PAX6 is a candidate gene for the proper development of human eye. The gene encompasses on the chromosome number 11p13. The expression of PAX6 gene occurs not only in eye but in many human organs like brain and pancreas. The present study focuses on the transcript analysis in ocular tissues and non-ocular tissues.
The total mRNA extracted from ocular tissues including lens
epithelium, retina, iris and the non-ocular tissues like Trabecular Meshwork
Cells, HeLa Cells, besides Aniridic patient as well as the normal person blood
samples. The isolated RNAs were subjected to convert into cDNA by RT-PCR along
with the house keeping genes (beta actin) used as a positive control in all the
reactions.
The post PCR analysis revealed 540bp amplicon in all the
isolated ocular and non-ocular tissues thus proved that the reaction setup was
correct. In the ocular tissues showed approximately 600bp in retina, 855bp in
iris and 1055bp in lens epithelium (PAX6 and PAX6 5a isoforms). The expression
analysis of PAX6 gene in Non-ocular tissues HeLa cells showed 1.2 kb
amplification however the mutant revealed showed unusual banding pattern of
190bp and 200bp. Sequencing of the unusual bands showed the irrelevant hypothetical
LOC441763 mRNA. RT-PCR amplification confirms the presence of various PAX6 mRNA
expressions in ocular and non-ocular tissues.
Lens epithelium, retina and iris tissues have significant
PAX6 expression while the retina contains an overwhelming majority of mRNAs
encoding crystalline and other genes but still express PAX6 isoforms at high
levels. HeLa is a fibroblast based cell line and it would not expect a
fibroblast to express a lot of PAX6 However PAX6 gene expression might be high
in these cells for some reason. The PAX6 gene partial expression was found in
mutant WBC and not in wild type WBC cells, the real fact for this reason is
unknown. Hence the exploration of the PAX6 gene should be further analyzed in
future.
Author(s) Details:
G. Neethirajan,
Department of Genetics, Aravind Medical Research Foundation, India and PG Research Centre in Biotechnology, MGR College, Hosur, TN, India.
Krishnadas S. R.,
Department of Glaucoma, Aravind Eye Hospital, India.
Vijayalakshmi P.,
Department of Pediatric Ophthalmology and Strabismus, Aravind Eye Hospital, India.
Shashikanth Shetty,
Department of Pediatric Ophthalmology and Strabismus, Aravind Eye Hospital, India.
Sundaresan P.,
Department of Genetics, Aravind Medical Research Foundation, India.
Please see the link here: https://stm.bookpi.org/NVMMS-V3/article/view/13954
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