Samantha Mirczuk

Perturbation of the T cell receptor repertoire occurs with increasing age in dogs

ABSTRACT Immunosenescence is the gradual deterioration in immune system function associated with ageing. This decline is partly due to involution of the thymus, which leads to a reduction in the output of naive T cells into the circulating lymphocyte pool. Expansion of existing naive and memory T cell populations, to compensate for the reduction in thymic output, can lead to reduced diversity in the T cell repertoire with increasing age, resulting in impairment of immune responses to novel antigenic challenges, such as during infection and vaccination. Since associations between T cell repertoire and age have only been examined in a limited number of species, to gain further insights into this relationship, we have investigated age‐related changes in the canine T cell receptor (TCR) repertoire. Blood samples were obtained from Labrador retriever dogs of varying ages and variation in the complementary determining region 3 (CDR3) of the T cell receptor beta (TCRB) chain was investigated. CDR3 size spectratyping was employed to evaluate clonal expansion/deletion in the T cell repertoire, allowing identification of profiles within individual variable (V) region families that skewed away from a Gaussian distribution. Older dogs (10–13 years) were found to have an increased number of TCRB V gene spectratypes that demonstrated a skewed distribution, compared with young dogs (≤3 years). Additionally, there was a reduction in the number of clonal peaks present in the spectratypes of old dogs, compared with those of young dogs. The study findings suggest that there is an age‐associated disturbance in the diversity of the T cell receptor repertoire in dogs. HighlightsThe diversity of the human T cell repertoire is known to reduce in old age.Canine T cell receptor repertoire diversity was analysed using CDR3 spectratyping.An age‐associated disturbance was identified in the T cell repertoire of dogs.

Regulation of glucocorticoid metabolism in the boar testis and caput epididymidis by the gonadotrophin-cAMP signalling pathway

In target tissues, cortisol is metabolised by two 11β-hydroxysteroid dehydrogenase (11βHSD) isoenzymes, namely 11βHSD1 and 11βHSD2, both of which are co-expressed in the boar testis and reproductive tract. The present study has assessed whether cortisol-cortisone metabolism in boar testis and caput epididymidis can be regulated via the gonadotrophin-cAMP signalling pathway. 11βHSD activities were measured by using a radiometric conversion assay in static tissue culture. In both testis and caput epididymidis, the net reduction of cortisone but not the net oxidation of cortisol, was significantly decreased by luteinising hormone (by 53 ± 20% and 45 ± 9%, respectively, P < 0.05), forskolin (by 60 ± 7% and 57 ± 9%, respectively, P < 0.01) and 8-bromo-cAMP (by 54 ± 4% and 64 ± 1%, respectively, P < 0.01). This suppression of 11-ketosteroid reductase activity in the boar testis by forskolin could be attenuated by the protein kinase A (PKA) inhibitor, H89. Hence, within the boar testis and the caput epididymidis, the local actions of glucocorticoids are modulated by gonadotrophin-cAMP-PKA signalling via their selective effects on the reductase activity of 11βHSD.

Pituitary pathology and gene expression in acromegalic cats

Abstract The prevalence of GH-secreting pituitary tumors in domestic cats (Felis catus) is 10-fold greater than in humans. The predominant inhibitory receptors of GH-secreting pituitary tumors are somatostatin receptors (SSTRs) and D2 dopamine receptor (DRD2). The expression of these receptors is associated with the response to somatostatin analog and dopamine agonist treatment in human patients with acromegaly. The aim of this study was to describe pathological features of pituitaries from domestic cats with acromegaly, pituitary receptor expression, and investigate correlates with clinical data, including pituitary volume, time since diagnosis of diabetes, insulin requirement, and serum IGF1 concentration. Loss of reticulin structure was identified in 15 of 21 pituitaries, of which 10 of 15 exhibited acinar hyperplasia. SSTR1, SSTR2, SSTR5, and DRD2 mRNA were identified in the feline pituitary whereas SSTR3 and SSTR4 were not. Expression of SSTR1, SSTR2, and SSTR5 was greater in acromegalic cats compared with controls. A negative correlation was identified between DRD2 mRNA expression and pituitary volume. The loss of DRD2 expression should be investigated as a mechanism allowing the development of larger pituitary tumors.

Seasonal and dietary influences on adipose tissue and systemic gene expression in control and previously laminitic ponies

&NA; The aims of the study were to determine whether adipose tissue global gene expression (1) differs between never laminitic (NL) and previously laminitic (PL) ponies, (2) is influenced by season and/or a diet designed to simulate spring grass, and (3) differences seen also occur systemically in peripheral blood mononuclear cells (PBMCs). Subcutaneous adipose tissue and PBMCs were obtained from six NL and six PL ponies on three occasions; summer, winter (season study), and in winter after consuming a diet simulating spring grass for 7 days (diet study). Adipose tissue global gene expression was determined using a 44K equine‐specific microarray, validated using multiplex quantitative real time polymerase chain reaction (qRT‐PCR), and analyzed using GeneSpring software and Ingenuity Pathway Analysis. PBMC gene expression was quantified using qRT‐PCR. The total number of genes whose expression differed (≥2‐fold change, P ≤ .01) between PL and NL ponies was greater in summer (192 genes) compared to winter (58 genes); 40/192 genes influenced by disease in the summer were also seasonally regulated and were predominantly associated with inflammation. The genes modified by dietary intervention and PBMC gene expression did not follow the same pattern as the season study. Thus, adipose tissue global gene expression differed between NL and PL ponies most in summer compared to winter, and these differentially expressed genes predominantly related to inflammation. HighlightsAdipose tissue global gene expression differs between normal and previously laminitic ponies.The differences are greater in summer compared to winter.The differences are not reproduced by feeding a diet mimicking spring grass in winter.Similar changes are not detected in peripheral blood mononuclear cells.

Raf kinase inhibitor protein1 is a myogenic inhibitor with conserved function in avians and mammals

Background: Raf Kinase Inhibitor Protein1 (RKIP) is a tumor suppressor that is present in several adult tissues. It functions as an inhibitor of both Raf/Mek/Erk and NFĸB signaling when unphosphorylated, but following phosphorylation the ability to inhibit Raf/Mek/Erk signaling is lost and RKIP becomes an activator of G‐protein coupled receptor signaling. In neonates and adults, RKIP is known to be expressed in muscle; however, its physiological function is currently unknown. Results: In this study, we show by in situ hybridization and immunofluorescence that RKIP is also expressed in developing chick embryonic muscle, and mouse C2C12 myoblasts. Furthermore, we demonstrate that, in these systems, it functions as an inhibitor of myogenesis: increased levels of RKIP suppress myotube differentiation whereas decreasing RKIP promotes differentiation. Additionally, we show that the ability of RKIP to inhibit myogenesis is dependent upon its phosphorylation state as only the nonphosphorylated form of RKIP suppresses myogenesis. Conclusions: This study, therefore, clearly demonstrates that RKIP has conserved functions as a myogenic inhibitor in both mammalian and avian muscle. Developmental Dynamics 245:902–912, 2016.