Inhibition of PIM1 kinase
Four male tree sparrows captured from LJX in 2022 were dosed daily by oral gavage at 100 mg/kg of SGI-1776, an experimental PIM1 kinase small molecule inhibitor, and the same numbers of LJX males were used as control group. SGI-1776 at 100 mg/kg was previously shown to be effective in abrogating tumor growth in mice (Chen et al., 2011). After 1 week of drug treatment was completed, all birds were euthanized and dissected. Total RNA was extracted from testis and used for sequencing. Their left seminal glomera were extracted and placed in 500 μL pre-warmed (40 ℃) Hank’s Balanced Salt Solution with an incision to obtain sperm suspensions (Yang et al, 2019). After 3 minutes dilution, the sperm suspensions were used to assess curvilinear velocity as an index of sperm velocity using a computer-assisted sperm analysis (CASA) system. Then the sperm suspensions were placed on a glass slide and smeared using another slide. The air-dried smeared slides were fixed with 5% formalin for 10 min and stained using Wright-Giemsa staining solution. The slides were used to assess sperm length at 400 × magnification.