Treatment with or Elimination of Vascular Endothelial Growth Factor A (VEGFA) Results in Altered mRNA Abundance of Undifferentiated Spermatogonia Niche, Renewal and Differentiation Factors

.............................................................................................................................79 Introduction ........................................................................................................................80 Materials and Methods .......................................................................................................83 Results ................................................................................................................................90 Discussion ..........................................................................................................................95 Literature Cited ................................................................................................................118 Summary .........................................................................................................................122 List of Figures Chapter 1 Figure 1. Genetic basis of sexual differentiation from the bipotential gonad ...................24 Figure 2. Murine steroidogenesis ......................................................................................25 Figure 3. Spermatogonial Stem Cell Niche, Renewal and Differentiation .......................26 Figure 4. Depiction of human splice variants of VEGFA proangiogenic isoforms and antiangiogenic isoforms at the mRNA and protein levels .................................................27 Figure 5. Signal transduction of VEGFA164 compared to VEGFA165b ........................28 Figure 6. NRP-1 binds proangiogenic isoforms of VEGFA .............................................29 Figure 7. Proposed effects of VEGFA isoforms on self-renewal and differentiation of undifferentiated spermatogonia .........................................................................................30 Chapter 2 Table 1. Quantitative PCR primers, probes and kits .........................................................62 Figure 1. Treatment scheme for injecting perinatal rat pups and early testis differentiation and germ cell development ........................................................................63 Figure 2. Testis seminiferous cord and interstitial areas for pups treated with 0.5 μg PBS, VEGFA164 or VEGFA165b..............................................................................................64 Figure 3. Testis seminiferous cord and interstitial areas for pups treated with 1 μg PBS, VEGFA164 or VEGFA165b..............................................................................................65 Figure 4. Testis seminiferous cord and interstitial areas for pups treated with 1 μg IgG or antiVEGFAxxxb ................................................................................................................66 Figure 5. VASA-positive germ cells per cord for pups treated with 0.5 μg PBS, VEGFA164 or VEGFA165b..............................................................................................67 Figure 6. VASA-positive germ cells per cord for pups treated with 1 μg PBS, VEGFA164 or VEGFA165b..............................................................................................68 Figure 7. VASA-positive germ cells per cord for pups treated with IgG or antiVEGFAxxxb ................................................................................................................69 Figure 8. Messenger RNA abundance of genes involved in survival and apoptosis ........70