Electron-microscopic localisation of thiol and disulphide groups by direct monomaleimido-nanogold labelling in the spermatozoa of a marsupial, the tammar wallaby (Macropus eugenii)

Lin, Mingwei; Sistina, Yulia; Rodger, John C.

doi:10.1007/bf00319119

Cited by 9 publications

(7 citation statements)

References 12 publications

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“…Monomaleimido-Nanogold (MMNA) MMNA is a reagent that specifically labels free thiol groups in proteins. It was used for the first time in sperm research by Lin et al (1995). In a previous work (Gimenez-Bonafé et al, 1999), we have demonstrated by MMNA labeling that free thiol groups disappear in E. cirrhosa ripe sperm nuclei.…”

Section: Labeling Withmentioning

confidence: 96%

“…In a previous work (Gimenez-Bonafé et al, 1999), we have demonstrated by MMNA labeling that free thiol groups disappear in E. cirrhosa ripe sperm nuclei. During these experiments, we observed that when the original procedure of Lin et al (1995) is slightly modified by limiting the labelling conditions, MMNA is an excellent marker for microtubules due to the large number of free sulfydryl (ÀSH) groups on cysteine residues of a and b tubulin. For the present work, small pieces of gonad were rinsed in phosphate-buffered saline (PBS), prefixed for 15 min in 0.5% glutaraldehyde/PBS, and rinsed several times with PBS.…”

Section: Labeling Withmentioning

confidence: 99%

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Evolution of octopod sperm I: Comparison of nuclear morphogenesis in Eledone and Octopus

Gimenez‐Bonafé¹,

Ribes

Zamora

et al. 2002

Molecular Reproduction Devel

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Morphogenesis of the Eledone cirrhosa sperm nucleus, as studied by electron microscopic techniques, is compared with that of Octopus vulgaris. Both species of cephalopods belong to the family Octopodidae. The results indicate that extensive nuclear helicoidization during E. cirrhosa spermiogenesis is brought about by modifications of the function of structural components already present in the late steps of O. vulgaris spermiogenesis. In particular, changes in the regulation of perinuclear microtubule contraction in E. cirrhosa spermatids, as well as a decrease in basicity of protamines, promote nuclear helicoidization. Disulphide bond formation between protamine molecules fixes the completely helicoidal shape of the nucleus in mature sperm of E. cirrhosa.

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Section: Labeling Withmentioning

confidence: 96%

Section: Labeling Withmentioning

confidence: 99%

Evolution of octopod sperm I: Comparison of nuclear morphogenesis in Eledone and Octopus

Gimenez‐Bonafé¹,

Ribes

Zamora

et al. 2002

Molecular Reproduction Devel

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“…It has been hypothesised that the acrosome is so located to maximise contact between its surface and that of the outer matrix of the zona pellucida, to facilitate spermzona binding and penetration (Bedford, 1991(Bedford, , 1996(Bedford, , 1998. Furthermore, in contrast to the sperm chromatin, the acrosome appears to be more stabilised than that of eutherians (Cummins, 1980, Mate & Rodger, 1991Sistina et al 1993) and it has been suggested that such stabilisation may be due to disulphide bonding of proteins within the acrosomal membranes and\or matrix (Lin et al 1995).…”

Section: mentioning

confidence: 99%

“…Conversely the acrosome of marsupial sperm is thought to be highly stabilised so that on air drying, it, unlike the nucleus, apparently does not undergo dispersion of its contents nor is it dispersed either after freezing or thawing or with ' moderate ' detergent treatment (Mate & Rodger, 1991 ;Sistina et al 1993). It has been suggested that the stability of the acrosomal membranes and\or matrix may be due to the presence of disulphide bonded proteins which, unlike in eutherians, are not present in the chromatin (Lin et al 1995).…”

Section: mentioning

confidence: 99%

The structural organisation of sperm head components of the wombat and koala (suborder: Vombatiformes): an enigma amongst marsupials

2001

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The sperm head structural organisation of the koala and hairy-nosed wombat, and the effects of varying concentrations of the ionic detergent, Triton X-100, on its component parts, were determined by electron microscopy. Although alike in form between the 2 species, the sperm nucleus of the former, but not the latter, had nuclear vacuoles and appeared to be more easily dispersed by the Triton X-100. The structure of the acrosome of the spermatozoon was similar between the 2 species and, in both, previously undescribed thin posterior and lateral segments were found to be present. It is suggested that this thin segment may facilitate sperm-zona and\or sperm-oolemma binding and fusion.

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“…On the basis of ultrastructural studies, this structure is thought to serve a passive stiffening role, strengthening the sperm midpiece, thereby modifying flagellar bending (Harding et al, 1975(Harding et al, , 1979Olsen, 1975). This structure is electron-dense and has been shown in the possum to be resistant to sodium dodecyl sulphate (SDS) detergent solubilisation (Temple-Smith and Bedford, 1976) due to extensive disulphide stabilisation (Mate et al, 1994;Lin et al, 1995). However, the molecular nature of this structure remains unknown.…”

Section: Introductionmentioning

confidence: 99%

Characterisation of fibrous sheath and midpiece fibre network polypeptides of marsupial spermatozoa with a monoclonal antibody

Harris

Rodger

1998

Mol. Reprod. Dev.

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In this study cytoskeletal antigens common to brushtail possum and tammar wallaby spermatozoa were characterised using a monoclonal antibody (PSA‐10). Using indirect immunofluorescence, the PSA‐10 antibody detected antigens predominantly associated with the midpiece and principal piece of mature, permeabilised marsupial spermatozoa. The principal piece determinant, shared by a variety of other species, was found to arise in the marsupial testis. Midpiece localisation of the PSA‐10 epitope was detected only in marsupial spermatozoa and shown to arise in the epididymis. Immunogold labelling demonstrated that the PSA‐10 antigens were predominantly associated with the fibrous sheath and midpiece fibre network of both possum and wallaby spermatozoa. Western blotting suggested that two major possum and wallaby sperm polypeptides of 158 and 182 kDa were associated with the midpiece fibre network, a cytoskeletal structure unique to marsupial spermatozoa. A 32 kDa polypeptide was associated with the principal piece fibre network and/or fibrous sheath. The finding that these marsupial sperm cytoskeletal proteins share a common linear epitope suggests that they share some sequence similarity. The midpiece fibre network of marsupial sperm, like the fibrous sheath, has been proposed to have a structural role in providing passive stiffening for the flagellum (Harding et al., 1975, 1979; Olsen, 1975). The PSA‐10 monoclonal antibody may provide a tool for comparative studies of mammalian sperm cytoskeletal proteins, particularly the marsupial midpiece fibre network. It may also allow the formation of this unique marsupial cytoskeletal structure, and its fate during the fertilisation process, to be followed by immunological means. Mol. Reprod. Dev. 50:461–473, 1998. © 1998 Wiley‐Liss, Inc.

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Electron-microscopic localisation of thiol and disulphide groups by direct monomaleimido-nanogold labelling in the spermatozoa of a marsupial, the tammar wallaby (Macropus eugenii)

Cited by 9 publications

References 12 publications

Evolution of octopod sperm I: Comparison of nuclear morphogenesis in Eledone and Octopus

Evolution of octopod sperm I: Comparison of nuclear morphogenesis in Eledone and Octopus

The structural organisation of sperm head components of the wombat and koala (suborder: Vombatiformes): an enigma amongst marsupials

Characterisation of fibrous sheath and midpiece fibre network polypeptides of marsupial spermatozoa with a monoclonal antibody

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