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

Labbé, Catherine; Loir, Maurice

doi:10.1007/bf02265153

Cited by 17 publications

(6 citation statements)

References 30 publications

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“…However, except for these ionic and water channels, there are still many types of membrane proteins unidentified in fish. Labbé and Loir (1991) isolated membrane proteins from spermatozoa of rainbow trout (O. mykiss) by four techniques (sonication, hypotonic shock, mechanical homogenisation after freeze-thawing, and nitrogen cavitation, in combination with continuous sucrose gradient centrifugation) and showed that most of the membrane proteins (with 42-and 30-kDa bands as the main proteins) were acidic to neutral.…”

Section: Membrane Proteinsmentioning

confidence: 99%

Sperm proteins in teleostean and chondrostean (sturgeon) fishes

Hulák

Linhart

2008

Fish Physiol Biochem

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Sperm proteins in the seminal plasma and spermatozoa of teleostean and chondrostean have evolved adaptations due to the changes in the reproductive environment. Analysis of the composition and functions of these proteins provides new insights into sperm motility and fertilising abilities, thereby creating possibilities for improving artificial reproduction and germplasm resource conservation technologies (e.g. cryopreservation). Seminal plasma proteins are involved in the protection of spermatozoa during storage in the reproductive system, whereas all spermatozoa proteins contribute to the swimming and fertilising abilities of sperm. Compared to mammalian species, little data are available on fish sperm proteins and their functions. We review here the current state of the art in this field and focus on relevant subjects that require attention. Future research should concentrate on protein functions and their mode of action in fish species, especially on the role of spermatozoa surface proteins during fertilisation and on a description of sturgeon sperm proteins.

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Section: Membrane Proteinsmentioning

confidence: 99%

Sperm proteins in teleostean and chondrostean (sturgeon) fishes

Hulák

Linhart

2008

Fish Physiol Biochem

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“…Plasma membrane extraction from sperm was carried out following an adapted protocol from de Curtis et al [40] and Labbé and Loir [41]. For total sperm, activated and nonactivated spermatozoa were recovered as above; homogenized in a buffer containing 20 mM Tris-HCl (pH 7.8), 3 mM MgCl 2 , 0.25 M sucrose, and protease inhibitors; and centrifuged at 1000 3 g for 20 min at 48C.…”

Section: Plasma Membrane Purification Of Intact Spermatozoa Head Anmentioning

confidence: 99%

Subcellular Localization of Selectively Permeable Aquaporins in the Male Germ Line of a Marine Teleost Reveals Spatial Redistribution in Activated Spermatozoa1

Chauvigné

Boj

Vilella

et al. 2013

Biology of Reproduction

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In oviparous vertebrates such as the marine teleost gilthead seabream, water and fluid homeostasis associated with testicular physiology and the external activation of spermatozoa is potentially mediated by multiple aquaporins. To test this hypothesis, we isolated five novel members of the aquaporin superfamily from gilthead seabream and developed paralog-specific antibodies to localize the cellular sites of protein expression in the male reproductive tract. Together with phylogenetic classification, functional characterization of four of the newly isolated paralogs, Aqp0a, -7, -8b, and -9b, demonstrated that they were water permeable, while Aqp8b was also permeable to urea, and Aqp7 and -9b were permeable to glycerol and urea. Immunolocalization experiments indicated that up to seven paralogous aquaporins are differentially expressed in the seabream testis: Aqp0a and -9b in Sertoli and Leydig cells, respectively; Aqp1ab, -7, and -10b from spermatogonia to spermatozoa; and Aqp1aa and -8b in spermatids and sperm. In the efferent duct, only Aqp10b was found in the luminal epithelium. Ejaculated spermatozoa showed a segregated spatial distribution of five aquaporins: Aqp1aa and -7 in the entire flagellum or the head, respectively, and Aqp1ab, -8b, and -10b both in the head and the anterior tail. The combination of immunofluorescence microscopy and biochemical fractionation of spermatozoa indicated that Aqp10b and phosphorylated Aqp1ab are rapidly translocated to the head plasma membrane upon activation, whereas Aqp8b accumulates in the mitochondrion of the spermatozoa. In contrast, Aqp1aa and -7 remained unchanged. These data reveal that aquaporin expression in the teleost testis shares conserved features of the mammalian system, and they suggest that the piscine channels may play different roles in water and solute transport during spermatogenesis, sperm maturation and nutrition, and the initiation and maintenance of sperm motility.

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“…Aliquots of sl-PC and sl-SM in chloroform were dried and resuspended by vigorous vortexing with SpM at 48C; 7.2 nmol of labeled lipid in SpM was added to the cell pellet containing about 2 3 10 9 sperm cells. Considering a membrane phospholipid content of about 375 nmol per 10 9 cells [51], the molar concentrations of sl-PC and sl-SM in the trout sperm membrane were about 1%. After 5 min at 188C, cells were washed by addition of 20 volumes SpM and then were centrifuged.…”

Section: Characterization Of Membrane Fluidity By Spin-labeled Lipidsmentioning

confidence: 99%

Characterization of Sperm Plasma Membrane Properties after Cholesterol Modification: Consequences for Cryopreservation of Rainbow Trout Spermatozoa1

Müller¹,

Müller²,

Pincemy³

et al. 2008

Biology of Reproduction

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During cryopreservation, the cell plasma membrane faces severe perils, including lipid phase separation, solute effects, and osmotic stresses associated with ice crystallization. How the initial biophysical properties of the plasma membrane can be modulated before cryopreservation in order to influence cellular resistance to the freeze-thaw stress is addressed in this study. Rainbow trout (Oncorhynchus mykiss) spermatozoa were chosen because the lack of an acrosome in this species suppresses potential interactions of cryopreservation with capacitation. Methyl-beta cyclodextrin-induced modulation of membrane cholesterol revealed the presence of a significant cholesterol exchangeable pool in the trout sperm plasma membrane, as membrane cholesterol content could be halved or doubled with respect to the basic composition of the cell without impairing fresh sperm motility and fertilizing ability. Biophysical properties of the sperm plasma membrane were affected by cholesterol changes: membrane resistance to a hypoosmotic stress increased linearly with membrane cholesterol whereas membrane fluidity, assessed with DPH (1,6-diphenyl-1,3,5-hexatriene) and with several spin-labeled analogues of membrane lipids, decreased. Phosphatidyl serine translocation between the bilayers was slowed at high cholesterol content. The increased cohesion of fresh trout sperm plasma membrane as cholesterol increased did not improve the fertilizing ability of frozen-thawed sperm whereas the lowest cholesterol contents impaired this parameter of sperm quality. Our study demonstrated that cholesterol induced a stabilization of the plasma membrane in rainbow trout spermatozoa, but this stabilization before cryopreservation brought no improvement to the poor freezability of this cell.

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

Cited by 17 publications

References 30 publications

Sperm proteins in teleostean and chondrostean (sturgeon) fishes

Sperm proteins in teleostean and chondrostean (sturgeon) fishes

Subcellular Localization of Selectively Permeable Aquaporins in the Male Germ Line of a Marine Teleost Reveals Spatial Redistribution in Activated Spermatozoa1

Characterization of Sperm Plasma Membrane Properties after Cholesterol Modification: Consequences for Cryopreservation of Rainbow Trout Spermatozoa1

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