Biomedical subjects

J Reinitz

Publications and source records attributed to J Reinitz.

16 recordsLinked to original sources

PubMed2002

Support vector regression applied to the determination of the developmental age of a Drosophila embryo from its segmentation gene expression patterns.

MOTIVATION: In this paper we address the problem of the determination of developmental age of an embryo from its segmentation gene expression patterns in Drosophila. RESULTS: By applying support vector regression we have developed a fast method for automated staging of an embryo on the basis of its gene expression pattern. Support vector regression is a statistical method for creating regression functions of arbitrary type from a set of training data. The training set is composed of embryos for which the precise developmental age was determined by measuring the degree of membrane invagination. Testing the quality of regression on the training set showed good prediction accuracy. The optimal regression function was then used for the prediction of the gene expression based age of embryos in which the precise age has not been measured by membrane morphology. Moreover, we show that the same accuracy of prediction can be achieved when the dimensionality of the feature vector was reduced by applying factor analysis. The data reduction allowed us to avoid over-fitting and to increase the efficiency of the algorithm.

Aging↗

PubMed2001

Thoracic patterning by the Drosophila gap gene hunchback.

Localized gene expression patterns are critical for establishing body plans in all multicellular animals. In Drosophila, the gap gene hunchback (hb) is expressed in a dynamic pattern in anterior regions of the embryo. Hb protein is first detected as a shallow maternal gradient that prevents expression of posterior gap genes in anterior regions. hb mRNA is also expressed zygotically, first as a broad anterior domain controlled by the Bicoid (Bcd) morphogen, and then in a stripe at the position of parasegment 4 (PS4). Here, we show that the PS4-hb stripe changes the profile of the anterior Hb gradient by generating a localized peak of protein that persists until after the broad domain has started to decline. This peak is required specifically for the formation of the mesothoracic (T2) segment. At the molecular level, the PS4-hb stripe is critical for activation of the homeotic gene Antennapedia, but does not affect a gradient of Hb repressive activity formed by the combination of maternal and Bcd-dependent Hb. The repressive gradient is critical for establishing the positions of several target genes, including the gap genes Kruppel (Kr), knirps (kni), and giant (gt), and the homeotic gene Ultrabithorax (Ubx). Different Hb concentrations are sufficient for repression of gt, kni, and Ubx, but a very high level of Hb, or a combinatorial mechanism, is required for repression of Kr. These results suggest that the individual phases of hb transcription, which overlap temporally and spatially, contribute specific patterning functions in early embryogenesis.

Animals↗

PubMed2001

Registration of the expression patterns of Drosophila segmentation genes by two independent methods.

MOTIVATION: To construct an integrated map of Drosophila segmentation gene expression from partial data taken from individual embryos. RESULTS: Spline and wavelet based registration techniques were developed to register Drosophila segmentation gene expression data. As ground control points for registration we used the locations of extrema on gene expression patterns, represented in 1D. The registration method was characterized by unprecedented high accuracy. A method for constructing the integrated pattern of gene expression at cellular resolution was designed. These patterns were constructed for 9 segmentation genes belonging to gap and pair-rule classes.

Algorithms↗

PubMed2000

HOX Pro: a specialized database for clusters and networks of homeobox genes.

It is now clear that the homeobox motif is well conserved across metazoan phyla. It has been established experimentally that a subset of genes containing this motif plays key roles in the orchestration of gene expression during development. Auto- and cross-regulatory functional interactions join homeobox genes into genetic networks. We have developed a specialized database HOX-Pro in order to arrange all available data on structure, function, phylogeny and evolution of Hox genes, Hox clusters and Hox networks. Its primary location is http://www.iephb.nw.ru/hoxpro. The database is also mirrored at http://www.mssm.edu/molbio/hoxpro. The HOX-Pro database is aimed at: (i) analysis and classification of regulatory and coding regions in diverse homeobox and related genes; (ii) comparative analysis of organization of 'Hox-based' genetic networks in the sea urchin Strongylocentrotus purpuratus, the fruit fly Drosophila melanogaster and the mouse Mus musculus; and (iii) analysis of phylogeny and evolution of homeobox genes and clusters.

Animals↗

PubMed1999

Transcriptional repression by the Drosophila giant protein: cis element positioning provides an alternative means of interpreting an effector gradient.

