Genes induced during the early developmental stages of the Cane Toad, Bufo (Chaunus) marinus

Halliday, Damien C.T.; Kennedy, Gavin; Hamilton, Nicholas H.R.; Tarmo, Suze; Alderman, James; Siddon, Nicole A.; Robinson, Anthony J.

doi:10.1016/j.gep.2008.04.005

Cited by 13 publications

(6 citation statements)

References 34 publications

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“…However, to confirm such co-distribution within smooth muscle, we used gastrokine-1 polyclonal antibody in other tissues and found, in agreement with recent related data on genes induced during early development stages [21] , that gastrokine was completely absent in skeletal muscle. However, also in agreement with this work, we found that gastrokine-1 was weakly present in bovine heart and, more particularly, targeted to Hiss fibers (personal data, not shown).…”

Section: Resultssupporting

confidence: 86%

“…Then, and as expected from recent studies on genes induced during early developmental stages [21] , gastrokine-1 was detected at E8 and E16 stages of the chick gizzard and duodenum. Similarly, the described gastrokine-1 localisation is in agreement with previous findings [1] , [12] but, in addition, there was clear co-localisation with specific smooth proteins such as smooth actin-filaments and caldesmon in the gizzard, but also in chick duodenum.…”

Section: Resultssupporting

confidence: 85%

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Biochemical Properties of Gastrokine-1 Purified from Chicken Gizzard Smooth Muscle

et al. 2008

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The potential role and function of gastrokine-1 (GNK1) in smooth muscle cells is investigated in this work by first establishing a preparative protocol to obtain this native protein from freshly dissected chicken gizzard. Some unexpected biochemical properties of gastrokine-1 were deduced by producing specific polyclonal antibody against the purified protein. We focused on the F-actin interaction with gastrokine-1 and the potential role and function in smooth muscle contractile properties.BackgroundGNK1 is thought to provide mucosal protection in the superficial gastric epithelium. However, the actual role of gastrokine-1 with regards to its known decreased expression in gastric cancer is still unknown. Recently, trefoil factors (TFF) were reported to have important roles in gastric epithelial regeneration and cell turnover, and could be involved in GNK1 interactions. The aim of this study was to evaluate the role and function of GNK1 in smooth muscle cells.Methodology/Principal FindingsFrom fresh chicken gizzard smooth muscle, an original purification procedure was used to purify a heat soluble 20 kDa protein that was sequenced and found to correspond to the gastrokine-1 protein sequence containing one BRICHOS domain and at least two or possibly three transmembrane regions. The purified protein was used to produce polyclonal antibody and highlighted the smooth muscle cell distribution and F-actin association of GNK1 through a few different methods.Conclusion/SignificanceAltogether our data illustrate a broader distribution of gastrokine-1 in smooth muscle than only in the gastrointestinal epithelium, and the specific interaction with F-actin highlights and suggests a new role and function of GNK1 within smooth muscle cells. A potential role via TFF interaction in cell-cell adhesion and assembly of actin stress fibres is discussed.

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Section: Resultssupporting

confidence: 86%

Section: Resultssupporting

confidence: 85%

Biochemical Properties of Gastrokine-1 Purified from Chicken Gizzard Smooth Muscle

et al. 2008

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“…It is well documented that a tadpole form of globin in anurans is replaced by adult globin during the course of metamorphosis [18,26,27] and here we demonstrate the existence of this switch for the first time in a Bufo species. We have previously demonstrated strong upregulation of adult globin genes during cane toad metamorphosis and that this was more pronounced than for any of the other genes induced at metamorphosis [17]. We have also previously shown that the recombinant BIV can be genetically modified to express adult globin in vitro [11] and confirmed here that this virus (rBIV/neo r /Adglob) is capable of infecting cane toad tadpoles.…”

Section: Discussionsupporting

confidence: 79%

“…Cane toad genes that are expressed in metamorphs but not in tadpoles therefore represent ideal targets to block and thus manipulate aspects of development. One documented example of the transition from a larval to an adult form in amphibians is haemoglobin [16] and we have used microarray analysis to establish that adult haemoglobin is significantly upregulated during cane toad metamorphosis [17]. Injecting tadpoles with adult globin interfered with expression of this protein in Rana catesbeiana, and induced changes in gene expression profiles of metamorphs [18].…”

