Mitotic microtubule development and histone H3 phosphorylation in the holocentric chromosomes of Rhynchospora tenuis (Cyperaceae).

In the present work we report the phosphorylation pattern of histone H3 and the development of microtubular structures using immunostaining techniques, in mitosis of Rhynchospora tenuis (2n = 4), a Cyperaceae with holocentric chromosomes. The main features of the holocentric chromosomes of R. tenuis coincide with those of other species namely: the absence of primary constriction in prometaphase and metaphase, and the parallel separation of sister chromatids at anaphase. Additionaly, we observed a highly conserved chromosome positioning at anaphase and early telophase sister nuclei. Four microtubule arrangements were distinguished during the root tip cell cycle. Interphase cells showed a cortical microtubule arrangement that progressively forms the characteristic pre-prophase band. At prometaphase the microtubules were homogeneously distributed around the nuclear envelope. Metaphase cells displayed the spindle arrangement with kinetochore microtubules attached throughout the entire chromosome extension. At anaphase kinetochoric microtubules become progressively shorter, whereas bundles of interzonal microtubules became increasingly broader and denser. At late telophase the microtubules were observed equatorially extended beyond the sister nuclei and reaching the cell wall. Immunolabelling with an antibody against phosphorylated histone H3 revealed the four chromosomes labelled throughout their entire extension at metaphase and anaphase. Apparently, the holocentric chromosomes of R. tenuis function as an extended centromeric region both in terms of cohesion and H3 phosphorylation.

Activation of p38 mitogen-activated protein kinase during normal mitosis in the developing retina.

The p38 member of the mitogen-activated protein kinase superfamily is engaged by phosphorylation in response to environmental stress signals, and may have either permissive or inhibitor roles upon cell proliferation. The cell cycle in the proliferative zone of the retina is tightly controlled and proceeds in synchrony with interkinetic migration of the neuroblast nuclei. We examined the association of p38 kinase activity with the cell cycle in the normal, non-stressed retina of the developing rat, maintained either in vivo or in vitro. Using immunohistochemistry, we show that mitotic profiles in the developing retina are highly enriched for phosphorylated p38. Blockade of p38 activity with the chemical inhibitor SB203580 for 4 h transiently arrested cells at the metaphase-anaphase transition and induced cell death after 20 h. p38 inhibition induced an aberrant mitotic profile, with chromosomes arranged in one side of the cell. The data show that p38 is active during normal mitosis and we suggest that p38 is required for the proper cell cycle progression during metaphase-anaphase transition in retinal neuroblasts.

Inducción de anafases anormales por polvos inorgánicos en cultivo de celulas BALBC/3T3 / Induction of abnormal anaphases by inorganic dusts in culture of BALBC/3T3 cells

Los polvos inorgánicos como el asbesto son capaces de inducir alteraciones cromosómicas in vitro. Estas alteraciones se han comprobado empleando fibras de longitud mayores de 5 µm. En el presente estudio se valoraron las anafases anormales inducidas por fibras de asbesto crisótilo y por hierro carbonilo con un tamaño menor de 5.0 µm. Se sembraron 10 células BALBC/3T3 en medio RPMI-1640 y se expusieron a las siguientes dosis de cada polvo: 0, 5, 10, 20, 40, 80 µg/ml durante 12 horas. Las células se fijaron y tiñeron con safranina-O para valorar las anafases anormales inducidas. Se observó que el asbesto crisótilo fue capaz de disminuir las anafases totales conforme de dosis de éste se elevó (de 36 por ciento a 8 por ciento), y el número máximo de anafases anormales fue de 5.6 por ciento con 40 µg/ml. A esta dosis, las anormalidades más comunes fueron los puentes anafásicos (4.3 por ciento) y las anafases multipolares (1.6 por ciento). El hierro carbolino no indujo disminución de las anafases y las anormales inducidas no fueron estadísticamente diferentes de las observadas en cultivos control. De nuestros resultados podemos concluir que las fibras de asbesto crisótilo menores de 5 µm de longitud inducen daño cromosómico directo e indirecto en cultivo celular