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C J Hogan

Publications and source records attributed to C J Hogan.

26 records · Page 2Linked to original sources

Developmental changes in tritium autoabsorption.

Developmental changes in autoabsorption of tritium emissions were examined in 30 brain regions in the rat at Postnatal Days 0, 4, 7, 10, 14, 21, and 28 and adulthood. Rats received tritiated 2-deoxyglucose in vivo. Alternate brain sections were extracted in chloroform, and autoradiographs were developed from extracted and nonextracted sections. The ratio of optical density values in extracted vs nonextracted sections was used to determine autoabsorption for each structure. Three principal temporal patterns in the development of adult levels of autoabsorption, determined by the optical density ratios, were identified: (1) a minimal increase pattern in which autoabsorption rose only slightly between birth and adulthood; (2) a plateau pattern in which a rapid early increase was followed by stable values; and (3) a late increase pattern in which autoabsorption remained relatively constant until Postnatal Day 28, with a large increase between Day 28 and adulthood. In addition, optical density ratios fluctuated during the second postnatal week in close to one-third of the structures. The data suggest that developmental events affecting the ratio of gray to white matter produce substantial local variations in the development of adult levels of autoabsorption that are distinct for each structure. To correct for autoabsorption effects in ontogenetic studies using tritium autoradiography, it is necessary to determine directly the degree of autoabsorption at a particular time point for the structure of interest. Our results indicate that the technique of in vivo administration of tritiated 2-deoxyglucose followed by chloroform extraction appears to be a sensitive and reproducible method for assessing autoabsorption at all ages.

Absorption↗

Physiological evidence for involvement of a kinesin-related protein during anaphase spindle elongation in diatom central spindles.

We have developed a new model system for studying spindle elongation in vitro using the pennate, marine diatom Cylindrotheca fusiformis. C. fusiformis can be grown in bulk to high densities while in log phase growth and synchronized by a simple light/dark regime. Isolated spindles can be attained in quantities sufficient for biochemical analysis and spindle tubulin is approximately 5% of the total protein present. The spindle isolation procedure results in a 10-fold enrichment of diatom tubulin and a calculated 40-fold increase in spindle protein. Isolated spindles or spindles in permeabilized cells can elongate in vitro by the same mechanism and with the same pharmacological sensitivities as described for other anaphase B models (Cande and McDonald, 1986; Masuda et al., 1990). Using this model, in vitro spindle elongation rate profiles were developed for a battery of nucleotide triphosphates and ATP analogs. The relative rates of spindle elongation produced by various nucleotide triphosphates parallel relative rates seen for kinesin-based motility in microtubule gliding assays. Likewise ATP analogs that allow discrimination between myosin-, dynein-, and kinesin-mediated motility produce relative spindle elongation rates characteristic of kinesin motility. Also, isolated spindle fractions are enriched for a kinesin related protein as identified by a peptide antibody against a conserved region of the kinesin superfamily. These data suggest that kinesin-like motility contributes to spindle elongation during anaphase B of mitosis.

Adenosine Triphosphate↗

Isolation of a sea urchin egg kinesin-related protein using peptide antibodies.

To understand the roles of kinesin and its relatives in cell division, it is necessary to identify and characterize multiple members of the kinesin superfamily from mitotic cells. To this end we have raised antisera to peptides corresponding to highly conserved regions of the motor domains of several known members of the kinesin superfamily. These peptide antibodies react specifically with the motor domains of kinesin and ncd protein, as expected, and they also react with several polypeptides (including kinesin heavy chain) that cosediment with microtubules (MTs) precipitated from AMPPNP-treated sea urchin egg cytosol. Subsequent fractionation of ATP eluates of these MTs yields a protein of relative molecular mass 330 x 10(3) that behaves as a complex of three polypeptides that are distinct from conventional kinesin subunits or fragments thereof. This complex contains 85 kDa and 95 kDa polypeptides, which react with our peptide antibodies, and a 115 kDa polypeptide, which does not. This triplet of polypeptides, which we refer to as KRP(85/95), binds to purified sea urchin egg tubulin in an AMPPNP-enhanced, ATP-sensitive manner and induces the formation of microtubule bundles. We therefore propose that the triplet corresponds to a novel sea urchin egg kinesin-related protein.

Adenosine Triphosphate↗

The mechanism of anaphase spindle elongation.

At anaphase chromosomes move to the spindle poles (anaphase A) and the spindle poles move apart (anaphase B). In vitro studies using isolated diatom spindles demonstrate that the primary mechanochemical event responsible for spindle elongation is the sliding apart of half-spindle microtubules. Further, these forces are generated within the zone of microtubule overlap in the spindle mid-zone.

Anaphase↗

Preprophase bands in a suspension culture of the monocot Spartina pectinata.

Continuous suspension cultures of the marsh grass Spartina pectinata grow as either unorganized colonies or files of cells. Immunofluorescence of tubulin revealed microtubule (MT) structures similar to those encountered in meristematic cells, including cortical microtubule (MT) bands in some interphase cells and in all prophase cells. These MT bands were judged to be preprophase bands (PPBs) on the basis of their temporal appearance in the cell cycle and their position and orientation relative to division planes. Although PPBs are widely thought to be associated with organized tissues and polarized divisions, there are reports of PPBs in suspension cultures of four dicot species. This is the first report of a PPB in suspension cultures of a monocot species.

Antibodies, Monoclonal↗