No abstract

Sperm motility can be maintained in vitro by incubation in a defined medium under specific conditions. In most studies, the exact role of various constituents of epididymal fluid, including calcium, has remained obscure. Most of the culture media have included millimolar concentrations of calcium, but previous reports have indicated that millimolar calcium inhibits sperm motility. In this present study, we sought the optimum concentration of extracellular calcium required for optimum sperm motility. This study showed that extracellular calcium has a concentration-dependent biphasic role in motility regulation. It promoted motility and velocity at lower (10 µM) concentration whereas notably inhibited it at higher concentrations. When external membrane-bound calcium was removed by ethylene glycol tetraacetic acid, motility decreased considerably. To confirm the motility-inhibiting role of calcium above 10 µM, a sperm motility-stimulating protein (MSP) recently reported from our laboratory was used which at 0.9 μM induces motility in 60-70 % cells. Calcium at 10 µM had no appreciable effect on the motility-promoting activity of the MSP but depressed the activity above 10 µM. Thus, our present results emphasize the biphasic role of extracellular calcium and the importance of its optimum concentration in different buffers and media used for sperm motility initiation.

Presently available contraceptives are mostly hormonal or detergent in nature with numerous side effects like irritation, lesion, inflammation in vagina, alteration of body homeostasis, etc. Antimicrobial peptides with spermicidal activity but without adverse effects may be suitable alternatives. In the present study, spermicidal activity of a cationic antimicrobial peptide VRP on human spermatozoa has been elucidated. Progressive forward motility of human spermatozoa was instantly stopped after 100 μM VRP treatment and at 350 μM, all kinds of sperm motility ceased within 20 s as assessed by the Sander-Cramer assay. The spermicidal effect was confirmed by eosin-nigrosin assay and HOS test. VRP treatment (100 μM) in human spermatozoa induced both the intrinsic and extrinsic pathways of apoptosis. TUNEL assay showed VRP treatment significantly disrupted the DNA integrity and changed the mitochondrial membrane permeability as evident from MPTP assay. AFM and SEM results depicted ultra structural changes including disruption of the acrosomal cap and plasma membrane of the head and midpiece region after treatment with 350 μM VRP. MTT assay showed after treatments with 100 and 350 μM of VRP for 24 hr, a substantial amount of Lactobacillus acidophilus (about 90% and 75%, respectively) remained viable. Hence, VRP being a small synthetic peptide with antimicrobial and spermicidal activity but tolerable to normal vaginal microflora, may be a suitable target for elucidating its contraceptive potentiality.

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