Morphological correlates of aquatic and terrestrial locomotion in a semi-aquatic frog, Rana esculenta: no evidence for a design conflict

Semi-aquatic frogs are faced with an unusual locomotory challenge. They have to swim and jump using the same apparatus, i.e. the hind limbs. Optimization of two tasks that require mutually incompatible morphologies or physiologies cannot occur simultaneously. In such cases, natural selection will result in some compromise, i.e. an intermediate phenotype that can perform both tasks reasonably well, but its performance will never match that of a specialized phenotype. We found no direct evidence for a trade-off between jumping and swimming performance nor for a coupled optimization. This could be due to the importance of overall quality, as suggested by the fact that some frogs possess greater overall muscularity than others, irrespective of their body size. Another explanation could be that some morphological characteristics have a positive effect on both locomotor modes and others show a trade-off effect. The net effect of these characteristics could result in an overall absence of correlation between the two locomotor performances. Size has a great influence on the morphological data and on jumping performance, but not if performance is expressed as velocity. The body shape of an anuran is conservative and scales mostly isometrically.     

Survival of Hoxa13 homozygous mutants reveals a novel role in digit patterning and appendicular skeletal development

The loss of HOXA13 function severely disrupts embryonic limb development. However, because embryos lacking HOXA13 die by mid‐gestation, the defects present in the mutant limb could arise as a secondary consequence of failing embryonic health. In our analysis of the mutant Hoxa13^GFP allele, we identified a surviving cohort of homozygous mutants exhibiting severe limb defects including: missing phalanx elements, fusions of the carpal/tarsal elements, and significant reductions in metacarpal/metatarsal length. Characterization of prochondrogenic genes in the affected carpal/tarsal regions revealed significant reduction in Gdf5 expression, whereas Bmp2 expression was significantly elevated. Analysis of Gdf5 mRNA localization also revealed diffuse expression in the carpal/tarsal anlagen, suggesting a role for HOXA13 in the organization of the cells necessary to delineate individual carpal/tarsal elements. Together these results identify Gdf5 as a potential target gene of HOXA13 target gene and confirm a specific role for HOXA13 during appendicular skeletal development. Developmental Dynamics 239:446–457, 2010. © 2009 Wiley‐Liss, Inc.     

Choline acetyltransferase immunoreactive cortical interneurons do not occur in all rodents: a study of the phylogenetic occurrence of this neural characteristic

The present study was designed to provide results aimed at testing whether the interneurons with choline acetyltransferase immunoreactivity (ChAT), probably representing GABA interneurons, found in the cerebral cortex of the rat represent a common feature of the order Rodentia. Initially we verified the presence of ChAT immunoreactive bipolar cell bodies, axons and terminal-like fibres in pigmented (Long-Evans) and non-pigmented (Sprague-Dawley) strains of Rattus norvegicus, confirming that the ChAT polyclonal antibodies (AB144P and AB143, Chemicon; VChAT, Sigma) with the immunohistochemical techniques used provided the same staining as previously described for this species. We then examined pigmented (AKR3) and non-pigmented (C3H) strains of Mus musculus, wild caught striped mice (Rhabdomys pumilio), bushveld gerbil (Tatera brantsii), greater canerat (Thryonomys swinderianus) and common molerat (Cryptomys hottentotus). The AB144P antibody revealed cortical interneurons in both strains of M. musculus and in R. pumilio, but not in the other species. In all species/strains cortical ChAT immunoreactive axons and terminal-like fibres were localized with the AB144P antibody. In the non-Rattus species/strains there was no evidence for localization of ChAT immunoreactivity in any cortical cell bodies using the AB143 and vesicular acetylcholine transporter (VChAT) antibodies despite extensive localization in axons and terminal-like fibres. It is concluded that bipolar cortical GABA interneurons in certain rodent species may develop ChAT immunoreactivity but not VChAT immunoreactivity making the cholinergic relevance of ChAT in the GABA interneurons uncertain and may exclude these neurons from being part of the traditionally defined cholinergic system.     

Detection of variation in the ribosomal RNA gene clusters by a modified fluorescencein situ hybridization method

Summary Physical mapping of genes by fluorescencein situ hybridization (FISH) has become routine using fluorescein isothiocyanate (FITC) for probe detection and propidium iodide (PI) for chromosome staining. We have modified this conventional FISH method in a way that utilizes Texas red (TR) for signal detection and quinacrine mustard (QM) for chromosome banding. Using this Texas red and quinacrine (TRQ) method, we were able to identify individual acrocentric chromosomes with varying degrees of ribosomal RNA gene clusters. Two acrocentric chromosomes were found to carry extremely small number of rRNA gene copies as compared to the other eight counterparts in human diploid lymphoblastoid cell line GM00130B. Thus, the TRQ method allows one to probe for a specific sequence while identifying individual chromosomes and will be powerful for the chromosomal localization of various genes.     

