Mitotic cytosol inhibits invagination of coated pits in broken mitotic cells

Receptor-mediated endocytosis is inhibited during mitosis in mammalian cells and earlier work on A431 cells suggested that one of the sites inhibited was the invagination of coated pits (Pypaert, M., J. M. Lucocq, and G. Warren. 1987. Eur. J. Cell Biol. 45: 23-29). To explore this inhibition further, we have reproduced it in broken HeLa cells. Mitotic or interphase cells were broken by freeze-thawing in liquid nitrogen and warmed in the presence of mitotic or interphase cytosol. Using a morphological assay, we found invagination to be inhibited only when mitotic cells were incubated in mitotic cytosol. This inhibition was reversed by diluting the cytosol during the incubation. Reversal was sensitive to okadaic acid, a potent phosphatase inhibitor, showing that phosphorylation was involved in the inhibition of invagination. This was confirmed using purified cdc2 kinase which alone could partially substitute for mitotic cytosol.     

Identification of a Golgi-associated protein that undergoes mitosis dependent phosphorylation and relocation

By means of a monoclonal antibody (BH3), we have identified a 57-kD protein (p57) that in interphase is restricted largely to the perinuclear region of the cell. Double label immunofluorescence microscopy suggests localization of p57 to the Golgi complex and associated membranous structures. Protease protection experiments and chemical extractability indicate that p57 is a peripheral membrane protein exposed to the cytoplasm. p57 displays unique behavior during mitosis. At the end of G2 or in early prophase, p57 leaves the perinuclear region and accumulates very rapidly within the nucleus, at a time when the nuclear envelope is still intact and before nuclear lamina disassembly. This relocation of p57 coincides with its hyperphosphorylation on serine and threonine residues. After nuclear envelope breakdown p57 becomes uniformly distributed throughout the mitotic cytoplasm until in late telophase when it returns to its perinuclear location and is once again excluded from the nucleus. The behavior of p57 during mitosis suggests that it may play a role in the cellular reorganization evident during mitotic prophase.     

A case of true hermaphroditism reveals an unusual mechanism of twinning

Traditionally twins are classified as dizygous or fraternal and monozygous or identical (Hall Twinning, 362, 2003 and 735-743). We report a rare case of 46,XX/46,XY twins: Twin A presented with ambiguous genitalia and Twin B was a phenotypically normal male. These twins demonstrate a third, previously unreported mechanism for twinning. The twins underwent initial investigation with 17-hydroxyprogesterone and testosterone levels, pelvic ultrasound and diagnostic laparoscopy. Cytogenetic analysis was performed on peripheral blood cells and skin fibroblasts. Histological examination and Fluorescence in situ hybridization studies on touch imprints were performed on gonadal biopsies. DNA analysis using more than 6,000 DNA markers was performed on skin fibroblast samples from the twins and on peripheral blood samples from both parents. Twin A was determined to be a true hermaphrodite and Twin B an apparently normal male. Both twins had a 46,XX/46,XY chromosome complement in peripheral lymphocytes, skin fibroblasts, and gonadal biopsies. The proportion of XX to XY cells varied between the twins and the tissues evaluated. Most significantly the twins shared 100% of maternal alleles and approximately 50% of paternal alleles in DNA analysis of skin fibroblasts. The twins are chimeric and share a single genetic contribution from their mother but have two genetic contributions from their father thus supporting the existence of a third, previously unreported type of twinning.     

Polarity and signalling in plant embryogenesis

The establishment of the apical-basal axis is a critical event in plant embryogenesis, evident from the earliest stages onwards. Polarity is evident in the embryo sac, egg cell, zygote, and embryo-suspensor complex. In the embryo-proper, two functionally distinct meristems form at each pole, through the localized expression of key genes. A number of mutants, notably of the model genetic organism Arabidopsis thaliana, have revealed new gene functions that are required for patterning of the apical-basal axis. There is now increasing evidence that two particular modes of signalling, via auxin and cell wall components, play important roles in co-ordinating the gene expression programmes that define determinative roles in the establishment of polarity.     

