The selenoprotein phospholipid hydroperoxide glutathione peroxidase (PHGPx) changes its physical characteristics and biological functions during sperm maturation PHGPx exists as a soluble peroxidase in spermatids but persists in mature spermatozoa as an enzymatically inactive, oxidatively cross-linked, insoluble protein In the midpiece of mature spermatozoa, PHGPx protein represents at least 50 percent of the capsule material that embeds the helix of mitochondria The role of PHGPx as a structural protein may explain the mechanical instability of the mitochondrial midpiece that is observed in selenium deficiency

https://science.sciencemag.org/content/sci/285/5432/1393.full.pdf

Dual Function of the Selenoprotein PHGPx During Sperm Maturation

The spermatozoa of most invertebrates (e.g., sea urchins) and nonmammalian vertebrates (e.g., fishes and amphibians) have full capacity to fertilize eggs upon leaving the testis. Testicular spermatozoa of mammals, on the other hand, do not possess the ability to do so. Their fertilizing capacity develops as they pass through the epididymis (Young, 1931; Nishikawa and Waide, 1952; Blandau and Rumery, 1964; Bedford, 1966; Orgebin-Crist, 1967). This process, apparently “unique” to mammals, is referred to as the epididymal maturation of spermatozoa. Even after their maturation, however, spermatozoa require an additional phase of maturation or capacitation within the female genital tract before they are able to fertilize eggs (Austin, 1951, 1952; Chang, 1951a). Thus, epididymal maturation and capacitation are two extra steps that mammaliam spermatozoa must take before they can effect fertilization. In this chapter, I will discuss how mammalian spermatozoa prepare themselves for fertilization and how the spermatozoa and eggs interact during fertilization. The process and mechanisms of sperm transport in the female genital tract will not be dealt with extensively here. Readers are referred to Bishop (1961, 1969), Blandau (1969), Bedford (1970b, 1972b), Thibault (1972, 1973a), Zamboni (1972), Blandau and Gaddum-Rosse (1974), Hafez and Thibault (1975), Overstreet and Katz (1977), Overstreet and Cooper (1978, 1979b), Overstreet et al. (1978), Shalgi and Kraicer(1978), Cooper et al. (1979) and Hunter (1975, 1980). The rejection or elimination of extra spermatozoa by the fertilized egg, one of the most fascinating events in fertilization, will not be discussed here. Instead, readers are referred to the chapter by Dr. Wolf in this volume. The physiology of egg activation has been described and discussed to some extent by Gwatkin (1977) and Yanagimachi (1978a). There are numerous reviews dealing with general aspects of mammalian fertilization. The following are recommended to aid in grasping the outline of mammalian fertilization: Austin and Bishop (1957), Austin and Walton (1960), Austin (1961, 1968), Blandau (1961), Piko (1969), Thibault (1969), Bedford (1970a,b, 1972b), Gwatkin (1976, 1977), Yanagimachi (1977, 1978a), Bedford and Cooper (1978) and Hunter (1980).

Mechanisms of Fertilization in Mammals

BACKGROUND: Sperm hyperactivation is critical to fertilization, because it is required for penetration of the zona pellucida. Hyperactivation may also facilitate release of sperm from the oviductal storage reservoir and may propel sperm through mucus in the oviductal lumen and the matrix of the cumulus oophorus. Hyperactivation is characterized by high amplitude, asymmetrical flagellar bending. METHODS: This is a review of the original literature on the mechanisms that regulate hyperactivation, including physiological factors and signaling pathways. RESULTS: Computer-assisted semen analysis systems can be used to identify hyperactivated sperm by setting minimum thresholds for curvilinear velocity (VSL) and lateral head movement and a maximum threshold for path linearity. Hyperactivation is triggered by a rise in flagellar Ca(2+) resulting from influx primarily through plasma membrane CatSper channels and possibly also by release of Ca(2+) from a store in the redundant nuclear envelope. It requires increased pH and ATP production. The physiological signals that trigger the rise in Ca(2+) remain elusive, but there is evidence that the increased Ca(2+) acts through a calmodulin/calmodulin kinase pathway. Hyperactivation is considered part of the capacitation process; however, the regulatory pathway that triggers hyperactivation can operate independently from that which prepares sperm to undergo the acrosome reaction. Hyperactivation may be modulated by chemotactic signals to turn sperm toward the oocyte. CONCLUSIONS: Little is known about exactly what triggers hyperactivation in human sperm. This information could enable clinicians to develop reliable fertility assays to assess normal hyperactivation in human sperm samples.

