Plasma membrane of trout spermatozoa : I. Isolation and partial characterization - INRAE - Institut national de recherche pour l’agriculture, l’alimentation et l’environnement Access content directly
Journal Articles Fish Physiology and Biochemistry Year : 1991

Plasma membrane of trout spermatozoa : I. Isolation and partial characterization

Abstract

The plasma membrane from spermatozoa of rainbow trout was isolated by four techniques: sonication, hypotonic shock, mechanical homogenization after freeze-thawing, and nitrogen cavitation, in combination with continuous sucrose gradient centrifugation. Nitrogen cavitation (900 PSI, 20 min equilibration at 4°C) was the most effective technique. Following nitrogen cavitation, four bands were recovered in the sucrose gradient at densities ≈ 1.03, 1.05, 1.09 and 1.15 g/ml. Electron microscopy revealed membrane vesicles of various sizes in bands 1 to 3, while enzyme analysis revealed a 3.9 to 5.5-fold enrichment in 5'-nucleotidase and little contamination by lactate dehydrogenase (cytosol) and succinic dehydrogenase (mitochondria). Lipid analysis of bands 1 and 2 indicated a 6 to 7-fold enrichment in cholesterol and a cholesterol: phospholipid ratio of 0.59–0.70. Seven classes of phospholipids were present in bands 1–3 with no significant differences observed among bands. These data indicate that the vesicles (in bands 1 and 2) obtained after nitrogen cavitation are primarily plasma membranes. Membranes in band 3 appear to be slightly contaminated with nuclear membranes. Most of the plasma membrane proteins were acidic to neutral. The 2 main membrane proteins were 42 and 30 Kilodaltons.

Keywords

Fichier principal
Vignette du fichier
Labb1991FPB_1.pdf (2.9 Mo) Télécharger le fichier
Origin : Publisher files allowed on an open archive
Loading...

Dates and versions

hal-02716127 , version 1 (01-06-2020)

Identifiers

Cite

Catherine Labbé, Maurice Loir. Plasma membrane of trout spermatozoa : I. Isolation and partial characterization. Fish Physiology and Biochemistry, 1991, 9, pp.325-338. ⟨10.1007/BF02265153⟩. ⟨hal-02716127⟩

Collections

INRA LPGP INRAE
8 View
98 Download

Altmetric

Share

Gmail Facebook X LinkedIn More