Correlative Light and Scanning Electron Microscopy for Observing the Three-Dimensional Ultrastructure of Membranous Cell Organelles in Relation to Their Molecular Components

Although the osmium maceration method has been used to observe three-dimensional (3D) structures of membranous cell organelles with scanning electron microscopy (SEM), the use of osmium tetroxide for membrane fixation and the removal of cytosolic soluble proteins largely impairs the antigenicity of molecules in the specimens. In the present study, we developed a novel method to combine cryosectioning with the maceration method for correlative immunocytochemical analysis. We first immunocytochemically stained a semi-thin cryosection cut from a pituitary tissue block with a cryo-ultramicrotome, according to the Tokuyasu method, before preparing an osmium-macerated specimen from the remaining tissue block. Correlative microscopy was performed by observing the same area between the immunostained section and the adjacent face of the tissue block. Using this correlative method, we could accurately identify the gonadotropes of pituitary glands in various experimental conditions with SEM. At 4 weeks after castration, dilated cisternae of rough endoplasmic reticulum (RER) were distributed throughout the cytoplasm. On the other hand, an extremely dilated cisterna of the RER occupied the large region of the cytoplasm at 12 weeks after castration. This novel method has the potential to analyze the relationship between the distribution of functional molecules and the 3D ultrastructure in different composite tissues.     

体外培养小鼠下颌下腺细胞生长特性的实验研究

目的：研究小鼠下颌下腺细胞的培养方法,探讨下颌下腺细胞培养条件及细胞生长特性,为研究干细胞转分化涎腺腺泡细胞以及涎腺再生研究奠定理论基础和技术支持。方法：取2周龄的小鼠,组织块法和消化法分别进行培养,用活细胞观察法和HE染色法记录细胞形态学特征;免疫荧光染色法鉴定;通过生长曲线和细胞倍增时间来比较两种方法对细胞增殖能力的影响。结果：组织块法和消化法均可以成功的培养下颌下腺细胞,（1）组织块法培养的细胞呈卵圆形或多边形,10天左右成铺路石样;消化法培养细胞亦为上皮样细胞,呈多边形,胞质丰富;（2）HE染色下颌下腺细胞呈多边形,胞核明显;（3）Cytokeratin13和AQP5表达阳性,Wimentin表达阴性;（4）组织块法培养细胞增殖相对较平缓,消化法培养细胞增长较迅速;（5）消化法培养倍增时间（1．3471±0．6071）天比组织块法倍增时间（2．1887±1．1503）明显缩短（P〈0．05）;结论：体外可以成功的培养下颌下腺细胞,但是同时证明下颌下腺细胞长期传代较困难,这为研究干细胞转分化涎腺细胞和涎腺再生体内实验提供了理论和实验基础。     

Vasoactive intestinal polypeptide stimulates nonovarian progesterone secretion in rabbits

Recent experiments conducted in this laboratory have shown that intravenous infusions of vasoactive intestinal polypeptide (VIP) induced significant increases in plasma progesterone (P) in female rabbits. The purpose of this study was to determine the organ source of this P and to clarify the mechanisms by which it is induced. Intravenous infusions of VIP (37.5, 75, and 150 pmol/kg per min for 60 min) produced acute dose-dependent increases in plasma P in intact estrous rabbits. In ovariectomized (OVX) animals, VIP infusion (75 pmol/kg per min) produced a P increase of the same magnitude. In animals both OVX and adrenalectomized (ADX), this VIP effect was eliminated. The only significant change noted in luteotropic hormone (LH) or follicle stimulating hormone (FSH) was a decrease in FSH immediately following VIP infusion (150 pmol/kg). VIP infusion significantly increased plasma cortisol in intact and OVX animals, but not in OVX/ADX animals. It is concluded that VIP primarily stimulates the adrenal component of P secretion in the rabbit, via mechanisms independent of LH or FSH.     

