Assessment of Sperm Function Under Fertilizing Conditions

Harrison, R. A. P.; Ashworth, P.J.C.; Miller, Nigel

doi:10.1111/j.1439-0531.1995.tb00002.x

Cited by 14 publications

(11 citation statements)

References 16 publications

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“…On this basis, 33.6% of the low-calcium live population had become refractory. It may be noted that a similar reduction in responsiveness to bicarbonate of stored spermatozoa was described by Harrison et al (13), and Guthrie and Welch (14), in terms of fewer plasma membraneintact spermatozoa exhibiting bicarbonate-induced plasmamembrane phospholipid disorder as detected by merocyanine-540 staining.…”

Section: Discussionmentioning

confidence: 76%

“…Special emphasis was put on the detection of possible changes in specific responses to bicarbonate because there have been indications that spermatozoa may lose their responsiveness to bicarbonate during storage (13,14). Our hypothesis was that this loss of response could prove to be a sensitive measure of storage-associated loss of semen quality (i.e., loss of fertilizing capacity).…”

Section: Introductionmentioning

confidence: 99%

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Bivalent response to long‐term storage in liquid‐preserved boar semen: A flow cytometric analysis

Henning

Petrunkina

Harrison³

et al. 2012

Cytometry Pt A

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The fertility of liquid-preserved boar semen declines during storage at 178C, insemination trials even indicating early losses in fertilizing ability within the first 24-48 h of storage. Standard semen parameters barely reflect these changes in semen quality, and new approaches for assessment of functional changes in stored spermatozoa are needed. Capacitation, the essential prefertilization step for spermatozoa in the female genital tract, is specifically induced in vitro by bicarbonate. Therefore, we have investigated changes in responsiveness of boar spermatozoa to bicarbonate during storage. Ejaculates of 14 boars were diluted in Beltsville thawing solution, cooled to 178C and stored for 12, 24, 72, 120, and 168 h before investigation. At each time, basic semen quality was characterized by sperm motility and viability. Subsequently, washed subsamples were incubated in variants of an in vitro fertilization (IVF) medium and assessed for kinetic changes of viability (plasma membrane integrity) and intracellular calcium concentration using flow cytometry in combination with propidium iodide and Fluo-3. By this means, it was possible to determine specific effects of bicarbonate and calcium on sperm subpopulations over incubation time. During storage, standard semen parameters remained on a high level. However, flow cytometric analysis of sperm responses to capacitating and control media revealed two opposing effects of storage. There was a loss of response to bicarbonate in part of the live sperm population but an increasing degree of instability in the rest. Assessment of response to capacitating media by flow cytometry appears a markedly more sensitive way of monitoring sperm functionality during storage than the standard semen parameters of motility and viability. ' 2012 International Society for Advancement of Cytometry

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Section: Discussionmentioning

confidence: 76%

Section: Introductionmentioning

confidence: 99%

Bivalent response to long‐term storage in liquid‐preserved boar semen: A flow cytometric analysis

Henning

Petrunkina

Harrison³

et al. 2012

Cytometry Pt A

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“…Not only the epithelium itself but also the intraluminal fluid fulfils important functions including the facilitation of sperm capacitation, final zona pellucida (ZP) maturation, fertilization and nourishment of the ovulated oocyte and the early embryo (Parrish et al, 1989a;McNutt & Killian, 1991;Kawakami et al, 1998;Rodriguez-Martinez, 2001;Rodriguez-Martinez et al, 2001Hunter & Rodriguez-Martinez, 2004). A variety of substances within ODF are considered to be sperm-capacitating agents, including bicarbonate (Harrison et al, 1996;Tienthai et al, 2004, Harrison & Gadella, 2005, glycosaminoglycans (GAGs) Kawakami et al, 2000, Tienthai et al, 2004, calcium (DasGupta et al, 1993;Visconti et al, 2002), norepinephrine (Way & Killian, 2002) and progesterone (Libersky & Boatman, 1995). A variety of substances within ODF are considered to be sperm-capacitating agents, including bicarbonate (Harrison et al, 1996;Tienthai et al, 2004, Harrison & Gadella, 2005, glycosaminoglycans (GAGs) Kawakami et al, 2000, Tienthai et al, 2004, calcium (DasGupta et al, 1993;Visconti et al, 2002), norepinephrine (Way & Killian, 2002) and progesterone (Libersky & Boatman, 1995).…”

Section: Introductionmentioning

confidence: 99%

“…The higher the degree of disorder among phospholipids in the sperm membrane, the higher the amount of merocyanine-540 that binds, thus increasing the amount of fluorescence it releases when exposed to a laser beam (Harrison et al, 1996). Early plasma membrane lipid changes may be detected by the hydrophobic dye merocyanine-540.…”

Section: Introductionmentioning

confidence: 99%

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In vitrocapacitation of bull spermatozoa by oviductal fluid and its components

Bergqvist

Ballester

Johannisson

et al. 2006

Zygote

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Sperm capacitation is crucial for fertilization. However, debate continues on exactly how, where and when capacitation is elicited in the bovine female genital tract. In this study we used merocyanine-540 and the chlortetracycline (CTC) assay to test how capacitation of bull spermatozoa is affected in vitro by exposure to oviductal fluid (ODF) collected in vivo, various glycosaminoglycans (GAGs) or bicarbonate. Following different durations of exposure, spermatozoa were stained with CTC or merocyanine-540, and evaluated with epifluorescent light microscopy or flow cytometry, respectively. Incubation time did not significantly affect capacitation. Exposure (30-120 min) to ODF capacitated (p < 0.05) bull spermatozoa as measured by either merocyanine-540 or CTC. Hyaluronan was the only GAG that induced a significant increase in B-pattern spermatozoa (capacitated; p = 0.012) compared with controls. Dermatan sulphate also induced capacitation (merocyanine-540 high fluorescence; p = 0.035). Exposure to bicarbonate-enriched media also yielded an increase in merocyanine-540 high fluorescence (p < 0.0001). When bicarbonate was added to the other treatments (ODF or GAGs) an equal increase in merocyanine-540 high fluorescence was noted (p < 0.0001), compared with before addition of bicarbonate and independent of the treatment before exposure. There was no significant difference in the number of B-pattern spermatozoa when bicarbonate was added, but an significant increase in spermatozoa with an acrosome-reacted (AR)-pattern (p < 0.0001) was observed. Exposure of spermatozoa to solubilized zonae pellucidae significantly increased the AR-pattern spermatozoa (p = 0.016). In conclusion, ODF was more potent in inducing capacitation of bull spermatozoa than the individual GAGs. Our results also indicate that bicarbonate is an effector of bull sperm capacitation.

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