Treadmill vs. overground trotting - a comparison of two kinetic measurement systems.

Force plates are considered the gold standard for kinetic gait analysis, benefiting from measuring three-dimensional ground reaction forces. Nevertheless, the major disadvantage is that many trials are required during overground locomotion to capture adequate single-limb contacts. Additionally, the dogs slightly change velocities during overground walking, influencing kinetic and kinematic gait parameters. An alternative is using an instrumented treadmill that benefits from capturing many steady-state gait cycles at a constant speed quickly. The goal of this study was (1) to compare overground with treadmill locomotion and (2) to compare the instrumented treadmill with force plates for dogs kinetics measurements. Twelve client-owned dogs were measured during treadmill trotting while the treadmill was placed on force plates. Additionally, the dogs were measured during trotting along an alley over eight force plates. Bland-Altman plots, Pearson's (r), and concordance correlation coefficients (rc) were computed to explore the relative and general agreement between the measurement methods and overground and treadmill trotting. Overground and treadmill trotting gave an excellent agreement in peak vertical forces and impulses (r > 0.9, rc > 0.9). The instrumented treadmill showed similar force-time curves in shape and size and provided an excellent congruity for all parameters compared to force plates (r > 0.8, rc > 0.8). As a reliable tool in measuring key gait parameters, the instrumented treadmill may benefit from fast and reproducible data comparable to overground trotting.

Exceptional Changes in Skeletal Anatomy under Domestication: The Case of Brachycephaly.

"Brachycephaly" is generally considered a phenotype in which the facial part of the head is pronouncedly shortened. While brachycephaly is characteristic for some domestic varieties and breeds (e.g., Bulldog, Persian cat, Niata cattle, Anglo-Nubian goat, Middle White pig), this phenotype can also be considered pathological. Despite the superficially similar appearance of "brachycephaly" in such varieties and breeds, closer examination reveals that "brachycephaly" includes a variety of different cranial modifications with likely different genetic and developmental underpinnings and related with specific breed histories. We review the various definitions and characteristics associated with brachycephaly in different domesticated species. We discern different types of brachycephaly ("bulldog-type," "katantognathic," and "allometric" brachycephaly) and discuss morphological conditions related to brachycephaly, including diseases (e.g., brachycephalic airway obstructive syndrome). Further, we examine the complex underlying genetic and developmental processes and the culturally and developmentally related reasons why brachycephalic varieties may or may not be prevalent in certain domesticated species. Knowledge on patterns and mechanisms associated with brachycephaly is relevant for domestication research, veterinary and human medicine, as well as evolutionary biology, and highlights the profound influence of artificial selection by humans on animal morphology, evolution, and welfare.

La braquicefalia generalmente se considera un fenotipo en el que el cráneo, específicamente el hocico, es notablemente acortado. Mientras que la braquicefalia es característica de algunas variedades domésticas y razas (p.e. Bulldog, gato persa, vaca ñata, cabra anglo nubiana, cerdo Middle White), también se puede interpretar como un fenotipo patológico. A pesar de que la braquicefalia tiene una apariencia semejante, por lo menos superficial, en estas variedades y razas, al examinarla más en detalle se descubre que la "braquicefalia" incluye una variedad de diferentes modificaciones del cráneo que probablemente tienen diferentes subyacentes genéticos y de desarrollo y que están relacionados con la historia de la raza. Revisamos las diferentes definiciones y propiedades relacionadas con la braquicefalia en varias especies domésticas. Describimos diferentes tipos de braquicefalia (tipo bulldog, "katantognático" y braquicefalia alométrica) y analizamos condiciones morfológicas relacionadas con la braquicefalia incluyendo enfermedades (p.e. síndrome obstructivo respiratorio). Además, examinamos los complejos procesos genéticos y de desarrollo subyacentes y los motivos culturales y de desarrollo por las que variedades braquicéfalas pueden ser más o menos prevalentes en ciertas especies domésticas. El conocimiento de patrones y mecanismos asociados a la braquicefalia son relevantes para la investigación sobre la domesticación, medicina veterinaria y humana, así como para la biología evolutiva y destaca la profunda influencia de la selección artificial sobre la morfología y bienestar de los animales y su evolución.

