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High concentration of growth differentiation factor-9 (GDF-9) to the ram semen had a negative effect on the sperm positive rheotaxis.

Herein, the microfluidic device technique was used to investigate the effects of GDF-9 concentrations and exposure time on the ram sperm positive rheotaxis (PR). Semen was collected from six rams and utilized for PR, motility, and sperm kinetic parameters analysis using computer-assisted sperm analysis with controlled flow velocity following 0, 10, 20, or 30 min of incubation at 37°C with GDF-9 (200 ng/mL, 400 ng/mL, or 600 ng/mL; semen sample without GDF-9 was used as a control). Results revealed that there was not an interaction between effects of GDF-9 concentrations and incubation duration on PR% (P = 0.457) and TM% (P = 0.099). A Simple main effects analysis showed that GDF-9 concentrations had an effect on PR% (P = 0.003). However, the incubation duration did not had an effect on PR% (P = 0.101). GDF-9 concentrations had not an effect on TM% (P = 0.817). In contrast, the incubation duration affected on TM% (P = 0.026). A higher PR% was found (P < 0.05) at 200 ng GDF-9 after 10 min (46.7±10.3), and 20 min (45.5±11.5) of incubation. After 30 min incubation, the PR% was lowest (P < 0.05) at 400 ng of GDF-9 (30.6±14.1), and 600 ng of GDF-9 (32.2±9.6). There was no difference (P > 0.05) in the sperm kinetic parameters between the four treatment groups. In conclusion, the ram sperms had the best rheotaxis properties after 10 and 20 min of incubation with 200 ng of GDF-9, and were sensitive to high concretions.