Cell cycles in the male accessory glands of mealworm pupae

During the pupal stage of Tenebrio molitor, the accessory reproductive glands of males grow by cell division. Within the secretory epithelium of the bean-shaped accessory glands (BAGs), cell numbers triple. In the tubular accessory glands (TAGs), the increase is 14-fold. There are two mitotic maxima in each gland. The first maximum occurs at 1-2 days while the second is at 4-5 days. The second maximum coincides with the major ecdysteroid peak described by Delbecque et al. [Dev. Biol. 64, 11-30 (1978)]. Nuclei were isolated from TAGs during the pupal mitotic bouts and during mitotic inactivity in the adult. After Feulgen or propidium iodide staining, the DNA content of these nuclear populations was measured by absorption cytophotometry or by fluorescence flow cytometry, respectively. The proportion of cells in each phase of the cycle was calculated using an iterative model. After mitoses have ended in the late pupa, the cells were arrested in G2. [3H]Thymidine was injected into 1- and 4-day pupae to pulse-label cells of the TAGs. After allowing various periods from 4 to 60 hr for cells to progress through G2 to reach mitosis, fractions of labelled mitoses were determined by autoradiography. From the combined cytometric and autoradiographic data, the duration of each phase of the cell cycle was calculated assuming the population was in exponential growth. Cell cycles in 4-day pupal TAGs take 48 hr. G1, S, G2, and, M lasted 13, 14, 17, and 4 hr, respectively.     

Ecdysteroids accelerate mitoses in accessory glands of beetle pupae

During the 9-day pupal period of Tenebrio molitor (the mealworm beetle), the cells of the male accessory glands undergo divisions for 7 days. There are two maxima in the mitotic activity in the glands in vivo, one at 1 day and the other at 4 days. The latter peak coincides with the large surge of ecdysterone occurring in the pupal stage. By the use of in vitro culture techniques, it has been demonstrated that the first bout of mitosis in both glands proceeds in basal medium, while the second bout of mitosis requires a physiological level of ecdysterone. Ecdysone was less effective than ecdysterone. Sensitivity to ecdysterone did not change significantly between Day 1 and Day 4 of the pupal stage. The results are discussed in relation to the effects of ecdysterone on cell division in mesodermal and ectodermal derivatives.     

Cloning and expression analysis of an ADP-ribosylation factor from Solanum tuberosum L.

Summary A cDNA clone encoding an ADP-ribosylation factor from potato (Solanum tuberosum L.) was isolated. The nucleotide and deduced amino acid sequences show high homology to known ADP-ribosylation factor sequences from Arabidopsis, yeast, cow and man. In northern blot experiments, all tissues analysed showed expression of the corresponding mRNA. Strongest expression was found, however, in potato tubers.Abbreviations ARF, ADP ribosylation factor - BSA bovine serum albumin - EDTA ethylenediaminetetraacetic acid - SDS sodium dodecyl sulfate - SSC sodium saline citrate     

The hormonal control of accessory reproductive gland development in female Schistocerca gregaria

Allatectomy of adult female Schistocerca gregaria prevents the normal development of the accessory reproductive glands and no secretion is produced. Development of the glands can be restored by the administration of synthetic juvenile hormone and the response is dose-dependent. A continuous supply of hormone is required for maintenance of secretory activity. In the normal developmental sequence the total protein content of the glands remains constant until the time at which vitellogenesis occurs in the terminal oöcytes. As maturation proceeds there is a linear increase in protein content of the glands. The initial increase occurs as a result of cellular changes in the glands and is then followed by an increase due to an accumulation of secretion in the lumina.     

Schisandra henryi C. B. Clarke in vitro cultures: a promising tool for the production of lignans and phenolic compounds

Plant Cell, Tissue and Organ Culture (PCTOC) - We initiated and optimized in vitro culture conditions of the endemic Chinese plant species—Schisandra henryi C. B. Clarke. Different types of...     

Identification and quantification of catecholamines in potato plants (Solanum tuberosum) by GC-MS

Dopamine, norepinephrine, and normetanephrine were identified by GC-MS in potato (Solanum tuberosum L.) plants, the latter was new for plants. The highest amount of catecholamines was found in leaves. A developmental stage dependent variation in potato leaf catecholamines accumulation was also observed with highest level in third leaves. Catecholamine contents decrease during cold storage of tubers to undetectable levels. Mechanical wounding of leaves led to a small increase in the level of catecholamines investigated.     

GC-MS analysis of amino acid enantiomers as their N(O,S)-perfluoroacyl perfluoroalkyl esters: application to space analysis

The target of the in-situ research of optical activity in extraterrestrial samples stimulated an extended investigation of a GC-MS method based on the derivatization of amino acids by using a mixture of perfluorinated alcohols and perfluorinated anhydrides. Amino acids are converted to their N(O,S)-perfluoroacyl perfluoroalkyl esters in a single-step procedure, using different combinations of the derivatization reagents trifluoroacetic anhydride (TFAA)-2,2,2-trifluoro-1-ethanol (TFE), TFAA-2,2,3,3,4,4,4-heptafluoro-1-butanol (HFB), and heptafluorobutyric anhydride (HFBA)-HFB. The derivatives obtained are analyzed using two different chiral columns: Chirasil-L-Val and gamma-cyclodextrin (Rt-gamma-DEXsa) stationary phases which show different and complementary enantiomeric selectivity. The mass spectra of the derivatives are studied, and mass fragmentation patterns are proposed: significant fragment ions can be identified to detect amino acid derivatives. The obtained results are compared in terms of the enantiomeric separation achieved and mass spectrometric response. Linearity studies and the measurement of the limit of detection (LOD) show that the proposed method is suitable for a quantitative determination of enantiomers of several amino acids. The use of the programmed temperature vaporiser (PTV) technique for the injection of the untreated reaction mixture is a promising method for avoiding manual treatment of the sample and decreasing the LOD.