"Cladus" and clade: a taxonomic odyssey.

The fate of "clade," both as concept and word, is reconstructed here beginning with its first appearance in 1866 as "Cladus," in Haeckel's Generelle Morphologie, continuing up to the present. Although central to phylogenetics, the concept of clade is paradoxical since it has been ambiguously understood or even misunderstood by its own promoters. Writings by Ernst Haeckel, Lucien Cuénot, and Julian Huxley, the three authors who discussed the notion of clade at length, are analyzed here in detail as a means of exploring this paradox. First conceived as a rank for a higher-level category, and later as a taxon, the clade is understood today in connection with Hennig's definition of a monophyletic group rather than through Huxley's successful but somehow ambiguous formalization. The inability of these authors to formulate a clear-cut exposition of the concept is considered here within three contexts: firstly, the burden of pre-Darwinian classifications based on similarity; secondly, the underestimation of Darwin's description of a genealogical group; and thirdly, the predominance of thinking in process (vs thinking in pattern), which was the basis of evolutionary systematics in the mid-twentieth century.

Three-dimensional movements of the pelvis and the lumbar intervertebral joints in walking and trotting dogs.

Current knowledge of the physiological range of motion (ROM) in the canine axial system during locomotion is relatively limited. This is particularly problematic because dogs with back-related dysfunction frequently present for routine consultations. To collect detailed kinematic information and describe the three-dimensional motions of the pelvis and the lumbar spine (i.e. intervertebral joints S1/L7-L2/L1), we recorded ventro-dorsal and latero-lateral X-ray videos of three walking and trotting dogs and reconstructed their pelvic and intervertebral motions using X-ray reconstruction of moving morphology and scientific rotoscoping. Pelvic roll displayed a monophasic motion pattern and the largest ROM with on average 13° and 11° during walking and trotting, respectively. Pelvic yaw had the smallest ROM with on average 5° (walk) and 6° (trot). A biphasic pattern was observed for pelvic pitch with a mean ROM of 8°. At both gaits, the greatest intervertebral motions occurred either in S1/L7 or L7/L6. The intervertebral motions were mono- or biphasic in the horizontal and the transverse body planes and biphasic in the sagittal plane. Cranial to L6/5, the ROM tended to decrease from 3° to <1.5° in all three planes. Our results confirm that pelvic displacement and intervertebral joint movements are tightly linked with pelvic limb action at symmetrical gaits. The overall small movements, particularly cranial to L5, are consistent with the epaxial musculature globally stabilising the spine against the external and internal limb forces acting on the pelvis and the trunk during walking and trotting.

Sagittal joint instability in the cranial cruciate ligament insufficient canine stifle. Caudal slippage of the femur and not cranial tibial subluxation.

OBJECTIVE: This in vivo study qualitatively describes the sagittal motion pattern of the cranial cruciate ligament (CrCL) insufficient canine stifle in operated and unoperated joints with cranio-caudal laxity on palpation. MATERIAL AND METHODS: Sagittal stifle kinematics were recorded in vivo in dogs (> 15 kg BW) with unilateral (n = 7) or bilateral (n = 6) complete CrCL rupture and positive cranial drawer test as well as two sound control dogs using uniplanar fluoroscopic kinematography with the dogs walking on a treadmill. Stifle stability and sagittal motion pattern of the femur and the tibia were determined by visual inspection of the fluoroscopic video sequences. RESULTS: Control dogs showed no cranio-caudal instability, identical to the contralateral stifles of the dogs with unilateral rupture. All unoperated stifles with CrCL rupture (n = 6) showed caudal slippage of the femur at the beginning of the stance phase. Of the 13 operated stifles (TightRope: n = 1, tibial tuberosity advancement, TTA: n = 6, tibial plateau leveling osteotomy, TPLO: n = 5, cranial closing wedge osteotomy, CCWO: n = 1) nine were unstable, showing the same motion pattern as the unoperated stifles. CONCLUSION: In the CrCL insufficient stifle with in vivo cranio-caudal instability caudal slippage of the distal femur at tow touch is the predominant motion pattern. CLINICAL SIGNIFICANCE: The discrepancy between in vivo motion pattern and in vitro simulation of CrCL insufficiency in which cranial tibial subluxation is the predominant sagittal motion pattern warrants further studies.