The adaptation of a reflex response to the ongoing phase of locomotion in fish

Summary The reflex response to stimulation of the tail fin has been studied in the swimming fish, by bilateral electromyographical (EMG) recordings in several segments along the body. The response varies with the phase of swimming. When the muscles on one side (segment) are active, a large response will occur on this side but no response on the contralateral side at the same level. When the other side becomes active an identical stimulus will cause an activation of this side but no response on the previously active side. When the movements were filmed a powerful mechanical effect was demonstrated with an augmentation of the ongoing movement, that would result in an instantaneous increase in speed. The stimulus causes in addition a shortening of the duration of the swimming cycle and its components. Most of the results were obtained on spinal dogfish, which also exhibits spontaneous locomotion after a spinal transection. Mainly electrical bipolar stimulation of the tail fin was used. Identical stimuli applied in different phases on an ongoing movement, thus give a reflex response that changes dramatically with the phase of the movement. This phase dependent reflex reversal is functionally meaningful; it is fast and due to spinal mechanisms.     

Incorporating BMP-2 and skeletal muscle to a semitendinosus autograft in an oversized tunnel yields robust bone tunnel ossification in rabbits: Toward single-step revision of failed anterior cruciate ligament reconstruction

Background

Bone tunnel widening after anterior cruciate ligament (ACL) reconstruction is a known complication that can lead to graft failure. Subsequent revision surgery typically involves a two-stage procedure. The aim of this study was to test a novel autologous tendon graft retaining muscle tissue combined with Human Recombinant Bone Morphogenetic Protein-2 (rh-BMP-2) leading to rapid ossification of the muscle tissue, simultaneously replenishing bone stock and producing a mechanically stable bone–tendon insertion.

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.     

Autoregulatory reactions of the skeletal muscle arterioles to a decrease in arterial pressure

Internal radius, wall thickness, and blood flow rate are measured in ratm. cremaster arterioles of different orders of branching under normal and stepwise decreased arterial pressure. Two types of reactions to a 20% decrease in arterial pressure are observed: active autoregulatory reaction (42% vessels) and passive reaction (58%). It is shown that relative changes in the diameter of arterioles in autoregulatory and passive reactions depend on the relative thickness of arteriolar wall and becomes more pronounced as this parameter increases. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 124, No. 9, pp. 267–270, September, 1997     

Poneratoxin,a new toxin from an ant venom,reveals an interconversion between two gating modes of the Na channels in frog skeletal muscle fibres

The effects of synthetic poneratoxin (PoTX), a new toxin isolated from the venom of the ant Paraponera clavata, were studied under current- and voltage-clamp conditions in frog skeletal muscle fibres. PoTX induces a concentration-dependent (10^–9 M–5×10^–6 M) prolongation of the action potentials and, at saturating concentration, a slow repetitive activity developing at negative potentials. PoTX specifically acts on voltage-dependent Na channels by decreasing the peak Na current (I _Na) and by simultaneously inducing a slow I _Na which starts to activate at –85 mV and inactivates very slowly. Both the fast and the slow components of I _Na are suppressed by tetrodotoxin and reverse at the same potential corresponding to the equilibrium potential for Na ions. The fast component of I _Na has voltage dependence, activation and steady-state inactivation almost similar to those of the control I _Na. The voltage dependence of the slow Na conductance is 40 mV more negative than that of the fast one. The results suggest that PoTX affects all the Na channels and that the fast and the slow I _Na components originate from a possible PoTX-induced interconversion between a fast and a slow operating mode of the Na channels.     

Immunity to type IX collagen in rodents: a study of type IX collagen for autoimmune and arthritogenic activities

Type IX collagen (CIX), a cartilage-specific glycoprotein, constitutes ≤ 10% of cartilage collagen. To ascertain whether CIX can induce arthritis as shown for type II and XI collagen (CII and CXI), outbred rats were sensitized with bovine, chick and human CIX; inbred rats, mice, and guinea pigs were sensitized with bovine CIX. Mice and guinea pigs proved resistant to arthritis, as did rats sensitized with CIX/Freund's incomplete adjuvant (FIA). Arthritis was seen in rats when 100 μg of Mycobacterium tuberculosis (Mtb) were added to FIA, but seldom with smaller doses of Mtb, suggesting the arthritis was adjuvant-induced. High levels of antibodies to rat CIX, containing complement-fixing subclasses, were detected in rat sera in addition to DTH and lymphocyte proliferation responses to rat CIX. Given the potential for CIX-induced disease, CIX-sensitized rats were injected intraperitoneally with lipopolysaccharide (LPS) to stimulate proinflammatory cytokine release, and intra-articularly with rat CIX to stimulate arthritis. LPS stimulation was ineffective; however, intra-articularly injected CIX produced transient synovitis. When rats with stable adjuvant arthritis were sensitized with CIX/FIA, significant increases in paw volume were measured compared with controls given CI/FIA. Immunohistochemical studies of actively and passively sensitized rats revealed deposits of CIX antibody, but not C3, at the joint margins where proteoglycan staining was weak. Together, these findings suggest that autoimmunity to CIX, in contrast to CII and CXI, is not directly pathogenic but may contribute to joint injury provided arthritis is initiated by an independent disease process.     