Early developmental patterning of the Drosophila embryo is driven by the activities of a diverse set of maternally and zygotically derived transcription factors, including repressors encoded by gap genes such as Krüppel, knirps, giant and the mesoderm-specific snail. The mechanism of repression by gap transcription factors is not well understood at a molecular level. Initial characterization of these transcription factors suggests that they act as short-range repressors, interfering with the activity of enhancer or promoter elements 50 to 100 bp away. To better understand the molecular mechanism of short-range repression, we have investigated the properties of the Giant gap protein. We tested the ability of endogenous Giant to repress when bound close to the transcriptional initiation site and found that Giant effectively represses a heterologous promoter when binding sites are located at -55 bp with respect to the start of transcription. Consistent with its role as a short-range repressor, as the binding sites are moved to more distal locations, repression is diminished. Rather than exhibiting a sharp 'step-function' drop-off in activity, however, repression is progressively restricted to areas of highest Giant concentration. Less than a two-fold difference in Giant protein concentration is sufficient to determine a change in transcriptional status of a target gene. This effect demonstrates that Giant protein gradients can be differentially interpreted by target promoters, depending on the exact location of the Giant binding sites within the gene. Thus, in addition to binding site affinity and number, cis element positioning within a promoter can affect the response of a gene to a repressor gradient. We also demonstrate that a chimeric Gal4-Giant protein lacking the basic/zipper domain can specifically repress reporter genes, suggesting that the Giant effector domain is an autonomous repression domain.

Animals↗

PubMed1999

Spatio-temporal registration of the expression patterns of Drosophila segmentation genes.

The application of image registration techniques resulted in the construction of an integrated atlas of Drosophila segmentation gene expression in both space and time. The registration method was based on a quadratic spline approximation with flexible knots. A classifier for automatic attribution of an embryo to one of the temporal classes according to its gene expression pattern was developed.)

Animals↗

PubMed1998

Stripe forming architecture of the gap gene system.

In this report, we show that gap genes encode exactly one set of pair-rule stripes, which occur in the native even-skipped position. The core of this work is a detailed analysis that shows how this conclusion follows from the arrangement of gap domains in the embryo. This analysis shows that: (1) pattern forming information is transmitted from gap to pair-rule genes by means of a nonredundant set of morphogenetic gradients, and (2) the stripe forming capability of the gap genes is constrained by the arrangement of these gradients and by the fact that each gap domain consists of a pair of correlated gradients. We also show that in the blastoderm, the regulatory sign of a transcriptional regulator is unlikely to change in a concentration dependent manner. The principal analytic tool used to establish these results is the gene circuit method. Here, this method is applied to examine hybrid data sets consisting of real gene expression data for four gap genes and hypothetical pair-rule expression data generated by translating native even-skipped data along the anterior-posterior axis. In this way, we are able to investigate the stripe forming capabilities of the gap gene system in the complete absence of pair-rule cross regulation. We close with an inference about evolutionary development. It is argued that the constraints on gap gene architecture identified here are a consequence of selective pressures that minimize the number of gap genes required to determine segments in long-germ band insects.

Animals↗

PubMed1998

Rapid preparation of a panel of polyclonal antibodies to Drosophila segmentation proteins.

We describe a method for rapidly raising a panel of high quality polyclonal antibodies from bacterially expressed proteins. Approximately 1(2)/3 days of preparation is required per protein. One step that speeds up the procedure is the visualization of purified bands by precipitated sodium dodecyl sulfate (SDS). Antigenicity of the purified recombinant proteins may be increased by precipitation in double-distilled water. The results of using the serums obtained for fluorescent staining of Drosophila embryos are shown.

Animals↗

PubMed1998

Automated assay of gene expression at cellular resolution.

We have recently developed an automated image processing method for obtaining quantitative values for average levels of gene expression at the resolution of a single cell. This method is described in the present paper. We place this method within a larger framework for the study of gene regulation in Drosophila, stressing that gene circuit models and improved data processing methods are mutually reinforcing approaches to this problem.

Animals↗

PubMed1995

Model for cooperative control of positional information in Drosophila by bicoid and maternal hunchback.

The blastoderm of the fruit fly Drosophila melanogaster is unusually well suited for analysis of fundamental questions in animal development. One such question is how genes specify the positional information which determines the developmental pathways (fate) of cells at appropriate spatial locations. In this paper we propose a dynamical model of gene regulation which explicitly describes how positional information is used in the blastoderm. The model is applied to analyze important experimental findings on the dependence of cell fate on the concentration of the Bicoid morphogen. The model shows that positional information in the presumptive middle body is cooperatively determined by maternal products of the bicoid and hunchback genes.

Animals↗

PubMed1995

Mechanism of eve stripe formation.

In this paper we analyze the formation of stripes of expression of the pair-rule gene eve. We identify detailed mechanisms which control the formation of stripes 2-5. Each stripe is formed as a result of generalized activation by bcd and ubiquitous transcription factors combined with localized repression by gap genes. Each of the eight stripe borders of these four stripes is shown to be under the control of a particular gap gene expression domain. Protein synthesis from eve and its controlling gap genes begins at the same time, but localized eve expression is substantially delayed relative to localized expression of gap domains. We show that this delay results from a change in the spatial balance between activation and repression due to the intensification and refinement of gap domains during cleavage cycle 14. eve stripe formation is ordered in time; stripe 2 appears earlier than stripes 3-5. We show that this happens because the formation of stripe 2 is less dependent on gap domain refinement than is the case for stripes 3-5: Each of stripes 3-5 is controlled by a pair of overlapping gap domains, whereas stripe 2 is controlled by a disjoint pair of gap domains. Finally, we observe that eve stripes do not form unless Eve protein has an extremely small diffusivity, and argue that this low diffusivity is a result of the apical localization of pair-rule message. This implies that localization of pair-rule message is required for stripe formation. The essential tool used to obtain these results is the method of gene circuits, which is a new approach to the analysis of gene expression data. Its purpose is to provide a way to use this data to infer how concentrations of products of a given gene change with time and how these changes are influenced by the activating or repressing effects of the products of other genes. The gene circuit method is based on three main ideas, explained in the paper. First is the choice of protein concentrations as state variables for the description of gene regulation. Second is the summary of chemical reaction kinetics by coarse-grained rate equations for protein concentrations. Third is the use of least squares fits to gene expression data to measure phenomenological parameters occurring in the gene circuit.

