A rare non-canonical splice site in Trema orientalis SYMRK does not affect its dual symbiotic functioning in endomycorrhiza and rhizobium nodulation.

BACKGROUND: Nitrogen-fixing nodules occur in ten related taxonomic lineages interspersed with lineages of non-nodulating plant species. Nodules result from an endosymbiosis between plants and diazotrophic bacteria; rhizobia in the case of legumes and Parasponia and Frankia in the case of actinorhizal species. Nodulating plants share a conserved set of symbiosis genes, whereas related non-nodulating sister species show pseudogenization of several key nodulation-specific genes. Signalling and cellular mechanisms critical for nodulation have been co-opted from the more ancient plant-fungal arbuscular endomycorrhizal symbiosis. Studies in legumes and actinorhizal plants uncovered a key component in symbiotic signalling, the LRR-type SYMBIOSIS RECEPTOR KINASE (SYMRK). SYMRK is essential for nodulation and arbuscular endomycorrhizal symbiosis. To our surprise, however, despite its arbuscular endomycorrhizal symbiosis capacities, we observed a seemingly critical mutation in a donor splice site in the SYMRK gene of Trema orientalis, the non-nodulating sister species of Parasponia. This led us to investigate the symbiotic functioning of SYMRK in the Trema-Parasponia lineage and to address the question of to what extent a single nucleotide polymorphism in a donor splice site affects the symbiotic functioning of SYMRK. RESULTS: We show that SYMRK is essential for nodulation and endomycorrhization in Parasponia andersonii. Subsequently, it is revealed that the 5'-intron donor splice site of SYMRK intron 12 is variable and, in most dicotyledon species, doesn't contain the canonical dinucleotide 'GT' signature but the much less common motif 'GC'. Strikingly, in T. orientalis, this motif is converted into a rare non-canonical 5'-intron donor splice site 'GA'. This SYMRK allele, however, is fully functional and spreads in the T. orientalis population of Malaysian Borneo. A further investigation into the occurrence of the non-canonical GA-AG splice sites confirmed that these are extremely rare. CONCLUSION: SYMRK functioning is highly conserved in legumes, actinorhizal plants, and Parasponia. The gene possesses a non-common 5'-intron GC donor splice site in intron 12, which is converted into a GA in T. orientalis accessions of Malaysian Borneo. The discovery of this functional GA-AG splice site in SYMRK highlights a gap in our understanding of splice donor sites.

Congenital malformations of the lumbosacral vertebral column are common in neurologically normal French Bulldogs, English Bulldogs, and Pugs, with breed-specific differences.

Congenital vertebral malformations are common findings on diagnostic imaging of the vertebral column in "screw-tailed" brachycephalic dogs. The aims of this study were to evaluate the prevalence and anatomical characteristics of lumbosacral congenital vertebral malformations in French Bulldogs, English Bulldogs, and Pugs presenting for problems unrelated to spinal disease, as well as possible associations with the degree of tail malformation, lumbosacral intervertebral disc herniation, or spondylosis deformans. In this retrospective cross-sectional study, CT scans of vertebrae L6 to S3 and of the coccygeal vertebrae were reviewed for type of congenital vertebral malformations (hemivertebrae, block vertebrae, lumbosacral transitional vertebrae, and spina bifida), lumbosacral intervertebral disc herniation, lumbosacral spondylosis deformans, and degree of tail malformation. In 76 (51.0%) of the 149 included dogs (53 French Bulldogs, 37 English Bulldogs, and 59 Pugs) at least one type of congenital vertebral malformations was found, with lumbosacral transitional vertebrae being the most common (34.2%). There was a significantly higher prevalence of lumbosacral transitional vertebrae (54.2%) and lower prevalence of hemivertebrae (1.7%) in Pugs compared to English (13.5% and 24.3%, respectively) and French Bulldogs (26.4% and 32.0%, respectively). Tail malformation was significantly more severe in dogs with evidence of hemivertebrae. Congenital vertebral malformations are a common finding in the lumbosacral vertebral column of French Bulldogs, English Bulldogs, and Pugs. These anatomical variances need to be considered when interpreting diagnostic studies and when planning for neurosurgical and neurodiagnostic procedures. Furthermore, this study suggests a possible association between the degree of tail malformation and lumbosacral hemivertebrae.

