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Analysis of the developing Xenopus tail bud reveals separate phases of gene expression during determination and outgrowth.

We have studied Xenopus tail development from the end of gastrulation to the commencement of outgrowth at the tail bud stage. We show that an early group of genes are expressed at the stage of tail bud determination, at the end of gastrulation, and a late group are expressed at around stage 27 just before tail bud outgrowth. Together, these genes define seven distinct regions of the tail bud as outgrowth commences. We have previously shown that formation of a tail bud depends on the interaction of three tissue regions, called N, M and C, at stage 13. Here we show that expression of the late group of genes is dependent on this NMC interaction. We describe molecular correlates of two of these regions, M and C, which were formerly unobservable and whose existence was inferred from embryological experiments.

Animals↗

Morphological diversity of dying cells during regression of the human tail.

During normal human development a number of transient structures form and subsequently regress completely. One of the most prominent structures that regress during development is the human tail. We report here a histological and ultrastructural study of cell death in the cranial and caudal (tail) parts of the neural tube in 4 to 6-week-old human embryos. Initially, the human tail is composed of tail bud mesenchyme which differentiates into caudal somites, secondary neural tube, notochord and tail gut. Later on, these structures gradually regress by cell death. During the investigated period, we observed two morphologically distinct types of dying cells. The well-described apoptotic type of cell death was observed only in the cranial neural tube that forms during primary neurulation. The other type of cell death characterized by necrotic morphology was observed in the tail mesenchyme and in the caudal neural tube that forms during secondary neurulation. This morphological diversity suggests that besides differences in origin and fate there are different mechanisms of developmental cell death between two parts of the human neural tube. We can speculate that the apoptotic type of cell death is associated with the precise control of cell numbers and that the other morphologically distinct type of cell death is responsible for the massive removal of transitory structures.

Abortion, Induced↗

Cyclosporin A inhibits thyroid hormone-induced shortening of the tadpole tail through membrane permeability transition.

Regression of the tadpole tail through muscule cell apoptosis is one of the most spectacular events in amphibian metamorphosis. Accumulated evidence has shown that mitochondrial membrane permeability transition (MPT) plays a crucial role in apoptosis. Previously we reported that cyclosporin A (CsA) suppressed 3,5,3'-triiodothyronine (T(3))-induced mitochondrial swelling, which was coupled with cytochrome c (Cyt.c) release through MPT [Comp. Biochem. Phys. 130 (2001) 411-418]. To further clarify the mechanism of tadpole metamorphosis, the present study investigates the effect of CsA on T(3) induced tadpole tail shortening. A low concentration of T(3) (5 x 10(-8) M) was found to induce a shortening of stage X Rana rugosa tadpole tails, accompanied by an increase in caspase-3- and -9 like protease activity, as well as an increase in DNA-fragmentation and ladder formation, while CsA was seen to suppress the effects of T(3). The stage X tadpole tail was found to express Bax mRNA and this expression was not affected by T(3) treatment. CsA, on the other hand, proved to have a slightly supressive effection on Bax expression. 20 microM T(3) as well as 50 microM Ca(2+) induced swelling in mitochondria isolated from the liver of R. rugosa resulting in the release of apoptosis related substances, and the released fraction activated cytosolic caspase-3 and -9 in the presence of dATP. This result indicated that Cyt.c might be released from mitochondria by treatment with T(3) through both direct and indirect action of T(3). From these results and other data it was concluded that mitochondrial MPT plays an important role in T(3)-induced apoptosis in the tadpole tail, resulting in tail shortening, and CsA was seen to suppress the effects of T(3).

Amino Acid Sequence↗

Endothelin-1-induced contraction is impaired in the tail artery of renal hypertensive rats.

