Extractions reveal specific argentophilic proteins in rat and bull sperm heads

Yagi, Ahmed; Paranko, Jorma

doi:10.1002/ar.1092390203

Cited by 4 publications

(4 citation statements)

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“…Regional distribution of actin in the perinuclear theca resembles that of calicin, a highly extraction resistant basic protein specific for the a b sperm head cytoskeleton, including the murine perforatorium (Longo et al, 1987;Paranko et al, 1988). Actin may also colocalize with a group of less well identified argentophilic constituents of the cytoplasm (Yagi and Paranko, 1994). Indeed, based on the specific distribution and apparent conformation to a three-dimensional packing of the perinuclear theca, these accumulations of proteins may play a structural role in maintaining the integrity of the sperm head.…”

Section: Discussionmentioning

confidence: 99%

“…On this background and in view of the present findings it is tempting to propose that the negative immunocytochemical results in the rat and mouse spermatozoa are, within limits of the antibody specificity, due to the structural masking of the antigens. In this regard, it is significant to note that anionic SDS which specifically dissolves the plasma and acrosomal membranes and some of the fibro-granular material of the perinuclear theca (O'Brien and Bellve, 1980;Bellve et al, 1990Bellve et al, , 1992Yagi and Paranko, 1994) does not only retain actin in the perinuclear theca but also effectively demasks cryptic actin epitopes.…”

Section: Discussionmentioning

confidence: 99%

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Immunocytochemical detection of actin and 53 kDa polypeptide in the epididymal spermatozoa of rat and mouse

Paranko¹,

Yagi²,

Kuusisto³

1994

Anat. Rec.

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Intense labeling of actin in the SDS-extracted rat and mouse spermatozoa was presumably due to the generated demasking of actin epitopes embedded in the perinuclear cytoplasm. The results are important in confirming that actin in the rat and mouse sperm head is not lost during spermiogenesis but apparently contributes to the three-dimensional packing of the mature perinuclear cytoplasm. This study further demonstrates the importance of the methods used in sample preparation and advantages of confocal microscopy when attempting to detect cytoskeletal proteins which, as in spermatozoa, may occur in small quantities.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Immunocytochemical detection of actin and 53 kDa polypeptide in the epididymal spermatozoa of rat and mouse

Paranko¹,

Yagi²,

Kuusisto³

1994

Anat. Rec.

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“…A rigid postacrosomal sheath is a putative site for actin in a number of mammalian spermatozoa (Clarke and Yanagimachi, 1978;Tamblyn, 1980;Clarke et al, 1982;Flaherty et al, 1986Flaherty et al, , 1988Yagi and Paranko, 1992), where it may interact with actin-binding proteins such as spectrin (Kann et al, 1993) and a group of basic proteins including calicin and multiple band proteins (Longo et al, 1987;Paranko et al, 19881, and with less well identified silver binding proteins (Yagi and Paranko, 1994).…”

Section: Lmmunolocalization Ofmentioning

confidence: 99%

Actin, α‐actinin, and spectrin with specific associations with the postacrosomal and acrosomal domains of bovine spermatozoa

Yagi

Paranko

1995

Anat. Rec.

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Destabilization of membranes with selected extractions induces changes in the acrosomal lamina mimicking acrosomal vesicle formation. The lateral prongs and posterior ring structure, respectively, may serve as anterior and posterior anchors for the extraction-resistant post-acrosomal sheath. The lateral prongs may also be a merger zone for actin, alpha-actinin, and spectrin with important implication on sperm function. The latter two proteins may be involved in acrosomal vesicle formation. It is apparent that extractions have a significant effect on the detectability of sperm cytoskeletal elements.

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“…SDS is a stronger detergent than Triton X-100, which can destroy the nucleus, so that it has additional property that melt the acrosomal membrane, if exceeded the optimum value, and finally makes disappear [19][20][21]. However, application of 0.05% SDS successfully separates the spermatid head-tail connection.…”

Section: Discussionmentioning

confidence: 99%

A New Preparation Protocol for Measurement of Testicular Sperm Production

Choi

Tsunekawa

Kanai‐Azuma

et al. 2008

Journal of Reproduction and Development

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Abstract. In studies of male reproductive toxicity, measuring daily sperm production is a quite important criterion. However, the accuracy of the values measured by the basic protocol is still controversial. In order to enhance the homogeneity of countable testicular sperm/spermatid heads, this report introduces a new enzymatic method with a subsequent detergent treatment. The testis of rat was firstly homogenized in phosphate-buffered saline. The homogenate (buffer mix) was then treated with collagenase and trypsin, and then sodium dodecyl sulfate (SDS) was added to produce detergent-resistant sperm/spermatid heads (detergent mix). After examination by hemocytometer, the coefficient of variation (CV) of the number of sperm/spermatid heads was compared with that obtained from the buffer mix. In addition, a MicroCell chamber was applied to the examination, and the CV was compared with other cases. In both examinations, homogeneity was improved by the detergent mix preparation. Counting with the hemocytometer showed an increased number of sperm/spermatid heads compared with that of the buffer mix (P<0.001), and the CV was decreased (P<0.05). In addition, when the MicroCell chamber was applied, the numbers increased about 3-hold compared with that of the buffer mix (P<0.001). The CV of the detergent mix was 23.7%, while that of the buffer mix was 38.9%. These results clearly demonstrate that the new preparation protocol generated in this study can provide more actual and accurate values when measuring daily sperm production. Key words: Agglutination, Collagenase, Daily sperm production, Homogeneity, Rat, Sodium dodecyl sulfate (SDS) (J. Reprod. Dev. 54: [90][91][92][93] 2008) perm analysis is an essential item for evaluation of male reproductive toxicity. This evaluation is generally categorized into sperm count, motility and morphology [1]. For sperm counting, methods using counting a chamber, computer-assisted sperm analysis systems (CASA) and automated systems (e.g., IVOS HTM-IDENT; HTM Integrated Visual Optical System semen analyzer) are widely used [2][3][4][5][6]. Among these methods, use of a counting chamber is problematic because of variation. CASA and HTM-IDENT are also inapplicable because of dependence on technical skill and high installation costs [2][3][4][5][6][7][8][9][10][11][12]. In addition to the problems concerning these measurement systems, the viscosity and agglutination of the sample (e.g., semen), the inhomogeneity of the sperm suspension distribution and the technician's carefulness, fatigue and skill in pipetting can cause problematic variations in analyses [1,[13][14][15][16].In animal experiments, epididymal and testicular sperm are usually used for sperm counting [17]. However, in the case of epididymal sperm, the condition and position (caput or cauda) of the sperm collection may cause variation [3]. In measurement of daily sperm production, testicular homogenization-resistant sperm/ spermatids, which contain mature spermatozoa after spermiation and step 17-19 spermatids in case of...

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Extractions reveal specific argentophilic proteins in rat and bull sperm heads

Cited by 4 publications

References 63 publications

Immunocytochemical detection of actin and 53 kDa polypeptide in the epididymal spermatozoa of rat and mouse

Immunocytochemical detection of actin and 53 kDa polypeptide in the epididymal spermatozoa of rat and mouse

Actin, α‐actinin, and spectrin with specific associations with the postacrosomal and acrosomal domains of bovine spermatozoa

A New Preparation Protocol for Measurement of Testicular Sperm Production

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