精原干细胞及其在转基因技术中的应用

睾丸内生殖细胞分化起源于不断更新的干细胞小池,继而首先引起生精细胞(精原细胞)分化。这些定型细胞然后遵循一个固定的程序进行演化,它们首先同步化地进行六次有丝分裂,然后进入第一次减数分裂期。以形态学和细胞学标准,鉴别精原干细胞,根据注入不育者睾丸的分化活力,进行功能检测。目前干细胞的纯化和鉴定研究取得了日新月异的发展。生殖系生物学基本理论和多个领域中的临床前研究将会从中得益,同时附植前的基因转移为转基因技术开辟一条崭新的途径。细胞特征:①源于初生睾丸生长捕获生殖母细胞的少量精原干细胞(2～3×104/成年睾丸),主要分布在生精小管的周围;②细胞遗传学分析首次鉴定表明,它们是缺乏异染色质的圆形小细胞;③通过移植入缺乏生殖细胞的受体睾丸,在功能上表现为具有重建永久精子发生的能力;④基于α6β1整合素和CD9表达,可以获得较高浓度的精原干细胞,在转基因启动子驱动下异源表面蛋白质得以表达,可进一步纯化产物;⑤逆转录病毒载体能使基因转移先于移植,转基因后代随后即可产生。     

Influence of Heat Stress on the Cortisol and Oxidant‐Antioxidants Balance During Oestrous Phase in Buffalo‐Cows (Bubalus bubalis): Thermo‐protective Role of Antioxidant Treatment

In the present study, the effect of heat stress, which is commonly observed in the animals of Upper Egypt area in summer, as well as the effect of antioxidant treatment as a thermo‐protective was examined. In this study, the animals (n = 120) were divided into winter group (n = 40, bred during winter) and summer group (n = 80, bred during summer) as well as, animals in the summer group were divided into first subgroup animals (n = 40) and injected with Viteselen intramuscularly (15 ml) twice weekly for 10 weeks and second subgroup animals (n = 40) were not treated (as control). Serum levels of progesterone (P4), oestradiol (E2), cortisol, superoxide dismutase (SOD), lipid peroxidase (LPO) and nitric oxide (NO) were measured. The pregnancy rate of all animals was detected rectally. The levels of oestradiol and the activity of the antioxidant SOD were decreased in serum of animals in behavioural oestrus during summer as compared with those in winter. During the same time period the levels of oxidants such as LPO and NO were increased in the serum of animals again in the phase of oestrus. In another group of animals treated by intramuscular injection with 15 ml viteselen (antioxidant) twice weekly for 6 weeks during hot months, the activities of serum SOD showed an increase and the levels of oxidants and cortisol decreased. Moreover, the levels of oestradiol were increased during the oestrous behaviour. The pregnancy rate was decreased in animals under heat stress and the pregnancy rate was enhanced dramatically when these animals received antioxidants during the heat stress. This means that the heat‐stress in Upper Egypt may affect the fertility of animals and pregnancy rate and this effect may be through an increased production of free radicals and decreased production of antioxidants as well as increased levels of cortisol. Treatment of animals or supplementation with antioxidants before the beginning of months of heat‐stress and also during the stress period may correct the infertility due to heat‐stress through the decrease in cortisol secretion and a decrease in the oxidative stress. These results resulted in an increase in pregnancy rate in treated animals.     

Effects of seasonal variation,group size and sex on the activity budget and diet composition of the addax antelope

Behaviour and diet composition are critical elements in conservation biology within the scope of reintroduction programs. Here we focused on addax (Addax nasomaculatus), a Critically Endangered antelope species, in the Jbil National Park, Tunisia. In this study, we advanced the hypothesis that season, day period, sex and social structure have an effect on the activity budgets and diet composition of addax. Three groups were selected (large group, adult pair and solitary male). Resting, moving, grazing, vigilance and other behaviours were recorded during two seasons (dry and wet) and in the morning and the afternoon. In addition, faecal samples were collected. The results indicate that season and day period were the main factors affecting the activity budgets. The resting behaviour increased from the wet to the dry season, and from morning to afternoon. The grazing and moving behaviours increased during the wet season and decreased from morning to afternoon. Addax select a mixed diet composed of perennial and herbaceous plant species notably during the wet season. In addition, we observed that addax modify their behaviour according to the season and forage availability. This represents an adaptive strategy to survive in a desert climate with a stochastic low-resource environment and depending on ambient climatic conditions.     

Histological and immunohistochemical investigations of ovarian interstitial glands during non‐breeding season in camels (Camelus dromedarius)

The aim of this was to investigate the histology and immunohistochemistry of interstitial glands during non‐breeding season in camel ovaries. A total of 21 mature, non‐pregnant and apparently healthy camels aged between 8 and 12 years were slaughtered. The ovaries were removed within 15 min, cleaned from adipose tissue, weighted and examined grossly. The histological preparation was made, and then, the blocks were cut at 3–5 microns thickness and stained by H&E for histological examinations. Moreover, some sections were stained with Sudan Black for lipid detection. Immunohistochemical analysis of paraffin‐embedded ovarian tissues was performed to detect the localization of S‐100, vimentin, progesterone receptors (PR) and oestrogen receptors (ER). Immunoreactive signals were detected using UltraVision Detection System. The results revealed that the interstitial glands were located in the cortical region and they were arranged in various arrangements either single, in couple or in groups rich in lipid droplet. All interstitial gland arrangements were enclosed by connective tissue capsules containing fibroblasts and collagenous fibres separated them from the surrounding ovarian structures. Both interstitial glands and their surrounding CT were penetrated by several blood vessels. There was a strong immunoreactive signal for S‐100 in the nuclei of interstitial cells, and no signals were detected either in cells of the interstitial glands or their connective tissue with PR. We could conclude that the interstitial gland is distinct in ovary of camel and further studies are needed to elucidate its rule in steroid synthesis.     

Effects of Exogenous Tumour Necrosis Factor-α on the Secretory Function of the Bovine Reproductive Tract Depend on Tumour Necrosis Factor-α Concentrations

The aim of study was to correlate tumour necrosis factor-α (TNF) infused doses used with the TNF concentrations achieved and with the secretory function of both the ovary and the uterus in cows. We evaluated the concentrations of progesterone (P4), prostaglandin (PG)F_2α, PGE₂ nitric oxide (NO) and TNF in the jugular vein and vena cava caudalis as parameters of exogenous TNF action on the female reproductive tract. Aortae abdominalis of cows (n = 18) were infused with saline or two doses of TNF (luteolytic – 1 μg or luteotrophic – 10 μg). In the peripheral blood, 1 μg TNF concentrations achieved within the range of 30–45 pg/ml, and 10 μg TNF provoked a sharp increase in achieved concentrations at a range of 250–450 pg/mL). The TNF concentrations achieved in vena cava caudalis were five to six times higher than that in peripheral blood (p < 0.001). One microgram TNF increased PGF_2α and NO (p < 0.001) and decreased P4 (p < 0.05). The higher TNF dose stimulated P4 and PGE₂ (p < 0.01). TNF infusion at luteolytic dose achieved its concentrations at the physiological range previously observed in cows. Luteotrophic TNF dose achieved the concentrations in vena cava caudalis that are much higher than physiological level and were previously noted in pathological circumstances (i.e. mastitis , metritis ).