/pdf/control-of-hyperactivation-in-sperm-1uizccscs2.pdf

Control of hyperactivation in sperm

▪ Abstract Mammalian selenium-containing proteins can be divided into three groups: proteins containing nonspecifically incorporated selenium, specific selenium-binding proteins, and specific selenocysteine-containing selenoproteins Selenoproteins with known functions identified so far include five glutathione peroxidases, two deiodinases, several thioredoxin reductases, and selenophosphate synthetase 2 Alternative splicing leads to a greater variety of selenoproteins, as was shown in the cases of a specific sperm nuclei glutathione peroxidase and some thioredoxin reductases Selenoprotein P, selenoprotein W, a 15-kDa selenoprotein, an 18-kDa selenoprotein, and several selenoproteins identified in silico from nucleotide sequence databases were found to contain selenocysteine but their functions are not known Gel electrophoretic separation of tissue samples from rats labeled in vivo with 75Se showed the existence of further selenium-containing proteins

Mammalian selenium-containing proteins.

Evidence for specific selenium target tissues and new biologically important selenoproteins

Evidence that selenium in rat sperm is associated with a cysteine-rich structural protein of the mitochondrial capsules

The motility of rabbit spermatozoa recovered from the vagina, endocervix, uterus, and four regions of the oviduct was assessed visually by phase-contrast microscopy at intervals from one minute to 16 hours after a single mating. The percentage of motile cells in each sample was dependent on the temperature of recovery, ie, 23° vs 37°C, but was not influenced by the temperature of observation. Spermatozoa in the lower isthmus of the oviduct were the most temperature sensitive population to recovery at 23°C. When all manipulations and observations were performed at 37°C, the percentage of spermatozoa with progressive motility varied according to the region sampled and interval after mating. Populations from the vagina, uterus and upper regions of the oviduct usually had a high proportion of progressively motile cells with vigorous flagellar activity. Fewer spermatozoa showed progressive movement on recovery from the endocervix and lower 2 cm of the tubal isthmus and their flagellar activity was generally depressed. The decrease in flagellar beat frequency noted in the latter regions may be a major factor limiting sperm ascent in the female tract. A unique pattern of “activated” motility was seen exclusively in populations taken from the oviducts at 6 to 16 hours after mating. This motility pattern, consisting of alternating episodes of linear progressive and vigorous nonprogressive movement, may be analogous to the activated motility described for capacitated rodent spermatozoa.

The motility of rabbit spermatozoa recovered from the female reproductive tract

Rabbit spermatozoa recovered from the reproductive tract of females 12–13 hr postcoitum resisted head/tail separation induced by n-butylamine and dithiothreitol, but not sodium dodecylsulfate. Stabilization of the sperm head/tail junction also developed in vitro at 37°C in physiological media and in saline-Tris-HCl (pH 7.4). Resistance to dithiothreitol occurred in motile, but not immotile spermatozoa. Only nonmotile spermatozoa developed resistance to sodium dodecylsulfate in vitro, whereas both motile and immotile spermatozoa became resistant to n-butylamine. Stabilization to n-butylamine was time and temperature dependent and was accelerated by Cu2+, Mg2+, and Zn2+, but not Mn2+. The resistance of hamster and rabbit spermatozoa to n-butylamine developed in physiological media over the same time intervals as required for capacitation and the acquisition of hyperactivated motility.



Reagents that react with sulfhydryl groups had no effect on the development of resistance to n-butylamine but inhibited stabilization to sodium dodecylsulfate, suggesting that the latter stabilization may result from the formation of disulfide crosslinks at the head/tail junction. Reduction of aldehyde groups by sodium cyanoborohydride did not prevent stabilization to sodium dodecylsulfate, but did reduce detachment by dithiothreitol. Aldehyde groups thus are not involved in the stabilization of the head/tail junction to sodium dodecylsulfate, but may participate in new crosslinks stabilizing the head/tail junction to dithiothreitol. Inhibitors of transglutaminase did not prevent development of resistance to n-butylamine, sodium dodecylsulfate, or dithiothreitol indicating that head/tail stabilization does not involve intermolecular γ-glutamyl-ϵ-dysine bonds.

George W. Cooper

Papers

Evidence that selenium in rat sperm is associated with a cysteine-rich structural protein of the mitochondrial capsules

The motility of rabbit spermatozoa recovered from the female reproductive tract

Stabilization in vivo and in vitro of the spermatozoon head and tail to detachment induced by primary amine, thiol, and detergent

Characterization of humoral antibodies reactive with spermatozoa, N-acetyl galactosamine, and a putative blood group antigen in seminal plasma.