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.