Topographical localization of iron in brains of the aged fat-tailed dwarf lemur (Cheirogaleus medius) and gray lesser mouse lemur (Microcebus murinus)

Iron deposits in the human brain are characteristic of normal aging but have also been implicated in various neurodegenerative diseases. Among nonhuman primates, strepsirhines are of particular interest because hemosiderosis has been consistently observed in captive aged animals. In particular, the cheirogaleids, because of their small size, rapid maturity, fecundity, and relatively short life expectancy, are a useful model system for the study of normal and pathological cerebral aging. This study was therefore undertaken to explore iron localization in the brain of aged cheirogaleids (mouse and dwarf lemurs) with histochemistry and magnetic resonance microscopy. Results obtained with both techniques were comparable. There was no difference between old animals in the two species. The young animals (3 years old) showed no iron deposits. In the old animals (8–15 years old), iron pigments were mainly localized in the globus pallidus, the substantia nigra, the neocortical and cerebellar white matter, and anterior forebrain structures, including the nucleus basalis of Meynert. This distribution agrees with previous findings in monkeys and humans. In addition, we observed iron in the thalamus of these aged nonhuman primates. Microscopic NMR images clearly reveal many features seen with the histochemical procedure, and magnetic resonance microscopy is a powerful method for visualizing age-related changes in brain iron. Am. J. Primatol. 45:291–299, 1998. © 1998 Wiley-Liss, Inc.     

HISTOPATHOLOGICAL AND HISTOCHEMICAL CHANGES IN HONEYBEE LARVAE (APIS MELLIFERA L.) AFTER INFECTION WITH BACILLUS LARVAE,THE CAUSATIVE AGENT OF AMERICAN FOULBROOD DISEASE

Morphological, histochemical and cytochemical changes were examined in honeybee larvae after infection with the bacterium Bacillus larvae. The results indicate cell necrosis in the midgut epithelium accompanied by increasing cell vacuolization and nuclear pyknosis following per os inoculation with B. larvae. Many autolysosomes were positive for acid phosphatase. Non-vacuolar acid phosphatase activity was also found in lysed cell compartments. No such activity was found in regenerative epithelial cells. Degradation of haemocytes, salivary glands and other tissues was also observed. Histochemical analyses after per cutaneous inoculation with B. larvae of three- and five-day-old honeybee larvae show intense non-vacuolar acid phosphatase activity followed by disintegration of infected salivary glands, epithelial cell cytoplasm and haemocytes.     

植物体细胞胚发生的细胞生物学研究进展

体细胞胚发生是植物界的一个普遍现象,本文从组织细胞学、超微结构、电镜细胞化学、分子生物学等方面对体细胞胚发生发育过程中的形态建成和细胞结构,Ca2 和ATPase的动态变化,淀粉和蛋白质代谢,核糖体、线粒体等对极性变化的影响,畸形胚的发生和体胚同步化的诱导调控,基因调控等方面进行了综述,为进一步深入了解体细胞胚的发生发育规律及基因调控机制以及植物高效再生体系的建立提供参考。     

hTERT cDNA片段的克隆及其单克隆抗体与喉癌的发病机制

端粒酶的激活及其调控机制至今不仍不清楚。为研究喉癌发生中端粒酶表达规律及激活的可能机制,我们克隆了hTERT cDNA片段并制备了抗hTERT单克隆抗体。应用此抗体对喉癌组织进行了免疫组化检测,发现喉癌分化程度降低与癌组织中hTERT阳性细胞率增高有关;而c-Myc表达与hTERT表达呈明显正相关,提示c-Myc可能对喉癌发生过程中端粒酶的激活起着重要作用。这些研究表明,喉癌的发生可能是由于c-Myc的过度表达使端粒酶表达上调,从而使喉鳞状上皮细胞达到永生化,这一机制不仅存在于喉癌发生的早期,而且贯穿于喉癌的发展过程。     

甘遂乳汁管的解剖学及营养器官中二萜类化合物分布的研究

利用石蜡切片、半薄切片及组织化学的方法对甘遂各器官中乳汁管的类型、分布和大小进行了研究。结果表明,甘遂乳汁管为无节分枝型,在各器官中主要分布在维管束韧皮部的外侧或周围,此外,在根的中柱鞘薄壁细胞、叶片的叶肉组织、果实的中果皮及胚乳细胞内也有少量乳汁管的分布。甘遂乳汁管的大小在各器官中略有不同,其中茎中直径最大为39.79μm,块根中直径居中为36.90μm,叶肉组织中直径最小,仅有8.94μm;乳汁管的密度在其营养器官中差异较大,依次为叶>茎>根>块根。组织化学实验结果显示二萜类化合物在甘遂的营养器官中分布广泛。根中柱鞘和韧皮部的薄壁组织细胞,茎皮层、维管形成层和韧皮部的薄壁组织细胞,叶的叶肉细胞以及叶脉中的厚角组织和薄壁组织细胞的腔中都显示不同程度的红色,而乳汁管中乳汁的显色较深。     

BMP Signaling and Podocyte Markers Are Decreased in Human Diabetic Nephropathy in Association With CTGF Overexpression