Fibre type composition in the lumbar perivertebral muscles of primates: implications for the evolution of orthogrady in hominoids.

The axial musculoskeletal system is important for the static and dynamic control of the body during both locomotor and non-locomotor behaviour. As a consequence, major evolutionary changes in the positional habits of a species are reflected by morpho-functional adaptations of the axial system. Because of the remarkable phenotypic plasticity of muscle tissue, a close relationship exists between muscle morphology and function. One way to explore major evolutionary transitions in muscle function is therefore by comparative analysis of fibre type composition. In this study, the three-dimensional distribution of slow and fast muscle fibres was analysed in the lumbar perivertebral muscles of two lemuriform (mouse lemur, brown lemur) and four hominoid primate species (white-handed gibbon, orangutan, bonobo, chimpanzee) in order to develop a plausible scenario for the evolution of the contractile properties of the axial muscles in hominoids and to discern possible changes in muscle physiology that were associated with the evolution of orthogrady. Similar to all previously studied quadrupedal mammals, the lemuriform primates in this study exhibited a morpho-functional dichotomy between deep slow contracting local stabilizer muscles and superficial fast contracting global mobilizers and stabilizers and thus retained the fibre distribution pattern typical for quadrupedal non-primates. In contrast, the hominoid primates showed no regionalization of the fibre types, similar to previous observations in Homo. We suggest that this homogeneous fibre composition is associated with the high functional versatility of the axial musculature that was brought about by the evolution of orthograde behaviours and reflects the broad range of mechanical demands acting on the trunk in orthograde hominoids. Because orthogrady is a derived character of euhominoids, the uniform fibre type distribution is hypothesized to coincide with the evolution of orthograde behaviours.

Kinematics and center of mass mechanics during terrestrial locomotion in northern lapwings (Vanellus vanellus, Charadriiformes).

Avian bipedalism is best studied in derived walking/running specialists. Here, we use kinematics and center of mass (CoM) mechanical energy patterns to investigate gait transitions of lapwings-migratory birds that forage on the ground, and therefore may need a trade-off between the functional demands of terrestrial locomotion and long distance flights. The animals ran on a treadmill while high-speed X-ray videos were recorded within the sustainable speed range. Instantaneous CoM mechanics were computed from integrating kinematics and body segment properties. Lapwings exhibit similar locomotor characteristics to specialized walking/running birds, but have less distinct gaits. At slow speeds no clear separation between vaulting (i.e., walking) and bouncing (i.e., running) energy patterns exists. Mechanical energy recovery of non-bouncing gaits correlates poorly with speed and suggests inefficient use of the inverted pendulum mechanism. Speed ranges of gaits overlap considerably, especially those of grounded running, a gait with CoM mechanics indicative of running but without an aerial phase, and aerial phase running, with no preferential gait at most speeds. Compliant limb morphology and grounded running in birds can be regarded as an evolutionary constraint, but lapwings effectively make use of advantages offered by this gait for a great fraction of their speed range. Thus, effective usage of grounded running during terrestrial locomotion is suggested generally to be a part of striding avian bipedalism-even in species not specialized in walking/running locomotion.

Rapid genetic diversification within dog breeds as evidenced by a case study on Schnauzers.

