Genetic screening of the inherited Ichtyosis causative mutation in Chianina cattle

Inherited Ichthyosis is a genetic disorder reported in both humans and animals, including bovines. Two inherited forms were reported in cattle and both are transmitted in an autosomal recessive manner: Ichthyosis Fetalis (IF) and Ichthyosis Congenita (IC). A causative mutation of IF in Chianina cattle was recently indentified in the ABC12 gene. This work reports the first genetic screening using this recently available genetic test on Chianina cattle. Tests were performed on both the population of farm breeding selected young bulls (131 samples randomly chosen) and high breeding value sires (16 samples). Results confirm a low total prevalence of carriers in the selected sire population (2/131; 1.5%) and the presence of the disease allele among the high value selected sires (1/16; 6.3%). This result strengthens the importance to continue the genetic screening program, particularly in performance tested bulls approved for use in AI or natural servic

STUDIO DI PATOLOGIE GENETICHE E CARATTERI DI INTERESSE A BASE EREDITARIA NEGLI ANIMALI D'AFFEZIONE

Primary hypertrophic cardiomyopathy (HCM) is the most common cardiac disease in cats and humans, characterized by an impressive phenotypic and genotypic heterogeneity. Autosomal dominant inheritance and two causative mutations in cardiac myosin binding protein C 3 gene (MYBPC3), at position A31P and R820W, were respectively identified in Maine Coon and Ragdoll cats with HCM. Beyond the reported Maine Coon and Ragdoll mutations, A74T, another single nucleotide polymorphism (SNP) in MYBPC3, was suspected to cause HCM in Maine Coon. In the present work, HCM-associated mutations have been genotyped by direct sequencing in 741 Italian cats examined through standardized ultrasound. The prevalence of the mutations and their correlation with the disease in Maine Coon and Ragdoll cats have been calculated. In one more focused cohort of 393 samples, the MYBPC3 region including A31P and A74T loci it has been analyzed more in detail. Multiple alignment have been performed clustering samples by different breeds and several \u201cgeno-variants\u201d have been recorded. Future research will be directed to apply high density array (75.000 SNPs) to the research on feline hypertrophic cardiomyopathy, aiming at identifying more mutations in the affected animals and at evaluating their association with the developments of the disease. X-linked progressive retinal atrophy 2 (XLPRA2) is a severe, early-onset, and progressive rod and cone disease of dogs, caused by a 2 bp microdeletion in RPGRORF15. This study provides a list of the relevant DE miRNAs associated with retinal degeneration and disease progression in XLPRA2-mutant retinas at the most relevant disease-related ages: before (3 wks), during (7 wks), and after (16 wks) the peak of photoreceptor death, to further examine their role in retinal development and in RPGRORF15 mutant retinas. Expression profiles of age-matched 3, 7, and 16 wks old normal and XLRPA2-mutant retinas (3/age/group) were analyzed using miRNA-specific Affymetrix microarrays (46,228 probes/7,815 probe sets/177 canine specific). To confirm the data obtained from the array, quantitative real-time PCR of miR-183, miR-155, miR-146a, miR-129, miR-29b, and miR-19a was performed. Future research will be directed to real-time PCR validation of additional DE miRNAs, in vitro analysis at the protein level, integration with recently published transcriptomic data10, and comparison with other similar retinal degenerative diseases (e.g. rcd1) in order to determine if the DE miRNAs are disease-specific, or common to several early onset retinal degenerations

Comparison of different procedures to isolate feline peripheral blood mononuclear cells (PBMCs) from small volumes of blood

Studies on leukocytes isolated from feline blood do not provide details on the performances of isolation techniques.1 From the few numerical data available, however, it can be assumed that the purity of isolated cell populations is high but their recovery rate is low.2 Therefore, large volumes of blood (difficult to collect from cats with spontaneous diseases) are required to obtain enough leukocytes for in vitro studies. The aim of this study was to assess the performances of isolation techniques on small volumes of feline blood. Blood samples (1 to 5 mls) were drawn from clinically healthy cats and placed in EDTA-coated tubes. Fifteen session of tests (10 using Ficoll, 5 using Percoll) were performed. In 9 cases cells were further separated by adherence on Petri dishes (PD) and in 5 cases using iron-labelled monoclonal antibodies against leukocyte antigens followed by magnetic sorting (MS). Cell purity (i.e. the percentage of each population) and recovery (i.e. the percentage of cells of each population recorded after isolation compared with blood) were then calculated. The purity of lymphocytes was significantly higher (P=0.015) with Ficoll (79.6 \ub1 3.3) than with Percoll (61.0 \ub112.0); the purity of monocytes was low, and significantly higher (P=0.015) with Percoll (32.7 \ub1 13.5) than with Ficoll (9.0 \ub1 1.6). The recovery rate of lymphocytes was low, and significantly higher (P<0.001) with Ficoll (54.7 \ub1 27.5) than with Percoll (12.0 \ub1 4.2). The recovery rates of monocytes recorded with Ficoll (59.4\ub1 34.7) or Percoll (38.5 \ub1 9.38) were not significantly different. The purity of cell types in PD was not morphologically determinable. However, assuming that at least 90% of adherent and non adherent cells were monocytes and lymphocytes, respectively, the recovery rate for both the populations was lower than 10% in most cats. The purity after MS was generally high, but the recovery rate was variable and the number of yielded cells very low. In conclusion, none of the techniques applied in this study provides good performances in terms of number of cells, purity and recovery rate, when applied to small volumes of blood. This suggest that the isolated cells could not be representative of the population in blood, and limits the use of these 3 techniques in cats with spontaneous diseases. Therefore, cell functions in spontaneous feline disease should be investigated in whole blood rather than on isolated cell populations