Ultrastructural observations on anther tapetum development of freeze-fixed Ledebouria socialis Roth (Hyacinthaceae)

The tapetal ultrastructure of high-pressure-frozen, freeze-substituted Ledebouria socialis Roth (Hyacinthaceae) is described from the tetrad stage up to microspore mitosis. Cytoplasmic degeneration of the tapeturn occurs after microspore mitosis. During the tetrad stage and the early free-microspore stage the tapetum cells appear to be meristematic; after callose dissolution they show an intense exocytosis of polysaccharides into the anther locule. Later, the tapetum cells are characterized by abundant endoplasmic reticulum (ER). Highly osmiophilic pollenkitt precursor substances accumulate within distinct, partly irregular shaped cytoplasmic domains (“osmiophilic bodies”), which are intimately associated with the ER. It remains to be verified whether or not these bodies are derived from the ER. Because of their preservation and staining patterns the contents of these bodies are tentatively interpreted as flavonoids, one of the main pollenkitt pigments in angiosperms. Apart from these pigment bodies, there exist four other kinds of lipophilic inclusion within the anther (cells). The general aspects of lipid preservation in freeze-substituted samples are discussed. Staining with hot alcoholic phosphotungstic acid yielded good contrast of the ER and other membranes, which are often difficult to visualize in freeze-substituted, resin-embedded samples.     

Comparative morphology of the epidermis of seven species of polyclad flatworms (Platyhelminthes: Rhabditophora)

The ultrastructure of the epidermis of seven species of polyclad flatworms (Phaenocelis medvedica, Phaenocelis peleca, Pleioplana atomata, Boninia divae, Pericelis orbicularis, Enchiridium periommatum, and Cycloporus variegatus) representing six families is described. In all seven species, the epidermis consists of a single layer of columnar cells that rests on a bipartite basement membrane. Epithelial cell surfaces are covered by numerous microvilli and cilia. Cilia contain microtubules arranged in the 9 + 2 pattern, and from their basal bodies two striated rootlets arise, a rostrally directed one running parallel to the apical cell membrane, and a vertical one at a right angle to the first rootlet. Numerous epithelosomes and mitochondria occupy the apical parts of the cells. The basal part of the cells is highly folded, forming a cell web that connects to the basement membrane. The basement membrane consists of a thin basal lamina and a thick, multilayered reticular lamina. The number of layers in the reticular lamina varies among the different species and appears to be correlated with body size. Numerous canals containing either pigment granules or nervous processes perforate the basement membrane. We have identified four different types of glands: rhabdite glands, rhabdoid glands, mucoid glands containing vacuoles filled with flocculent material, and mucoid glands resembling thread cells of hagfish slime glands. The latter have been found only in P. orbicularis. Pigment cells were found in all species examined with the exception of C. variegatus, which takes its coloration from its ascidian prey. Our results further support the unique taxonomic status of Boniniidae.     

高脂血症大鼠下颌下腺内AQP1和AQP5表达及意义

目的 观察高脂血症大鼠下颌下腺内AQP1和AQP5表达的变化.方法雄性 S D大鼠 20只,随机分为 2组,对照组(C组)给予全价颗粒饲料喂养;高脂饮食组(H组)给予高脂饮食 (饲料成分为胆固醇 2%、猪油10%、基础饲料 88% )连续喂养2个月,各组动物均不限制饮水.2个月成模后,取血检测血脂;取大鼠下颌下腺组织,进行免疫组织化学染色(SP法) 和计算机图像分析.结果 ①血脂检测结果:C组TG与H组TG比较有显著性差异(P<0.05);C组TC和H组TC比较有显著性差异(P<0.05).②免疫组化结果:C组大鼠下颌下腺AQP1平均光密度值与 H组AQP1平均光密度值比较有差异性(P<0.05);C组大鼠下颌下腺AQP5平均光密度值与 H组AQP5平均光密度值比较有差异性(P<0.05).结论 高脂血症大鼠下颌下腺导管上皮细胞内AQP1和AQP5的表达减少,为探讨高脂血症导致下颌下腺分泌功能降低的病理机制提供了形态学依据.     

