Functional lifespan of the corpus luteum (CL) in non-pregnant dogs exceeds that of pregnant animals and may last for more than 80 days. Prolactin and LH act luteotropic, however, luteolytic mechanisms are poorly understood. Other than in life stock there is no uterine luteolysine and it was postulated that local paracrine/autocrine mechanisms might play a major role. In following this hypothesis the present investigations have clearly demonstrated that up-regulation of prostaglandin synthesis in the CL as indicated by the expression of cyclo-oxygenase II occurs with its formation and not regression, pointing towards a luteotropic rather than luteolytic action. Throughout dioestrus luteal and other cells of the CL express the oestrogen (ERalpha) and progesterone receptor (PR). While ERalpha expression was not cycle-related, PR concentrations were high in the early and late-luteal phase and a regulatory role of both steroids on CL-function is assumed. As in other species also in the dog the immune system seems to participate in the mechanisms regulating CL-function as an increased presence of lymphocytes within the CL could be detected at the beginning [CD4- CD8-, major histocompatibility complex (MHC)II-antigen expressing cells] and during the latter half of dioestrus (CD8- and MHCII-antigen expressing cells). Thus, leucocyte-derived cytokines may be important and the expression of the mRNA for interleukin (IL)8, IL10, IL12, tumour necrosis factor (TNF)alpha and transforming growth factor (TGF) beta1 was observed throughout dioestrus. Electron microscopy confirmed the slow process of luteolysis; first distinct signs of degeneration were seen on day 60, accompanied by some apoptotic events. From these data it is concluded that luteal regression as monitored by the gradual decrease of systemic progesterone concentrations in the dog is not an actively regulated but rather a permissive process. Immune-mediated events may play a key role. Changes in the vascular supply, as indicated by the expression of endoglin, seem to be of lower importance.

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BackgroundThe uterine response to the presence of embryos is poorly understood in the domestic dog (Canis familiaris). The intimate embryo-maternal cross-talk, which begins following the hatching of blastocysts and embryo attachment leads to strong structural and functional remodelling of the uterus. A part of this process is decidualisation, comprising morphological and biochemical changes that result in formation of maternal stroma-derived decidual cells. These are an integral part of the canine placenta materna, which together with the maternal vascular endothelium are the only cells of the canine endotheliochorial placenta able to resist trophoblast invasion. These cells are also the only ones within the canine placenta expressing the progesterone receptor (PGR). Understanding the decidualisation process thus appears essential for understanding canine reproductive physiology.MethodsHere, we investigated the capability of canine uterine stromal cells to decidualise in vitro, thereby serving as a canine model of decidualisation. A dbcAMP-mediated approach was chosen during a time course of 24 - 72 h. Tissue material from six (n = 6) healthy, dioestric bitches was used (approximately 2 weeks after ovulation). Cells were characterized by differential staining, nearly 100 % of which were vimentin-positive. Scanning and transmission electron microscope analyses were applied, and morphological changes were recorded with a live cell imaging microscope. Expression of several decidualisation markers was investigated.ResultsThe in vitro cultured stromal cells acquired characteristics of decidual cells when incubated with 0.5 mM dbcAMP for 72 h. Their shape changed from elongated to rounded, while ultrastructural analysis revealed higher numbers of mitochondria and secretory follicles, and an increased proliferation rate. Elevated expression levels of IGF1, IGF2, PRLR and ERα were observed in decidualised cells; PRL and ERβ remained mostly below the detection limit, while PGR remained unaffected. The expression of smooth muscle α actin (αSMA), another decidualisation marker, was strongly induced. Among prostaglandin system members, levels of COX2 (PTGS2) and of PGE2-synthase (PTGES) were upregulated. Expression of the PGE2 receptors, PTGER2 and PTGER4, was clearly detectable.ConclusionAn in vitro decidualisation model with canine uterine stromal cells was successfully established, allowing future, more detailed studies to be undertaken on the underlying molecular and endocrine mechanisms of canine decidualisation.Electronic supplementary materialThe online version of this article (doi:10.1186/s12958-015-0066-4) contains supplementary material, which is available to authorized users.

OBJECTIVE: To investigate the effect of weight loss on GLUT 4 content of insulin sensitive tissues of obese mice. SUBJECTS: Mice were made obese by neonatal treatment with monosodium glutamate (MSG). In addition, one group of obese animals was submitted to a caloric restriction to promote 20% weight loss (MSG-L). Both groups were compared to age-matched control mice. MEASUREMENTS: Anthropometric data, glycaemia and insulinaemia were measured. The GLUT 4 protein was assessed by Western blotting analysis in white (WAT) and brown (BAT) adipose tissue, and skeletal (SM) and cardiac (CM) muscles. RESULTS: The MSG mice were very obese according to their morphometric analysis, showing moderate hyperglycaemia with severe hyperinsulinaemia, and reduced (P`0.001) glucose/insulin (G/I) ratio. The procedure for weight loss promoted a signi®cant (P`0.001) reduction of both glycaemic and insulinaemic levels, and an increase in G/I ratio. Compared to control animals, the GLUT 4 content in obese MSG mice, was decreased by 30% (P`0.05) in SM and CM, by 80% (P`0.001) in BAT and in different subcellular membrane fractions of WAT. On the other hand, transporter protein content was restored to normal levels in MSG-L animals. CONCLUSION: The reduced GLUT 4 content of insulin sensitive tissues from MSG-treated obese mice is recovered by a 20% loss in weight. This mechanism can be involved in the observed increase of insulin sensitivity.