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Fig. 3. Distribution of the newly-born Ngn3- and Kit-positive
spermatogonia, and the seminiferous epithelial cycle prepattern.
(A-H) Double-staining ISH of testes sections at the indicated ages with
the indicated genes. Overlaid images of Kit or Ngn3 (purple,
bright field) and galectin 1 (red fluorescence) are shown in A,C,E,G; galectin
1 signals in the same field are shown in B,D,F,H. Arrows in A and C indicate
the Kit- and Ngn3-positive spermatogonia, respectively. E,F
and G,H are adjacent sections. Note the variable intensities of galectin 1
signals within the seminiferous tubule cross-sections and their relationship
with Kit- or Ngn3-positive spermatogonia (see Results). Asterisks (E-H)
represent typical segments with a high galectin 1 mRNA signal;
Kit-positive, but not Ngn3-positive, spermatogonia
preferentially localize to these segments. Segments marked by hearts show
lower galectin 1 signals and exhibit a preference for Ngn3-positive
spermatogonia. Scale bars: 100 µm, in D for A-D, in H for E-H. (I)
Quantification of Kit- or Ngn3-positive spermatogonia
localized in tubule segments categorized by different levels of galectin 1
mRNA (high, medium or low). Tubule cross-sections marked with H, M or L in B
and D are examples of segments with high, medium and low levels of galectin 1
mRNA, respectively. Data are represented as `preference' to each category of
tubule segment. a and b represent significant deviation with P-values
of <0.0001 and <0.05, respectively. See Materials and methods and Tables
S2-S7 in the supplementary material. The number of Ngn3-positive
spermatogonia at P3 and P4 was too low for statistical analyses. (J)
Model for the generation of spermatogonia subpopulations. One area of
seminiferous tubules is represented to align tubules with increasing age. The
galectin 1 mRNA level is shown by the gradient: black, highest; white, lowest.
As galectin 1 expression increases and reduces in cycles (black to gray to
white to black, etc.), its expression domain shifts leftward in a wave-like
manner. Kit-positive spermatogonia (red ovals) appear specifically at the
galectin 1-high segments (red bands). By contrast, Ngn3-positive spermatogonia
(green ovals) are generated separately from Kit-positive spermatogonia around
the segments of medium level of galectin 1 expression (green bands).
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