Allele Frequency of APAF1 Mutation in Holstein Cattle in Brazil.

Lukas Garrido Albertino,Julia Franco Ferreira,João Pedro Marmol Oliveira,Alexandre Secorun Borges,Ana Luísa Holanda Albuquerque,José Paes Oliveira-Filho,Thais Helena Constantino Patelli

doi:10.3389/fvets.2022.822224

Lukas Garrido Albertino, Julia Franco Ferreira + Show 5 more

Open Access

https://doi.org/10.3389/fvets.2022.822224

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Abstract

APAF1 is an autosomal recessive inherited mutation, associated with Holstein haplotype 1 (HH1) and characterized by a substitution of cytosine for a thymine (c.1741C>T) in chromosome 5. The mutation causes fetal and embryonic loss, between 60 and 200 days of gestation, and reduced conception rate. The ARMS-PCR is considered a simple and low-cost method to determine single nucleotide polymorphism (SNP) with no need for genetic sequencing of the animal genome. This study aimed to verify the allelic frequency of APAF1 mutation in Brazilian Holstein cattle. A total of 248 Holstein DNA samples (210 cows and 38 bulls) were analyzed, and synthetic genes were manufactured to validate the primers developed by the authors. All animals assessed in this study were classified as wild-type for APAF1 mutation. The primers and protocol developed for the ARMS-PCR technique work with 100% specificity and efficiency since the amplicon formations are as expected according to the genotypes. In conclusion, the mutation responsible for APAF1 was not detected in the Brazilian Holstein cattle population assessed in this prevalence study, although it is not possible to affirm that APAF1 does not occur in Brazilian Holstein animals. The tetra-primer ARMS-PCR protocol for APAF1 mutation that has been validated here may be a relatively simple and economical method to determine the animals' genotype.

Highlights

The APAF1 protein is an important molecule in the cell apoptosis process [1] and central nervous system development during embryogenesis [2]
The ARMS-polymerase chain reaction (PCR) primers and protocol developed here worked in the classification and determination of wild-type animals, forming amplicons according to their preview fragments [430 bp for outer primers and 286 bp for inner (C)]
The genetic basis of the Holstein Brazilian herd has been formed by animals, semen, and embryos imported from Europe and North America [13] and previous studies in other countries have indicated the importance and impact of this mutation in the breed [3,4,5, 8,9,10,11,12], the APAF1 mutation has not been described in Brazil yet

Summary

Introduction

The APAF1 (apoptotic protease activating factor-1) protein is an important molecule in the cell apoptosis process [1] and central nervous system development during embryogenesis [2]. In Holstein cattle, the mutation causes fetal and embryonic loss between 60 and 200 days of gestation, and reduced conception rate, and was estimated to cause more than 500,000 abortions and a loss of US$450 million to the dairy industry [3]. This mutation was associated with a common ancestor and carrier, the bull Pawnee Farm Arlinda Chief and its descendants, i.e., Walkway Chief Mark and Millu Betty Invahoe Chief. The ARMS-PCR is APAF1 Mutation in Brazilian Holstein Cattle considered a simple and low-cost method to determine SNP with no need for genetic sequencing of the animal genome [7], and it has been recently validated to determine the HH1 in Holstein cattle [6]

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