Gene expression of six major milk proteins in primary bovine mammary epithelial cells isolated from milk during the first twenty weeks of lactation

Sigl, Tanja; Meyer, H. H. D.; Wiedemann, S.

doi:10.17221/6347-cjas

Cited by 30 publications

(59 citation statements)

References 48 publications

(58 reference statements)

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“…Similar results were obtained in cows, where extracted quantity of pBMEC mRNA did not vary during experimental timeframe [18]. In the present study, the RNA yield and pBMEC in milk of Surti buffaloes were positively and significantly correlated (0.76).…”

Section: Resultssupporting

confidence: 89%

“…[18] with slight modifications. The recovered pBMECs further used for recovering RNA for down step quantification of major milk lipogenic gene expression.…”

Section: Resultsmentioning

confidence: 99%

“…[18]. After extraction, RNA was purified using miRNeasy Mini kit (Qiagen, Germany) and followed by on-column digestion with the RNase-free DNase (Qiagen, Germany) RNA was quantified by Nanodrop spectrophotometric (Thermo Scientific ND 2000C, USA).…”

Section: Methodsmentioning

confidence: 99%

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Relative gene expression of fatty acid synthesis genes at 60 days postpartum in bovine mammary epithelial cells of Surti and Jafarabadi buffaloes

Janmeda¹,

Kharadi²,

Pandya³

et al. 2017

Vet World

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Aim:Aim of the study was to study the relative gene expression of genes associated with fatty acid synthesis at 60 days postpartum (pp) in bovine mammary epithelial cells (MECs) of Surti and Jafarabadi buffaloes.Materials and Methods:A total of 10 healthy Surti and Jafarabadi buffaloes of each breed were selected at random from Livestock Research Station, Navsari and Cattle Breeding Farm, Junagadh, Gujarat, respectively, for this study. Milk sample was collected from each selected buffalo at day 60 pp from these two breeds to study relative gene expression of major milk fat genes using non-invasive approach of obtaining primary bovine MECs (pBMEC) from milk samples.Results:In this study overall, the relative expression of the six major milk lipogenic genes butyrophilin subfamily 1 member A1 (BTN1A1), stearoyl-CoA desaturase (SCD), lipoprotein lipase (LPL), glycerol-3-phosphate acyltransferase mitochondrial (GPAM), acetyl-coenzyme A carboxylase alpha (ACACA), and lipin (LPIN) did not show changes in expression patterns at 60th day of lactation in both Surti and Jafarabadi buffaloes.Conclusion:The pBMEC can be successfully recovered from 1500 ml of milk of Surti and Jafarabadi buffaloes using antibody-mediated magnetic bead separation and can be further used for recovering RNA for down step quantification of major milk lipogenic gene expression. The relative expression of the six major milk lipogenic genes BTN1A1, SCD, LPL, GPAM, ACACA, and LPIN did not show changes in expression patterns in both Surti and Jafarabadi buffaloes, suggesting expression levels of lipogenic genes are maintained almost uniform till peak lactation without any significant difference.

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Section: Resultssupporting

confidence: 89%

“…[18] with slight modifications. The recovered pBMECs further used for recovering RNA for down step quantification of major milk lipogenic gene expression.…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Relative gene expression of fatty acid synthesis genes at 60 days postpartum in bovine mammary epithelial cells of Surti and Jafarabadi buffaloes

Janmeda¹,

Kharadi²,

Pandya³

et al. 2017

Vet World

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“…Then, the genomic approach (Schopen et al 2009;Pribyl et al 2010Pribyl et al , 2012Pribyl et al , 2013Matejickova et al 2013;Szyda et al 2013) could be completed by the known major genes concerning milk performance and by the biometric approach (Sigl et al 2012;Meszaros et al 2013;Zavadilova and Stipkova 2013;Zavadilova and Zink 2013). When different effects depending on the breed are found, e.g.…”

Section: Resultsmentioning

confidence: 99%

Effect of DGAT1 polymorphisms on the estimated breeding values of Czech Simmental sires

