Modulation of angiogenesis by topical application of leptin and high and low molecular heparin using the Japanese quail chorioallantoic membrane model.

Pathological angiogenesis characterized by uncontrollable vessel growth is an accompanying feature of many diseases. The avian embryo chorioallantoic membrane (CAM) is an excellent model for angiogenesis research. In our study we used a less common Japanese quail CAM model for the testing of angiogenic potential of leptin, high-molecular (heparin sodium) andlow-molecular (nadroparin calcium) heparins. Heparins play a significant role in vascular endothelial cell function, and they are able to modulate the activities of angiogenic growth factors. On embryonic day 7 leptin (5 µg per CAM), heparin sodium (75 IU per CAM) and nadroparin calcium (47.5 IU per CAM) in 500 µl PBS were applied on the CAM surface. After 24 h the fractal dimension (Df) of the vasculature was evaluated. Samples from each group were histologically analyzed and VEGF-A and Quek1 expression were detected by qPCR. Df was significantly increased in the leptin group. A moderate stimulatory effect of heparin sodium and an inhibitory effect of nadroparin calcium were observed. Both leptin and heparin sodium caused a noticeable increase in the CAM thickness compared to the control and nadroparin calcium groups. We observed an increased number of blood vessels and accumulation of fibroblasts. There was no significant impact on gene expression of VEGF-A and Quek1 24 h after treatment, however, trends similar to the changes in Df and CAM thickness were present. The resulting effect of nadroparin administration on Quek1 levels was exactly the opposite to that of leptin (p < 0.05).

Effect of rooibos tea (Aspalathus linearis) on Japanese quail growth, egg production and plasma metabolites.

1. Birds have been proposed as a suitable model for studies on ageing because of their long life in comparison with similar-sized mammals. However, some weak fliers, such as Galliformes, are the exception to this rule. The aim of the present study was to determine the effects of the treatment with rooibos tea (Aspalathus linearis), a natural source of flavonoid antioxidants and compounds with phyto-oestrogenic activity, on postnatal development and egg production of aged Japanese quail (Coturnix coturnix japonica). 2. Substitution of drinking water with traditional rooibos tea or diet supplementation with ground rooibos tea affected body weight of Japanese quail up to 100 d of age. The body weight of males drinking rooibos tea or eating rooibos-supplemented food decreased significantly. There was a trend toward increased body weight of tea drinking females and a significant increase in the body weight of hens fed the rooibos-supplemented diet. Although rooibos treatment did not significantly increase egg production in young hens, the decrease in egg production of rooibos-treated aged hens (360 d of age) was significantly reduced, regardless of the egg production levels (high - 80%; low - 20%) before the treatment. 3. The results suggest that treatment with rooibos tea positively affected body weight and egg production in quail hens and prolonged the productive period of aged animals. Further studies would be needed to address the question whether these effects are due to the antioxidant or phyto-oestrogenic activities of rooibos.

Role of leptin in farm animals: a review.

The discovery of hormone leptin has led to better understanding of the energy balance control. In addition to its effects on food intake and energy expenditure, leptin has now been implicated as a mediator of diverse physiological functions. Recently, leptin has been cloned in several domestic species. The sequence similarity suggests a common function or mechanism of this peptide hormone across species. Leptin receptors are expressed in most of tissues, which is consistent with the multiplicity of leptin functions. The main goal of this review was to summarize knowledge about effect of leptin on physiology of farm animals. Experiments point to a stimulatory action of leptin on growth hormone (GH) secretion, normal growth and development of the brain. Surprisingly, leptin is synthesized at a high rate in placenta and may function as a growth factor for fetus, signalling the nutritional status from the mother to her offspring. Maturation of reproductive system can be stimulated by leptin administration. Morphological and hormonal changes, consistent with a major role of leptin in the reproductive system, have also been described, including the stimulation of the release of luteinizing hormone (LH), follicle-stimulating hormone (FSH) and prolactin. Leptin has a substantial effect on food intake and feeding behaviour in animals. Administration of leptin reduces food intake. Its level decrease within hours after initiation of fasting. Leptin also serves as a mediator of the adaptation to fasting, and this role may be the primary function for which was the molecule evolved.

Effect of in ovo leptin administration on the development of Japanese quail.

Potential changes in the activity of endocrine axes related to growth as a result of leptin administration during embryonic development of birds were evaluated in the Japanese quail as a model bird with fast growth and development. On day 5 of incubation, 0.1 microg or 1 microg of recombinant mice leptin in 50 microl of phosphate buffered saline were injected into the albumen of eggs. Animals from each group were killed by decapitation on day 0, 2, 5, 7, 14, 21, 28, 35, 42, 49 and 56 of life. Plasma concentrations of triiodothyronine (T(3)), thyroxin (T(4)), corticosterone, testosterone, total lipids, triacylglycerols, cholesterol, glucose and alkaline phosphatase activity were measured. Quail treated by leptin hatched earlier (5-24 hours) and had a higher body weight than the control group (P<0.05-0.001). Mean body weight across the whole observed period was higher in both treated groups as compared to the control group (P<0.05). Leptin in ovo administration was accompanied by changes of endocrine and metabolic parameters during postembryonic development. The most prominent changes appeared immediately after hatching (T(3), T(4), total lipids, triacylglycerols) and before sexual maturity. It is suggested that leptin acts as a general signal of low energy status to neuroendocrine systems in birds which improves utilization of nutrients.