Apoptosis of neuronal cells in the brains of postembryonic silkworms Bombyx mori (Lepidoptera: Bombycidae)

Kim, Mi Young; Min, Kang; Choi, Chang Ok; Song, Hoyoung; Seok, Chang Lee; Kim, Bong Woo; Kang, Pil Don; Jeon, Soung Hoo; Cho, Byung Pil; Bae, Yeon Jae; Ko, Young-Gyo; Hee, Bong Lee

doi:10.14411/eje.2009.041

Cited by 5 publications

(7 citation statements)

References 61 publications

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“…Taken together, these findings suggest that most of the neurons that died in the ventral ganglia tested underwent apoptosis in response to an increase in the titre of 20E in the haemolymph, as reported here for silkworm SOG and AG6 to TAG, silkworm brain (Kim et al, 2009) and M. sexta APR(5) and APR(6) (Zee & Weeks, 2001), but some neurons, such as those in TG3 and AG4, do not.…”

Section: Discussionsupporting

confidence: 78%

“…In our previous study of neuronal apoptosis in the brains of postembryonic stages of silkworm (Kim et al, 2009), peak numbers of apoptotic neurons were detected on day-4 in 4th instar larvae and day-9 in 5th instar larvae and day-4 in pupae. No apoptotic neurons were detected on day-1 in 1st instar larvae, on day-7 in pupae or on day-1 in adults.…”

Section: Discussionmentioning

confidence: 83%

“…5, most of the neurons that normally showed apoptosis in SOG, TG1, TG3, and AG4 to AG8 of day-1 and day-4 pupae exhibited no apoptosis after ActD or CHX was injected into day-8 5th instar larvae and day-2 pupae. This suggests that the neuronal apoptosis triggered in these ganglia by the increase in 20E may be induced by the synthesis of a new protein initiated by 20E, as in the brain of silkworm (Kim et al, 2009).…”

Section: Discussionmentioning

confidence: 99%

“…Day-5 and day-9 5th instar larvae were used to monitor the pattern of neuronal apoptosis and for inducing or inhibiting it using 20E or other chemicals. For monitoring the pattern of neuronal apoptosis or the effects of in vivo injection of 20E and other chemicals, larvae, pupae and adults 12 h after hatching or at a particular ecdysis were selected from the rearing stock (Kim et al, 2009). …”

Section: Experimental Animalsmentioning

confidence: 99%

“…The results of this study on neuronal apoptosis in silkworm VG were compared with those obtained in a previous study (Kim et al, 2009) using the brains of postembryonic silkworms in order to define the mechanisms of CNS development during the postembryonic life of insects.…”

Section: Introductionmentioning

confidence: 99%

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Characterization of neuronal apoptosis in the ventral ganglia of postembryonic silkworms Bombyx mori (Lepidoptera: Bombycidae)

Song¹,

Choi²,

Kim³

et al. 2009

Eur. J. Entomol.

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Abstract.A fluorescent triple staining method was developed to stain the cytoplasm of neurons red, the nuclei of all kinds of cells, including neurons, blue and the nuclei of apoptotic neurons in cyan in the twelve ventral ganglia (VG) of the Bombyx mori ventral nerve cord. This differential staining method was used to distinguish between apoptotic and normal neurons in the suboesophageal ganglion (SOG), thoracic ganglia (TG)1 to TG3 and abdominal ganglia (AG)1 to AG8 and also determine the changes in the numbers of apoptotic neurons that occur during postembryonic development. In most of the VG tested, neuronal apoptosis was most marked during the period from the end of larval life to the mid pupal stage. The greatest number of apoptotic neurons was found in SOG of day-5 pupae, TG1 to TG3 and AG1 to AG4 of day-1 pupae, and AG5 to AG8 of day-4 pupae. In vivo injection of 20-hydroxyecdysone (20E) into day-8 5th instar larvae resulted in both a considerable increase in the number of apoptotic neurons and cleavage of procaspase-3 into caspase-3, which induced neuronal apoptosis in SOG and AG6 to AG8 in day-1 pupae, and a slight increase in the number of apoptotic neurons in TG1. In TG3 and AG4, however, it had little effect on the number of apoptotic neurons or cleavage of procaspase-3. Treatment of the VG of both day-8 5th instar larvae and day-2 pupae with protein synthesis inhibitors by in vivo injection triggered a significant inhibition of neuronal apoptosis and procaspase-3 cleavage in most of these ganglia in day-1 pupae and day-4 pupae, but not TG3 and AG4, in which there was little procaspase-3 and caspase-3. In vivo injection of caspase-8 and -3 inhibitors into day-8 5th instar larvae and day-2 pupae led to a substantial inhibition of neuronal apoptosis and of procaspase-3 cleavage in SOG, AG6 and TAG, but not in TG3 or AG4 of day-1 pupae and day-4 pupae. These findings suggest that neurons that die in SOG, TG1 and AG6 to AG8 in day-1 and -4 pupae may undergo apoptosis induced by the synthesis of a new protein and caspase-8-and -3-implicated signal transduction by the increase in titre of 20E in the haemolymph but not the neuronal aopotosis in TG3 and AG4. This study provides neurobiologists with valuable information and a means of studying neuronal apoptosis in the nervous system of insects.

