Proteomics analysis of adult testis from <i><scp>B</scp>ombyx mori</i>

Zhang, Yan; Dong, Zhaoming; Gu, Peiming; Zhang, Weiwei; Wang, Dandan; Guo, Xiaomeng; Xia, Qingyou

doi:10.1002/pmic.201300507

Cited by 7 publications

(5 citation statements)

References 26 publications

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“…The luminal proteins (60 μg) were digested according to previously reported methods 7 32 60 61 . The resulting tryptic peptides were recovered by centrifugation in the ultrafiltration tube, lyophilized, and resuspended in 35 μL of 0.1% formic acid.…”

Section: Methodsmentioning

confidence: 99%

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Analysis of proteome dynamics inside the silk gland lumen of Bombyx mori

Dong

Song

Zhang

et al. 2016

Sci Rep

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The silk gland is the only organ where silk proteins are synthesized and secreted in the silkworm, Bombyx mori. Silk proteins are stored in the lumen of the silk gland for around eight days during the fifth instar. Determining their dynamic changes is helpful for clarifying the secretion mechanism of silk proteins. Here, we identified the proteome in the silk gland lumen using liquid chromatography–tandem mass spectrometry, and demonstrated its changes during two key stages. From day 5 of the fifth instar to day 1 of wandering, the abundances of fibroins, sericins, seroins, and proteins of unknown functions increased significantly in different compartments of the silk gland lumen. As a result, these accumulated proteins constituted the major cocoon components. In contrast, the abundances of enzymes and extracellular matrix proteins decreased in the silk gland lumen, suggesting that they were not the structural constituents of silk. Twenty-five enzymes may be involved in the regulation of hormone metabolism for proper silk gland function. In addition, the metabolism of other non-proteinous components such as chitin and pigment were also discussed in this study.

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Section: Methodsmentioning

confidence: 99%

“…Peptide searches were performed with the Andromeda search algorithms 63 . The search parameters were set as reported previously 7 32 60 61 . A minimum of one unique peptides was required for the identified protein.…”

Section: Methodsmentioning

confidence: 99%

Analysis of proteome dynamics inside the silk gland lumen of Bombyx mori

Dong

Song

Zhang

et al. 2016

Sci Rep

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“…Hence, the proteomics study is very important to focus on the active products in the cells rather than gene expression study by using microarray and RT-PCR analysis 6,7 . Two-dimensional gel electrophoresis combined with mass spectrometry analysis has been frequently used in classical proteomics research to identify and functional characterization of the specific proteins from different organs and tissues of Bombyx mori such as collateral glands 8 , haemolymph [9][10][11] , fat body 12,13 , silk glands 14,15 , protharasic gland 16 , endocrine organs 17 , larval head, diapauses and non-diapauses egg and embryo [18][19][20] , integument, trachea and adult scale 21 , midgut and peritrophic membrane [22][23][24][25] , larval gonads and testis 26,27 , Malpighian tubules 28 , wing discs 29 and newly hatched larva 30 . Further, proteome profiling of several organs and tissues analysis revealed that these proteins play a crucial role in the nervous system development, digestion, protein synthesis, transport of ions and O 2 , reproduction, stress response and apoptosis in silkworm Bombyx mori.…”

Section: Introductionmentioning

confidence: 99%

Comparative Proteomic Analysis of Larva and Adult Heads of Silkworm, Bombyx mori (Lepidoptera: Bombycidae)