Hanusová¹,

Mikova²,

Večerek³

et al. 2014

CZECH J ANIM SCI

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ABSTRACT:The aim was to evaluate the effect of polymorphisms in the promoter and in the coding region of the DGAT1 gene on the estimated breeding values (EBV) of Czech Simmental sires. The K232A polymorphism (n = 191) in the coding region was genotyped by PCR/RFLP, and the KU and SA polymorphisms in the promoter (n = 203) were identified in an automatic sequencer. In the K232A polymorphism, the frequency of the genotype AA (Alanine) was greater than that of the genotype KA, the homozygous genotype KK (Lysine) was not found. Similarly, the allele A predominated over the K allele (0.945 and 0.055). The EBV for milk performance have been assigned to the genotypes, and the associations quantified. For the AA genotype and A allele, positive association with EBV of milk yield and protein yield was found, and negative association with the breeding values of fat percentage and yield, and protein percentage, but only the value of fat content was found to be significant. The positive non-significant association of the A variant with the protein yield was caused by the high milk yield. In the KU polymorphism, the CC genotype was associated significantly with lower EBV for the fat percentage, both the C allele and the CC genotype were associated with higher EBV for milk yield, so both the fat and the protein yield were non-significantly increased. For the diplotypes K232A/KU, there was a significant association with the fat percentage. The AACC combination seemed to have some breeding potential. The K232A polymorphism explained maximum of 6.2% of EBV variability, the KU polymorphism of 4.4%, and the SA polymorphism of 4.2%. The diplotypes K232A/KU explained maximum of 7.4% of variability. The highest proportion of variability was explained for fat percentage. The results confirmed the important role of the BTA14 region in controlling milk performance.

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“…Secretion of casein is the hallmark of the bovine mammary epithelial cells (Fox & McSweeney, 2006). The genes those are responsible for most of milk proteins are regulated by the intricate interplay of peptide and steroid hormones, predominantly the lactogenic hormones such as prolactin, hydrocortisone, insulin, cell-cell and cell-substratum interactions (Sigl, Meyer, & Wiedemann, 2012). We used protein induction media consisted basal growth media supplemented with lactogenic hormones such as hydrocortisone, prolactin, and β-estradiol and progesterone as mammary epithelial cell proliferating hormone.…”

Section: Discussionmentioning

confidence: 99%

Differentiation dynamics of mammary epithelial stem cells from Korean holstein dairy cattle under ECM-free conditions

Sharma

Kim

Sodhi

et al. 2015

Journal of Biomolecular Structure and Dynamics

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The "stem cells" are commonly defined as "cells capable of self-renewal through replication and differentiating into specific lineages". The mammary gland contains functional stem/progenitor cells. The current study was planned with the objectives to study the differentiation dynamics of Korean Holstein mammary epithelial stem cells (KHMESCs) under the optimum culture conditions. Lineage negative KHMESCs isolated from mammary tissue of lactating cows have shown the typical differentiation dynamics with formation of lobulo-alveolar structures in in vitro culture. This suggests the existence of bipotential mammary epithelial stem cells in the mammary gland. The strong mRNA expression of pluripotency factors indicates stemness, whereas expression of milk protein genes and epithelial cell-specific gene indicate their differentiation capabilities. Further, immunostaining results have shown the differentiation capabilities of KHMESCs into both luminal and basal lineages under the extracellular matrix (ECM, matrigel) free environment. However, under matrigel, the differentiation process was comparatively higher than without matrigel. Immunostaining results also suggested that differentiated cells could secrete milk proteins such as β-casein. To our knowledge, these data represent the first report on the differentiation dynamics and establishment of mammary epithelial stem cells from Korean Holstein with typical stemness properties. It was observed that isolated KHMESCs had normal morphology, growth pattern, differentiation ability, cytogenetic and secretory activity even without ECM. Therefore, it is concluded that established KHMESCs could be used for further studies on Korean Holstein dairy cows related to lactation studies, as non-GMO animal bioreactors and stem cell-based management of bovine mastitis including post-mastitis damage.

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Gene expression of six major milk proteins in primary bovine mammary epithelial cells isolated from milk during the first twenty weeks of lactation

Cited by 30 publications

References 48 publications

Relative gene expression of fatty acid synthesis genes at 60 days postpartum in bovine mammary epithelial cells of Surti and Jafarabadi buffaloes

Relative gene expression of fatty acid synthesis genes at 60 days postpartum in bovine mammary epithelial cells of Surti and Jafarabadi buffaloes

Effect of DGAT1 polymorphisms on the estimated breeding values of Czech Simmental sires

Differentiation dynamics of mammary epithelial stem cells from Korean holstein dairy cattle under ECM-free conditions

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