Molecular characterization and expression analysis of two peptidoglycan recognition proteins (CcPGRP5, CcPGRP6) in larvae ontogeny of common carp Cyprinus carpio L. and upon immune stimulation by bacteria

Zhang, Fumiao; Shan, Shijuan; Xu, Xiaoyang; Wang, Yao; Zhang, Yonghuan; Yin, Miao; Gao, Yang

doi:10.1186/s12917-018-1744-1

Cited by 19 publications

(9 citation statements)

References 47 publications

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“…The quality and concentration of all total RNA samples were assessed using a NanoDrop Spectrophotometer (Thermo Scientific, USA). First-strand cDNA was synthesized from 2 µg total RNA with a FastQuant RT Kit (with gDNase) (Tiangen, China) according to the manufacture’s instructions [ 28 , 29 ]. Total RNA from the collected samples was extracted following the procedure above, and then the cDNA was stored at -80 ℃ until use for real-time quantitative PCR (RT-qPCR).…”

Section: Methodsmentioning

confidence: 99%

Molecular characterization of a new IgZ3 subclass in common carp (Cyprinus carpio) and comparative expression analysis of IgH transcripts during larvae development

Zhang

et al. 2021

BMC Vet Res

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Background Immunoglobulins (Igs) distributed among systemic immune tissues and mucosal immune tissues play important roles in protecting teleosts from infections in the pathogen-rich aquatic environment. Teleost IgZ/IgT subclasses with different tissue expression patterns may have different immune functions. Results In the present study, a novel secreted IgZ heavy chain gene was cloned and characterized in common carp (Cyprinus carpio). This gene exhibited a different tissue-specific expression profile than the reported genes IgZ1 and IgZ2. The obtained IgZ-like subclass gene designated CcIgZ3, had a complete open reading frame contained 1650 bp encoding a protein of 549 amino acid residues. Phylogenetic analysis revealed that CcIgZ3 was grouped with carp IgZ2 and was in the same branch as IgZ/IgT genes of other teleosts. Basal expression detection of the immunoglobulin heavy chain (IgH) in healthy adult common carp showed that CcIgZ3 transcripts were widely expressed in systemic immune tissues and mucosal-associated lymphoid tissues. CcIgZ3 was expressed at the highest levels in the head kidneys, gills, and gonads, followed by the spleen, hindgut, oral epithelium, liver, brain, muscle, foregut, and blood; it was expressed at a very low level in the skin. The transcript expression of CcIgZ3 in leukocytes isolated from peripheral blood cells was significantly higher than that in leukocytes isolated from the spleen. Different groups of common carp were infected with Aeromonas hydrophila via intraperitoneal injection or immersion. RT-qPCR analysis demonstrated that significant differences in CcIgZ3 mRNA levels existed between the immersion and injection groups in all the examined tissues, including the head kidney, spleen, liver, and hindgut; in particular, the CcIgZ3 mRNA level in the hindgut was higher in the immersion group than in the injection group. The different routes of A. hydrophila exposure in common carp had milder effects on the IgM response than on the CcIgZ3 response. Further study of the relative expression of the IgH gene during the development of common carp showed that the tissue-specific expression profile of CcIgZ3 was very different from those of other genes. RT-qPCR analysis demonstrated that the CcIgZ3 mRNA level increased gradually in common carp during the early larval development stage from 1 day post fertilization (dpf) to 31 dpf with a dynamic tendency similar to those of IgZ1 and IgZ2, and IgM was the dominant Ig with obviously elevated abundance. Analyses of the tissue-specific expression of IgHs in common carp at 65 dpf showed that CcIgZ3 was expressed at mucosal sites, including both the hindgut and gill; in contrast, IgZ1 was preferentially expressed in the hindgut, and IgZ2 was preferentially expressed in the gill. In addition to RT-qPCR analysis, in situ hybridization was performed to detect CcIgZ3-expressing cells and IgM-expressing cells. The results showed that CcIgZ3 and IgM transcripts were detectable in the spleens, gills, and hindguts of common carp at 65 dpf. Conclusions These results reveal that CcIgZ3 gene transcripts are expressed in common carp during developmental stage not only in systemic tissues but also in mucosal tissues. CcIgZ3 expression can be induced in immune tissues by A. hydrophila challenge via immersion and intraperitoneal injection with significantly different expression profiles, which indicates that CcIgZ3 is involved in the antimicrobial immune response and might play an important role in gut mucosal immunity.

