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1.
J Invertebr Pathol ; 132: 157-164, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26455997

RESUMO

Balance between reactive oxygen species (ROS) and the antioxidant (AO) defense mechanisms is vital for organism survival. Insects serve as an ideal model to elucidate oxidative stress responses as they are prone to different kinds of stress during their life cycle. The present study demonstrates the modulation of AO enzyme gene expression in the insect pest, Achaea janata (castor semilooper), when subjected to different oxidative stress stimuli. Antioxidant enzymes' (catalase (Cat), superoxide dismutase (Sod), glutathione-S-transferase (GST) and glutathione peroxidase (Gpx)) partial coding sequences were cloned and characterized from larval whole body. Tissue expression studies reveal a unique pattern of AO genes in the larval tissues with maximum expression in the gut and fat body. Ontogeny profile depicts differential expression pattern through the larval developmental stages for each AO gene studied. Using quantitative RT-PCR, the expression pattern of these genes was monitored during sugar-induced (d-galactose feeding), infection-induced (Gram positive, Gram negative and non-pathogenic bacteria) and pesticide-induced oxidative stress (Bt Cry toxin). d-Galactose feeding differentially modulates the expression of AO genes in the larval gut and fat body. Immune challenge with Escherichia coli induces robust upregulation of AO genes when compared to Bacillus coagulans and Bacillus cereus in the larval fat body and gut. Cry toxin feeding predominantly induced GST upregulation in the gut. The current study suggests that though there are multiple ways of generation of oxidative stress in the insect, the organism tailors its response by insult- and tissue-specific recruitment of the antioxidant players and their differential regulation for each inducer.


Assuntos
Mariposas/fisiologia , Estresse Oxidativo , Sequência de Aminoácidos , Animais , Antioxidantes/metabolismo , Catalase/genética , Catalase/metabolismo , Clonagem Molecular , Escherichia coli/imunologia , Regulação da Expressão Gênica , Glutationa Peroxidase/genética , Glutationa Peroxidase/metabolismo , Glutationa Transferase/genética , Glutationa Transferase/metabolismo , Mariposas/genética , Mariposas/imunologia , Espécies Reativas de Oxigênio/metabolismo , Alinhamento de Sequência , Análise de Sequência de Proteína , Superóxido Dismutase/genética , Superóxido Dismutase/metabolismo
2.
Comp Biochem Physiol C Toxicol Pharmacol ; 158(4): 199-206, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23973827

RESUMO

Thyroid hormones play crucial role in several biological processes including reproduction. Disruption of normal thyroid status by environmental contaminants can cause severe impairment in reproductive functions. In our previous study, we reported down-regulation of a protein in seminal vesicular fluid of air-breathing catfish, Clarias gariepinus during experimentally induced hyperthyroidism. N-terminal amino acid sequence analysis followed by search in sequence database denoted it to be lipocalin-type prostaglandin D2 synthase (ptgds-b). In the present study, we cloned full-length cDNA of ptgds-b based on the N-terminal amino acid sequence. Surprisingly, Northern blot as well as RT-PCR analysis demonstrated the presence of ptgds-b transcript predominantly in seminal vesicles and developing testis. Further, ptgds-b mRNA significantly decreased in seminal vesicles following L-thyroxine overdose while there was an increased expression of ptgds-b after depletion of thyroid hormone by thiourea and withdrawal of the treatments reverted this effect. Treatment of catfish with human chorionic gonadotropin and estradiol significantly reduced ptgds-b expression. Taken together, we report ptgds-b as a thyroid hormone regulated protein in the seminal vesicles in addition to gonadotropin and estradiol. Further studies might explain the exclusive presence of ptgds-b in seminal vesicles and developing testis yet present data evaluated it as a putative biomarker for thyroid hormone disruption.


Assuntos
Proteínas de Peixes/genética , Oxirredutases Intramoleculares/genética , Lipocalinas/genética , Glândulas Seminais/metabolismo , Tiroxina/farmacologia , Transcriptoma/efeitos dos fármacos , Sequência de Aminoácidos , Animais , Sequência de Bases , Northern Blotting , Gonadotropina Coriônica/farmacologia , Estradiol/farmacologia , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Humanos , Oxirredutases Intramoleculares/classificação , Lipocalinas/classificação , Masculino , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Glândulas Seminais/enzimologia , Glândulas Seminais/crescimento & desenvolvimento , Homologia de Sequência de Aminoácidos , Testículo/enzimologia , Testículo/crescimento & desenvolvimento , Testículo/metabolismo , Tioureia/farmacologia , Glândula Tireoide/efeitos dos fármacos , Glândula Tireoide/metabolismo
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