Mapping of an autosomal dominant gene for Dupuytren's contracture to chromosome 16q in a Swedish family

Mapping of an autosomal dominant gene for Dupuytren's contracture to chromosome 16q in a Swedish family.Dupuytren's contracture (DC) (OMIM 126900) is the most common connective tissue disease of mankind and has both heritable and sporadic forms. The inherited form is most frequently observed among the xanthochroi peoples of Northern Europe where its most common manifestations are thickening of the palmar fascia and contracture of the fingers. We ascertained a five-generation Swedish family in which DC is inherited in an autosomal dominant manner with high, but incomplete, penetrance by the end of the fifth decade. Blood was collected from all affected and informative unaffected family members for the performance of a genome-wide scan at a resolution of approximately 8 cM for all autosomes. Linkage was established to a single 6 cM region between markers D16S419 and D16S3032 on chromosome 16. A maximal two-point logarithm of odds (LOD) score of 3.18 was achieved at microsatellite marker D16S415 with four other markers in the region producing LODs of >1.5.     

Heterogeneity in the occurrence of a subset of autoantibodies to glutamic acid decarboxylase in autoimmune polyendocrine patients with islet cell antibodies

Glutamic acid decarboxylase-65 (GAD-65) is a major target for autoantibodies and autoreactive T cells in patients with insulin-dependent diabetes mellitus (IDDM). Autoantibodies to GAD are also found in patients with stiff man syndrome (SMS) or polyendocrine autoimmunity (PE). The epitope specificities of autoantibodies to GAD in IDDM and SMS have been well documented, but the locations of autoantibody epitopes of GAD in PE patients have not been mapped. Thus, the properties of anti-GAD antibodies in PE patients (with or without diabetes) were investigated. The ability of PE serum antibodies to inhibit the binding of the mouse monoclonal antibody, GAD-6, to native GAD in ELISA was determined. For PE patients without diabetes, levels of inhibition of GAD-6 binding ranged from 0% to almost 70% and were unrelated to the level of binding of serum antibodies to GAD (P = 0.351) or to the functional affinities of these antibodies. This suggests differences in the epitope specificities of anti-GAD antibodies in different patients. Levels of inhibition were also unrelated to clinical condition. SMS antibodies showed similar levels of inhibition of GAD-6 binding. Similar analysis was applied to PE patients with diabetes and levels of inhibition of GAD-6 binding to GAD were determined. These ranged from 0% to 80%, and levels of inhibition were similar in samples taken before or after diabetes onset. There was no significant difference between anti-GAD antibodies from PE patients with or without diabetes in the range of abilities to inhibit GAD-6 binding to GAD, although the highest levels of inhibition were given by sera from non-diabetic patients. This raises the possibility of differential expression of subsets of anti-GAD antibodies in progressive versus slow or non-progressive anti-islet autoimmune responses. Serum antibodies of PE and SMS patients did not inhibit the binding of antibodies specific for the extreme C-terminus of GAD, indicating that this is not the site of the epitopes for the patients' antibodies or for GAD-6.     

Pathomorphological signs of damage to capillary endothelium in skeletal muscle after traumatic injury to or replantation of a limb

Several months after an extensive crush injury to or replantation of a limb in rats, capillary endotheliocytes of its skeletal muscle are seen to undergo ultrastructural changes of both “dark” and “light” types. Destructive/degenerative changes of the “light” type are accompanied by intracellular edema and cytoplasmic homogenization and can eventually result in monocellular colliquative necrosis. Changes of the “dark” type are characterized by signs of functional overstrain in the endotheliocytes, succeeded by destructive/degenerative cytoplasmic changes, increased electron density of intracellular organelles, loss of distinct boundaries by the nucleus and intracellular structures, rupture of cell membranes, endothelial desquamation, and some other changes, which eventually lead to coagulation necrosis, followed by breakdown of the cell into small fragments. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 118, N^no 10, pp. 378–383, October, 1994     

Detection of distant metastasis to skeletal muscle by 18F-FDG-PET in a case of intrahepatic cholangiocarcinoma

