Identification of candidate genes for congenital splay leg in piglets by alternative analysis of DNA microarray data

Abstract

The congenital splay leg syndrome in piglets is characterized by a temporarily impaired functionality of the hind leg muscles immediately after birth. Etiology and pathogenetic mechanisms for the disease are still not well understood. We compared genome wide gene expression of three hind leg muscles (M. adductores, M. gracilis and M. sartorius) between affected piglets and their healthy littermates with the GeneChip® Porcine Genome Array (Affymetrix) in order to identify candidate genes for the disease. Data analysis with standard algorithms revealed no significant differences between both groups. By application of an alternative approach, we identified 63 transcripts with differences in two muscles and 5 genes differing between the groups in three muscles. The expression of six selected genes (SQSTM1, SSRP1, DDIT4, ENAH, MAF, and PDK4) was investigated with SYBRGreen RT - Real time PCR. The differences obtained with the microarray analysis could be confirmed and demonstrate the validity of the alternative approach to microarray data analysis. Four genes with different expression levels in at least two muscles (SQSTM1, SSRP1, DDIT4, and MAF) are assigned to transcriptional cascades related to cell death and may thus indicate pathways for further investigations on congenital splay leg in piglets.

Prevalence of Swimming Puppy Syndrome in 2,443 Puppies during the Years 2006–2012 in Thailand

“…In the case of pigs, splay leg has been proven to be a hereditary disorder [10, 11]. Maak and others [11] pinpointed the candidate genes for splay leg in piglets using DNA microarray data, comparing the genome-wide gene expression of three hindlimb muscles between affected and healthy piglets.…”

“…Maak and others [11] pinpointed the candidate genes for splay leg in piglets using DNA microarray data, comparing the genome-wide gene expression of three hindlimb muscles between affected and healthy piglets. They found 63 transcripts with differences in two muscle groups and 5 gene differences in three muscle groups.…”

Exclusion of sequence polymorphisms in the porcine ITGA5 and MIR148B loci as causal variation for congenital splay leg in piglets

“…Such materials are peer-reviewed and may be re-organized for online delivery, but are not copy-edited or typeset. 3 Recently, microRNAs (MIR) have been shown to modulate the expression of a large number of genes during myogenesis and postnatal growth in swine. Background: Integrin alpha 5 (ITGA5) encodes the alpha chain of the heterodimeric fibronectin receptor.…”

The Splay Leg Syndrome in Piglets: A Review

“…The existence of one or more major genes for congenital splay leg seems possible (Maak et al, 1999(Maak et al, , 2001a2001b). Among the fragments strongly displayed in the splay leg muscle, are identified the porcine CDKN3 gene (cyclin-dependent kinase inhibitor 3) (Maak et al, 2002).…”

“…CDKN3 belongs to a family of dual-specificity protein phosphatases and is involved in cell cycling (Gyuris et al, 1993;Hannon et al, 1994;Maak et al 2003). Recently, Maak et al (2009) (Ward and Bradley, 1980;Jirmanova, 1983;Ooi et al, 2006;Boettcher et al, 2007;2008). Ooi et al (2006) described congenital splay leg as muscle fiber atrophy characterized by an increased expression of the atrophy marker FBXO32 (atrogin, MAFbx) and histological signs of a generalized muscle fiber hypoplasia in skeletal muscles of splay leg piglets (Ooi et al, 2006).…”