Biomedical subjects

Melanie Ziman

Publications and source records attributed to Melanie Ziman.

6 recordsLinked to original sources

PubMed2006

A multiphasic role for Pax7 in tectal development.

The optic tectum differentiates from the mesencephalic alar plate and matures into a characteristically laminated structure. Evidence presented here suggests a role for Pax7 in all stages of development of tectal architecture, from regionalisation to specification of neurons and tectal topography. Analysis of Pax7 expression profiles over a range of developmental stages (E2-E12) suggests a biphasic role for Pax7: initially Pax7 expressing cells in the proliferative neuroepithelial layer establish tectal polarity whereas later Pax7 is expressed in neurons of the retino-recipient precursor stratum griseum et fibrosum superficiale (sgfs) laminae where graded levels may establish tectal topography. Furthermore, co-localisation immunofluorescence confirmed that Pax7 is initially expressed in the majority of proliferative neuroepithelial cells and later in a subset of neurons of the sgfs laminae.

Age Factors↗

PubMed2005

Multiplex analysis of intracellular signaling pathways in lymphoid cells by microbead suspension arrays.

Phosphorylation analysis of signaling proteins is key for examining intracellular signaling pathways. Conventional biochemical approaches, e.g. immunoprecipitation, Western blot, and ELISA, have played a major role in elucidation of individual signaling events. However, these methods are laborious, time-consuming, and difficult to adapt for high throughput analysis. A multiplex approach to measure phosphorylation state of multiple signaling proteins simultaneously would significantly enhance the efficiency and scope of signaling pathway analysis for mechanistic studies and clinical application. This report describes a novel multiplex microbead suspension array approach to examine phosphoproteomic profiles in lymphoid cells. In the Jurkat T-cell leukemia line, the multiplex assay enabled targeted investigation of phosphorylation kinetics of signal transduction from receptor proximal events (tyrosine phosphoproteins CD3, Lck, Zap-70, and linker for T-cell activation) to cytosolic events (serine/threonine phosphoproteins Erk and Akt) to transcription factors (serine/threonine phosphorylated Rsk, cyclic AMP-response element-binding protein, and STAT3). To broaden the application of the multiplex analysis, signaling pathways were also studied in B-cell lymphoid tumor lines that included chronic lymphocytic leukemia lines. In these cell lines, multiplex suspension array enabled phosphoproteomic analysis of signaling cascade mediated by Syk, a homolog of Zap-70. Results obtained by multiplex analysis were confirmed by immunoprecipitation and Western blot methods. The examples of T-cell and B-cell signaling pathway analyses in this report demonstrate the utility of the multiplex suspension arrays to investigate phosphorylation dynamics and kinetics of several signaling proteins simultaneously in signal transduction pathways.

B-Lymphocytes↗

PubMed2005

Simultaneous serodetection of 10 highly prevalent mouse infectious pathogens in a single reaction by multiplex analysis.

Under current practices of mouse colony maintenance, sera from mice are analyzed for antibodies against several widespread infectious pathogens by conventional immunoassays, generally enzyme-linked immunosorbent assay (ELISA). To test for multiple agents, these methods consume large volumes of mouse serum and are laborious and time-consuming. More efficient immunoassays, using small amounts of sample, are therefore needed. Accordingly, we have developed a novel multiplex diagnostic system that employs fluorescent microbeads, coated with purified antigens, for simultaneous serodetection of 10 mouse infectious agents. Individually identifiable, fluorescent microbeads were coated with antigens from Sendai virus, mouse hepatitis virus, Theiler's mouse encephalomyelitis virus/GDVII strain, mouse minute virus, mouse cytomegalovirus, respiratory enteric orphan virus (Reo-3 virus), mouse parvovirus, calf rotavirus for epizootic diarrhea virus of infant mice, vaccinia virus for ectromelia virus, and Mycoplasma pulmonis. Standard sera, singly positive for antibodies to individual infectious agents, were generated by inoculation of BALB/cj and C57BL/6j mice. Sera from these experimentally infected mice, as well as sera from naturally infected mice, were analyzed using a mixture of microbeads coated with antigens of the 10 infectious agents listed above. Results demonstrated that the multiplex assay was at least as sensitive and specific as ELISA for serodetection. Importantly, the multiplex assay required only 1 microliter of serum for simultaneous serodetection of the 10 mouse infectious agents in one reaction vessel. Thus, this multiplex microbead assay is a reliable, efficient, and cost-effective diagnostic modality that will impact serosurveillance of mice used in research.

Animals↗

PubMed2004

Expression profiles suggest a role for Pax7 in the establishment of tectal polarity and map refinement.

The role for Pax7 in establishing tectal polarity and map refinement was authenticated by gene expression studies in vivo and in vitro. Throughout development (stages E2-E12 were examined) a rostral(low)-caudal(high) and dorsal(high)-ventral(low) Pax7 expression gradient was detected immunohistochemically in the chick optic tectum, indicating a role for Pax7 in establishing tectal polarity. Chick retino-recipient tectal cells positive for Pax7 also co-expressed ephrin-A2, a molecule involved in the establishment and refinement of the retinotopic map. In vitro, PAX7 up-regulated ephrin-A2 when transfected into undifferentiated P19 cells; cells became negative for both Pax7 and ephrin-A2 protein following treatment with anti-sense oligonucleotides. These results suggest that in addition to being involved in the early establishment of tectal polarity, Pax7 plays a later role in retino-tectal map formation and refinement.

Animals↗

PubMed2004

Haemoproteus spp. and Leukocytozoon spp. in a captive raptor population.

Raptors are commonly infected with two blood parasites of the family Haemoproteidae, Haemoproteus spp. and Leukocytozoon spp. To determine if age or length of time in captivity influence prevalence of Haemoproteus spp. and Leukocytozoon spp. infection in captive raptors, blood samples were collected from 55 birds from April 1999 to May 2000. Blood smears were examined for parasitemia and influence of age and length of time in captivity at the time of sample collection were compared. We found juvenile and adult birds were more likely to be infected with Leukocytozoon spp. than were nestlings (P = 0.006) and birds present for > 365 days were more likely to be infected with Haemoproteus spp. and/or Leukocytozoon spp. than were birds captive for < 365 days.

Age Factors↗

PubMed2003

A dorso-ventral gradient of Pax6 in the developing retina suggests a role in topographic map formation.

Expression of the transcription factor Pax6 was assessed immunohistochemically in embryonic chick retina during retino-tectal map formation. A low dorsal to high ventral gradient was found that correlated with expression of the axonal guidance cue EphB2. Furthermore, transfection of Pax6 into undifferentiated P19 cells up-regulated EphB2. The results raise the possibility that Pax6 is upstream of EphB2 and that its graded expression defines the dorso-ventral axis of the retino-tectal projection.

Animals↗