Molecular Characterization of Cytidine Monophospho-N-Acetylneuraminic Acid Hydroxylase (CMAH) Gene and Frequency of Blood Types in Stray Cats of İzmir, Turkey

Abstract

Cytidine monophospho-n-acetylneuraminic acid hydroxylase (CMAH) gene associated with blood groups in cats encodes CMAH enzyme that converts Neu5Ac to Neu5Gc. Although variations in CMAH gene of pedigree cats have been revealed, the presence/lack of them in non-pedigree stray cats is unknown. Therefore, the present study aimed to investigate the variations in CMAH gene and the quantity of Neu5Ac and Neu5Gc on erythrocytes of non-pedigree stray cats (n:12) living in İzmir, Turkey. Meanwhile, these 12 cats were typed using the mitochondrial DNA control region. Also, the frequency of blood types was determined in 76 stray cats including 12 cats that were used for CMAH and Neu5A/Neu5Gc analysis. In total, 14 SNPs were detected in 5’UTR as well as in exon 2, 4, 9, 10, 11 and 12 of CMAH gene. Among these SNPs, -495C>T in 5’UTR was detected for the first time as heterozygous in type A and AB cats, and homozygous and heterozygous in type B cats. The remaining 13 that have been detected in previous studies were also found as homozygous or heterozygous. Homozygous form (T/T) of the -495C>T polymorphism was found among only type B cats. Among the polymorphisms previously determined in the literature, homozygous form of the -371C>T polymorphism was found among only type B cats whereas heterozygous form (A/C) of the 327A>C polymorphism was detected in only type AB cats. Both Neu5Gc and Neu5Ac were detected in type A and AB cats. In type B cats, only Neu5Ac was detected. Among two type AB cats, the level of Neu5Ac was found higher in cat carrying heterozygous form (T/C) of 1392T>C. Mitotypes A, A6, D, E and 1 were detected among stray cats analysed for the characterisation of CMAH gene. The prevalence of type B cats (67.1%) was higher than others. As a result, the presence of a new SNP as well as previous SNPs indicates that more variations can be found in stray cats with a more comprehensive study in the future. Also, the high prevalence of type B cats demonstrates the high risk of neonatal isoerythrolysis among stray cats living in İzmir, Turkey.

Genetic epidemiology of blood type, disease and trait variants, and genome-wide genetic diversity in over 11,000 domestic cats

“…However, while there are several studies confirming genotypes associated with blood type AB [ 20 , 23 ], the genetic blood type determination and immunological blood type determination for two cats were unconcordant. In a recent study [ 77 ], type AB cats which carried one copy of a variant resulting in blood type B, had a higher level of Neu5Ac present that could depending on the technologies used impact results. This provides further justification for the supportive role of the DNA-based blood type determination approaches in breeding and veterinary care, provided that the appropriate carefully validated genetic variants are assayed.…”

“…This provides further justification for the supportive role of the DNA-based blood type determination approaches in breeding and veterinary care, provided that the appropriate carefully validated genetic variants are assayed. However, additional genetic variants of the CMAH gene have been identified that may play a role in blood type determination in domestic shorthair and stray cats [ 23 , 77 ], and further investigations are warranted.…”

N-glycolylneuraminic acid as a carbohydrate cancer biomarker

“… ND in spleen and liver tissues [ 19 , 80 ] Cat Type A (mainly express Neu5Gc) contain two intact A alleles and type B (express Neu5Ac only) and AB (express both antigens) carried two mutated alleles with distinctive mutations which are linked to the CMAH protein expression and its activity Detected in various tissues Detected in the cell lysates from two feline cells The erythrocytes in type A cats express Neu5Gc with low levels of Neu5Ac and type B cats only express Neu5Ac, while type AB cats express both antigens at similar levels. [82] , [83] , [84] , [85] , [86] European dog The well-reserved CMAH gene but with SNPs Detected in low to absent levels in canine cell lines ND in the cell extracts of two western canine cell lines ND in glycolipid extractions from erythrocytes [ 19 , 81 , 82 , 84 , 87 , 88 ] …”