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Section: Methodsmentioning

confidence: 99%

Molecular characterization of a new IgZ3 subclass in common carp (Cyprinus carpio) and comparative expression analysis of IgH transcripts during larvae development

Zhang

et al. 2021

BMC Vet Res

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“…The introduction of each new antibiotic is accompanied by the emergence of antibiotic-resistant strains. In contrast, innate immunity provides the host with immediate protection from infection and has maintained its antimicrobial effect for millions of years without the frequent emergence of drugresistant strains (Zhao et al, 2017;Zhang et al, 2019). Then the concept of biological control based on immune system has been put forward gradually, therefore, it has a great significance to study the immune defense response of OFM for biological control in the future.…”

Section: вісник сумського національного аграрного університетуmentioning

confidence: 99%

Молекулярна Ідентифікація Білка Пептидоглікану Короткого Типу, GMPGRP-Sc Від Grapholitha Molesta

Cao¹,

Cao²,

Zhu³

et al. 2022

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Білок розпізнавання пептидогліканів (PGRP) є важливим рецептором розпізнавання паттернів, який міститься як у безхребетних, так і у хребетних. Він відіграє важливу роль в антибактеріальному імунітеті, завдяки своїй помітній здатності виявляти та усувати збудника інфекції. Однак PGRP, в основному ідентифіковано з Drosophila melanogaster та Bombyx mori, а щодо інших сільськогосподарських комах повідомлень мало про їхні функції та механізм. У цьому дослідженні короткотиповий ген PGRP під назвою GmPGRP-SC був ідентифікований у Grapholitha molesta – східної плодожерки (OFM) – на основі аналізу бази даних груп транскрипції OFM з нашої лабораторії та Національного центру інформації з біотехнологій (https://www.ncbi.nlm.nih.gov/). GmPGRP-SC містить домен PGRP і має найтісніший генетичний зв'язок з геном PGRP Leguminivora glycinivorella, відповідно до послідовності та філогенетичного аналізу. Метод ПЛР у реальному часі було використано для аналізу картини його експресії на стадіях розвитку OFM та в різних тканинах личинки OFM. Наразі, відносно рівні експресії гена GmPGRP-SC в OFM були проаналізовані після інфікування Beauveria bassiana. Результати показали, що загальна кДНК від GmPGRP-SC становила 3221 bp (базових, основних пар), а кодуючі області – 2268 bp, що кодують 756 амінокислотних залишків. Рівень експресії GmPGRP-SC був найвищим у стадії лялечки OFM. Водночас, у різних тканинах OFM його відносна експресія була вищою в епітелії та гемоциті, тоді як іншим стадіям та тканинам притаманна порівняно нижча та з невеликою різницею. Рівень експресії GmPGRP-SC був істотно різним, коли суспензія спор B. bassiana становить 105 конідій/мкл, інфікованих через 48 годин. Тоді, коли суспензія спор B. bassiana становить 107 конідій/мкл, рівень експресії GmPGRP-SC також був різним. Усі ці результати закладуть основу для вивчення ролі та функцій GmPGRP-SC у вродженому імунітеті OFM, а також сприятимуть подальшому вивченню молекулярної взаємодії між OFM та B. bassiana. Результати досліджень можуть допомогти знайти потенційні молекули-мішені та забезпечити наукову основу для розробки нових біогенних пестицидів та реалізації Зеленої боротьби зі шкідниками (GPM).

