ALBERT

Notes: The developmental cycle of the teeth in Plethodon cinereus is analyzed on morphological grounds using alizarin preparations. All the stages in development do not occupy the same proportion of the life cycle time. Functional teeth and germs at an early stage in development occupy a large proportion of the life cycle time, whereas the processes of tooth shedding and ankylosis occur very quickly. The time during which any locus does not bear a functional tooth, and is therefore a non-functional locus, is reduced to a minimum. P. cinereus has a basic pattern of tooth replacement which is consistent with Zahnreihen which are 2.0 tooth spaces apart. Variations in the replacement pattern are common and these are produced by relatively small fluctuations in the spacing of the Zahnreihen around the „mean„ of 2.0. Localized disturbances which produce breaks in the replacement pattern and cause waves to cross also occur. These may be due to the failure of tooth germs to develop, the fusion of tooth germs, or may be the result of the inherent variability in a complex biological system. This variability causes individual tooth germs to develop too slowly or too quickly and hence assume an „abnormal“ position thus causing breaks in the replacement pattern. Tooth replacement may be controlled by an intra-local mechanism(s) rather than by stimuli which travel along the jaw.

Notes: A recently presented model of tongue projection dynamics is used to generate a series of predictions concerning morphologies to be expected under selection for increased distance of projection, increased speed of projection, and increased directional versatility. A general understanding of biomechanical events and the model are used as points of departure for making specific predictions concerning details of structure in skeletal, muscular and connective tissue components of the tongue and associated structures. Comparative methods are used to examine these predictions in the genera of plethodontid salamanders. These salamanders are known to project their tongues to different degrees, and this knowledge is used to test the hypotheses concerning morphological specialization. Three distinct groups of plethodontid salamanders have evolved specializations for long distance projection, and these genera differ from one another in important ways in respect to specific character complexes. For example, the tropical genera and Hydromantes use CBII as the major force transmission element in the skeleton, while Eurycea and its allies use CBI in this role. Hydromantes differs from both in having a uniquely proportioned and structured hyobranchial skeleton and associated musculature. Less extreme specializations for tongue projection are found in different combinations in three other groups. Finally, two distinct groups of generalized species having only limited tongue projection capabilities are recognized, each having a unique complex of inter-related features. Each of these eight groups is recognized and characterized as a functional mode, and hypotheses concerning the biomechanical meaning of the character complexes of each are formulated.

Notes: Basal tail constriction occurs in about two-thirds of the species of plethodontid salamanders. The constriction, which marks the site of tail autotomy, is a result of a reduction in length and diameter of the first caudal segment. Gross and microscopic anatomical studies reveal that many structural specializations are associated with basal constriction, and these are considered in detail. Areas of weakness in the skin at the posterior end of the first caudal segment, at the attachment of the musculature to the intermyotomal septum at the anterior end of the same segment, and between the last caudosacral and first caudal vertebrae precisely define the route of tail breakage. During autotomy the entire tail is shed, and a cylinder of skin one segment long closes over the wound at the end of the body.It is suggested that specializations described in this paper have evolved independently in three different groups of salamanders.Experiments and field observations reveal that, contrary to expectations, frequency of tail breakage is less in species with apparent provisions for tail autotomy than in less specialized species. The tail is a very important, highly functional organ in salamanders and it is suggested that selection has been for behavioral and structural adaptations for control of tail loss, rather than for tail loss per se.

Notes: Regional variation in the vertebral column of several species of salamanders (families Ambystomatidae, Salamandridae and Plethodontidae) is analyzed. Measurements of three dimensions, centrum length, prezygapophyseal width, and transverse process length, provide the data. Ontogenetic, interspecific, intergeneric and interfamilial patterns of positional variation are diagrammed and discussed. Distinctive patterns of variation characterize the families, genera, and to a lesser extent, the species. The patterns of ambystomatid salamanders are the most generalized, and probably reflect derivation from a primitive ancestral stock. The most specialized conditions occur in the fully terrestrial plethodontids, a group generally considered to be highly derived. Data such as those presented here will aid in the identification of fossils.The patterns described have functional significance. For example, species which have an aquatic larval stage and which return to aquatic breeding sites have vertebrae which taper in length and width behind the pelvis. This is a feature associated with production of a traveling wave in the tail which is necessary for propulsion in water. Fully terrestrial species do not have a tapering column. In them, standing waves, such as occur in the trunk region of all species, typically occur in the tail. The caudal vertebrae of terrestrial species are rather uniform in dimensions for some distance, and the tail is cylindrical in form. Other functionally important features include the narrowing and shortening of some anterior vertebrae, associated with the development of a neck in some species with tongue feeding mechanisms. In contrast, species which use their heads as wedges during locomotion have broadened anterior vertebrae which serve as sites of origin for hypertrophied neck muscles.

