Purpose: To investigate the existence of dipeptide transporter PepT1 at the mRNA level in the pigmented rabbit conjunctiva. Methods: Total RNA was isolated from the conjunctival and the jejunal (positive control) epithelial cells using the guanidine hvdrochloride-chloroform-isopropanol method. The amount of total RNA was determined and the integrity of RNA was analyzed. Complementary DNA was made by using total RNA and reverse transcriptase, which was then amplified using polymerase chain reaction. Two primers (bases 31 to 50 and 1031 to 1050) were synthesized based on the rabbit jejunal oligopeptide transporter PepT1 cDNA sequence. The length of PCR product was predicted to be 1.0 kb. The RT-PCR products were analyzed with 1% agarose gel and stained with ethidium bromide. Results: RT-PCR products revealed a positive band at 1.0 kb for both the conjunctival and the jejunal epithelial samples. The signal intensity for the band from the conjunctival epithelial sample, however, was lesser than that from the jejunal sample when equal amounts of RNA were used. This suggests a lower level of expression for PepT1 in the conjunctiva compared to the jejunum. Conclusions: Previously we have demonstrated H+-coupled dipeptide transport properties in the pigmented rabbit conjunctiva. Although the expression level at the mRNA level is lower in the conjunctiva compared to the jejunum, these results coupled with our earlier findings suggest the possibe existence of dipeptide transporter in the pigmented rabbit conjunctiva.
|Original language||English (US)|
|Journal||Investigative Ophthalmology and Visual Science|
|State||Published - Feb 15 1996|
ASJC Scopus subject areas
- Sensory Systems
- Cellular and Molecular Neuroscience