Animals↗

PubMed1994

Computer-assisted restriction mapping: an integrated approach to handling experimental uncertainty.

Building a map of restriction sites from double-digest gel data can be a complex and frustrating task, especially when many DNA fragments are detected or when the gel results are ambiguous. 'Double Digester' is an interactive, graphical computer program which helps researchers understand and resolve such data. It explicitly represents the experimental data, the associated uncertainties, the researcher's hypotheses and possible map interpretations. Alternative solutions are frequently possible, and the differences between them may help determine which additional experiments might resolve ambiguities. Initial use has confirmed the benefits of this approach, and has suggested ways in which it can be refined and extended. Double Digester meets the need for a practical tool to help build restriction maps, and also illustrates how a computer-based tool can confront experimental uncertainty in an integrated fashion.

Algorithms↗

PubMed1991

A connectionist model of development.

We present a phenomenological modeling framework for development. Our purpose is to provide a systematic method for discovering and expressing correlations in experimental data on gene expression and other developmental processes. The modeling framework is based on a connectionist or "neural net" dynamics for biochemical regulators, coupled to "grammatical rules" which describe certain features of the birth, growth, and death of cells, synapses and other biological entities. We outline how spatial geometry can be included, although this part of the model is not complete. As an example of the application of our results to a specific biological system, we show in detail how to derive a rigorously testable model of the network of segmentation genes operating in the blastoderm of Drosophila. To further illustrate our methods, we sketch how they could be applied to two other important developmental processes: cell cycle control and cell-cell induction. We also present a simple biochemical model leading to our assumed connectionist dynamics which shows that the dynamics used is at least compatible with known chemical mechanisms.

Animals↗

PubMed1990

Theoretical and experimental analysis of the phage lambda genetic switch implies missing levels of co-operativity.

The behavior of the cro-repressor switch in phage lambda, a temperate phage of Escherichia coli, is described at the organismal level within a dynamical system framework. The molecular biology of the switch has been well characterized up to the level of the regulation of transcription initiation. In this paper we construct a description of a lysogen whose prophage is mutated in certain genes, so that the switch is functionally isolated from the rest of the phage genome. Such a lysogen has two stable epigenetic states, and comparison of the theory with measurements of intracellular cro concentration corresponding to one of these states suggest that additional levels of regulation, not included in the current description of the switch, should exist for this system. In addition, we suggest a new method for the measurement of intracellular concentrations.

Bacteriophage lambda↗

PubMed1990

Control of the initiation of homeotic gene expression by the gap genes giant and tailless in Drosophila.

The process of segmentation in Drosophila is controlled by both maternal and zygotic genes. Members of the gap class of segmentation genes play a key role in this process by interpreting maternal information and controlling the expression of pair-rule and homeotic genes. We have analyzed the pattern of expression of a variety of homeotic, pair-rule, and gap genes in tailless and giant gap mutants. tailless acts in two domains, one anterodorsal and one posterior. In its anterior domain tailless exerts a repressive effect on the expression of fushi tarazu, hunchback, and Deformed. In its posterior domain of action, tailless is responsible for the establishment of Abdominal-B expression and demarcating the posterior boundary of the initial domain of expression of Ultrabithorax. giant is an early zygotic regulator of the gap gene hunchback: in giant- embryos, alterations in the anterior domain of hunchback expression are visible by the beginning of cycle 14. giant also regulates the establishment of the expression patterns of Antennapedia and Abdominal-B. In particular, giant is the factor that controls the anterior limit of early Antennapedia expression.

Animals↗

PubMedDate not supplied

Prediction of mutant expression patterns using gene circuits.

Networks of interacting transcription factors, or gene circuits, form an essential part of the metabolic pathways controlling macromolecular synthesis. This paper conveys two new results about gene circuits. We first show how a gene circuit for mutant phenotypes can be constructed from the wild type gene circuit for the same organism. We then present results of computational studies that show that mutant expression patterns can be correctly predicted using gene circuits whose parameters have been determined from wild type data only. Further computational studies demonstrate that this property is insensitive to errors as large as a factor of two in the input data. Together, these results show that gene circuits can be used to identify the regulatory mechanisms governing an entire family of genotypes from a knowledge of the wild type genotype alone. It is argued that this fact forms the basis for a new paradigm in genetics.

Gene Expression↗