In vitrostudy on the effects of lead and mercury on porcine ovarian granulosa cells

Abstract

The heavy metals lead (Pb) and mercury (Hg) pose potential risks to sustainability of environment and thus to our future generations. General objective of this in vitro study was to examine the secretory activity of porcine ovarian granulosa cells after Pb and Hg administration and to outline the potential intracellular mediators of its effects. For this purpose, release of insulin-like growth factor I (IGF-I) and steroid hormone progesterone (P(4)), expression of proliferation- related (cyclin B1) and apoptosis-related (caspase-3) peptides was examined in porcine ovarian granulosa cells after heavy metals administration. Obtained data indicate Pb-induced inhibition of IGF-I release at lower doses (0.063 mg/mL and 0.046 mg/mL) by ovarian granulosa cells. However, P(4) release was not influenced by Pb addition, while the expression of cyclin B1 and caspase-3 was induced by Pb addition. These results indicate that Pb can affect the pathway of proliferation and apoptosis of porcine ovarian granulosa cells through intracellular substances such as cyclin B1 and caspase-3. On the other hand, the P(4) release by ovarian granulosa cells of pregnant gilts was stimulated by experimental Pb administration at doses of 0.25 mg/mL and 0.063 mg/mL and experimental Hg administration at doses 0.25 mg/mL and 0.083 mg/mL. P(4) release by ovarian cells of pregnant gilts was not influenced by a combinatory dose of FSH (1.0 ng/mL) + Pb (0.083 mg/mL) + Hg (0.083 mg/mL) but it was inhibited by experimental administration of FSH (10 ng/mL) + Pb (0.25 ng/mL) + Hg (0.25 ng/mL). Possible involvement of heavy metals - Pb and Hg and pituitary hormone FSH, in the regulation of P(4) release by porcine ovarian granulosa cells of pregnant gilts was noted. Data obtained from in vitro studies suggest the dose dependent association of heavy metals administration with the hormonal release by porcine ovarian granulosa cells. This association also depended on pregnancy of the gilts.

In vitroassessment of molybdenum-induced secretory activity, proliferation and apoptosis of porcine ovarian granulosa cells

“…Whether Mo exerts its effect centrally or directly on the pituitary has not been established. [20] The molecular pathways stimulated by lead [5] and cobalt, [6] the expression of cyclin B1 and caspase-3 were found in previous experiments. The expression of proliferation related peptide cyclin B1 in ovarian granulosa cells was affected by Mo addition at all the doses used (1.0 -0.09 mg/mL) which is in accordance with our previous study with trace metal Co. [6] Caspase-3 expression was also stimulated by Mo addition (1.0 and 0.5 mg/mL).…”

“…[42] In our study Mo was able to induce changes in porcine ovarian granulosa cells as it was published previously with chicken ovarian granulosa cells. [12] From earlier investigations we reported the effects of lead, mercury [4,5] and cobalt (Co) [6] on porcine ovarian granulosa cells. In the present study IGF-I release was decreased by Mo addition at the doses 1.0 mg/mLand 0.5 mg/mL.…”

Lead induced alterations in rabbit spermatozoa motility and morphology in vitro

“…After processing, the semen samples were incubated in the laboratory together with lead (lead chloride PbCl 2 ; Sigma-Aldrich, St. Louis, USA) in sterilized air-tight test tubes under three different temperature regimes -at laboratory temperature (22°C), in refrigerator (4°C), and in incubator (37°C). For in vitro exposure, PbCl 2 was diluted immediately before use and in five experimental concentrations was added to semen samples except for the control group based on our previous studies (Massanyi et al 2003(Massanyi et al , 2004(Massanyi et al , 2007Slivkova et al 2009;Kolesarova et al 2010;Tvrda et al 2013). The experimental concentrations of Pb were: 0 µg/ml (control group KF), 0.45 µg/ml (group 5F), 1.79 µg/ml (group 4F), 4.02 µg/ml (group 3F), 7.15 µg/ml (group 2F), and 11.17 µg/ml (group 1F), as presented in detail in Table 1.…”

“…This apparent specificity depends on the dose and the duration of action of the compounds (Massanyi et al 2007;Kolesarova et al 2010;Roychoudhury and Massanyi 2014). Previous studies by our group described the Pb concentrations in testis and semen of animals, human semen, and their correlations to spermatozoa quality (Massanyi et al 2003(Massanyi et al , 2004(Massanyi et al , 2007Slivkova et al 2009;Tvrda et al 2013).…”

Multi-Element Preconcentration/Separation of Some Metal Ions in Environmental Samples by Using Co-precipitation

“…However, heavy metals like zinc, iron, and copper are among the key nutrients that must be provided by dietary sources (9)(10)(11). The levels of these elements in environmental samples are generally in the ppb range and their determinations are influenced by highly saline environmental media (12)(13)(14)(15)(16). Prior to instrumental determinations of heavy metals at trace levels, an enrichmentseparation step is necessary (17)(18)(19)(20)(21)(22).…”