Fertility and Sterility

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Fertility and Sterility Hyaluronic acid binding by human sperm indicates cellular maturity, viability, and unreacted acrosomal status Gabor Huszar, M.D., Ciler Celik Ozenci, Ph.D., Sevil Cayli, M.S., Zoltan Zavaczki, M.D., Eleonora Hansch, M.D., Lynne Vigue, M.S. Fertility and Sterility Volume 79, Pages 1616-1624 (June 2003) DOI: 10.1016/S0015-0282(03)00402-3

FIGURE 1 Phase contrast microscopy of sperm in semen binding to an HA spot. Please observe the sperm bound to the perimeter of the HA with a head first orientation and the sperm tails pointing away from the HA. Some of the sperm in the field did not bind to the HA. Huszar. Hyaluronic acid-bound human sperm. Fertil Steril 2003. Fertility and Sterility 2003 79, 1616-1624DOI: (10.1016/S0015-0282(03)00402-3)

FIGURE 2 Time-related binding of sperm to HA in 15 semen samples (mean ± SEM). The percentage of HA-bound sperm was established by videotaping the process for 2-minute periods up to 22 minutes. Maximum binding occurred by the 8–10 minute period. Huszar. Hyaluronic acid-bound human sperm. Fertil Steril 2003. Fertility and Sterility 2003 79, 1616-1624DOI: (10.1016/S0015-0282(03)00402-3)

FIGURE 3 Hyaluronic acid binding by sperm in semen and by washed sperm after a 10-minute incubation period (○, 10 individual patients; ■, mean; N = 10 men studied). Huszar. Hyaluronic acid-bound human sperm. Fertil Steril 2003. Fertility and Sterility 2003 79, 1616-1624DOI: (10.1016/S0015-0282(03)00402-3)

FIGURE 4 (Left) CK-B immunocytochemistry of the initial semen and HA-bound sperm fractions for visualization of clear mature sperm, and sperm with various degrees of cytoplasmic retention. (a) Sperm originating in the initial semen fraction, (b) HA-bound sperm. The CL, I, and DK letters designate clear (mature), intermediate, and dark (diminished maturity) sperm. (Right) Aniline blue staining of sperm in the initial semen and HA-bound sperm fraction for detection of spermatozoa with various levels of persistent histone content. (c) Sperm originating in the initial semen. (d) HA-bound sperm. Huszar. Hyaluronic acid-bound human sperm. Fertil Steril 2003. Fertility and Sterility 2003 79, 1616-1624DOI: (10.1016/S0015-0282(03)00402-3)

FIGURE 5 The proportion of mature (□ clear), intermediate (▨), and immature (■ dark) sperm in the semen and HA-bound fractions. Brown bars: CK-B immunocytochemistry; Green bars: HspA2 immunocytochemistry; Blue bars: aniline blue stain. Huszar. Hyaluronic acid-bound human sperm. Fertil Steril 2003. Fertility and Sterility 2003 79, 1616-1624DOI: (10.1016/S0015-0282(03)00402-3)

FIGURE 6 Sperm viability and HA binding. Sperm treated with the FertiLight kit were applied to HA-coated slides, and after 15 minutes the slides were gently rinsed. (a) Semen sperm fraction. (b) Hyaluronic acid–bound sperm fraction after the rinsing step. Huszar. Hyaluronic acid-bound human sperm. Fertil Steril 2003. Fertility and Sterility 2003 79, 1616-1624DOI: (10.1016/S0015-0282(03)00402-3)

FIGURE 7 Hyaluronic acid–bound sperm fraction treated with fluorescein isothiocyanate conjugate–labeled lectin. (a) The fluorescence microscopy image indicates that all HA-bound sperm exhibit either bright (BR) or intact slightly patchy (SP) capacitating acrosomal membrane patterns. Sperm in which the capacitation pattern was more advanced did not bind to the HA-coated surfaces. (b) In the light microscopy picture of the same field, each sperm head is well identifiable. Huszar. Hyaluronic acid-bound human sperm. Fertil Steril 2003. Fertility and Sterility 2003 79, 1616-1624DOI: (10.1016/S0015-0282(03)00402-3)