supporting sperm penetration into the dimple by orienting VE bundles and exposing sugar residues such as fucose. In 'in vitro' tests, VE binding of sperm occurs only if sperm are exposed to A23187, in agreement with previous data (Campanella et al., 1997: Mol Reprod Dev 47:323-333). Sperm binding occurs all over the VE. Accordingly, extracts of the VE covering the animal or the vegetal hemisphere have the same affinity to lectins (DBA, DSA, GNA, MAA, SBA, SNA, UEA-I, WGA). The CE contains six main glycoproteins. Peptide mapping indicated that during CE transformation into VE, gp 42 shifts to an apparent M(r) of 40 and gp 61 is converted to an apparent M(r) of 63 kDa. Lectin blot analyses showed extensive changes in cross-reactivity of most glycoproteins during the CE-->VE transition. The fact that DBA and UEA-I stain gp 63 rather than gp 61 and that this change is related only to gp 63, suggested that O-glycosylation and terminal fucose might be acquired by gp 63 in preparation of fertilization. Gp 63 has recently been cloned (Vaccaro et al., submitted) and shown to exhibit high homology to Xenopus gp 69/64, a VE sperm ligand (Tian et al., 1997a: J. Cell Biol. 136: 1099-1108; Tian et al., 1997b: Dev Biol 187:143-153), and to ZP2 of mammals.
Caputo
M
M
Dipartimento di Biologia Evolutiva e Comparata, Universita' di Napoli, Napoli, Italy.
Infante
V
V
Talevi
R
R
Vaccaro
M C
MC
Carotenuto
R
R
Campanella
C
C
D000818:Animals; D001001:Anura / Q000502:physiology*; D016764:Cell Polarity; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005306:Fertilization / Q000502:physiology*; D006023:Glycoproteins / Q000378:metabolism; D037102:Lectins / Q000378:metabolism; D008297:Male; D010057:Oviducts / Q000378:metabolism; D010063:Ovum / Q000502:physiology* / Q000648:ultrastructure; D010449:Peptide Mapping; D013094:Spermatozoa / Q000378:metabolism*; D014817:Vitelline Membrane / Q000737:chemistry* / Q000378:metabolism
D006023:Glycoproteins; D037102:Lectins