Dissection and Staining of &lt;em&gt;Drosophila&lt;/em&gt; Larval Ovaries

Maimon, Iris; Gilboa, Lilach

doi:10.3791/2537-v

Cited by 2 publications

(2 citation statements)

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“…Larvae were collected at 24-48h (first instar) and 72-120h (third instar) and dissected in ice cold 1X PBS (Teixeira et al, 2017). Larval tissue was fixed in 4% formaldehyde for 20 minutes, washed three times in PBT (1X PBS with 1% Triton X-100) and blocked in PBTB for 1 hour (Maimon and Gilboa, 2011). Adult ovaries were dissected in cold 1X PBS and fixed in 4% formaldehyde for 20 minutes.…”

Section: Drosophila Genetics and Husbandrymentioning

confidence: 99%

Tolerance thresholds underlie responses to DNA damage during germline development

Jansen,

Gebert,

Kumar

et al. 2024

Preprint

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Selfish DNA modules like transposable elements (TEs) are particularly active in the germline, the lineage that passes genetic information across generations. New TE insertions can disrupt genes and impair the functionality and viability of germ cells. However, we find that in P-M hybrid dysgenesis in Drosophila, a sterility syndrome triggered by the P-element DNA transposon, germ cells harbour unexpectedly few new TE insertions, despite accumulating DNA double-strand breaks (DSBs) and inducing cell cycle arrest. Using an engineered CRISPR-Cas9 system, we show that generating DSBs at silenced P-elements or other non-coding sequences is sufficient to induce germ cell loss independently of gene disruption. Indeed, we demonstrate that both developing and adult mitotic germ cells are sensitive to DSBs in a dosage-dependent manner. Following the mitotic-to-meiotic transition, however, germ cells become more tolerant to DSBs, completing oogenesis regardless of the accumulated genome damage. Our findings establish DNA damage tolerance thresholds as crucial safeguards of genome integrity during germline development.

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Section: Drosophila Genetics and Husbandrymentioning

confidence: 99%

Tolerance thresholds underlie responses to DNA damage during germline development

Jansen,

Gebert,

Kumar

et al. 2024

Preprint

View full text Add to dashboard Cite

show abstract

“…Pupae were sexed by the presence of sex-combs (Flagg, 1988). Third-instar larvae migrating on the side of the vial were selected and sexed (Maimon and Gilboa, 2011).…”

Section: Strains and Maintenancementioning

confidence: 99%