Upregulation of Urinary Alpha‐2‐HS Glycoprotein During Pregnancy of Murrah Buffaloes

ABSTRACT Urine samples were systematically collected from inseminated Murrah buffaloes (Bubalus bubalis) on days 0, 7, 14, 21 and 28 (with day 0 representing the day of artificial insemination). Following confirmation of pregnancy via trans rectal palpation 45 days of insemination, the animals were...

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Published in:Reproduction in domestic animals 2024-12, Vol.59 (12), p.e70001-n/a
Main Authors: Prasada Rao, T., Paramesh, V., Pranavi, S., Manasa, V., Kumar, T. V. Sai, Padmaja, K.
Format: Article
Language:English
Subjects:
alpha-2-HS-Glycoprotein - analysis
alpha-2-HS-Glycoprotein - metabolism
Animals
Artificial insemination
Buffalo
Buffaloes - urine
early pregnancy
Female
fetuin‐A
Fingerprinting
Glycoproteins
Insemination, Artificial - veterinary
MALDI‐TOF
Mass spectrometry
Mass spectroscopy
Murrah buffaloes
Peptide mapping
Pregnancy
Pregnancy, Animal - urine
Proteins
Up-Regulation
Urine
Western blotting
Wheat germ
Wheat germ agglutinin
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Summary:ABSTRACT Urine samples were systematically collected from inseminated Murrah buffaloes (Bubalus bubalis) on days 0, 7, 14, 21 and 28 (with day 0 representing the day of artificial insemination). Following confirmation of pregnancy via trans rectal palpation 45 days of insemination, the animals were categorised into pregnant and non‐pregnant groups (n = 10 each). The urine samples on 0, 7, 14, 21 and 28 days of pregnant and one sample from non‐pregnant preferably collected on 28th day was used for SDS‐PAGE after diafiltration. We focused on urine samples obtained on 28th day of insemination as it could contain high fetuin‐A levels associated with pregnancy. Each of the two urine samples from both the pregnant and non‐pregnant groups underwent purification through wheat germ agglutinin (WGA) affinity separation. The resulting fraction/elute was subsequently employed for the determination and validation of fetuin‐A through 1D SDS‐PAGE, western blotting and MALDI‐TOF analysis. The samples were used for measuring the concentration of fetuin‐A by ELISA. Fetuin‐A was significantly higher (1020.50 ± 19.75 mg/L) in pregnant Murrah buffalo urine than their non‐pregnant counterparts (86.5 ± 6.33 mg/L), with a p 
ISSN:0936-6768
1439-0531
1439-0531
DOI:10.1111/rda.70001