Assessment of menace response in neurologically and ophthalmologically healthy cats.

OBJECTIVES: Assessment and interpretation of menace response (MeR) in cats can be challenging. The prevalence of abnormal MeR in healthy cats is unknown. The aim of this study was to prospectively evaluate MeR in visually healthy cats. METHODS: Fifty cats without history or clinical evidence of neurological or ophthalmological disease were assessed by two examiners: standing behind the cat (mode A), in front of the cat (mode B), and in front of the cat, covering the contralateral eye (mode C). MeR was scored from 1-5 (absent, weak, moderate, strong, complete). Examination modes were compared concerning presence and score (descriptive statistic, 95% confidence interval, χ2 test). This was compared to a three-level scoring system (negative, reduced, positive). Score reproducibility between the two examiners was assessed (Cohen's kappa [κ] test). Video footage allowed self-re-evaluation and evaluation of the second examiner (κ analysis). Learning/tiring effect (McNemar test), influence of age, body weight (Spearman's rho test), skull type (χ2 test) and being an indoor or outdoor cat (Mann-Whitney U-test) were evaluated. RESULTS: MeR was always elicited with at least one technique. Comparable results were obtained with the five- and three-level scoring systems. Mode A achieved strong/complete (positive) MeR in 84.5%, mode B in 82% and mode C in 60%. Exact score reproducibility between the two examiners was slight to fair (κ = 0.208-0.281). Intrarater agreement for video self-assessment (κ = 0.544-0.639), as well as inter-rater agreement (extrinsic video assessment), was moderate to substantial (κ = 0.584-0.645). No learning/tiring effect ( P = 0.530) or association with body weight ( P = 0.897), age ( P = 0.724), skull type ( P >0.05) and being an indoor/outdoor cat ( P = 0.511) were evident. CONCLUSIONS AND RELEVANCE: The majority of visually healthy cats revealed a strong/complete MeR when the contralateral eye remained uncovered, but 40% failed when the contralateral eye was covered. The most reliable examination mode was achieved standing behind the cat.

Caudal articular process dysplasia of thoracic vertebrae in neurologically normal French bulldogs, English bulldogs, and Pugs: Prevalence and characteristics.

The aims of this study were to evaluate the prevalence and anatomical characteristics of thoracic caudal articular process dysplasia in French bulldogs, English bulldogs and Pugs presenting for problems unrelated to spinal disease. In this retrospective cross-sectional study, computed tomography scans of the thoracic vertebral column of these three breeds were reviewed for the presence and location of caudal articular process hypoplasia and aplasia, and compared between breeds. A total of 271 dogs met the inclusion criteria: 108 French bulldogs, 63 English bulldogs, and 100 Pugs. A total of 70.4% of French bulldogs, 84.1% of English bulldogs, and 97.0% of Pugs showed evidence of caudal articular process dysplasia. Compared to French and English bulldogs, Pugs showed a significantly higher prevalence of caudal articular process aplasia, but also a lower prevalence of caudal articular process hypoplasia, a higher number of affected vertebrae per dog and demonstrated a generalized and bilateral spatial pattern more frequently. Furthermore, Pugs showed a significantly different anatomical distribution of caudal articular process dysplasia along the vertebral column, with a high prevalence of caudal articular process aplasia between T10 and T13. This area was almost completely spared in French and English bulldogs. As previously suggested, caudal articular process dysplasia is a common finding in neurologically normal Pugs but this also seems to apply to French and English bulldogs. The predisposition of clinically relevant caudal articular process dysplasia in Pugs is possibly not only caused by the higher prevalence of caudal articular process dysplasia, but also by breed specific anatomical characteristics.