EurekaMag.com logo
+ Site Statistics
References:
52,725,316
Abstracts:
28,411,598
+ Search Articles
+ Subscribe to Site Feeds
EurekaMag Most Shared ContentMost Shared
EurekaMag PDF Full Text ContentPDF Full Text
+ PDF Full Text
Request PDF Full TextRequest PDF Full Text
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter
Follow on Google+Follow on Google+
Follow on LinkedInFollow on LinkedIn

+ Translate

Identification of capacitation in boar spermatozoa by chlortetracycline staining


Theriogenology 45(2): 373-381
Identification of capacitation in boar spermatozoa by chlortetracycline staining
The functional status of boar spermatozoa undergoing capacitation in vitro was investigated. Two fluorescent stains were used: chlortetracycline (CTC) and a FITC-conjugated lectin (FITC-PSA). The first has been used for the direct identification of the capacitated boar spermatozoa, while the second, based on the identification of capacitated spermatozoa by their ability to undergo zona-induced acrosome reaction (AR), was used to confirm and validate the CTC assay in this species. Spermatozoa obtained from 5 different boars was washed and incubated under capacitating conditions. Aliquots of spermatozoa were collected at 0, 90 and 180 min of incubation and then stained with CTC or FITC-PSA. After CTC staining, 3 different fluorescent patterns were observed: Pattern A with the fluorescence uniformly distributed on the sperm head, Pattern B with the fluorescence concentrated in the post-acrosomial region, and Pattern C with the fluorescence concentrated in the acrosomial region, The percentage of spermatozoa displaying fluorescent Pattern A decreased throughout the incubation while that of spermatozoa with Pattern C showed a concomitant progressive increase. Pattern B fluorescence remained unchanged throughout the maturation period. Exposure to zonae pellucidae (ZP) brought back the levels of Pattern C fluorescence to basal values. Since only the capacitated spermatozoa are believed to react to ZP, this observation together with the rising incidence of Pattern C throughout maturation suggests that fluorescence in the acrosomial region identifies capacitated spermatozoa. The analysis of acrosome integrity carried out with FITC-PSA showed that the proportion of zona-induced AR was nearly the same as that of spermatozoa displaying Pattern C, thus confirming that CTC staining is suitable for the detection of boar sperm capacitation. In the second part of this study, CTC was used to investigate the effects of sperm origin and storage on the capacitation process. Our finding demonstrates that capacitation kinetics show wide variations in sperm samples derived from different boars- moreover, capacitation is also affected by sperm storage. While fresh semen showed a progressive increase in capacitated spermatozoa, ranging from low levels at the beginning of the culture to 46 % at the end of incubation, the refrigerated semen had a relatively high percentage of capacitated spermatozoa at the beginning of culture, but this proportion increased only slightly during the following 90 to 180 min of treatment. These data indicate that CTC can be used to identify capacitated boar spermatozoa, and, because of its rapid and easy execution, it can be used routinely to identify the optimal capacitation time for different sperm samples.

Accession: 002863359

PMID: 16727801

DOI: 10.1016/0093-691x(96)81099-5

Download PDF Full Text: Identification of capacitation in boar spermatozoa by chlortetracycline staining



Related references

Capacitation-like alterations in cooled boar spermatozoa: assessment by the chlortetracycline staining assay and immunodetection of tyrosine-phosphorylated sperm proteins. Animal Reproduction Science 73(3-4): 197-209, 2002

In vitro capacitation of dog spermatozoa as assessed by chlortetracycline staining. Theriogenology 52(4): 617-628, 2000

Staining patterns of bull, ram and boar spermatozoa before and after in vitro capacitation in different media. Farbeverhalten von Bullen , Schafbock und Eber Spermien vor und nach in vitro Kapazitation in verschiedenen Medien: 144 pp., 1995

Effects of extender, incubation temperature, and added seminal plasma on capacitation of cryopreserved, thawed boar sperm as determined by chlortetracycline staining. Animal Reproduction Science 90(3-4): 347-354, 2005

Lectin histochemistry during in vitro capacitation and acrosome reaction in boar spermatozoa: new lectins for evaluating acrosomal status of boar spermatozoa. Acta Histochemica 98(1): 93-100, 1996

Cooling of boar spermatozoa before freezing and post-thawing quality and evaluation of plasma membranes using chlortetracycline staining. Deutsche Tierarztliche Wochenschrift 104(8): 302-306, 1997

Cooling of boar spermatozoa prior to freezing and post thaw quality and evaluation of membrane state using chlortetracycline (CTC) staining. Dtw. Deutsche Tierarztliche Wochenschrift 104(8): 302-306, 1997

Cooling of boar spermatozoa prior to freezing and post thaw quality and evaluation of the membrane state using Chlortetracycline (CTC)-staining. DTW (Deutsche Tieraerztliche Wochenschrift) 104(8): 302-306, 1997

Changes in tyrosine phosphorylation associated with true capacitation and capacitation-like state in boar spermatozoa. Molecular Reproduction and Development 71(1): 88-96, 2005

Comparison of the capacitation-like state of cooled boar spermatozoa with true capacitation. Reproduction 122(6): 889-898, 2001