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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.

Biotechnology of the mammary gland - perspectives of genetic engineering of milk proteins

100%

Zwierzchowski L.

Biotechnologia

1998

issue 2

33-56

In this article, state of art and perspectives in mammary gland biotechnology are reviewed. Recent progress in recombinant DNA technology as well in embryo manipulation and transfer has made the introduction of specific genes into the germline of animals relatively easy. With appropriate genetic constructs, the expression of the inserted genes in transgenic animals can be controlled in a tissue-specific and in a differentiation-specific manner. Thus, it is now possible to consider alteration of the composition of milk produced by lactating animals in a variety of ways. There is a growing list of foreign milk proteins that have been expressed, and one can envisage placing almost any protein gene of interest under the control of promoter of a milk protein gene. Many human proteins of a potential pharmaceutical use may be now produced in the mammary glands of laboratory or farm animals. Modification of milk composition can be extended not only to produce proteins of commercial value but also, by manipulation of key metabolic enzymes, to fat, lactose, and other components of milk. Many alternations in ruminants' milk composition, including 'humanization' of cow's milk, are planned, however, these manipulations must avait the development of totipotent embryonic cell lines (ESC) of farm animals, cells that enable gene manipulation by homologous recombination. In spite of a great progress, many obstacles and difficulties still exist on the way to economical production of human paharmaceuticals in farm 'transgenic bioreactoractors'. These dificulties are discussed in detail.

Mast cells in the mouse mammary gland ? correlation with the development of lactiferous structures

88%

Szewczyk G., Szukiewicz D., Zaczek R., MaslinskA D.

Folia Biologica

2000

vol. 48

issue 1-2

13-17

Specimens of mouse mammary glands obtained from animals being in different phases of reproductive cycle were collected. After staining MCN, the total mast cell area (TMC), mean mast cell area (MMC), and lactiferous structure density index (LDI) were examined in sections, using a computer image analysis system. Statistically significant (P<0.05) results were: 1. An increase in MCN observed in Group I (4-5 and 9-10 days of pregnancy), a decrease in MCN observed in Group II (2nd and 10th day of lactation); 2. Changes of TMC fully corresponding to changes of MCN; 3. Increase in MMC observed in Group I at mid (days 9-10) and at the end (days 18-19) of pregnancy, decrease in MMC observed in Group IIB (10th day of lactation); 4. LDI (%) higher at the end of pregnancy (Group IC) and during lactation (groups IIA, IIB), compared with control (23.5 ? 4.12, 37.6 ? 3.24, 71.0 ? 4.33 vs. 3.8 ? 0.39). The observed changes in the number and size of MC strictly correspond to physiological phenomena leading to alternation of the mouse mammary gland functional status by development/involution of the lactiferous structures.

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Search results

Immunorelevant gene expression in LPS-challenged bovine mammary epithelial cells

Biotechnology of the mammary gland - perspectives of genetic engineering of milk proteins

Mast cells in the mouse mammary gland ? correlation with the development of lactiferous structures