Section: Introductionmentioning

confidence: 99%

Assessment of Virally Vectored Autoimmunity as a Biocontrol Strategy for Cane Toads

et al. 2011

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BackgroundThe cane toad, Bufo (Chaunus) marinus, is one of the most notorious vertebrate pests introduced into Australia over the last 200 years and, so far, efforts to identify a naturally occurring B. marinus-specific pathogen for use as a biological control agent have been unsuccessful. We explored an alternative approach that entailed genetically modifying a pathogen with broad host specificity so that it no longer caused disease, but carried a gene to disrupt the cane toad life cycle in a species specific manner.Methodology/Principal FindingsThe adult beta globin gene was selected as the model gene for proof of concept of autoimmunity as a biocontrol method for cane toads. A previous report showed injection of bullfrog tadpoles with adult beta globin resulted in an alteration in the form of beta globin expressed in metamorphs as well as reduced survival. In B. marinus we established for the first time that the switch from tadpole to adult globin exists. The effect of injecting B. marinus tadpoles with purified recombinant adult globin protein was then assessed using behavioural (swim speed in tadpoles and jump length in metamorphs), developmental (time to metamorphosis, weight and length at various developmental stages, protein profile of adult globin) and genetic (adult globin mRNA levels) measures. However, we were unable to detect any differences between treated and control animals. Further, globin delivery using Bohle iridovirus, an Australian ranavirus isolate belonging to the Iridovirus family, did not reduce the survival of metamorphs or alter the form of beta globin expressed in metamorphs.Conclusions/SignificanceWhile we were able to show for the first time that the switch from tadpole to adult globin does occur in B. marinus, we were not able to induce autoimmunity and disrupt metamorphosis. The short development time of B. marinus tadpoles may preclude this approach.

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“…(2018) ; the anti-inflammatory IL-10, the proinflammatory IFN-γ, and the complement protein C1s, developed by our research group in Brazil, using R. marina transcriptome provided by Gardner et al. (2018) ; and the housekeeping gene β-actin developed by Halliday et al. (2008) .…”

Section: Methodsmentioning

confidence: 99%

Lipopolysaccharide Regulates Pro- and Anti-Inflammatory Cytokines, Corticosterone, and Melatonin in Toads

Ferreira

Neto

Titon

et al. 2021

Integrative Organismal Biology

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Glucocorticoids and melatonin (MEL) show integrated and complex immunomodulatory effects, mostly described for endotherms, yet underexplored in amphibians. In this context, the RT-qPCR of molecules mediating inflammatory processes in amphibians is a valuable tool to explore the relationships among molecular biology, endocrine mediators, and immune response in these animals. In this study, toads (Rhinella diptycha) received an intraperitoneal saline injection or lipopolysaccharide (LPS; 2 mg/kg). Six hours post-injection, we analyzed plasma corticosterone (CORT) and MEL levels and pro and anti-inflammatory molecules (IL-1β, IL-6, IL-10, IFN-γ, and C1s). We found increased CORT and decreased MEL levels in response to LPS. Also, IL-1β, IL-6, and IL-10 were upregulated in LPS-injected toads compared with saline-injected. Overall, our results demonstrate an LPS-induced inflammatory response with endocrine and immune modulation in R. diptycha toads, exhibiting expected patterns for an inflammatory stimulus within this timeframe (6 h post-injection). Toads were responsive to LPS by secreting different cytokines, such as proinflammatory cytokines IL-1β and IL-6, related to immune cell attraction to inflammatory sites and the anti-inflammatory cytokine IL-10, which limits the rate of leukocyte infiltration, inflammation, and downregulates the expression of proinflammatory cytokines. Increased circulating CORT levels are probably associated with the activation of the hypothalamus-pituitary-interrenal axis by the LPS and the endocrine actions of IL-6. Furthermore, decreased circulating MEL levels are likely due to inhibited MEL secretion by the pineal gland by inflammatory stimuli, indicating the activation/existence of the immune-pineal axis in amphibians.

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Genes induced during the early developmental stages of the Cane Toad, Bufo (Chaunus) marinus

Cited by 13 publications

References 34 publications

Biochemical Properties of Gastrokine-1 Purified from Chicken Gizzard Smooth Muscle

Biochemical Properties of Gastrokine-1 Purified from Chicken Gizzard Smooth Muscle

Assessment of Virally Vectored Autoimmunity as a Biocontrol Strategy for Cane Toads

Lipopolysaccharide Regulates Pro- and Anti-Inflammatory Cytokines, Corticosterone, and Melatonin in Toads

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