Influence of Inseminate Components on Porcine Leucocyte Migration In Vitro and In Vivo after Pre‐ and Post‐Ovulatory Insemination

Abstract

A post-breeding migration of leucocytes (PMN) into the uterus is considered to be an important reason for sperm losses. Minimizing such effects may be necessary for successful insemination with low sperm numbers, as required with sex-sorted spermatozoa. We examined the magnitude of PMN influx 3 h after pre- or post-ovulatory insemination with various combinations of seminal plasma (SP), semen extender Androhep (AH; Minitüb, Tiefenbach, Germany) and sperm preparations (S). Pre-ovulatory inseminations with preparations containing 98% AH caused a massive influx of PMN, independent of whether spermatozoa were present (628 +/- 189 x 10(6) leucocytes/uterine horn) or not (580 +/- 153 x 10(6)). Post-ovulatory, 98% AH caused a comparable immigration only in the absence of sperm cells (AH: 569 +/- 198 x 10(6), AH+S: 162 +/- 102 x 10(6)). The presence of SP significantly dampened the numbers of recruited uterine leucocytes. The reaction to all inseminates containing 98% SP both with and without spermatozoa, used before ovulation (SP: 14 +/- 6 x 10(6), SP+S: 73 +/- 27 x 10(6)) and after ovulation (SP: 60 +/- 32 x 10(6), SP+S: 51 +/- 33 x 10(6)) did not differ significantly from controls using phosphate buffered saline (PBS) (pre-ovulatory: 1 +/- 1 x 10(6), post-ovulatory: 11 +/- 9 x 10(6)). Quantitative in vitro transmigration assays with blood-derived PMN proved that AH-induced leucocyte migration into the uterus to be not as a result of direct chemotaxis, because, on account of the chelator citrate, AH significantly inhibited the transmigration towards recombinant human Interleukin-8 (rhCXCL8) (AH: 14 +/- 5% migration rate vs controls: 37 +/- 6%, p < 0.05). Supernatants of spermatozoa incubated in PBS for 1, 12 or 24 h showed neither chemoattractive nor chemotaxis-inhibiting properties. SP at > or =0.1% [v/v] significantly inhibited the in vitro transmigration of PMN. With respect to in vivo migration of neutrophils, the striking difference in the results between semen extender and seminal plasma suggests that adaptation of extender composition is needed to reflect more closely the in vivo regulatory potential of natural seminal plasma.

Effects of Heat Stress on the Well-Being, Fertility, and Hatchability of Chickens in the Northern Guinea Savannah Zone of Nigeria: A Review

“…In mammals, spermatozoa's binding with uterine epithelial cells is a strong index of spermatozoa viability and fertilizing capacity, implying that spermatozoa attachment to uterine epithelial cells is indicative of normal ultrastructure and mitochondrial membrane potential [57–59]. Töpfer-Petersen et al [60] and Ardón et al [61] showed that bound spermatozoa have both prolonged lifespan and better chromatin integrity than unbound spermatozoa cells.…”

Selection against spermatozoa with fragmented DNA after postovulatory mating depends on the type of damage

“…Two mechanisms could be responsible of this effect. On one hand, immune cells present in the uterine mucosa [41-43] such as neutrophils could extrude their nuclear DNA next to associated proteins to form neutrophil extracellular traps (NETs) [44] that retain spermatozoa with disturbed motility or plasma membrane. During NETs disaggregation, DNAses present in the seminal plasma [45,46] can act removing connections between neutrophils, attached sperm and proteins, allowing an efficient cleaning of the reproductive tract.…”

Methods for Improving In Vitro and In Vivo Boar Sperm Fertility

“…; Taylor et al. ). Although there is a hypothesis that the role of phagocytosis is to selectively remove sperm that have finished being functional, such as post‐capacitated cells (Eisenbach ), the incidences of PMNs phagocytized dead sperm in all treatments appear to be similar to those of PMNs phagocytized live cells (Matthijs et al.…”