Arginine acts as a protective and reversal agent against glycolytic inhibitors in spermatozoa.

It is known that the amino acid arginine stimulates sperm motility and glycolytic activity. We have earlier studied its efficacy as a stimulator of glycolysis in goat spermatozoa under anaerobic conditions. Here, we have assessed the influence of arginine in reversing the impairment caused by glycolytic inhibitors, iodoacetamide and iodoacetic acid. Glycolysis has been monitored by measuring the consumption of 13C labeled glucose and the amount of 13C labeled lactate produced under different experimental conditions, using 13C NMR. It is observed that both L- and D-arginine are able to prevent and reverse the inhibitory action of glycolytic inhibitors. The reversal effect of arginine gives rise to about eight times higher metabolic activity as compared to the inhibited cells while structurally related amino acids such as nitro-arginine, homo-arginine, lysine and ornithine are ineffective. The energetics of spermatozoa as measured by 31P NMR show a reduction in ATP level in cells incubated with iodoacetamide. Treatment of these cells with both L- and D-arginine restores the ATP level. The results may have significance in the treatment of male infertility.

Distinct endoplasmic reticulum signaling pathways regulate apoptotic and necrotic cell death following iodoacetamide treatment.

Environmental stress induces the synthesis of glucose-regulated proteins (Grps) in the endoplasmic reticulum (ER) and heat shock proteins (Hsps) in the cytoplasm. Iodoacetamide (IDAM), a prototypical alkyating agent, induces both Grp and Hsp synthesis in renal epithelial cells and causes necrosis which is prevented by prior activation of the ER stress response (pre-ER stress) [Liu, H., et al. (1997) J. Biol. Chem. 272, 21751-21759]. In this study, we examined the biochemical pathways leading to IDAM-induced apoptosis and investigated the role of the ER stress response in apoptotic cell death. The antioxidant N,N'-diphenyl-p-phenylenediamine (DPPD) prevented necrosis after IDAM treatment, but the cells went on to die with hallmarks of apoptosis, i.e., cell detachment, caspase-3 activation, cleavage of poly(ADP-ribose)polymerase (PARP), and DNA-ladder formation, all of which were blocked by the general caspase inhibitor zVAD. As with IDAM-induced necrosis, dithiothreitol protected against apoptosis, but cell permeable calcium chelators did not, suggesting that distinct biochemical pathways mediate these two forms of cell death. Pre-ER stress, but not heat shock, prevented IDAM-induced apoptosis. pkASgrp78 cells are deficient in Grp78 induction due to expression of a grp78 antisense RNA and are more sensitive to necrosis. However, these cells were resistant to IDAM-induced apoptosis and had increased basal levels of Grp94 and a KDEL-containing protein of about 50 kDa. Thus, the expression of grp78 antisense perturbs ER functions and activates expression of other ER stress genes accounting for the resistance to apoptosis. Taken together, the data describe functionally distinct signaling pathways through which the ER regulates apoptosis and necrosis caused by chemical toxicants.

Glutathione: a protective agent in Salmonella typhimurium and Escherichia coli as measured by mutagenicity and by growth delay assays.

Cultures of some aerobically grown strains of Salmonella typhimurium and Escherichia coli contain up to 24 microM extracellular glutathione (GSH) [Owens RO, Hartman PE (1985): Environ Mutagen 7(Suppl 3): 47] in addition to having intracellular GSH concentrations in the millimolar range. The addition of 26 microM GSH to cultures of Salmonella typhimurium strain TA1534 partially protected the bacteria from the toxic effects causing growth delay by 54 microM N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). When MNNG was preincubated with equimolar GSH, the mutagenicity of the MNNG was neutralized. The addition of micromolar GSH to cultures of an Escherichia coli GSH- strain protected the cells from growth inhibition by micromolar concentrations of mercuric chloride, methyl mercuric chloride, silver nitrate, cisplatin, cadmium chloride, cadmium sulfate, and iodoacetamide. In the cases of mercuric chloride, cisplatin, MNNG, silver nitrate, and iodoacetamide, reaction products with GSH were detected by paper chromatography. In contrast to reduced GSH, micromolar concentrations of oxidized glutathione (GSSG) provided little or no protection and formed no detectable reaction products. Export of GSH by enteric bacteria may provide an important defense mechanism against exogenous toxic agents otherwise active in the micromolar range.