Mol. Hum. Reprod. Advance Access originally published online on March 25, 2004
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Molecular Human Reproduction, Vol. 10, No. 5, pp. 365-372, 2004
© European Society of Human Reproduction and Embryology 2004
Cellular maturity and apoptosis in human sperm: creatine kinase, caspase-3 and Bcl-XL levels in mature and diminished maturity sperm
1Sperm Physiology and 2IVF Laboratories, Department of Obstetrics and Gynecology, Yale University School of Medicine, New Haven, CT 06525 USA and 3Department of Histology and Embryology, Akdeniz University, Antalya, 07070, Turkey
4 To whom correspondence should be addressed at: Sperm Physiology Laboratory, Department of Obstetrics and Gynecology, Yale School of Medicine, 333 Cedar Street, New Haven, CT 06510, USA. e-mail: gabor.huszar{at}yale.edu
The relationship between human sperm maturity and apoptosis is of interest because of the persistence of immature sperm in ejaculates in spite of various apoptotic processes during spermatogenesis. We assessed sperm maturity by HspA2 chaperone levels, and plasma membrane maturity by sperm binding to immobilized hyaluronic acid (HA). We also utilized objective morphometry. Sperm were stained with three antibody combinations: active caspase-3/creatine kinase (CK, a marker of cytoplasmic retention), caspase-3/the antiapoptotic Bcl-XL, and CK/Bcl-XL. In semen, 13% of sperm stained with CK, caspase-3 or Bcl-XL, and 28% had stained with two markers. In the mature HA-bound sperm fraction, <4% were single- or double-stained. Regarding sperm regions, CK staining, whether alone or as double staining, occurred in the head and midpiece (15–20%), whereas caspase-3 and Bcl-XL were primarily (>80% of sperm) in the midpiece. Morphometrical attributes of clear, single- and double-stained sperm, in line with their more pronounced maturation arrest, showed an incremental increase in head size (due to cytoplasmic retention) and shorter tail length. We hypothesize that during faulty sperm development, three alternatives may occur: (i) elimination of aberrant germ cells by apoptosis; (ii) in surviving immature cells, caspase-3 is activated, and in response the antiapoptotic Bcl-XL, and perhaps HspA2, provide protection; (iii) in a third type of immature sperm, in addition to the CK, caspase-3 and Bcl-XL expression, there are related manifestations of increased head size and shorter tail length. Thus, immature sperm may vary in the type of developmental arrest and in protection mechanisms for apoptosis. These variations are likely to explain the persistence of immature sperm in the ejaculate.
Key words: Key words: apoptosis/hyaluronic acid/maturity/morphometry/spermatogenesis/HspA2
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