Flow cytometry evaluation of lead and cadmium effects on mouse spermatogenesis

Abstract

Flow cytometry (FCM) is a powerful tool to evaluate cell DNA content and ploidy levels. We have assessed the accuracy of two protocols of nuclei isolation from paraffinized samples (P1 and P2) by comparing FCM results with those obtained using fresh material (F1-F3). After isolation, nuclei were stained with propidium iodide and quantitatively analysed by FCM for changes in germ cell ratios. Results obtained with Protocol P2 were similar to those obtained using the protocol that gave best results for fresh tissues (F2). Protocol P2 was then applied to paraffin embedded testicular samples from ICR-CD1 mice exposed to 1, 2 and 3 mg CdCl(2)/kg bw by single subcutaneous injection, and to 74 and 100 mg PbCl(2)/kg bw administered in four repeated doses. The highest doses of CdCl(2) decreased the number of haploid (1C) cells and increased the number of diploid (2C), S phase and tetraploid (4C) cells. Treatment with PbCl(2) did not induce significant changes in testicular cells subpopulations. These results support the usefulness of FCM in evaluating the effect of toxic substances on mouse spermatogenesis, using both fresh and paraffinized material.

Evaluating the ameliorative efficacy of Spirulina platensis on spermatogenesis and steroidogenesis in cadmium-intoxicated rats

“…Similar result was obtained by Oliveira et al (2006). The decreased sperm motility in Cd-treated rats could be attributed to the ability of Cd to bind with calmodulin instead of calcium inhibiting its role in sperm motility (Schlingmann et al 2007).…”

Lead chloride affects sperm motility and acrosome reaction in mice

“…Previous work from our laboratory, investigating the effects of lead chloride (at 74 e 100 mg PbCl 2 /kg bw) on mouse spermatogenesis, showed an increase in the percentage of cells in S-phase, suggesting that this compound might have a mitogenic effect (Oliveira et al 2006). In this context, spermatogenesis and sperm function could also be affected by exposure to these concentrations of lead.…”

“…The subcutaneous route was selected since it allows a slow absorption of lead and allows a higher control of the experiment and has been previously used in other studies with lead (Graça et al 2004;Oliveira et al 2006) and other metals [e.g. cadmium (Zhou et al 2004)].…”

“…cadmium (Zhou et al 2004)]. Three groups of ten mice each were subcutaneously injected with 74 and 100 mg PbCl 2 /kg bw for four consecutive days, as described previously (Oliveira et al 2006). Control groups of ten mice each were injected with saline vehicle (0.9% NaCl).…”

The protective effect of ascorbic acid and thiamine supplementation against damage caused by lead in the testes of mice

“…Adhikari et al [1] also reported that Pb could induce apoptosis in the germ cells within the semineferous tubules. Although adverse effects of lead compounds on the testis have long been studied [5] , effective drugs reversing the toxicity of lead on male reproduction are still scarce and insufficient.…”