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.
Silk production quantity is the most economically important characteristic of the domesticated silkworm moth, Bombyx mori. It is controlled by multiple loci. The cocoon and silk production quantity of silkworm strains Jingsong and Lan10 have significantly diverged. A backcross population (BC1) was bred using Jingsong and Lan10 as parents to identify quantitative trait loci (QTLs) for silk quality. In this research, a genetic linkage map of the silkworm was constructed using the BC1 mapping population. The map contained 85 sequence-tagged site markers, 80 simple sequence repeat markers, and 16 single nucleotide polymorphisms. A linkage map was constructed from the data, which consisted of 181 markers distributed over 28 expected linkage groups and spans 2147.1 cM in total length. Fourteen QTLs were detected for cocoon filament length, whole cocoon weight, pupae weight, filament weight, and cocoon shell weight. The 14 QTLs were distributed in 5 linkage groups (linkage groups 1, 14, 18, 23 and 25) based on the constructed linkage map. In addition, five QTLs, which had the highest logarithm (base 10) of odds (LOD) values, were located on the first chromosome, three of which located at the same region in linkage group 1. These results represent an important foundation for the map-based cloning of QTLs and marker-assisted selection for improving the silk quality of economically important silkworm strains., Bing Li ... [et al.]., and Obsahuje seznam literatury
Steroid hormone 20-hydroxyecdysone and the sesquiterpenoid juvenile hormone are the main regulators of insect development; however, it is unclear how they interact in the regulation of metamorphic events. Using the silkworm, Bombyx mori, we show that the juvenile hormone analogue fenoxycarb affects the cascade of ecdysone regulated genes that control the programmed cell death in the larval midgut. Morphological changes that occur during cell death were investigated by studying cross-sections of the midgut stained with hematoxylin and eosin. Apoptosis-specific DNA fragmentation was detected using TUNEL assay. Expression patterns of genes ATG8 and ATG12, which were used as indicators of autophagy, and genes of the ecdysone-regulated gene cascade were examined using real-time quantitative polymerase chain reaction. Fenoxycarb application on day 0 of the 5th larval instar extended the feeding period and postponed programmed cell death in mature larval midgut. This effect was probably due to a delay in ecdysone secretion and associated changes in gene expression were mostly not a direct response to the fenoxycarb. However, differences in the gene expression patterns in the control and fenoxycarb treated insects during the prepupal and early pupal stages indicated that fenoxycarb may also exert a more direct effect on some genes of the ecdysone regulated gene cascade., Ebru Goncu, Ramazan Uranli, Osman Parlak., and Obsahuje bibliografii