Expr1746

The distribution of TRA-1 in the germlines of wild-type XX L4 hermaphrodites, tra-2(lf) and tra-2(lf); fem-1(lf) animals was also analyzed. Similar to the intestine, the distribution of TRA-1 is dependent on the sexual state of the tissue. In L4 hermaphrodite and tra-2(lf) germlines, which make only sperm, TRA-1 staining is similar to adult males; TRA-1 nuclear levels are high only in the mitotic region. However, in tra-2(lf); fem-1(lf) germlines, which make only oocytes, TRA-1 nuclear levels are high across the germline. The fact that TRA-1 nuclear levels are high throughout the germline suggests that fem-1 may regulate nuclear TRA-1 levels in the meitotic pachytene region of wild-type adult hermaphrodites.

Expr9997

spe-5 mRNA is dramatically up regulated in wild type (wt) males as compared to spe-5 mutant males and, also, in fem-3 gain of function (gf; female soma, male germline) as compared to fem-3 loss of function (lf; female soma, female germline) worms. Adult wt hermaphrodites continue to show significant spe-5 expression after they have completed spermatogenesis, as do fem-3 loss of function (lf; female soma, female germline) worms. These data indicate that, while spe-5 expression is up regulated during spermatogenesis, it is also expressed in one or more other tissues.

Expr13404

cdc-25.3 mRNA is highly expressed in N2 hermaphrodites at the young-adult stage, when oogenesis actively occurs, and in feminized fem-1(lf) hermaphrodites, but not in adult males or in masculinized fem-3(gf) hermaphrodites. These results indicate that cdc-25.3 is preferentially expressed during oogenesis.

Expr3675

Compared to wild-type expression, ccf-1 is present at a higher level (148%) in fem-1(lf) adults that produce oocytes but no sperm. ccf-1 expression is reduced in fem-3(gf) (52%) and glp-4(lf) (31%) which produce sperm but no oocytes and have a severely reduced and undifferentiated germline, respectively. The higher expression of ccf-1 in fem-3(gf) than in glp-4 adults could suggest a weak expression of ccf-1 in the male germline. Altogether these results suggest that ccf-1, although present at all stages, is expressed at a higher level in embryos and adults, in which expression is dependant upon the germ line.

Expr10041

In situ hybridizations of dissected gonads were performed to ascertain the precise spatial pattern of spe-44 expression. The fem- 3(gf) and fem-1(lf) mutations were used to restrict hermaphrodite gametogenesis to sperm or oocyte production, respectively. Abundant expression was first detected in fem-3(gf) animals during the L3 stage in the early meiotic germline. DAPI staining of the chromatin indicated that expression coincides with early pachytene of meiosis I. By contrast, spe-44 expression was undetectable in fem-1(lf) germlines at any stage of development. Thus, spe-44 expression is restricted to the sperm-producing germline at a time and place consistent with the regulation of sperm gene expression.

Expr12068

An egl-46 transcriptional reporter confirmed the previously published expression pattern for egl-46 in the FLP, HSN and PVD neurons and other neurons in the head. In addition, the egl-46 reporter colocalized with egl-13::gfp reporter (expressed in BAG, URX, AQR and PQR) in the BAG neurons at the L2 and L3 stages of larval development; suggesting that egl-46 is expressed during short temporal window in the BAG neurons, as has been observed in other neurons previously (WU et al. 2001). Note: we only rarely observed colocalization with the BAG marker, suggesting that the egl-46 promoter drives expression in the BAG neurons in a transient manner.

Expr1941

spe-6 mRNA could be amplified from fem-3(q23gf) hermaphrodite mRNA (this mutant has hermphrodite soma but makes only sperm germline), but not not from fem-1(hc17ts) hermaphrodite mRNA (mutant hermaphrodites make only oocytes, no sperm); cDNA clone ca01c03 (GenBank accesion nos. AW057333 and AW057136) are from spermatogenesis-specific libraries; SPE-6 protein only detected in extracts from sperm-producing worms (i.e. fem-3(gf), not fem-1(lf) on western blots.

Expr11093

egl-1 appears to be specifically expressed in cells destined to die during embryogenesis, including the NSM sister cells. The NSM sister cells normally die during a stage called the 1.5-fold stage of embryogenesis. At this stage of development it is difficult to identify cells on the basis of their position within the embryo. The authors therefore analysed the expression of Pegl-1his24-gfp in the background of the ced-3 lf mutation n717. ced-3(n717) blocks programmed cell death, including the death of the NSM sister cells. They found that the activity of EGL-1 is regulated at the transcriptional level in the NSMs and NSM sister cells and, hence, that the cell-death fate of these cells is specified by egl- 1 expression.

Expr8942

cdc-25.2 mRNA was hardly detectable in glp-1(q224) hermaphrodites compared with N2 hermaphrodites, indicating that cdc-25.2 is preferentially expressed in the germ line. In N2 hermaphrodites, expression of cdc-25.2 mRNA was relatively low until L4 larval stage, but the expression increased significantly in young adult (YA; just after the L4 to adult molting but before egg laying has started) and adult (Ad; egg-laying gravid adult worms) stages, when oogenesis actively occurs in hermaphrodite worms. Furthermore, although cdc-25.2 mRNA was expressed in feminized fem-1(hc17lf) hermaphrodites, it was barely detectable in N2 males and in masculinized fem-3(q20gf) hermaphrodites. These results indicate that cdc-25.2 is expressed primarily in oocytes and not in sperm. However, compared with the robust expression in the N2 hermaphrodite germ line, the expression was much weaker in the feminized fem-1(lf) germ line, suggesting that the presence of sperm or fertilization positively affect the cdc-25.2 expression level. In conclusion, quantitative real-time RT-PCR showed that cdc-25.2 is predominantly expressed in oocytes of adult hermaphrodites.