Gerd B. Müller

Evo-devo : extending the evolutionary synthesis

Evolutionary developmental biology (evo–devo) explores the mechanistic relationships between the processes of individual development and phenotypic change during evolution. Although evo–devo is widely acknowledged to be revolutionizing our understanding of how the development of organisms has evolved, its substantial implications for the theoretical basis of evolution are often overlooked. This essay identifies major theoretical themes of current evo–devo research and highlights how its results take evolutionary theory beyond the boundaries of the Modern Synthesis.

Epigenetic Mechanisms of Character Origination

The close mapping between genotype and morphological phenotype in many contemporary metazoans has led to the general notion that the evolution of organismal form is a direct consequence of evolving genetic programs. In contrast to this view, we propose that the present relationship between genes and form is a highly derived condition, a product of evolution rather than its precondition. Prior to the biochemical canalization of developmental pathways, and the stabilization of phenotypes, interaction of multicellular organisms with their physicochemical environments dictated a many-to-many mapping between genomes and forms. These forms would have been generated by epigenetic mechanisms: initially physical processes characteristic of condensed, chemically active materials, and later conditional, inductive interactions among the organisms constituent tissues. This concept, that epigenetic mechanisms are the generative agents of morphological character origination, helps to explain findings that are difficult to reconcile with the standard neo-Darwinian model, e.g., the burst of body plans in the early Cambrian, the origins of morphological innovation, homology, and rapid change of form. Our concept entails a new interpretation of the relationship between genes and biological form.

High-resolution episcopic microscopy: a rapid technique for high detailed 3D analysis of gene activity in the context of tissue architecture and morphology

We describe a new methodology for rapid 2D and 3D computer analysis and visualisation of gene expression and gene product pattern in the context of anatomy and tissue architecture. It is based on episcopic imaging of embryos and tissue samples, as they are physically sectioned, thereby producing inherently aligned digital image series and volume data sets, which immediately permit the generation of 3D computer representations. The technique uses resin as embedding medium, eosin for unspecific tissue staining, and colour reactions (β-galactosidase/Xgal or BCIP/NBT) for specific labelling of gene activity and mRNA pattern. We tested the potential of the method for producing high-resolution volume data sets of adult human and porcine tissue samples and of specifically and unspecifically stained mouse, chick, quail, frog, and zebrafish embryos. The quality of the episcopic images resembles the quality of digital images of true histological sections with respect to resolution and contrast. Specifically labelled structures can be extracted using simple thresholding algorithms. Thus, the method is capable of quickly and precisely detecting molecular signals simultaneously with anatomical details and tissue architecture. It has no tissue restrictions and can be applied for analysis of human tissue samples as well as for analysis of all developmental stages of embryos of a wide variety of biomedically relevant species.

External marker‐based automatic congruencing: A new method of 3D reconstruction from serial sections

Computer‐based three‐dimensional (3D) visualizations reconstructed from sectional images represent a valuable tool in biomedical research and medical diagnosis. Particularly with those imaging techniques that provide virtual sections, such as CT, MRI, and CLSM, 3D reconstructions have become routine. Reconstructions from physical sections, such as those used in histological preparations, have not experienced an equivalent breakthrough, due to inherent shortcomings in sectional preparation that impede automated image‐processing and reconstruction. The increased use of molecular techniques in morphological research, however, generates an overwhelming amount of 3D molecular information, stored within series of physical sections. This valuable information can be fully appreciated and interpreted only through an adequate method of 3D visualization.

Computer-based three-dimensional visualization of developmental gene expression.

A broad understanding of the relationship between gene activation, pattern formation and morphogenesis will require adequate tools for three-dimensional and, perhaps four-dimensional, representation and analysis of molecular developmental processes. We present a novel, computer-based method for the 3D visualization of embryonic gene expression and morphological structures from serial sections. The information from these automatically aligned 3D reconstructions exceeds that from single-section and whole-mount visualizations of in situ hybridizations. In addition, these 3D models of gene-expression patterns can become a central component of a future developmental database designed for the collection and presentation of digitized, morphological and gene-expression data. This work is accompanied by a web site (http://www.univie.ac.at/GeneEMAC).

The Paraclinoid Carotid Artery: Anatomical Aspects of a Microneurosurgical Approach

&NA; The paraclinoid area is investigated anatomically for possible microneurosurgical approaches to the C3 segment of the internal carotid artery and to structures in the vicinity of the anterior siphon knee. Removal of the anterior clinoid process reveals a tight connective tissue ring that fixes the internal carotid artery to the surrounding osseous structures at the point of its transdural passage. Transection of this fibrous ring opens a microsurgical pathway to the carotid C3 segment. The artery is surrounded by a loose connective tissue layer that allows blunt preparation along the C3 segment, without compromising the cranial nerves and without damaging venous compartments of the cavernous sinus. This approach provides neurosurgical access to paraclinoidal aneurysms, to partly intracavernous aneurysms, and to carotid‐ophthalmic aneurysms, allowing control of the proximal aneurysm neck and of the parent artery itself. In cases of tumors involving the medial sphenoid ridge, the apex of the orbit, or the cavernous sinus, the pericarotid connective tissue can serve as a guide layer for access along the internal carotid artery.