Diabetic nephropathy is characterized by decreased expression of bone morphogenetic protein-7 (BMP-7) and decreased podocyte number and differentiation. Extracellular antagonists such as connective tissue growth factor (CTGF; CCN-2) and sclerostin domain-containing-1 (SOSTDC1; USAG-1) are important determinants of BMP signaling activity in glomeruli. We studied BMP signaling activity in glomeruli from diabetic patients and non-diabetic individuals and from control and diabetic CTGF^+/+ and CTGF^+/− mice. BMP signaling activity was visualized by phosphorylated Smad1, -5, and -8 (pSmad1/5/8) immunostaining, and related to expression of CTGF, SOSTDC1, and the podocyte differentiation markers WT1, synaptopodin, and nephrin. In control and diabetic glomeruli, pSmad1/5/8 was mainly localized in podocytes, but both number of positive cells and staining intensity were decreased in diabetes. Nephrin and synaptopodin were decreased in diabetic glomeruli. Decrease of pSmad1/5/8 was only partially explained by decrease in podocyte number. SOSTDC1 and CTGF were expressed exclusively in podocytes. In diabetic glomeruli, SOSTDC1 decreased in parallel with podocyte number, whereas CTGF was strongly increased. In diabetic CTGF^+/− mice, pSmad1/5/8 was preserved, compared with diabetic CTGF^+/+ mice. In conclusion, in human diabetic nephropathy, BMP signaling activity is diminished, together with reduction of podocyte markers. This might relate to concomitant overexpression of CTGF but not SOSTDC1. (J Histochem Cytochem 57:623–631, 2009)     

A spatiotemporal analysis of enzymatic activities associated with carbon metabolism in wild-type and mutant embryos of Arabidopsis using in situ histochemistry

Arabidopsis as a molecular genetic model offers many advantages for the study of seed development, but these do not extend to biochemical and enzymatic studies, which are often compromised by the limited amount of material available from the small developing embryos. A set of assays based on the coupling of an enzymatic reaction to the reduction of NAD, NADP or FAD, and subsequent reduction and precipitation of a tetrazolium salt, have been adapted to investigate 18 enzyme activities associated with carbon metabolism in developing Arabidopsis embryos. The use of organelle-specific marker enzymes demonstrates the utility of the method for detection of activities in mitochondria, plastids and peroxisomes as well as the cytosol. The temporal staining patterns obtained allow classification of the activities into three main categories based on whether they peak in the early, intermediate or late stages of maturation. An interesting switch from ATP to pyrophosphate consuming pathways occurs at the onset of the maturation phase, which involves key steps in primary carbon metabolism such as phosphofructokinase. This spatiotemporal characterization of carbon metabolism has also been applied to various mutants disrupted in embryo development including gnom (gn), acetyl-CoA carboxylase1 (acc1), schlepperless (slp), and wrinkled1 (wri1). The data obtained demonstrate that the extent to which carbon metabolism is affected in mutants is not necessarily correlated to the severity of the mutation considered. Through the advanced characterization of trehalose-6-P synthase1 (tps1) embryos, this approach finally provides new insight into the regulatory role played by trehalose metabolism in embryo development.     

肾上腺髓质素在正常大鼠脑中的分布一免疫组化及原位杂交分析

目的验证大鼠脑内是否存在ADM及其mRNA。方法取10只健康雄性SD大鼠,体重200-250g,应用免疫组织化学法和原位杂交组织化学法检测正常大鼠脑内ADM及其mRNA的表达情况。结果在大鼠脑内ADM及其mRNA阳性细胞主要表达在大脑皮质、海马结构、齿状回、丘脑、室旁组织、脉络丛、室管膜细胞、基底节、血管内皮细胞,其中脉络丛、室旁组织、丘脑为高表达区,其次为大脑皮质、海马。在大鼠大脑内ADMmRNA的表达与ADM阳性细胞的表达相一致。结论ADM及其mRNA在大脑内广泛表达,提示ADM在中枢神经系统内具有重要的作用。     

Improvement in Double Staining With Fluoro-Jade C and Fluorescent Immunostaining: FJC Staining Is Not Specific to Degenerating Mature Neurons

Fluoro-Jade C (FJC) staining has been used to detect degenerating neurons in tissue sections. It is a simple and easy staining procedure and does not depend on the manner of cell death. In some experiments, double staining with FJC and fluorescent immunostaining (FI) is required to identify cell types. However, pretreatment for FJC staining contains some processes that are harsh to fluorophores, and the FI signal is greatly reduced. To overcome this issue, we improved the double staining protocol to acquire clear double-stained images by introducing the labeled streptavidin–biotin system. In addition, several studies indicate that FJC can label non-degenerating glial cells, including resting/reactive astrocytes and activated microglia. Moreover, our previous study indicated that degenerating mesenchymal cells were also labeled by FJC, but it is still unclear whether FJC can label degenerating glial cells. Acute encephalopathy model mice contained damaged astrocytes with clasmatodendrosis, and 6-aminonicotinamide-injected mice contained necrotic astrocytes and oligodendrocytes. Using our improved double staining protocol with FJC and FI, we detected FJC-labeled degenerating astrocytes and oligodendrocytes with pyknotic nuclei. These results indicate that FJC is not specific to degenerating neurons in some experimental conditions: