146 SPERM STORAGE: EXPRESSION OF PROGESTERONE RECEPTORS, STRUCTURAL PROTEINS, AND HEAT SHOCK PROTEINS IN THE AVIAN OVIDUCT

Abstract

In avian species, sperm is stored for a long period in sperm storage tubules (SST) located in the utero-vaginal junction. It has been shown that progesterone (P4) regulates the release of sperm from SST for fertilization [Ito et al. 2011 Endocrinology 152, 3952–3621] in quails. In the same context, heat shock proteins (HSP) such as the HSP70 family activate sperm motility in quails [Hiyama et al. 2013 Reproduction 147, 167–178] and may participate in sperm release. In addition, we can hypothesise that at least some structural proteins from SST cells could play a role during sperm storage and release by interacting with sperm. Our objective was to study the potential effect of P4, HSP, and structural proteins on sperm storage potential, by analysing the expression of P4 receptors (PR and mPRα), HSPA8 and HSPB1, CNN1, TAGLN and DES genes in the avian genital tract. Uterus, vagina, and utero-vaginal junctions were collected from 2 divergent lines of hens. One lines displays a long period of sperm storage (21 days, line F+, n = 5), whereas the second displays a shorter period of sperm storage (10 days, line F–, n = 6). Total RNA was extracted and reverse transcribed, and then mRNA coding for PR, mPRα, HSPA8, HSPB1, CNN1, TAGLN and DES were assessed by qPCR (CFX96 Touch Real-Time PCR Detection System, Bio-Rad Laboratories, Hercules, CA, USA). Quantification was performed by using the relative standard curve method. Results were normalized by the geometric mean value of 2 reference genes, GAPDH and S17. Relative amounts of mRNA in each tissue were compared between lines by a Mann-Whitney test. Differences were considered significant when P < 0.05. No significant difference was observed between lines for both P4 receptors, as well as for HSPA8, regardless of the tissue. Expression of CNN1, TAGLN, and DES was significantly higher in the utero-vaginal junction of line F– than line F+ (P < 0.01). Expression of HSPB1 was significantly higher in utero-vaginal junction of line F– than of line F+ (P < 0.05). In conclusion, our data suggest that HSPB1 and structural proteins CNN1, TAGLN, and DES are involved in the regulation of sperm storage in the utero-vaginal junction in hens.