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ARS Home » Northeast Area » Beltsville, Maryland (BARC) » Beltsville Agricultural Research Center » Animal Biosciences & Biotechnology Laboratory » Research » Publications at this Location » Publication #352082
Title: Characterization of gene expression in turkey sperm storage tubules from non-inseminated hens over the course of reproduction

Author
item KRASNEC, KATINA - US Department Of Agriculture (USDA)
item Long, Julie

Submitted to: Poultry Science Association Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 7/23/2018
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Sperm storage in the female reproductive tract is a known biological feature for a wide range of species. In the turkey, sperm residing in the hen's sperm storage tubules (SST) retain fertilizing ability for 10 weeks after a single insemination; however, the molecular and cellular mechanisms are largely undefined. A subset of data from a larger study involving non-inseminated, sham-inseminated, and sperm-inseminated hens was analyzed to evaluate SST gene expression over time in the non-inseminated hens. Reproductive tracts were recovered at the onset (day 1, n=3), peak (day 30, n=4), and end (day 90, n=3) of reproduction. The region of the utero-vaginal junction (UVJ) containing SSTs was dissected and flash frozen. Laser capture microdissection of cryosections was used to isolate and remove SST cells from the surrounding vaginal epithelial tissue for RNA extraction. Approximately 25-30 million fragments of 150bp paired-end reads per sample were generated using RNASeq (Illumina NextSeq). Sequences were aligned to the turkey reference genome and annotations were used to identify gene transcripts. Differential gene expression was assessed using pairwise comparisons (day 1 vs day 30; day 1 vs day 90; day 30 vs day 90) to identify genes that were significantly (+/- 1.5-fold change; p<0.05) up- or down-regulated. Differentially expressed genes were broadly categorized as: SST microenvironment; immune function; vascularization; cell structure; cell migration/proliferation; and apoptosis. Genes involved with lipid synthesis, metabolism, and peroxidation (e.g. CAT, CH25H, LPL, PLCD1) were up-regulated at peak reproduction compared with onset and end. The majority of genes associated with potassium and calcium binding or channel formation (e.g. CALCRL, FBLN2, KCHJ15, KCNK5, TMTC1) were up-regulated at the onset of reproduction; however at peak reproduction, KCNMB1, TRPV4 and TRPC4 were up-regulated. Cell migration and proliferation genes (EPHBG, FSTL1, POSTN, PRKX) were up-regulated at onset and declined at peak through the end of reproduction, except for CTGF which was up-regulated at peak and declined at the end. Genes involved in apoptosis (ELMO1, AEN) were only found to be up-regulated at the end of reproduction. Results indicate that specific genes are associated with the molecular function of the SST during the reproductive lifespan of commercial turkey hens. These insights, paired with examination of gene expression from the SSTs of inseminated hens, may enable a greater understanding of SST function that would help improve in vitro semen storage methods and/or fertility.