As a result of strong artificial selection, the domesticated dog has arguably become one of the most morphologically diverse vertebrate species, which is mirrored in the classification of around 400 different breeds. To test the influence of breeding history on the genetic structure and variability of today's dog breeds, we investigated 12 dog breeds using a set of 19 microsatellite markers from a total of 597 individuals with about 50 individuals analysed per breed. High genetic diversity was noted over all breeds, with the ancient Asian breeds (Akita, Chow Chow, Shar Pei) exhibiting the highest variability, as was indicated chiefly by an extraordinarily high number of rare and private alleles. Using a Bayesian clustering method, we detected significant genetic stratification within the closely related Schnauzer breeds. The individuals of these three recently differentiated breeds (Miniature, Standard and Giant Schnauzer) could not be assigned to a single cluster each. This hidden genetic structure was probably caused by assortative mating owing to breeders' preferences regarding coat colour types and the underlying practice of breeding in separate lineages. Such processes of strong artificial disruptive selection for different morphological traits in isolated and relatively small lineages can result in the rapid creation of new dog types and potentially new breeds and represent a unique opportunity to study the evolution of genetic and morphological differences in recently diverged populations.

Arboreal locomotion in small new-world monkeys.

The postural and locomotor activity and its relation to substrates was observed in 3 Saguinus oedipus, and 3 Saimiri sciureus for comparison, during a period of 10 h for each individual. The animals moved freely in cages of 3.40 m x 3.40 m x 2.40 m (height) on rather diverse substrates. Observations were made according to the focal-animal-method, with combined instantaneous and continuous sampling. They were protocoled in schematic form and video-recorded. In addition, 3 further Saguinus oedipus were subjected to an X-ray cinematographic study on a modified treadmill to unveil metric parameters of the locomotor pattern preferred on slender and compliant ("arboreal") substrates, the walk. Independent from the substrates, the postures of the two species differed in details, as do the preferred substrates. Horizontal, comfortable substrates are favored most. Walking ranked top in frequency, followed by jumping and galloping (in a strict sense). All other locomotor modes described for primates played a minor role or lacked entirely, like the trot. Average distance of leaps was only 0.60 m, landings were mainly on the same level as take-offs. In Saguinus, the movements of both limbs, including the shoulder blade, followed the pattern common to small mammals in general: At the end of the stance phase, humerus and tibia are nearly parallel to the substrate, while just before touchdown ulna and femur are in this position. The walk in both species was surprisingly fast (1 m/s), reaching the speed of much larger cursorial animals, like humans.

Three-dimensional reconstruction of the anal striated musculature in a human fetus.

Conflicting opinions in the recent literature indicate that the morphological organization and function of the anorectal continence organ has, up to now, not been clearly understood. But a clear imagination of the spatial arrangements of this compound muscle system is of clinical relevance for the pediatric surgeon performing reconstructive surgery. We analyzed 18-microm sections of the pelvic region of 4 human fetuses in order to describe the individual components of this muscle complex. A series of 630 Azan-stained sections was the base for the computer-assisted 3D reconstruction of the levator ani and the external sphincter complex in a male human fetus (14th week p.c.). In this context, special attention was paid to the intermediate muscle layer of the puborectalis which develops ventrally from the funnel-shaped levator ani and joins the tripartite ring system of the sphincter muscle dorsally. Our findings lead to a clear imagination of the spatial arrangement of this intermediate layer and characterize the anorectal muscle complex as an integrated ensemble in which the puborectalis holds a key position.

Spatiotemporal surface EMG characteristics from rat triceps brachii muscle during treadmill locomotion indicate selective recruitment of functionally distinct muscle regions.

