Effects of starvation and sucrose feeding on corpora allata of last instar larvae of Manduca sexta: in vitro activity, size, and cell number

Abstract Larvae of the tobacco hornworm moth Manduca sexta starved for the first 3 days of the last (fifth) stadium undergo a supernumerary moult. If they are provided with sucrose during the starvation period, they develop into normal pupae although pupation is delayed. The activities of the corpora allata (CA) from normal, starved, and sucrose fed larvae were followed through the fifth stadium with a radiochemical assay for Juvenile Hormone (JH) biosynthesis. An attempt was made to correlate CA-activity with CA cell number, size, and protein content.
In CA of normally fed larvae the rate of JH synthesis declined to undetectable levels by day 4 which was also the time of exposure of the dorsal vessel. In CA of starved larvae, the rate of JH synthesis at first decreased but began to increase on day 3 and reached a peak value by day 7 , at which time head capsule slippage occurred. In CA of sucrose fed larvae, the rate of biosynthesis declined as in normal larvae but the decline was extended over a longer period. Exposure of the dorsal vessel was delayed in the same manner and occurred on days 7–9. The major JH in all cases was JH-II.
The CA comprise c. 150 cells in the early fifth stadium, and this number remained constant during the fifth stadium in all three feeding regimens. In normal larvae, CA size and protein content increased several-fold during the stadium whereas in starved and sucrose-fed larvae they increased slowly and in agreement with the altered timing of developmental events. In none of the groups was the CA activity pattern correlated with morphometric changes of the CA. The rates of JH biosynthesis were not closely correlated with published JH titre curves. The in vivo mechanisms for regulation of JH production remain to be elucidated.     

Rad52 recruitment is DNA replication independent and regulated by Cdc28 and the Mec1 kinase

Recruitment of the homologous recombination machinery to sites of double‐strand breaks is a cell cycle‐regulated event requiring entry into S phase and CDK1 activity. Here, we demonstrate that the central recombination protein, Rad52, forms foci independent of DNA replication, and its recruitment requires B‐type cyclin/CDK1 activity. Induction of the intra‐S‐phase checkpoint by hydroxyurea (HU) inhibits Rad52 focus formation in response to ionizing radiation. This inhibition is dependent upon Mec1/Tel1 kinase activity, as HU‐treated cells form Rad52 foci in the presence of the PI3 kinase inhibitor caffeine. These Rad52 foci colocalize with foci formed by the replication clamp PCNA. These results indicate that Mec1 activity inhibits the recruitment of Rad52 to both sites of DNA damage and stalled replication forks during the intra‐S‐phase checkpoint. We propose that B‐type cyclins promote the recruitment of Rad52 to sites of DNA damage, whereas Mec1 inhibits spurious recombination at stalled replication forks.