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Immunorelevant gene expression in LPS-challenged bovine mammary epithelial cells

100%

Pareek R., Wellnitz O., Van_ D. R., Burton J., Kerr D.

Journal of Applied Genetics

2005

vol. 46

issue 2

171-177

Infection of the bovine mammary gland, in addition to causing animal distress, is a major economic burden of the dairy industry. Greater understanding of the initial host response to infection may lead to more accurate selection of resistant animals or to novel prophylactic or therapeutic intervention strategies. The epithelial cell plays a role in the host response by alerting the immune system to the infection and providing a signal as to where the infection is located. To understand this process better, a cDNA microarray approach was used to search for potential signals produced by mammary epithelial cells in response to exposure to Escherichia coli lipopolysaccharide (LPS). Total RNA from separate cultures of epithelial cells from 4 Holstein cows was harvested 6 h after LPS challenge or control conditions. For each cow, RNA from control or LPS-exposed cells was transcribed to cDNA and labeled with Cy3 or Cy5, then pooled and applied to a bovine total leukocyte (BOTL) microarray slide containing 1278 unique transcripts. Dye reversal was used so that RNA from two of the control cultures was labeled with Cy3 while RNA from the other two control cultures was labeled with Cy5. From the resulting microarray data we selected 4 of the 9 genes significantly (P < 0.02) induced (>1.25-fold) in response to LPS exposure for more detailed analysis. The array signal intensity for 3 of these genes, RANTES/CCL5, IL-6 and T-PA, was relatively low, but quantitative real-time RT-PCR (Q-RT-PCR) analysis revealed that they were induced 208-fold, 10-fold and 3-fold, respectively. The gene that showed the greatest fold induction by microarray analysis (2.5-fold) was CXCL5. This gene had a relatively strong signal intensity on the array and was easily detected by northern blot analysis, which indicated a 10-fold induction. This cell culture model system provides evidence for an important role of the mammary epithelial cell in initiating the innate response to infection.

Polymerase chain reaction-heteroduplex (PCR-HD) polymorphism within the bovine STAT5A gene

88%

Flisikowski K., Zwierzchowski L.

Journal of Applied Genetics

2003

vol. 44

issue 2

185-189

Signal transducers and activators of transcription (STATs) are transcription factors mediating signals of various hormones and cytokines. STAT5A, previously known as the mammary gland factor (MGF), mediates the action of prolactin on milk protein gene expression in mammary epithelial cells. We used polymerase chain reaction-heteroduplex (PCR-HD) and sequencing methods for detection of nucleotide sequence polymorphism in intron 15 of the bovine STAT5A gene. A 281-bp gene fragment, from nt 12525 to nt 12806 (GenBank AJ 237937), was amplified with PCR, denatured and subjected to polyacrylamide gel electrophoresis to detect PCR-HD polymorphism. Three genotypes and two alleles were identified. DNA samples derived from homozygotes AA and BB were sequenced. A trinucleotide CCT deletion was found in the variant B of the STAT5A gene at position 12549. In a group of 72 beef bulls of various breeds and 49 Friesian bulls genotyped by PCR-HD mostly the AA genotype was found (from 83 to 61% depending on breed). The frequency of allele A varied between 0.91 and 0.77. Animals of genotype BB were found in Charolaise and Limousine breeds only.

Detecting a deletion in the coding region of the bovine bone morphogenetic protein 15 gene (BMP15)

75%

Zhang L.-P., Gan Q.-F., Zhang X.-H., Li H.-D., Hou G.-Y., Li Y.-J., Gao X., Ren H.-Y., Chen J. B., Xu S.-Z.

Journal of Applied Genetics

2009

vol. 50

issue 2

145-148

The aim of this study was to detect polymorphism in the bovine bone morphogenetic protein 15 (BMP15) gene. On the basis of PCR-SSCP and DNA sequencing, a 4-bp deletion was identified in the coding region of the gene. Sequence analysis revealed that the deletion altered the reading frame and introduced a stop codon at position 264. Eight breeds (Luxi, Qinchuan, Nanyang, Jinnan, Bohai Black, Menggolian, Holstein, and Simmental) were genotyped by PCR-SSCP. No cows homozygous for this mutation were observed in these breeds. Heterozygous cows were detected in Luxi, Qinchuan, Nanyang, Jinnan and Bohai Black cattle. Fecundity was not increased in heterozygous individuals.

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3 Journal of Applied Genetics

1 2009

1 2005

1 2003

1 Burton J.

1 Chen J. B.

1 Flisikowski K.

1 Gan Q.-F.

1 Gao X.

1 Hou G.-Y.

1 Kerr D.

1 Li H.-D.

1 Li Y.-J.

1 Pareek R.

1 Ren H.-Y.

1 Van_ D. R.

1 Wellnitz O.

1 Xu S.-Z.

1 Zhang L.-P.

1 Zhang X.-H.

1 Zwierzchowski L.

Search results

Immunorelevant gene expression in LPS-challenged bovine mammary epithelial cells

Polymerase chain reaction-heteroduplex (PCR-HD) polymorphism within the bovine STAT5A gene

Detecting a deletion in the coding region of the bovine bone morphogenetic protein 15 gene (BMP15)