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Section: Discussionsupporting

confidence: 78%

Section: Discussionmentioning

confidence: 83%

Section: Discussionmentioning

confidence: 99%

Section: Experimental Animalsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Characterization of neuronal apoptosis in the ventral ganglia of postembryonic silkworms Bombyx mori (Lepidoptera: Bombycidae)

Song¹,

Choi²,

Kim³

et al. 2009

Eur. J. Entomol.

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SILKWORM 30K PROTEIN INHIBITS ECDYSONE‐INDUCED APOPTOSIS BY BLOCKING THE BINDING OF ULTRASPIRACLE TO ECDYSONE RECEPTOR‐B1 IN CULTURED Bm5 CELLS

Kim

Song

Kim

et al. 2012

Arch Insect Biochem Physiol

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This study investigates the mechanism through which increased 30K protein inhibits ecdysone-induced apoptosis in the Bm5 silkworm ovarian cell line. Treatment of Bm5 cells with 20-hydroxyecdysone (20E) after transfection with the pIZT/V5-His control vector triggered apoptosis, but 20E treatment did not trigger apoptosis in Bm5 cells transfected with the pIZT/30K/V5-His vector. To confirm its inhibitory effect on apoptosis, 30K protein was first purified from Escherichia coli transformed with a 30K expression vector and used to generate specific antibodies in mice. Anti-30K antiserum was used to confirm synthesis of the 30K protein in pIZT/30K/V5-His-transfected Bm5 cells and to detect 30K protein binding to the ecdysone receptor-B1 (EcR-B1). Anti-30K antiserum was used to immunoprecipitate protein complexes containing 30K from Bm5 cells transfected with pIZT/30K/V5-His vector and treated with 20E. We observed that 30K proteins bound primarily to the EcR-B1 and not to ultraspiracle (USP). Reciprocal immunoprecipitation of EcR-B1-containing complexes from Bm5 cells transfected with control pIZT/V5-His vector and treated with 20E showed that EcR-B1 bound to USP in the absence of 30K but did not bind to USP in pIZT/30K/V5-His-transfected Bm5 cells. These results demonstrate that 30K proteins block USP binding to EcR-B1 through formation of a 30K/EcR-B1 complex, resulting in inhibition of 20E-induced Bm5 cell apoptosis.

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Increase of 30K protein in identified motoneurons by hemolymph results in inhibition of programmed cell death in silkworm, Bombyx mori (Lepidoptera, Bombycidae)

Song

Kim

et al. 2012

Journal of Insect Physiology

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Apoptosis of neuronal cells in the brains of postembryonic silkworms Bombyx mori (Lepidoptera: Bombycidae)

Cited by 5 publications

References 61 publications

Characterization of neuronal apoptosis in the ventral ganglia of postembryonic silkworms Bombyx mori (Lepidoptera: Bombycidae)

Characterization of neuronal apoptosis in the ventral ganglia of postembryonic silkworms Bombyx mori (Lepidoptera: Bombycidae)

SILKWORM 30K PROTEIN INHIBITS ECDYSONE‐INDUCED APOPTOSIS BY BLOCKING THE BINDING OF ULTRASPIRACLE TO ECDYSONE RECEPTOR‐B1 IN CULTURED Bm5 CELLS

Increase of 30K protein in identified motoneurons by hemolymph results in inhibition of programmed cell death in silkworm, Bombyx mori (Lepidoptera, Bombycidae)

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