Abstract
Really interesting new gene (RING) finger protein is a type of zinc-binding motif found in a large family of functionally distinct proteins. RING finger proteins are involved in diverse cellular processes including apoptosis, DNA repair, cell cycle, signal transduction, tumour suppressor, vesicular transport, and peroxisomal biogenesis. RING finger protein 38 (RNF38) is a member of the family whose functions remain unknown. To gain insight into the putative effects of RNF38 in the central nervous system, we localised its expression. The aim of this study was to identify the neuroanatomical location(s) of rnf38 mRNA and its peptide, determine the type of RNF38-expressing cells, and measure rnf38 gene expression in the brain of male tilapia. The distributions of rnf38 mRNA and its peptide were visualised using in situ hybridisation with digoxigenin-labelled RNA antisense and immunocytochemistry, respectively. Both were identically distributed throughout the brain, including the telencephalon, preoptic area, optic tectum, hypothalamus, cerebellum, and the hindbrain. Double-labelling immunocytochemistry for RNF38 and the neuronal marker HuC/D showed that most but not all RNF38 protein was expressed in neuronal nuclei. Quantitative real-time polymerase chain reaction showed the highest level of rnf38 mRNA in the midbrain, followed by the preoptic area, cerebellum, optic tectum, telencephalon, hindbrain and hypothalamus. These findings reveal a differential spatial pattern of RNF38 in the tilapia brain, suggesting that it has potentially diverse functions related to neuronal activity.
Highlights
Interesting new gene (RING) finger domain is a zinc-binding motif found in a large family of functionally distinct proteins (Eisenberg et al, 2002)
The identified partial length of rnf38 cDNA consisted of 300 nucleotides spanning 2 exons when compared to the tilapia genome (Figure 1A)
The identified partial sequence of rnf38 cDNA cloned from Nile tilapia showed 100% homology to the predicted sequence in the GenBank (XM 005467588.3) (Figure 1B), and there was a high degree of homology between the partial sequence with the rnf38 genes of other species such as astatotilapia (Haplochromis burtoni), medaka (Oryzias latipes), mouse (Mus musculus), rat (Rattus norvegicus), and human (Homo sapiens) (Figure 1B)
Summary
Interesting new gene (RING) finger domain is a zinc-binding motif found in a large family of functionally distinct proteins (Eisenberg et al, 2002). Proteins containing the RING finger motif are capable of mediating ubiquitination and act as E3 ubiquitin ligases. Rnf mRNA is widely expressed in the brain, heart, liver, spleen, placenta, and testis (Eisenberg et al, 2002), and its protein is a nuclear protein that can bind to and ubiquitinate tumour-suppressor protein p53 (Sheren and Kassenbrock, 2013) through its E3 ubiquitin ligase activity (Sheren and Kassenbrock, 2013; Buetow et al, 2015). Rnf is up-regulated in the hypothalamic area during ageing (Fu et al, 2006) and concomitantly deleted during the lymphoid blast transformation of chronic myeloid leukaemia (Nacheva et al, 2010)
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