The contraction induced by endothelin-1 (ET-1) was evaluated in tail arteries from normotensive two-kidney (2K) and hypertensive two-kidney-one-clip (2K-1C) rats. Since the maximal effect induced by ET-1 (0.1-30 or 100 nmol/l) was lower in 2K-1C (1.11 +/- 0.10 g) than in 2K (1.46 +/- 0.14 g) tail arteries, we evaluated the possible mechanisms involved in this blunted response. The sensitivity and efficacy of ET-1 were not affected by endothelium removal in either group. ET-1 failed to induce contraction of 2K and 2K-1C arteries in Ca(2+)-free medium. The contractile response induced by 10 nmol/l ET-1 was similarly inhibited by 0.1 microM nifedipine in arteries from 2K (81.6 +/- 3.3%) and 2K-1C (81.3 +/- 3.8%) rats. The effect of nifedipine was not potentiated by 10 mumol/l SK&F 96365. The cytosolic Ca2+ concentration ([Ca2+]c) was similarly increased by 30 nmol/l ET-1 in smooth muscle cells isolated from tail arteries of 2K (30.80 +/- 11.94 nmol/l) and 2K-1C (54.06 +/- 10.98 nmol/l) rats. In conclusion, the blunted contraction induced by ET-1 in 2K-1C tail arteries was not dependent on the endothelium or on decreased Ca2+ influx through channels sensitive to nifedipine or SK&F 96365. Since the increase of [Ca2+]c upon stimulation with ET-1 was similar in 2K and 2K-1C tail artery cells, probably the sensitivity to Ca2+ is decreased in 2K-1C tail arteries.

Animals↗

The molecular nature of the zebrafish tail organizer.

Based on grafting experiments, Mangold and Spemann showed the dorsal blastopore lip of an amphibian gastrula to be able to induce a secondary body axis. The equivalent of this organizer region has been identified in different vertebrates including teleosts. However, whereas the graft can induce ectopic head and trunk, endogenous and ectopic axes fuse in the posterior part of the body, raising the question of whether a distinct organizer region is necessary for tail development. Here we reveal, by isochronic and heterochronic transplantation, the existence of a tail organizer deriving from the ventral margin of the zebrafish embryo, which is independent of the dorsal Spemann organizer. Loss-of-function experiments reveal that bone morphogenetic protein (BMP), Nodal and Wnt8 signalling pathways are required for tail development. Moreover, stimulation of naive cells by a combination of BMP, Nodal and Wnt8 mimics the tail-organizing activity of the ventral margin and induces surrounding tissues to become tail. In contrast to induction of the vertebrate head, known to result from the triple inhibition of BMP, Nodal and Wnt, here we show that induction of the tail results from the triple stimulation of BMP, Nodal and Wnt8 signalling pathways.

Animals↗

Comparison of telemetric and tail-cuff blood pressure monitoring in adrenocorticotrophic hormone-treated rats.

1. The aim of the present study was to validate a telemetric blood pressure (BP) monitoring system against tail-cuff blood pressure in both adrenocorticotrophic hormone (ACTH)- and sham-treated rats. In the statistical analyses, we first tested whether there was a detectable effect on systolic blood pressure (SBP) of 10 days treatment with ACTH compared with saline. Second, we compared results of telemetered and tail-cuff measurements and, third, we developed a novel method for estimating the relative power of the two techniques. 2. Twenty-three male Sprague-Dawley rats were randomly divided into two groups: (i) ACTH (100 microg/kg per day, s.c; n = 12) treated; or (ii) sham treated (0.9% saline, s.c; n = 11). Systolic BP was measured by the telemetric system (sampled for 10 s every 2 min) continuously for 4 h (n = 16) or for 30 min (n = 23) and also by the indirect tail-cuff method daily (n = 23). Data were compared within and between groups; ordinary least products (OLP) regression analysis was then performed to test for bias between the two methods. Sample size/power estimations were also performed. 3. Adrenocorticotrophic hormone treatment raised telemetered SBP by 11 mmHg (P < 0.001) compared with 14 mmHg (P < 0.001) using the tail-cuff method. There was no fixed or proportional bias between the two methods of measurement, as shown by regression analysis. Power calculations indicate that a minimum sample size of six gives a power of telemetered to tail-cuff of 0.84/0.86 = 0.98. The power of 4 h versus 30 min BP measurements was 0.99/0.82 = 1.2. 4. Telemetry gave very similar results to the tail-cuff method. Telemetry allows for a longer period of measurement, giving greater power to the study so that fewer animals are needed.

Adrenocorticotropic Hormone↗

Transfer of the rat tail: an experimental model for free osteocutaneous transfer.