Notes: The first description of vertebral development in a plethodontid salamander is presented. Eurycea bislineata has larvae that hatch at a rather early stage of development. Somites and the notochord appear early. Somitic differentiation is slight, and no distinct sclerotome can be found. As a result, there is no clear primary segmentation of the skeletogenous tissue. No evidence of a sclerocoele can be found. The amount of sclerotomal cells surrounding the notochord is very low, relative to other tetrapods. Yet discrete perichordal rings of cells do form, in nearly midsegmental positions, and these give rise to the intervertebral cartilages. Osteogenesis of the centra is initiated prior to hatching and is coincidental with ossification of the neural arch. There is no sign of a neurocentral suture. The centrum forms as a thin shell of bone directly from sclerotomal cells. The notochord is a prominent feature of the vertebral column throughout life, retaining its integrity until late in life when some disintegration occurs locally. The notochord is filled with cartilage midvertebrally in late larval stages, and some additional cartilage forms later in life. The intervertebral cartilage enlarges greatly in late larval life. An opisthocoelous joint forms in this cartilage, apparently as a result of differential changes in the cells of the perichordal ring rather than by an invasion of cells from an external source. The intervertebral cartilage is a dominant structural and highly important functional feature of the adult vertebra. In metamorphosed individuals it may become extensively mineralized, and it consists of many different structural kinds of cartilage.The cranio-vertebral joint seems to form in a single segment, contrary to the condition reported by some early investigators. It is complex, and consists of articulations between the odontoid process of the atlas and the occipital arch, as well as between the occipital condyles and atlantal cotyles. The notochord plays a dominant role in the early development of the odontoid, but then changes radically and is absent in the adult process.The anterior trunk region seems to be much more conservative than posterior parts of the column. The patterns of nerve routes and nature of development of the ribs and rib bearers differs greatly from conditions elsewhere in the column. The rib patterns are similar to presumed ancestral conditions. Rib development on the sacral and caudosacral vertebrae is in some ways more similar to that of the anterior vertebrae than of the central trunk vertebrae.Quantitative aspects of variation in the vertebrae of adult salamanders are presented. There is more regional variation and less site variation than would be expected from literature reports.Evolutionary aspects of the origin of the cranio-vertebral joint, transverse process and ribs, patterns of segmentation, and centrum development are considered in the light of the new information on Eurycea. There is no evidence that more than one vertebra is involved developmentally or evolutionarily in the cranio-vertebral joint. The most generalized condition of rib bearers in living salamanders is one in which the dorsal and ventral bearers are in cartilaginous continuity during development. There are many variations on this theme in living species. It is inappropriate to speak of a resegmentation of the sclerotome in Eurycca, even though the adult vertebra is a transsegmental structure, because there is no primary segmentation of the scanty sclerotome. The important feature found in vertebral development in all tetrapods is the perichordal tube and its subsequent differentiation. Questinos concerning precise homologies of the salamander vertebral centrum with those of other vertebrates cannot be answered by data from development sequences with currently used criteria of homology. On the other hand, it appears that all centra, regardless of subdivision, are homologous in all tetrapods.

Notes: Innervation of the tongue and associated musculature in plethodontid salamanders was studied using Palmgren stained sectioned materials, fresh dissection, and whole mounts of experimental specimens treated with horseradish peroxidase (HRP). Species studied were chosen to represent modes of tongue projection recognized by Lombard and Wake ('77). Special attention was given to species of the genera Plethodon, Batrachoseps, Pseudoeurycea, and Hydromantes, but representatives of other genera were investigated. As expected we found that cranial nerves IX and X and spinal nerve 1 supplied the muscles involved in tongue movement. The peripheral courses of the nerves were traced, and both functionally related and phylogenetically determined routes were found. As relative projection length increases, the nerves supplying the tongue tip also increase in length. When the tongue is at rest the long nerves are stored in coils. The coil of ramus lingualis lies between the ceratobranchials, but that of ramus hypoglossus is more variable, although constant within a species. Ramus hypoglossus bifurcates into separate branches to tongue and anterior musculature of the floor of the mouth. In generalized, presumably primitive, modes the bifurcation and coiling are far anterior. In most of the tongue projection modes bifurcation is relatively posterior, but in one, bifurcation is anterior, but coiling is relatively posterior in position. The most unusual condition is in Hydromantes, in which bifurcation is relatively posterior and a coiled ramus hypoglossus joins a coiled ramus lingualis to form a unique, coiled common ramus to the tongue tip. Hydromantes has the greatest projection distance of any salamander.