Dufour gland of the digger wasp<Emphasis Type="Italic"> Liris niger:</Emphasis> structure and developmental and biochemical aspects

The tubiform Dufour gland in the digger wasp species Liris niger is about 1.0 mm long ( 0.15 mm). An alternating arrangement of longitudinal and circumferential bundles of striated muscle fibers surrounds the gland. The Dufour gland, together with the venom gland, enters the sting base and terminates in the sting. The glandular epithelium is monolayered. Glands about 3 day after imaginal ecdysis have an empty lumen but a thick lining epithelium. The gland cells are characterized by a well-developed vesicular smooth endoplasmic reticulum, sparse rough ER and numerous free ribosomes. They also exhibit several electron-lucent vesicles and autophagic vacuoles. Secretion of electron-dense material via the gland cuticle into the gland lumen is apparent. Glands more than 20 days after imaginal ecdysis display a large lumen and a thin epithelium. The cells show signs of degeneration with numerous cytolytic inclusions. Dufour gland liquid contains numerous polypeptides of molecular weights ranging from 14 to about 200 kDa. In addition the secretion consists predominantly of straight-chain hydrocarbons, accompanied by small amounts of esters. The major hydrocarbons are pentadecane and (Z)-8-heptadecene. Dufour gland secretion may have several functions: (1) the polypeptides might be involved in the gluing process of the eggs, while (2) the hydrocarbon oils may function as lubricants for the lancets and (3) might soften the secretion, thus allowing easier application of the glue. The lipophilic volatile material (4) might also be involved in pheromonal signaling.     

Negative Regulation of Human Growth Hormone Gene Expression by Insulin Is Dependent on Hypoxia-inducible Factor Binding in Primary Non-tumor Pituitary Cells

Insulin controls growth hormone (GH) production at multiple levels, including via a direct effect on pituitary somatotrophs. There are no data, however, on the regulation of the intact human (h) GH gene (hGH1) by insulin in non-tumor pituitary cells, but the proximal promoter region (nucleotides −496/+1) responds negatively to insulin in transfected pituitary tumor cells. A DNA-protein interaction was also induced by insulin at nucleotides −308/−235. Here, we confirmed the presence of a hypoxia-inducible factor 1 (HIF-1) binding site within these sequences (−264/−259) and investigated whether HIF-1 is associated with insulin regulation of “endogenous” hGH1. In the absence of primary human pituitary cells, transgenic mice expressing the intact hGH locus in a somatotroph-specific manner were generated. A significant and dose-dependent decrease in hGH and mouse GH RNA levels was detected in primary pituitary cell cultures from these mice with insulin treatment. Increasing HIF-1α availability with a hypoxia mimetic significantly decreased hGH RNA levels and was accompanied by recruitment of HIF-1α to the hGH1 promoter in situ as seen with insulin. Both inhibition of HIF-1 DNA binding by echinomycin and RNA interference of HIF-1α synthesis blunted the negative effect of insulin on hGH1 but not mGH. The insulin response is also sensitive to histone deacetylase inhibition/trichostatin A and associated with a decrease in H3/H4 hyperacetylation in the proximal hGH1 promoter region. These data are consistent with HIF-1-dependent down-regulation of hGH1 by insulin via chromatin remodeling specifically in the proximal promoter region.     

