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Supplementary Material for: Expression Profile of NOTCH3 in Mouse Spermatogonia
<p>Stable and sustainable spermatogenesis is supported by the strict
regulation of self-renewal and differentiation of spermatogonial stem
cells (SSC), which are a rare population of undifferentiated
spermatogonia. It has been revealed that some signaling factors regulate
the self-renewal of SSC; however, the molecular mechanism of SSC
maintenance is still not completely understood. Notch signaling is an
evolutionarily conserved juxtacrine signaling that plays important roles
in the cell fate determination of various tissue stem cells. Recently,
analyses of loss- and gain-of-function suggested that Notch signaling
was necessary for normal spermatogenesis. However, the expression of
Notch signal components in spermatogonia is still unclear. Here, we
analyzed the distribution of NOTCH3-expressing spermatogonia and the
target genes. Double immunostaining with differentiation markers
revealed that NOTCH3 was expressed in some undifferentiated and
differentiated spermatogonia in mouse testes. To define the target gene
of Notch3 signaling in spermatogonia, we analyzed the mRNA expression
pattern of <i>Hes</i> and <i>Hey</i> family genes during testis development. <i>Hes1</i> abundance was decreased during testis development, suggesting that spermatogonia may express <i>Hes1</i>.
Immunohistochemical analysis showed that HES1 was expressed in
prepubertal spermatogonia, whereas it was expressed predominantly in
adult Sertoli cells and weakly in adult spermatogonia. Furthermore,
NOTCH3-HES1 double-positive spermatogonia were in pup and adult testes.
These results suggest that Notch3 signaling in spermatogonia could
promote <i>Hes1</i> expression.</p