Notes: Each of the paired salivary glands of third instar larvae of the humpbacked fly Megaselia scalaris is a bag-like structure with a short neck region from which a single duct emerges. The two ducts form a common duct that empties into the ventral region of the pharynx near the mouthparts. The wall of the glands and ducts consists of a simple squamous epithelium that rests upon a connective tissue layer. Cells in the neck are less flattened than those found elsewhere. The basal surfaces of the cells are infolded most deeply in the neck and the least in the duct. The apical surfaces of the cells possess microvilli except in the duct where the apices of the cells are covered by a complex extracellular layer. This layer displays circularly arranged folds that accommodate a thread-like supportive structure resembling taenidial threads of tracheae. Elaborate junctional complexes are associated with the lateral surfaces of the cells. Elements of these complexes include a zonula adherens, a series of pleated septate desmosomes, and conventional desmosomes. The cytoplasm of the glandular cells is filled with RER and other organelles normally seen in cells that export proteins and mucosubstances. Secretory material found in the lumens of the glands reacts only moderately with the PAS procedure but more strongly with alcian blue and methods that demonstrate proteins. The nuclei of the glandular cells contain single large nucleoli and polytene chromosomes whose banding is rather indistinct. Treatment with EDTA produces detrimental effects on all of the foregoing ultrastructural features of the glands and ducts.

Notes: In artificial fluid, the spermatozoa move as linear cells or round up and rotate, propelled by spontaneous bending of their tails. Both linear and rounded cells can move forward and backward, but usually they move forward. The tails of all cells display, simultaneously, small primary bends and fewer, much larger secondary bends. Rounded cells form single secondary bends that remain unchanged as the cells rotate. They also form “node-like” primary bends that travel posteriorly or anteriorly as the cells rotate forward or backward, respectively. Linear cells move their anterior regions into and out of focus in a cyclic fashion. They form rather prominent primary bends, as well as two to four secondary bends that travel posteriorly as the cells move forward. Secondary bends change in shape continuously and are not sinusoidal. The cells follow approximately linear trajectories, but the distances traveled per cycle, speeds, and secondary bending patterns are variable. When methyl cellulose is added to artificial fluid, linear movement is improved, and forward speeds are approximately tripled. The movement of spermatozoa in natural fluid of the female reproductive tract is remarkably less stereotyped than that of cells in artificial fluid. The cells, usually resembling straight lines or arcs, are very flexible and active. They lack obvious cyclic activity and double bending patterns. They are capable of moving both forward and backward and of adjusting their bending activity and speed within rather wide limits. Their average forward speed is about nine times faster than that of cells in artificial fluid.

Notes: Plethodontid salamanders capture prey by projecting the tongue from the mouth. An analysis of theoretical mechanics of the hyobranchial skeleton is used to formulate a working hypothesis of tongue movements. Predictions that the skeletal elements of the tongue are included in the projectile and that the hyobranchial skeleton is folded during projection are central to the analysis, When decapitated in a particular way, salamanders project the tongue, and it is not retracted. When these heads are fixed and sectioned, examination confirms the predictions, In turn, these observations are used to refine the working hypothesis and to generate a general model of tongue dynamics for plethodontids. Muscles performing the major roles of projection (subarcualis rectus I) and retraction (rectus cervicis profundus) are identified. The skeleton is folded passively along a morphological track having the form of a tractrix, Predictions concerning the shape of the track and the exact configuration of the folded skeleton are confirmed by study of sectioned material. The skeleton unfolds along the track during retraction and is spread into the resting state, The model developed herein will be used as a basis for predictions concerning selection patterns in the family and for analytical purposes in comparative and evolutionary studies.