Extracellular Ca2+ sensing in salivary ductal cells

Ca(2+) is secreted from the salivary acinar cells as an ionic constituent of primary saliva. Ions such as Na(+) and Cl(-) get reabsorbed whereas primary saliva flows through the salivary ductal system. Although earlier studies have shown that salivary [Ca(2+)] decreases as it flows down the ductal tree into the oral cavity, ductal reabsorption of Ca(2+) remains enigmatic. Here we report a potential role for the G protein-coupled receptor, calcium-sensing receptor (CSR), in the regulation of Ca(2+) reabsorption by salivary gland ducts. Our data show that CSR is present in the apical region of ductal cells where it is co-localized with transient receptor potential canonical 3 (TRPC3). CSR is activated in isolated salivary gland ducts as well as a ductal cell line (SMIE) by altering extracellular [Ca(2+)] or by aromatic amino acid, l-phenylalanine (l-Phe, endogenous component of saliva), as well as neomycin. CSR activation leads to Ca(2+) influx that, in polarized cells grown on a filter support, is initiated in the luminal region. We show that TRPC3 contributes to Ca(2+) entry triggered by CSR activation. Further, stimulation of CSR in SMIE cells enhances the CSR-TRPC3 association as well as surface expression of TRPC3. Together our findings suggest that CSR could serve as a Ca(2+) sensor in the luminal membrane of salivary gland ducts and regulate reabsorption of [Ca(2+)] from the saliva via TRPC3, thus contributing to maintenance of salivary [Ca(2+)]. CSR could therefore be a potentially important protective mechanism against formation of salivary gland stones (sialolithiasis) and infection (sialoadenitis).     

N-Sulfation of Heparan Sulfate Regulates Early Branching Events in the Developing Mammary Gland

Branching morphogenesis, a fundamental process in the development of epithelial organs (e.g. breast, kidney, lung, salivary gland, prostate, pancreas), is in part dependent on sulfation of heparan sulfate proteoglycans. Proper sulfation is mediated by biosynthetic enzymes, including exostosin-2 (Ext2), N-deacetylase/N-sulfotransferases and heparan sulfate O-sulfotransferases. Recent conditional knockouts indicate that whereas primary branching is dependent on heparan sulfate, other stages are dependent upon selective addition of N-sulfate and/or 2-O sulfation (Crawford, B .E., Garner, O. B., Bishop, J. R., Zhang, D. Y., Bush, K. T., Nigam, S. K., and Esko, J. D. (2010) PLoS One 5, e10691; Garner, O .B., Bush, K. T., Nigam, S .K., Yamaguchi, Y., Xu, D., Esko, J. D., and Nigam, S. K. (2011) Dev. Biol. 355, 394–403). Here, we analyzed the effect of deleting both Ndst2 and Ndst1. Whereas deletion of Ndst1 has no major effect on primary or secondary branching, deletion of Ndst2 appears to result in a mild increase in branching. When both genes were deleted, ductal growth was variably diminished (likely due to variable Cre-recombinase activity), but an overabundance of branched structures was evident irrespective of the extent of gland growth or postnatal age. “Hyperbranching” is an unusual phenotype. The effects on N-sulfation and growth factor binding were confirmed biochemically. The results indicate that N-sulfation or a factor requiring N-sulfation regulates primary and secondary branching events in the developing mammary gland. Together with previous work, the data indicate that different stages of ductal branching and lobuloalveolar formation are regulated by distinct sets of heparan sulfate biosynthetic enzymes in an appropriate growth factor context.     

Fine structure of the urnulae of Balaustium mites (Actinotrichida: Erythraeidae) representing peculiar defense organs

The urnulae, until now the enigmatic paired dorsal protrusions on idiosoma dorsum in active postlarval forms of Balaustium mites, were studied using electron microscopy. They consist of walls made of unmodified integument, which form a cylinder covered by a roof of thin cuticle. At the posterior border of the urnula, the roof has a crescent slit. On its inner surface, a rather large muscle inserts with several tendons. The roof forms a flap under which the modified columnar epidermal cells containing numerous lipid inclusions are located. These lipids are probably secreted through pore canals of the overlying cuticle. Materials mainly originating from an extensive vesicular tissue situated underneath the columnar cells of the urnula and under the adjacent unmodified epidermis are extruded through the mentioned slit. Our results support previous studies that have suggested a function of the urnulae as defensive organs. Our study further suggests that the agent that provides the repellent effect comes mainly from the vesicular tissue, whereas the columnar cells with their lipid secretions are likely to restore the external secretion layer of the epicuticle after its destruction during the repellent release. Further structural and functional details are discussed and compared with other putative defensive secretory organs.