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Ano1 is a better marker than c-Kit for transcript analysis of single interstitial cells of Cajal in culture

100%

Loera-Valencia R., Wang X.-Y., Wright G. W. J., Barajas-Lopez C., Huizinga J. D.

Cellular and Molecular Biology Letters

2014

tom 19

nr 4

The interstitial cells of Cajal (ICC) drive the slow wave-associated contractions in the small intestine. A commonly used marker for these cells is c-Kit, but another marker named Ano1 was recently described. This study uses single-cell RT-PCR, qPCR and immunohistochemistry to determine if Ano1 could be reliably used as a molecular marker for ICC in single-cell mRNA analysis. Here, we report on the relationship between the expression of c-Kit and Ano1 in single ICC in culture. We observed that Ano1 is expressed in more than 60% of the collected cells, whereas c-Kit is found only in 22% of the cells (n = 18). When we stained ICC primary cultures for c-KIT and ANO1 protein, we found complete co-localization in all the preparations. We propose that this difference is due to the regulation of c-Kit mRNA in culture. This regulation gives rise to low levels of its transcript, while Ano1 is expressed more prominently in culture on day 4. We also propose that Ano1 is more suitable for single-cell expression analysis as a marker for cell identity than c-Kit at the mRNA level. We hope this evidence will help to validate and increase the success of future studies characterizing single ICC expression patterns.

Polymorphisms in intron 1 of the porcine POU1F1 gene

72%

Song C. Y., Gao B., Teng S. H., Wang X. Y., Xie F., Chen G. H., Wang Z. Y., Jing R. B., Mao J. D.

Journal of Applied Genetics

2007

tom 48

nr 4

371-374

This study was conducted to detect polymorphisms in intron 1 of porcine POU1F1 (POU domain, class 1, transcription factor 1, Pit 1, renamed as POU1F1) by comparative sequencing. Within the intron, 23 sites of variation were identified, including 16 single-nucleotide substitutions, 4 single-nucleotide indels, 2 short (3-bp and 17-bp), and one long (313-bp) indels. Several important regulatory motifs were found within the 313-bp indel by in silico analysis. The 313-bp indel was next genotyped in 11 Chinese native pig breeds and 4 western meat-type pig breeds. The appearance of genotypes varied between breeds: among Chinese native breeds, no AA and AB genotypes were found in Tibetan, Lingao, Min, Rongchang, and Songliao Black pigs, no AA genotype was found in Fenjing and Leping Spotted pigs, whereas in Pietrain and Landrace there were no BB genotypes, and all 19 Duroc pigs were A A homozygotes. The western meat-type pigs had high A allele frequencies and the Chinese pigs had more B alleles, except Jianquhai pigs. A positive association of the AA genotype with birth weight was observed in a commercial pig line. This paper demonstrated that the genetic variation in intron 1 of the pig POU1F1 gene was high and these polymorphisms may provide useful makers for QTL analysis.

Ograniczanie wyników

1 Cellular and Molecular Biology Letters

1 Journal of Applied Genetics

1 Barajas-Lopez C.

1 Chen G. H.

1 Gao B.

1 Huizinga J. D.

1 Jing R. B.

1 Loera-Valencia R.

1 Mao J. D.

1 Song C. Y.

1 Teng S. H.

1 Wang X. Y.

1 Wang X.-Y.

1 Wang Z. Y.

1 Wright G. W. J.

1 Xie F.

1 2014

1 2007

Wyniki wyszukiwania

Ano1 is a better marker than c-Kit for transcript analysis of single interstitial cells of Cajal in culture

Polymorphisms in intron 1 of the porcine POU1F1 gene