Intrahepatic cholangiocarcinoma is a rare malignancy that originates from the epithelial cells of the intrahepatic bile ducts. Intrahepatic cholangiocarcinoma can metastasize in lymphatic chains, including the hepatoduodenal ligament, and it often invades adjacent organs or metastasizes to other visceral organs such as the lungs, bones, adrenal glands, and brain. However, distant skeletal muscle metastasis is very rare. Moreover, a metastatic skeletal muscle tumor rarely shows specific symptoms, making it difficult to identify in a routine examination. A 45-year-old man with a chief complaint of right upper quadrant abdominal pain was admitted to our hospital. Abdominal ultrasound and computed tomography with contrast enhancement showed a malignant mass in the right hepatic lobe, and 2-[¹⁸F] fluoro-2-deoxy-D-glucose positron-emission tomography revealed distant skeletal muscle metastases in the thorax and buttock. The patient underwent an ultrasound-guided percutaneous needle biopsy for the metastatic low-echo masses in the skeletal muscle.     

Suggestive linkage of ADHD to chromosome 18q22 in a young genetically isolated Dutch population

Attention deficit/hyperactivity disorder (ADHD) is a common, highly heritable, neuropsychiatric disorder among children. Linkage studies in isolated populations have proved powerful to detect variants for complex diseases, such as ADHD. We performed a genome-wide linkage scan for ADHD in nine patients from a genetically isolated population in the Netherlands, who were linked to each other within 10 generations through multiple lines of descent. The genome-wide scan was performed with a set of 400 microsatellite markers with an average spacing of ±10–12 cM. We performed multipoint parametric linkage analyses using both recessive and dominant models. Our genome scan pointed to several chromosomal regions that may harbour ADHD susceptibility genes. None exceeded the empirical genome-wide significance threshold, but the Log of odds (LOD) scores were >1.5 for regions 6p22 (Heterogenetic log of odds (HLOD)=1.67) and 18q21–22 (HLOD=2.13) under a recessive model. We followed up these two regions in a larger sample of ADHD patients (n=21, 9 initial and 12 extra patients). The LOD scores did not increase after increasing the sample size (6p22 (HLOD=1.51), 18q21–22 (HLOD=1.83)). However, the LOD score on 6p22 increased to 2 when a separate analysis was performed for the inattentive type ADHD children. The linkage region on chromosome 18q overlaps with the findings of association of rs2311120 (P=10⁻⁵) and rs4149601 (P=10⁻⁴) in the genome-wide association analysis for ADHD performed by the Genetic Association Information Network consortium. Furthermore, there was an excess of regions harbouring serotonin receptors (HTR1B, HTR1E, HTR4, HTR1D, and HTR6) that showed a LOD score >1 in our genome-wide scan.     

Arteriolar reactivity of rat skeletal muscle to noradrenalin after whole-body gamma-irradiation in a dose of 1 Gy

Institute of Physicotechnical Problems, Moscow. Moscow Roentgeno-Radiologic Research Institute. (Presented by Academician of the Russian Academy of Medical Sciences A. D. Ado.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 114, No. 10, pp. 355–357, October, 1992.     

Flattened cortical maps of cerebral function in the rat: a region-of-interest approach to data sampling, analysis and display

We describe a method for the measurement, analysis and display of cerebral cortical data obtained from coronal brain sections of the adult rat. In this method, regions-of-interest (ROI) are selected in the cortical mantle in a semiautomated fashion using a radial grid overlay, spaced in 15 degrees intervals from the midline. ROI measurements of intensity are mapped on a flattened two-dimensional surface. Topographic maps of statistical significance at each ROI allow for the rapid viewing of group differences. Cortical z-scores are displayed with the boundaries of brain regions defined according to a standard atlas of the rat brain. This method and accompanying software implementation (Matlab, Labview) allow for compact data display in a variety of autoradiographic and histologic studies of the structure and function of the rat brain.     

Murine tyrosine hydroxylase maps to the distal end of chromosome 7 within a region conserved in mouse and man

The locus of the structural gene encoding tyrosine hydroxylase, Th, the rate limiting enzyme for catecholamine biosynthesis, was mapped to the distal end of mouse Chromosome (Chr) 7. A DNA probe of genomic origin of rat tyrosine hydroxylase was used to detect restriction fragment length variants among 8 inbred mouse strains. The strain distribution pattern of 77i allelic variants in 3 sets of recombinant inbred mouse strains was determined. Comparison of the strain distribution patterns of Th alleles with those of previously typed loci suggested Th was located on Chr 7. The Chr 7 assignment for Th was confirmed by analyzing 108 mice produced from an (NZB × SM)F1 × NZB backcross. Moreover, the Th locus was positioned distally on Chr 7. Mouse Chr 7 and human Chr 1 lp (the location of the human tyrosine hydroxylase gene) are known to share several homologous loci. With the addition of Th, the homology between the distal 2/3 of mouse Chr 7 and human Chr lip appears extensive.