Microtubule‐associated protein tau in human prostate cancer cells: Isoforms,phosphorylation, and interactions

Tau is a microtubule‐associated protein whose function has been investigated primarily in neurons. Recently, tau expression has been correlated with increased drug resistance in various cancers of non‐neuronal tissues. In this report, we investigate the tau expressed in cancerous prostate lines ALVA‐31, DU 145, and PC‐3. Prostate cancer tau is heat‐stable and highly phosphorylated, containing many of the modifications identified in Alzheimer's disease brain tau. RT‐PCR and phosphatase treatment indicated that all six alternatively spliced adult brain tau isoforms are expressed in ALVA‐31 cells, and isoforms containing exon 6 as well as high molecular weight tau isoforms containing either exon 4A or a larger splice variant of exon 4A are also present. Consistent with its hyperphosphorylated state, a large proportion of ALVA‐31 tau does not bind to microtubules, as detected by confocal microscopy and biochemical tests. Finally, endogenous ALVA‐31 tau can interact with the p85 subunit of phosphatidylinositol 3‐kinase, as demonstrated by co‐immunoprecipitations and in vitro protein‐binding assays. Our results suggest that tau in prostate cancer cells does not resemble that from normal adult brain and support the hypothesis that tau is a multifunctional protein. J. Cell. Biochem. 108: 555–564, 2009. © 2009 Wiley‐Liss, Inc.     

Intake compensates for resting metabolic rate variation in female C57BL/6J mice fed high-fat diets

Objective: The literature is divided over whether variation in resting metabolic rate (RMR) is related to subsequent obesity. We set out to see whether the effect of RMR on weight gain in mice could be revealed with high‐fat feeding. Research Methods and Procedures: Female C57BL/6J mice received a low‐ (10 kcal%fat n = 47), medium‐ (45 kcal%fat n = 50), or high‐fat diet (60 kcal%fat n = 50) for 12 weeks. Pre‐treatment RMR was measured by indirect calorimetry. Body composition was estimated using DXA before and after treatment. Results: Mice on the high‐fat diet gained 39% of body mass, whereas control animals gained 3.5%. There was no interaction between RMR and dietary type on weight gain, and there was no association between weight gain and RMR for any of the treatments. RMR accounted for 2.4% of the variation in pre‐treatment food intake corrected for initial body mass; however, the gradient of this relationship indicated that variations in RMR were, on average, compensated for by adjustments in food intake. Discussion: Individual variations in RMR did not predispose mice to weight gain independent of the dietary treatment. Deviations from the relationship between RMR and food intake were not associated with weight gain. This suggests that variations in energy expenditure, caused by RMR and physical activity, are closely linked to dietary intake, and, therefore, well compensated. Individual variations in the strength of this association may underpin individual variability in the responses to diet.     

A quantitative assessment of bone area increase due to ornamentation in the Crocodylia

Bone ornamentation, in the form of highly repetitive motives created by pits and ridges, is a frequent feature on vertebrate skull roofs and osteoderms. The functional significance of this character remains a matter of controversy and speculation. The many diverging hypotheses proposed to explain it all share a common logical prerequisite: bone ornamentation should increase significantly the surface area of the bones that bear it. In order to test this assumption in the Crocodylia, we developed a method for quantifying the gain in area due to ornamentation using a three‐dimensional‐surface scanner. On crocodylian osteoderms, the gain in area can be up to 40%, and on the cranial table, it ranges between 10 and 32% in adult specimens (in both cases, it shows substantial differences between the adults of the various species included in the sample). Area gain on the snout is lesser (0–20% in adults), and more variable between species. In general, bone ornamentation is less pronounced, and results in fewer area gains in juvenile specimens. The main morphometric results yielded by this study are discussed in reference to the few comparative data available hitherto, and to the functional interpretations proposed by previous authors. J. Morphol. 276:1183–1192, 2015. © 2015 Wiley Periodicals, Inc.