Expression patterns of immune associated genes in Euoniticellus intermedius and characterization of the embryonic cell line
Year: 2013
Type: dissertation
Abstract: As bacteria are becoming resistant to conventional antibiotics, researchers are looking for \nnew ways to combat microbial infection. We have begun to adopt genetic and functional genomic \napproaches to define the molecular determinants of pathogen resistance in the dung beetle, \nEuoniticellus intermedius. This dung beetle survives microbe-rich environments such as dung. This \nability makes it a potential model for the study of infectious agents and ecological damage. To date, \nE. intermedius has not been studied at the molecular level. In this study, a range of complimentary \nanalytical techniques were used to characterize the E. intermedius embryonic cell line established in \nour laboratory. These techniques characterize morphology, growth characteristics, karyotype, \nisoenzyme patterns and embryonic development. Complete characterization of the E. intermedius \ncell line is essential for the cell banks and for the regulatory requirements in biopharmaceutical \nproduction. \nThis study followed gene sequences and their comparisons for both adult and cell line to \nconfirm that the E. intermedius (EISA08) cell line is originated from the embryonic E. intermedius \ndung beetle. cDNA was synthesized from mRNA isolated from E. intermedius adult beetles and cell \nline (EISA08) was sequenced using GS (FLX) technology by a commercial facility, Inqaba \nBiotechnical Industries (Pty) Ltd, South Africa. In addition to characterization of the cell line, two \ngenes, namely hopscotch and ribosomal protein S9 (RpS9), were selected from the Flylab genome \ndata base. The E. intermedius database is a web-based system for the genome and transcriptome of \nthe dung beetle to evaluate the immune system of the dung beetle (http://Flylab.wits.ac.za/). \nhopscotch was selected because it is believed to be involved in the JAK-STAT signalling pathway \nfor anti-viral response, embryonic development and cell growth. Rsp9 was chosen as a loading \ncontrol because it is expected to be a housekeeping gene. The conserved molecular signalling pathway JAK-STAT is used by E. intermedius (as in other insects and humans) for immune defence \nand early embryonic development. The project followed hopscotch and Rsp9 gene expression in all \nthe E. intermedius life cycle developing stages; adult, pupae, larvae, embryo, and cell line cell \ngrowth, life cycle developing stages and embryonic development has was monitored. \nE. intermedius embryonic development is described as short germ-band. E. intermedius \nembryogenesis is regarded as basal and is observed in most arthropods. The study revealed that \nE. intermedius hopscotch is over expressed in the early developing stages, embryo, larvae, and \npupae and in the newly established cell line EISA08. The results from this study lead to the \nsuggestion that E. intermedius JAK-STAT pathway is activated early and has an important role in \nembryonic development, cell proliferation and immune defence. Studies of E. intermedius could \nprovide more insight into the properties and evolution of innate immunity and embryonic \ndevelopment.
Author: Mohammed Alaouna
Language: English
Subfield: Immunology
Field: Immunology and Microbiology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W2280449207