A new experimental model for free transfer of the rat tail has been developed. The vascular and nerve anatomy of the tail was studied in ten Wistar rats. The caudal artery (average diameter: 0.8 mm) provides the principal arterial supply to the tail. The main venous drainage is from bilateral dorsolateral subcutaneous veins (average diameter: 1.2 mm). The inferior caudal trunk establishes the cutaneous nerve branches to the skin of the tail. Free transfer of the tail to the carotid artery and external jugular vein, with the attachment of the tail to the dorsum, allows for clinical monitoring of the transfer, without risk of autophagia. This transfer is easy to perform, simple to monitor, and provides consistent yet challenging microsurgery for the beginning microvascular surgeon. In addition to its utility as a training model, it may also have an application as an easy research model for composite tissue transfer.

Anastomosis, Surgical↗

Bacteriophage P22 tail accessory factor GP26 is a long triple-stranded coiled-coil.

P22 is a well characterized tailed bacteriophage that infects Salmonella enterica serovar Typhimurium. It is characterized by a "short" tail, which is formed by five proteins: the dodecameric portal protein (gp1), three tail accessory factors (gp4, gp10, gp26), and six trimeric copies of the tail-spike protein (gp9). We have isolated the gene encoding tail accessory factor gp26, which is responsible for stabilization of viral DNA within the mature phage, and using a variety of biochemical and biophysical techniques we show that gp26 is very likely a triple stranded coiled-coil protein. Electron microscopic examination of purified gp26 indicates that the protein adopts a rod-like structure approximately 210 angstroms in length. This trimeric rod displays an exceedingly high intrinsic thermostability (T(m) approximately 85 degrees C), which suggests a potentially important structural role within the phage tail apparatus. We propose that gp26 forms the thin needle-like fiber emanating from the base of the P22 neck that has been observed by electron microscopy of negatively stained P22 virions. By analogy with viral trimeric coiled-coil class I membrane fusion proteins, gp26 may represent the membrane-penetrating device used by the phage to pierce the host outer membrane.

Amino Acid Sequence↗

Outer dense fibers serve as a functional target for Cdk5.p35 in the developing sperm tail.

Cdk5 is ubiquitously expressed in all tissues, but its activators, p35 and p39, are principally found in brain, and Cdk5 activity has mostly been associated with brain development, particularly neuronal differentiation and migration. Here we show that the p35 transcript and protein are also present in the testis, and an active Cdk5.p35 complex exists in this tissue as well. Cdk5 and p35 are prominently observed in elongating spermatid tails, particularly over the tail outer dense fibers (ODF). The appearance of Cdk5.p35 proceeds from the proximal to the distal end of elongating spermatids, coinciding with the proximal to distal assembly of ODF along the length of the tail axoneme. Incidentally, increased Cdk5.p35 activity is observed in isolated elongating spermatids and at a time when elongating spermatids appear in the developing testis, suggesting a role for Cdk5.p35 in spermiogenesis. The presence of Cdk5 and p35 in ODF isolated from rat sperm tails implies a strong association among these proteins. In vitro ODF phosphorylation by Cdk5.p35 and decreased in vivo sperm tail ODF phosphorylation in p35-deficient mice indicate that Cdk5.p35 is an integral component of ODF and that ODF is a functional Cdk5.p35 target in the testis. Our results demonstrate for the first time that Cdk5.p35 may participate in the regulation of sperm tail development via a mechanism involving ODF phosphorylation. Apparently, as in brain development, Cdk5.p35 plays a part in testis development.

Animals↗

Mathematical circulation model for the blood-flow-heat-loss relationship in the rat tail.

A mathematical model for the heat-loss-blood-flow relationship is developed for the rat tail. When supplied with experimental values of heat loss and blood flow, the model allows one to compute the distribution of flow in deep and cutaneous vessels as a function of body core and tail temperature and to determine the savings in heat loss that result from alterations in the pattern of circulation and from counter-current heat transfer. Blood flow in the cutaneous and deep lying veins of the tail is controlled by both central and local temperatures and increases fairly linearly with deep body temperature. However, the distribution of blood flow in the tail is controlled only by local tail temperature and is independent of deep body temperature. The change in venous distribution of flow has a great impact on the conservation of heat and can reduce the heat loss from the circulating blood by more than 50% when venous return is directed to deep lying veins. On the other hand, counter-current heat transfer is of only minor importance in the control of heat loss from the tail, resulting at most in a 10% saving of heat loss, and that only at the smallest rate of blood flow.

