Journal: Proceedings of the National Academy of Sciences of the United States of America
Article Title: A gene family required for human germ cell development evolved from an ancient meiotic gene conserved in metazoans
Figure Lengend Snippet: BOULE expression is distinct from that of DAZ and DAZL. DAZ is expressed in prenatal primordial germ cells, spermatogonial stem cells, and spermatocytes. DAZL is expressed in both the male and female germ line. BOULE is expressed in the cytoplasm of pachytene spermatocytes, persists through meiosis, and decreases in early spermatids. ( A ). ( B ). ( C ) Human testis section stained using antisera that recognize DAZ only. DAZ is expressed in spermatogonia and early spermatocytes, but is absent from late spermatocytes or postmeiotic cells. ( D ). ( E ) A Northern blot with polyadenylated RNA from different human tissues. Blot was hybridized with human BOULE cDNA that detects two testes specific transcripts. Lanes: 1, spleen; 2, thymus; 3, prostate; 4, testis; 5, ovary; 6, small intestine; 7, colon; and 8, leukocyte. ( F ) A Northern blot with polyadenylated RNA from mouse tissues. Blot was hybridized with human BOULE cDNA that detects three testes-specific transcripts. Lanes: 1, heart; 2, brain; 3, spleen; 4, lung; 5, liver; 6, muscle; 7, kidney; and 8, testis. ( G ) Anti-BOULE antisera detects a single 32-kDa protein in mouse testes ( b ) and a similar size protein in human testes ( a ) but not in other human or mouse tissues (data not shown), nor does it recognize DAZ protein expressed in yeast strain, RRY618 ( c ). The 50-kDa band in human testes is nonspecific as it is detected by preimmune also. ( H ) BOULE staining in human testis section is also restricted to cytoplasm of spermatocytes; no staining of spermatogonial stem cells is observed. ( I ) Stage III seminiferous tubules. BOULE is expressed in round spermatids (Spd) and secondary spermatocytes but not in spermatogonia (Spg) or primary spermatocytes (Spc). ( J ) Stage VII seminiferous tubule. BOULE is expressed in the cytoplasm of pachytene spermatocytes. There is no staining in spermatogonia and spermatids. ( K ) Stage X–XI seminiferous tubules. BOULE expression peaks in late pachytene stage spermatocytes. ( L and M ) Lower-magnification view of staining with preimmune and anti-BOULE antisera. ( L ) Preimmune of BOULE antisera (×200). ( M ).
Article Snippet: Northern hybridization was done on polyadenylated RNA blots (CLONTECH) using human and mouse cDNA clones.
Techniques: Expressing, Staining, Northern Blot