Multichannel surface EMG recordings of a multiheaded skeletal muscle during cyclic locomotion combined with cineradiography were analysed in a chronic experiment. The resulting detailed two-dimensional activation pattern from the long and lateral triceps brachii heads of the rat during treadmill locomotion were combined with gait characteristics and fibre typing of the muscle. Shortly before ground contact of the forelimb, maximum muscle activity was found in the proximal part of the long head of the muscle. During the stance phase maximum activity was observed in the proximal part of the lateral head. The frequency dependent behaviour of cross-covariance functions over both muscle heads confirmed this selective shift in activation. In the lateral triceps brachii head of the investigated rats, exclusively type II fibres were found. In the long head the frequency of type I fibres was the highest in the deep muscle layers, proximally more than distally, whereas type II fibres were dominant in more superficial muscle layers. A combination of physiological and histological findings supports an anticipating mechanism whereby fine-tuning of the vertical foot down manoeuvre is mainly achieved by the (type I fibre dominated) proximal deep compartment of the biarticular long triceps brachii head and force generation is predominantly executed by the monoarticular lateral triceps brachii head.

Surface EMG-recordings using a miniaturised matrix electrode: a new technique for small animals.

A new method for multichannel surface-EMG measurements in small animals is presented. The underlying scientific aim is the characterisation of the spreading and the co-ordination of skeletal muscle activation between different muscles or muscle parts, depending on various motor tasks. The myoelectrical signals were recorded monopolarly by a 16-channel matrix electrode on the muscle surface directly under the skin on the fascia of the investigated muscle, without damaging the muscle. Surface-EMG's were recorded for at least 5 days after surgery without electrical interferences. During defined motor tasks, the projection of the myoelectrical activation of the different parts of the M. triceps brachii of rats (Rattus norvegicus), pikas (Ochotona rufescens) and cuis (Galea musteloides) or the M. anconeus of toads (Bufo marinus) on the muscle surface was mapped. The locomotion of the investigated animals was monitored by a three-dimensional kinematic analysis (video and/or high-speed cineradiography). There was no perceptible influence from application of EMG matrix electrode. The miniaturised matrix electrode seemed practicable in gaining insight into changes in myoelectrical activation patterns (EMG mapping). This allows a characterisation of the intramuscular co-ordination processes corresponding to the actual morphofunctional state of the investigated animals.

Accelerated contractile function and improved fatigue resistance of calf muscles in newborn piglets with IUGR.

Asymmetrical intrauterine growth restriction is denoted by disproportional reduction of muscle mass compared with body weight reduction. However, effects on contractile function or tissue development of skeletal muscles were not studied until now. Therefore, isometric force output of serial-stimulated hindlimb plantar flexors was measured in thiopental-anesthetized normal weight (NW) and intrauterine growth-restricted (IUGR) 1-day-old piglets under conditions of normal, reduced (aortic cross clamping), and reestablished (clamp release) blood supply (measured by colored microspheres technique). Furthermore, muscle fiber type distribution was determined after histochemical staining, specific muscle force of the plantar flexors [quotient from absolute force divided by muscle mass (N/g)] was calculated, and glycogen content and morphometric data of the investigated muscles were estimated. Regional blood flow of hindlimb muscles was similar in NW (6 +/- 2 ml. min(-1). 100 g(-1)) and IUGR piglets (8 +/- 1 ml. min(-1). 100 g(-1)). Isometric muscle contractions induced a marked increase in regional blood flow of 4.1-fold in NW and 5-fold in stimulated hindlimb muscles of IUGR piglets (baseline blood flow). Specific force of NW piglet muscles (5.2 +/- 0.2 N/g) was significantly lower than IUGR piglet muscles (6.1 +/- 0.6 N/g; P < 0.05). Isometric muscle contractions (NW: 32.7 +/- 4.7 N; IUGR: 21.7 +/- 4.0 N) resulted in a higher rate of force decrease in the calf muscles of NW animals compared with IUGR piglets (8 +/- 2 vs. 3 +/- 1%; P < 0. 01). Functional restoration of contractile performance after hindlimb recirculation was nearly complete in IUGR piglets (98 +/- 1%), whereas in NW piglets a deficit of 9 +/- 3% was found (P < 0. 01). Muscle fiber type estimation revealed an increased proportion of type I fibers in flexor digitalis superficialis and gastrocnemius medialis in IUGR piglets (P < 0.05). These data clearly indicate that contractile function is accelerated in newborn IUGR piglets.

