TABLE 2.
Penetrance of him-17 high-temperature phenotypes
| Incidence of observed phenotype
|
|||||
|---|---|---|---|---|---|
| Genotype | Altered spatial organizationa | Interspersed meiotic and mitotic nucleib | Excess mitotic nuclei in distal germlineb | Ectopic H3S10p-positive figuresc | Mog phenotyped |
| him-17(e2707) | 10/10 | ||||
| him-17(me9) | 26/26 | 14/14e | 11/14 | Observed but not quantified | |
| him-17(me24) | 50/50 | 9/9 | 7/9 | 15/26 | 5/72 |
| him-17(ok424) | 16/16 | 7/7 | 6/7 | Observed but not quantified | |
| him-17(RNAi) | 16/16 | ||||
For experiments examining the effects of high temperature on him-17 hermaphrodite germlines, him-17(me24) or him-17(ok424) m+z− homozygous mutant hermaphrodites (m, maternal contribution; z, zygotic contribution) were selected as non-Unc progeny of him-17/nT1Unc heterozygotes. For him-17(me9) or him-17(e2707), m+z− homozygous mutant hermaphrodites were identified on the basis of their Him phenotype among the progeny of him-17/+ mothers. Their m−z− progeny were shifted to 25° for subsequent analysis. him-17 RNA interference was performed as in Reddy and Villeneuve (2004).
Numbers are combined totals of worms stained with DAPI only and germlines stained with DAPI and either anti-HIM-3 or anti-H3S10p.
Germlines were fixed and stained with DAPI and HIM-3 antibody; HIM-3-positive nuclei were scored as meiotic and HIM-3-negative nuclei were scored as mitotically cycling. In normal gonads, nuclei in the proximal half of the distal proliferative region are located adjacent to the gonad periphery, surrounding a central cytoplasmic core that is largely devoid of nuclei; in gonads that were scored as having excess mitotic nuclei in the distal proliferative region, the central core was instead filled with tightly packed HIM-3-negative nuclei.
Germlines were fixed and stained with DAPI and H3S10p antibody. H3S10p-positive figures were scored as ectopic if they were located in positions outside of the normal proliferative zone (the distal-most 20 rows of nuclei).
In gonads that were scored as Mog, there were no obvious signs of oocyte production and sperm nuclei were present in vast excess over the normal number, filling the proximal gonad and extending past the loop into the distal gonad (see Figure 1D). The Mog phenotype was also observed but not quantified in him-17(me9) and him-17(ok424) mutants.
Of 14, 1 had a clear second zone of mitotic proliferation in the proximal arm of the gonad.