Arunprasan¹,

Kannan²,

Anbalagan³

et al. 2016

J. of Entomology

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Background: The complete understanding of feeding and reproduction strategies during metamorphosis of silkworm Bombyx mori is very essential, for that a comparative proteomics analysis was used to investigate the proteins extracted from the head of the larva and adult of Bombyx mori. Materials and Methods: Proteins were separated and identified by using 2D-PAGE and MALDI-TOF-TOF-MS analysis respectively. The functions of each proteins were annotated using STRAP GO analysis. The expression level of each protein spots between larvae and adult were analyzed using 2D image platinum 7. Results: Using 2-DE gels stained with coomassie staining revealed, 315±3 and 296±3 proteins spots with pI ranges of 4-10 and 3.5-8 in the larva and adult head, respectively. Thirty proteins were identified based on differential and unique expression in the head of the larva and adult of B. mori. Among the 30 identified proteins, comparatively 10 proteins were up regulated and 6 proteins were down regulated in the larvae, whereas 6 proteins were up regulated and 10 were down regulated in the adult. In addition, 6 and 8 unique proteins expressions were observed in the larvae and adult, respectively. Database search combined with STRAP GO analysis revealed that few up regulated and unique protein in the larvae such as odorant binding proteins, argonaute protein, apolipophorin III precursor, enolase and iron-ion transport proteins were localized to regulate the feeding activity of larva. Furthermore, some proteins are highly and uniquely expressed from adult head such as glycoprotein hormone, N-acetyltranferase, cytochrome p450, DNA binding and intracellular transport protein, clathrin coat assembly protein and metabolic enzyme which may regulate adult ecolsion and reproduction. Conclusion: The results suggest and confirmed that the protein expression in each stage could alter the behaviors of the insect related to larval feeding, growth and reproduction in the adult. Further the characterization and functions of other proteins identified are discussed. This study will help to improve the silk production which leads to economy improvement in sericulture industry.

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“…In this study, female Ochsp70 expression was also induced through mating, which combined with the high expression levels of male Ochsp70 in the mature testis, suggests that Ochsp70 may functions as a male mating factor in O. communa . Similarly, 32 HSPs constitute a group of most abundant proteins in the adult testis proteomics of Bombyx mori , a 94.4 kDa Hsp70was also included ( Zhang et al, 2014 ), which were considered to be associated with spermatogenesis, reproduction, mitosis, and fertilization. This phenomenon is even more comprehensible in mammals ( Boelens et al, 2004 ; Jha et al, 2013 ; Zhang et al, 2014 ), wherein several hsp70 genes are expressed specifically in male germ cells ( Dix, 1997 ; Neuer et al, 2000 ; Carreira and Santos, 2020 ).…”

Section: Discussionmentioning

confidence: 99%

“…Similarly, 32 HSPs constitute a group of most abundant proteins in the adult testis proteomics of Bombyx mori , a 94.4 kDa Hsp70was also included ( Zhang et al, 2014 ), which were considered to be associated with spermatogenesis, reproduction, mitosis, and fertilization. This phenomenon is even more comprehensible in mammals ( Boelens et al, 2004 ; Jha et al, 2013 ; Zhang et al, 2014 ), wherein several hsp70 genes are expressed specifically in male germ cells ( Dix, 1997 ; Neuer et al, 2000 ; Carreira and Santos, 2020 ). Testicular sperms are the most diverse of all cell types, so it is not surprising that spermatogenesis is accompanied by the expression of hsp gene different expression.…”

Section: Discussionmentioning

confidence: 99%

The hsp70 new functions as a regulator of reproduction both female and male in Ophraella communa

Zhang

Ma²,

Ma³

et al. 2022

Front. Mol. Biosci.

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Heat shock proteins (Hsps) function as molecular chaperones that enable organisms to withstand stress and maintain normal life activities. In this study, we identified heat shock protein 70 (encoded by hsp70), which exhibits a higher expression in the mature male testis than in the unmature testis of Ophraella communa. Tissue expression profile revealed that Ochsp70 levels in males were highest in the testis, whereas those in females were highest in the head. Moreover, the expression of Ochsp70 was found to be significantly induced in female bursa copulatrix after mating. Double-stranded RNA dsOchsp70 was injected into males to performance RNA interference, which significantly decreased the male Ochsp70 expression levels within 20 d post-injection, whereas no effect was observed on the Ochsp70 expression level in the females after mating with dsOchsp70-injected males. However, significant downregulation of female fertility was marked simultaneously. Furthermore, knockdown of female Ochsp70 expression also led to a significant reduction in fertility. Finally, comparative transcriptomic analysis identified glucose dehydrogenase and insulin-like growth factor binding protein as putative downstream targets of Ochsp70. Overall, we deduced that Ochsp70 is an indispensable gene and a potential male mating factor in O. communa, which regulates reproduction.

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Proteomics analysis of adult testis from Bombyx mori

Cited by 7 publications

References 26 publications

Analysis of proteome dynamics inside the silk gland lumen of Bombyx mori

Analysis of proteome dynamics inside the silk gland lumen of Bombyx mori

Comparative Proteomic Analysis of Larva and Adult Heads of Silkworm, Bombyx mori (Lepidoptera: Bombycidae)

The hsp70 new functions as a regulator of reproduction both female and male in Ophraella communa

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