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“…The quantity and quality of the total RNA were assessed using the NanoDrop Spectrophotometer (Thermo Scienti c, USA) on all samples. The rst-strand cDNAs were synthesized from 2 µg total RNA according to the manufacture's instructions of FastQuant RT Kit (With gDNase) (Tiangen, China) [27,28]. Total RNA from the collected samples was extracted following the procedure above and then the cDNA was stored at -80 ℃ until used in real-time quantitative PCR (RT-qPCR).…”

Section: Molecular Cloning and Analysis Of Common Carp Ccigz3mentioning

confidence: 99%

Molecular Characterization of A New IgZ3 Subclass from Common Carp (Cyprinus Carpio) and Comparative Expression Analysis of IgH Transcripts During Ontogeny of Larvae

Zhang

et al. 2020

Preprint

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Background Immunoglobulins distributing among systemic immune tissues and mucosal immune tissues play an important role in teleost to protect them from infections in a pathogen-rich aquatic environment. Teleost IgZ/IgT subclasses with different tissue expression patterns may have varied immune functions.Results In the present study, a novel secreted IgZ heavy chain gene was cloned and characterized in common carp (Cyprinus carpio) which was different from the reported IgZ1 and IgZ2 in tissue-specific expression profile. The obtained IgZ-like subclass was designated as CcIgZ3 which complete open reading frame contained 1650 bp encoding a protein of 549 amino acid residues. The phylogenetic analysis revealed that CcIgZ3 was grouped with carp IgZ2 and was in the same branch as other teleosts IgZ/IgT. Basal expression detection of IgH in healthy adult common carp showed that CcIgZ3 transcripts were widely expressed in systemic immune tissues and Mucosal-associated lymphoid tissues. It was expressed at a higher level in the head kidney, gill, and gonad, followed by spleen, hindgut, oral epithelium, liver, brain, muscle, foregut, and blood, but at a very low level in the skin. The transcript level of CcIgZ3 mRNA in isolated leukocytes from peripheral blood cells was significantly higher than that of isolated leukocytes from the spleen. Different groups of common carp were infected with Aeromonas hydrophila via intraperitoneal injection and immersion respectively. The qRT-PCR analysis demonstrated that a significant difference in CcIgZ3 mRNA level between immersion and injection groups existed in all the detected tissues including head kidney, spleen, liver, and hindgut, especially in hindgut CcIgZ3 mRNA level was higher in immersion than in the injection group. Different routes for Aeromonas hydrophila challenge to common carp had comparatively less effect on IgM response. Further study on the relative expression of the IgH gene during the ontogeny of common carp presented that the tissue-specific expression profile of CcIgZ3 was very different from the others. In the early larval development stage of common carp from 1 dpf to 31 dpf, the qRT-PCR analysis demonstrated that the CcIgZ3 mRNA level increased gradually with a similar dynamic tendency of IgZ1 and IgZ2 and IgM was the dominant Ig with obviously higher abundance. The results of tissue-specific expression of IgH at 65 dpf of common carp showed that CcIgZ3 was expressed at mucosal sites including both hindgut and gill but IgZ1 was preferentially expressed in hindgut and IgZ2 in gill. Except for qRT-PCR analysis, the detection of the CcIgZ3-expressing cells and IgM-expressing cells by in situ hybridization was performed. The results showed that CcIgZ3 and IgM transcripts could be detected in the spleen, gill, and hindgut of the common carp at 65dpf. Conclusions These results revealed that CcIgZ3 gene transcript occurred in the early development stage of common carp not only in systemic tissues but also in mucosal tissues. CcIgZ3 could be induced with significantly different expression profiles at immune tissues after the challenge by Aeromonas hydrophila immersion and intraperitoneal injection, which indicated that CcIgZ3 might play a more important role in mucosal immunity than in systemic immunity.

show abstract

Molecular characterization and expression analysis of two peptidoglycan recognition proteins (CcPGRP5, CcPGRP6) in larvae ontogeny of common carp Cyprinus carpio L. and upon immune stimulation by bacteria

Cited by 19 publications

References 47 publications

Molecular characterization of a new IgZ3 subclass in common carp (Cyprinus carpio) and comparative expression analysis of IgH transcripts during larvae development

Molecular characterization of a new IgZ3 subclass in common carp (Cyprinus carpio) and comparative expression analysis of IgH transcripts during larvae development

Молекулярна Ідентифікація Білка Пептидоглікану Короткого Типу, GMPGRP-Sc Від Grapholitha Molesta

Molecular Characterization of A New IgZ3 Subclass from Common Carp (Cyprinus Carpio) and Comparative Expression Analysis of IgH Transcripts During Ontogeny of Larvae

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