Cineradiographic study of forelimb movements during quadrupedal walking in the brown lemur (Eulemur fulvus, Primates: Lemuridae).

Movements of forelimb joints and segments during walking in the brown lemur (Eulemur fulvus) were analyzed using cineradiography (150 frames/sec). Metric gait parameters, forelimb kinematics, and intralimb coordination are described. Calculation of contribution of segment displacements to stance propulsion shows that scapular retroversion in a fulcrum near the vertebral border causes more than 60% of propulsion. The contribution by the shoulder joint is 30%, elbow joint 5%, and wrist joint 1%. Correlation analysis was applied to reveal the interdependency between metric and kinematic parameters. Only the effective angular movement of the elbow joint during stance is speed-dependent. Movements of all other forelimb joints and segments are independent of speed and influence, mainly, linear gait parameters (stride length, stance length). Perhaps the most important result is the hitherto unknown and unexpected degree of scapular mobility. Scapular movements consist of ante-/retroversion, adduction/abduction, and scapular rotation about the longitudinal axis. Inside rotation of the scapula (60 degrees -70 degrees ), together with flexion in the shoulder joint, mediates abduction of the humerus, which is not achieved in the shoulder joint, and is therefore strikingly different from humeral abduction in man. Movements of the shoulder joint are restricted to flexion and extension. At touch down, the shoulder joint of the brown lemur is more extended compared to that of other small mammals. The relatively long humerus and forearm, characteristic for primates, are thus effectively converted into stride length. Observed asymmetries in metric and kinematic behavior of the left and right forelimb are caused by an unequal lateral bending of the spinal column.

Three-dimensional reconstruction of histological sections using modern product-design software.

BACKGROUND: Computer-based, three-dimensional reconstruction of histological sections is necessary for solving a diversity of questions in morphology and anatomy. Programming software for this purpose is difficult and time-consuming. Therefore, we tested if commercially available product-design software is useful for reconstructing anatomical virtual models. METHODS: We used Alias Wavefront Studio software on a Silicon Graphics workstation. Sections were projected with a newly developed microslide projector on a digitizing board and digitized interactively. Alternatively, pictures of sections were digitized on screen. The resulting set of contours was used as scaffold around which a surface was created manually. RESULTS: Repeated creation of the same object and subsequent measurements gave results that will be sufficiently reliable for many purposes. The visual presentation of objects satisfy highest demands. Little time is needed to reconstruct simple objects, and the time used for complex virtual models is acceptable. Manipulation of objects on screen is nearly in real time and rendering speed is high. CONCLUSIONS: Product-design software packages are a readily available and useful option for anatomists who want to do virtual reconstructions quickly without programming software themselves.

Professional practice model research collaboration. Issues in longitudinal, multisite designs.

Implementing and evaluating professional practice models in dynamic healthcare settings is difficult and poses unique problems for nurse researchers and administrators. The authors describe issues associated with collaborating during the conduct of longitudinal studies.

[The ontogenesis of the tympanic region of the Procaviidae (Mammalia: Hyracoidea)]. / Zur Ontogenese der Tympanalregion der Procaviidae (Mammalia: Hyracoidea).