Changes in weight of the pharyngeal gland and haemolymph titres of juvenile hormone,protein and vitellogenin in worker honey bees

Four physiological parameters (haemolymph-juvenile hormone titre, protein concentration, vitellogenin concentration, and pharyngeal gland dry weight) were examined in the following categories of queenright adult worker bees: summer bees 1–40 days old, winter bees 80–130 days old, 12–100-day old bees at the beginning of winter, 100–195-day old bees at the end of winter, and 1–100-day old bees experimentally induced to live longer in summer.In contrast to the continuously increasing titre of juvenile hormone in ageing summer bees, winter bees kept a constant low level. In bees at the beginning of winter, the hormone titre never reached high values. However, at the end of winter it rose from a low to a high level, comparable with the high titre of 24–40-day old summer bees. In experimentally induced longlived bees in summer, the juvenile hormone titre did not increase as in normal summer bees but remained low as in bees at the beginning of winter. Among the known natural juvenile hormones, only juvenile hormone III was present in the haemolymph of winter bees.The results support the hypothesis of polyphenism being regulated by the titre of juvenile hormone in the haemolymph.     

褛裳夜蛾的交配行为及雄蛾对性腺提取物的反应节律

详细观察了褛裳夜蛾成虫的交配行为,利用风洞、触角电位技术研究了雄蛾对性腺提取物的反应节律,通过林间诱蛾试验进行了验证,旨在为褛裳夜蛾性信息素的精确提取及性信息素组份分离、鉴定提供依据。研究结果表明:成虫的求偶、交配行为均发生在暗期,有一定的节律性:雌蛾在羽化3d以后开始求偶,1周左右表现最为强烈,3-4日龄雌蛾在暗期4-6h开始求偶,最大求偶率在暗期6-8h;5-9日龄在进入暗期就开始求偶,最大求偶率出现在暗期4-6h。风洞试验表明,3-7日龄的雄蛾对性腺提取物均有明显的性行为反应,5日龄雄蛾在暗期4-6h对性腺提取物的反应最为强烈。3-6日龄的成虫在暗期4-6h开始交配,而7-8日龄的成虫于暗期2-4h开始交配,6日龄的成虫交配率最高,交配高峰在暗期4-8h。雄蛾对性腺提取物的触角电位反应也有一定的节律性:雄蛾对4日龄雌蛾性腺提取物开始有电生理反应,对6-7日龄暗期5h性腺提取物反应最为强烈。林间诱蛾试验测定了性腺提取物的引诱活性,7日龄雌蛾性腺提取物林间诱蛾量最高,引诱高峰在暗期4-6h,该结果也验证了褛裳夜蛾雄蛾对性腺提取物的反应节律。     

长爪沙鼠和布氏田鼠汗腺的分布与密度

哺乳动物受到高温胁迫时可通过分泌汗液调节体温。一般认为啮齿类在高温时不出汗,但有关野生啮齿类汗腺的资料不多。我们以长爪沙鼠和布氏田鼠为研究对象,采用苏木精—伊红染色方法,观察了身体各部位皮肤的形态学特征、汗腺的分布和密度。结果显示:(1)两种鼠的头部、口角、胸部、腋窝、腹部、背部和后腿部的皮肤都无汗腺分布,只在不被毛的前足与被毛的后足有外泌汗腺分布;(2)长爪沙鼠前、后足皮肤汗腺的密度分别为(2.40±0.49)个/mm2和(0.15±0.03)个/mm2,前足显著多于后足;布氏田鼠前、后足皮肤汗腺的密度分别为(0.37±0.05)个/mm2和(0.21±0.08)个/mm2,前、后足无显著差异。两种鼠前后足皮肤的汗腺密度可能与被毛状态相关,有被毛则汗腺密度低。推测长爪沙鼠和布氏田鼠前后足的汗腺可能具有辅助散热和运动时增加摩擦力等功能。