Electroporation of cell membranes.

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

Loss of forward motility of rabbit epididymal spermatozoa in high K phosphate buffer is in- hibited by taurine, hypotaurine, epinephrine and bovine serum albumin. Pyruvate and lactate also show this effect. The rate of lipid peroxidation in these spermatozoa, as measured by rate of formation of malonaldialdehyde, is also inhibited by these agents. A close linear correlation be- tween percent inert spermatozoa and malondialdehyde was found, which was independent of the rate of peroxidation. Complete cessation of motility was observed at 0.5 nmol malondialdehyde/ iocells in the absence or presence of these agents, which is the same value found in other sus- pending media in a previous study LAlvarez and Storey (1982) Biol. Reprod. 27:1102-11081. Albumin was the most effective agent in preventing loss of motility and inhibiting lipid peroxida- tion. Hypotaurine was the next most effective, followed by taurine, epinephrine, pyruvate and lactate. Hypotaurine reduces the amount of rate of superoxide production, as measured by the rate of reduction of acetylated ferricytochrome c by 0, from rabbit sperm under these conditions and concomitantly reduces inactivation of the superoxide dismutase in these cells. Since superoxide seems to be the major inducer of lipid peroxidation in rabbit sperm, the protective effect of hypo- taurine, which should be readily permeant to the plasma membrane, may be ascribed to scavenging of intracellular superoxide. The mechanism of the protective action of albumin is not known. Rabbit epididymal spermatozoa lose motility over time if Ca2� or Mg2 are omitted from the suspending medium. This loss is not correlated with the rate of lipid peroxidation, which is un- affected by the absence or presence of these ions. The linear correlation between loss of motility and lipid peroxidation no longer holds in the absence of Ca2* and Mg2�, implying motility loss by a different mechanism.

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Taurine, hypotaurine, epinephrine and albumin inhibit lipid peroxidation in rabbit spermatozoa and protect against loss of motility.

Electrification in developed countries has progressively increased the mean level of extremely low-frequency electromagnetic fields (ELF-EMFs) to which populations are exposed; these humanmade fields are substantially above the naturally occurring ambient electric and magnetic fields of approximately 10(-4) Vm(-1) and approximately 10(-13) T, respectively. Several epidemiological studies have concluded that ELF-EMFs may be linked to an increased risk of cancer, particularly childhood leukemia. These observations have been reinforced by cellular studies reporting EMF-induced effects on biological systems, most notably on the activity of components of the pathways that regulate cell proliferation. However, the limited number of attempts to directly replicate these experimental findings have been almost uniformly unsuccessful, and no EMF-induced biological response has yet been replicated in independent laboratories. Many of the most well-defined effects have come from gene expression studies; several attempts have been made recently to repeat these key findings. This review analyses these studies and summarizes other reports of major cellular responses to EMFs and the published attempts at replication. The opening sections discuss quantitative aspects of exposure to EMFs and the incidence of cancers that have been correlated with such fields. The concluding section considers the problems that confront research in this area and suggests feasible strategies.

Biological responses to electromagnetic fields

During the past decade considerable evidence has accumulated demonstrating that nonthermal exposures of cells of the immune system to extremely low-frequency (ELF) electromagnetic fields (< 300 Hz) can elicit cellular changes that might be relevant to in vivo immune activity. A similar responsiveness to nonionizing electromagnetic energy in this frequency range has also been documented for tissues of the neuroendocrine and musculoskeletal system. However, knowledge about the underlying biological mechanisms by which such fields can induce cellular changes is still very limited. It is generally believed that the cell membrane and Ca(2+)-regulated activity is involved in bioactive ELF field coupling to living systems. This article begins with a short review of the current state of knowledge concerning the effects of nonthermal levels of ELF electromagnetic fields on the biochemistry and activity of immune cells and then closely examines new results that suggest a role for Ca2+ in the induction of these cell...

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Electromagnetic field effects on cells of the immune system: the role of calcium signaling.

Increased levels of hsp70 transcripts induced when cells are exposed to low frequency electromagnetic fields

Enhancement of cytochrome oxidase activity in 60 Hz magnetic fields

Alternating currents can increase or decrease the ATP-splitting activity of the membrane enzyme Na,K-ATPase. Either change depends on the AC frequency, and the greatest effect appears to be in the ELF range at about 100 Hz. The threshold for enzyme inhibition by AC was determined, and it is estimated to be an internal electric field circa 5 μV/cm. The corresponding current-density threshold approximates 8 nA/cm2. 1992 Wiley-Liss, Inc.

Threshold for inhibition of Na, K-ATPase by ELF alternating currents

The Moving Charge Interaction (MCI) model proposes that low frequency electromagnetic (EM) fields affect biochemical reactions through interaction with moving electrons. Thus, EM field activation of genes, and the synthesis of stress proteins, are initiated through EM field interaction with moving electrons in DNA. This idea is supported by studies showing that EM fields increase electron transfer rates in cytochrome oxidase. Also, in studies of the Na,K-ATPase reaction, estimates of the speed of the charges accelerated by EM fields suggest that they too are electrons. To demonstrate EM field effects on electron transfer in a simpler system, we have studied the classic oscillating Belousov--Zhabotinski (BZ) reaction. Under conditions where the BZ reaction oscillates at about 0.03 cycles/sec, a 60 Hz, 28 microT (280 mG) field accelerates the overall reaction. As observed in earlier studies, an increase in temperature accelerates the reaction and decreases the effect of EM fields on electron transfer. In all three reactions studied, EM fields accelerate electron transfer, and appear to compete with the intrinsic chemical forces driving the reactions. The MCI model provides a reasonable explanation of these observations.

Lily Soo

Papers

Increased levels of hsp70 transcripts induced when cells are exposed to low frequency electromagnetic fields

Enhancement of cytochrome oxidase activity in 60 Hz magnetic fields

Threshold for inhibition of Na, K-ATPase by ELF alternating currents

Electromagnetic acceleration of electron transfer reactions.

The threshold for Na,K-ATPase stimulation by electromagnetic fields