Faucette, Azure (2012-05). Changes in Gene Expression of Goat Developing Testes and Sperm During Breeding and Non-breeding Season. Doctoral Dissertation.
Testicular function is fundamental to male fertility, since testicular cells act in collaboration with each other to signal sex differentiation, the initiation of puberty and spermatogenesis. Complications that can be influenced by many factors will affect sperm number, morphology, motility, chromatin quality and acrosomal integrity. The purpose of these studies was to analyze the changes in gene expression in the developing testes and analyze the seasonal changes in gene products in sperm of mature bucks. In the first experiment, testes were harvested from five Alpine bucks at 0, 2, 4, 6, and 8 months of age. Northern and in situ hybridization indicated that the largest change in gene expression occurred during the first 4 months of goat testes development. Sex determining region Y-box 9 (SOX9) and Heat Shock protein A8 (HSPA8) peaked at 2 months of age, and were expressed in Sertoli cells and spermatogonium, respectively. At 4 months, expression of Stimulated by Retinoic Acid gene 8 (STRA8), Protamine1 (PRM1) and Outer Dense Fiber protein 2 (ODF2) was strongly up-regulated in early and maturing germ cells, respectively. In the second experiment, RNA from ejaculated spermatozoa collected from mature Alpine bucks in peak (October) and non-peak (April) breeding season were analyzed on a 4 x 44K Agilent bovine microarray. One thousand three hundred and eighteen gene products were differentially expressed 2-fold or more (p <= 0.05 ) was expressed in mature goat sperm collected October and April. To eliminate the likelihood of false positives, the cut off was set to fold change of 3 or more at p <= 0.01 which narrowed the list of genes to 50 transcripts. Real time PCR results confirmed the expression of Sperm Adhesion Molecule 1 (SPAM1) in April, and the expression of Glycerol kinase 2(GK2) and Myc Binding Protein 2 (MYCBP2) in October. Based on the results from both experiments, it can be concluded that: SOX9 and HSPA8 expression play an important role in tubular formation and germ cell maintenance; two months after SOX9 and HSPA8 expression, genes that are associated with spermatogenesis initiation and completion are upregulated; and validation of the seasonal changes in sperm mRNA levels may provide additional insight to testicular events as they relate to breeding and non-breeding season.