UNLABELLED: Lindahl (1948) has described the early ontogenesis of 14 fetal stages of Procavia capensis. The oldest fetus of his series has had a headlength of 20 mm. It is the series with which the present account on the development of the tympanic region in hyracoids starts. Further middle to late fetuses have been studied by means of serial sections or macroscopic preparations. First 5 different ontogenetic stages are described, and then we follow the development of different organs in their ontogenesis. RESULTS: 1. The development of the tympanic region is almost completed at birth, except for the external meatus. The bulla tympani is fully ossified. 2. The entotympanic forms the medial and rostral part of the Bulla tympani. Its anlage is first met at a headlength of 31 mm. From the very beginning, it is fused with the tubal cartilage. Hyracoids do not have a caudal entotympanic. The cartilaginous mass-taken for a caudal entotympanic by Van der Klaauw (1926)-is actually a tympanic process of the petrosal. Extensive discussion reveals that the structures named entotympanics in some eutherians are most likely not homologous. 3. It is almost impossible to separate the entotympanic from the tympanic in neonates already. This is due to the surprising fact that the tympanic produces secondary cartilage at the suture between the 2 elements. 4. A gonial is lacking in hyracoids; instead the malleus is fixed in a peculiar way at the tympanic. 5. According to Lindahl (1948), the stapedial artery exists till a fetal headlength of 14 mm. Then the artery obliterates but the crura stapedis with the stapedial foramen persist. 6. Hyracoids possess a tympanohyal which changes its relation to the facial nerve during ontogenesis. 7. The development of the Eustachian sac (Diverticulum tubae auditivae) is described and compared with its development in the horse, based on the few data available in literature. 8. The internal carotid artery changes its course during its ontogenesis. Up to late stages it passes over the caudal part of the Eustachian sac and along the medial side of the Bulla tympani. A sulcus caroticus might still be seen in neonates. It is only in postnatal development that it looses the course along the bulla and passes along the mediocaudal side of the Eustachian sac. 9. The formation of the Recessus meatus acustici externi in exactly takes place the way described only once by Hammar (1902). The cavitation of the recessus occurs independently of the cavity of the external meatus.

Constructional morphology: the analysis of constraints in evolution dedicated to A. Seilacher in honour of his 60. birthday.

Evolutionary change is opportunistic, but its course is strongly constrained in several fundamental ways. These constraints (historical/phylogenetic, functional/adaptive, constructional/morphogenetic) and their dynamic relationships are discussed here and shown to constitute the conceptual framework of Constructional Morphology. Notwithstanding recent published opinions which claim that the "discovery" of constraints renders Neodarwinian selection theory obsolete, we regard the insights of Constructional Morphology as being entirely consistent with this theory. As is shown here in the case of the Hyracoidea, formal analysis of the constraints which have framed the evolution of various characters extends our understanding of the evolution of a taxon.

Nuclear ribonucleoprotein complexes of amphibian liver. I. Characterization of the complex and its small molecular weight RNA moiety.

Nuclear RNA-protein complexes containing small molecular weight RNAs were isolated from hepatic nuclei of Rana catesbeiana tadpoles and frogs according to a procedure normally used for the isolation of heterogeneous nuclear ribonucleoprotein complexes from other eukaryotic tissues. Preliminary characterization of the tadpole nuclear RNP indicated a particle size of 50--70 S in sucrose density gradients and a buoyant density of 1.40 gm/ml in CsCl gradients. When analyzed on SDS-polyacrylamide gels, this complex was observed to contain at least 40 polypeptides ranging in molecular weight from 15,000 to 200,000. Nuclear RNA-protein complexes were also isolated from adult frog hepatic nuclei by the same protocol and the RNA moiety which had been purified from the frog complex was compared with the nuclear RNA isolated from the tadpole particles. Electrophoretic analysis of the nuclear RNA-protein-associated RNA revealed minor qualitative and quantitive differences in the more than 25 discrete bands (4--9 S) associated with each particle. Base analysis of tadpole and frog nuclear RNA revealed a nucleotide composition of approximately 50% adenosine plus uridine nucleotides, with an unusually high content of cytosine residues (approximately 30%). Comparison of the two RNA samples demonstrated a large increase in the adenosine content of frog unclear RNA, and the presence of a minor base in frog nuclear RNA which was absent in the tadpole sample. These results indicated that changes in the RNA content of the amphibian nuclear RNP complex had occurred during bullfrog development.