Ontogeny of the syndesmosis tibiofibularis and the evolution of the bird hindlimb: a caenogenetic feature triggers phenotypic novelty

SummaryThe underlying theme of this study is the contribution of developmental mechanisms to the generation of morphological novelty in evolution. The syndesmosis tibiofibularis, an important structural and functional link between the two zeugopod bones of the bird hindlimb, is used as a model for evolutionary novelty. We analyze the structural, developmental and adaptive aspects of its origin in a combined descriptive, experimental, and comparative approach.The ontogeny of the syndesmosis in the chick embryo involves several developmental steps, including the formation of a separate cartilage rudiment that in turn stimulates the formation of an osseous crest on the tibia, which with eventually replace the cartilage element itself. Some of the epigenetic requirements for the formation of the cartilage element and the osseous crest are demonstrated by experimentally increasing the distance between the two zeugopod bones, an operation that results in the absence of both cartilage and crest. Although a syndesmosis tibiofibularis associated with an osseous crest on the tibiotarsus is unique to birds in extant vertebrates, the presence of a distinct crest at the corresponding location in theropod dinosaurs indicates that a syndesmosis also existed in this group of archosaurs.The results of the study suggest that in the case of the syndesmosis tibiofibularis phenotypic evolutionary novelty is based on a caenogenetic feature, i.e. a feature that initially arose in response to changing developmental conditions. In conclusion we propose a model for the stepwise evolutionary modification of the sauropsid hindlimb, integrating adaptive trends and developmental mechanisms that interactively determine the transformations of skeletal limb morphology. The syndesmosis tibiofibularis and the mechanisms of its formation are not only shown to have played a key-role in this process, but its presence in theropod dinosaurs also points towards the origin of birds.

Evolution evolves: physiology returns to centre stage

Denis Noble, Eva Jablonka, Michael J. Joyner, Gerd B. Muller and Stig W. Omholt University of Oxford, Department of Physiology, Anatomy and Genetics, Oxford, UK Tel Aviv University, Cohn Institute for the History andPhilosophyof Science and Ideas, Ramat Aviv, Israel Mayo Clinic, Rochester, MN, USA University of Vienna, Department of Theoretical Biology, Vienna, Austria Norwegian University of Science and Technology, Faculty of Medicine, Trondheim, Norway

Embryonic motility: environmental influences and evolutionary innovation

Embryos do not passively await hatching from their eggs or amnionic containments but begin active movement very early on in their development. The first muscle contractions in the chick embryo start on the third day of incubation and subsequently assume a characteristic pattern of increasing and decreasing motor activity (Fig. 1) (Hamburger 1963; Bekoff 1981, 1992). It is long known that embryonic motility represents an important epigenetic component of development. In vertebrates, active movement of the embryo is required for the correct development of cartilage, bone, and joints; of muscles, tendons and ligaments; and of connectivities in the central nervous system. The intrauterine disturbance of embryonic movements leads to severe malformations and functional disorders, such as pathological motor patterns and neurological deficits that are retained throughout adult life (Bos et al. 2001). It is equally known that embryonic activity depends on environmental conditions, both chemical and physical, and that changes of environmental parameters can strongly affect the motility patterns. However, the effect of such environmental influences on development, via the alteration of embryonic activity, is little explored. Even less is known about the possible evolutionary consequences of this kind of environment–development interaction. This article provides an outline of the relevance of environment dependent embryonic activity for evolutionary developmental biology.

A new episcopic method for rapid 3-D reconstruction: applications in anatomy and embryology

Abstract The topographic relations of complex structures and the morphogenesis of organ systems can only be fully understood in their three-dimensional context. Three-dimensional (3-D) reconstruction of physically sectioned specimens has become an indispensable tool in modern anatomical and embryological research. Teaching also makes increasingly use of 3-D representations, in particular in the case of embryonic systems that undergo complicated transformations of form and shape. At present no cheap and simple technique is available that generates accurate 3-D models of sectioned objects. In this study we describe a novel technique that rapidly provides faithful 3-D models of sectioned specimens. The images are captured directly from the cutting surface of the embedding block after each sectioning and ”on block” staining step. Automatic image processing generates a stack of binary images of the specimen contour. Binary images of internal structures are obtained both by automatic segmentation and manual tracing. Since these image series are inherently aligned, they can be reconstructed three-dimensionally without time-consuming alignment procedures. The quality and the flexibility of the method are demonstrated by reconstructing three kinds of specimens of different histological composition and staining contrast: a 4 mm mouse embryo together with several of its inner organs, a cavernous sinus region of a human infant, and a segment of a human carotid artery. Very short processing times and the faithful representation of complex structural arrangements recommend this technique for routine use in morphological research and for creating embryologic teaching models or 3-D embryonic staging series.