Animals↗

Genetic evidence for two t complex tail interaction (tct) loci in t haplotypes.

The t complex on chromosome 17 of the house mouse is an exceptional model for studying the genetic control of transmission ratio, gametogenesis, and embryogenesis. Partial haplotypes derived through rare recombination between a t haplotype and its wild-type homolog have been essential in the genetic analysis of these various properties of the t complex. A new partial t haplotype, which was derived from the complete tw71 haplotype and which is called tw71Jr1, was shown to have unexpected effects on tail length and unique recombination breakpoints. This haplotype, either when homozygous or when heterozygous with the progenitor tw71 haplotype, produced short-tailed rather than normal-tailed mice on certain genetic backgrounds. Genetic analysis of this exceptional haplotype showed that the recombination breakpoints were different from those leading to any other partial t haplotype. Based on this haplotype, a model is proposed that accounts for genetic interactions between the brachyury locus (T), the t complex tail interaction (tct) locus, and their wild-type homolog(s) that determine tail length. An important part of this model is the hypothesis that the tct locus, which enhances the tail-shortening effect of T mutations, is in fact at least two, genetically separable genes with different genetic activities. Genetic analysis of parental and recombinant haplotypes also suggests that intrachromosomal recombination involving an inverted duplicated segment can account for the variable orientation of loci within an inverted duplication on wild-type homologs of the t haplotype.

Animals↗

Outer dense fibre proteins from human sperm tail: molecular cloning and expression analyses of two cDNA transcripts encoding proteins of approximately 70 kDa.

The outer dense fibres (ODF) are a main cytoskeletal structure of the sperm tail. Despite their importance in the morphology and function of the sperm tail, their constituents are poorly described. Here we investigate the protein composition of human outer dense fibres. Our results suggest that human ODF consist of about 10 major and of at least 15 minor proteins, where all major proteins are ODF1, ODF2 or ODF2-related proteins. From a human testis cDNA library, we isolated two slightly different cDNAs encoding ODF2 proteins of approximately 70 kDa. Human ODF2 cDNAs and their encoded proteins are very similar to those isolated from rat and mouse pointing to a high evolutionary pressure residing on these proteins. Transcription of ODF2 is restricted to testis tissue and more specifically to round spermatids as was demonstrated by a non-radioactive in-situ hybridization. ODF2 proteins were detected in the sperm tail. Their distribution along the length of the sperm tail shows that the ODF normally extend to about half the principal piece of the sperm tail. The former result opens the possibility for a screening regarding the distribution of sperm tail proteins related to motility disorders.

Amino Acid Sequence↗

Apoptosis in Xenopus tadpole tail muscles involves Bax-dependent pathways.

Apoptosis is a fundamental mechanism implicated in normal development. One of the most spectacular developmental events involving apoptosis is tail regression during amphibian metamorphosis. We analyzed how thyroid hormone (3, 5, 3'-triiodothyronine, T3), the orchestrator of metamorphosis, affects expression and function of the proapoptotic gene Bax in the tail muscle of free-living Xenopus tadpoles. During natural metamorphosis Bax mRNA was expressed in tail muscles and was spatially correlated with apoptosis. Precocious treatment of tadpoles with T3 induced Bax expression and apoptosis. To verify that Bax expression was causally related to apoptosis we used a naked DNA gene transfer method to express Bax in the dorsal tail muscle. This induced apoptosis, and the process was exacerbated by T3 treatment. To determine whether T3 effects on Bax expression involved transcriptional regulation, we injected a Bax promoter sequence into dorsal and caudal tail muscles. In the dorsal muscle, T3 treatment did not affect transcription from the Bax promoter. However, in the caudal muscle, T3 treatment significantly increased Bax transcription. We conclude that T3-induced apoptosis in Xenopus tadpole tail muscles involves Bax-activating and Bax-synergis tic mechanisms. These programs are induced in spatially and temporally distinct manners.

Animals↗

Vane emargination of outer tail feathers improves flight manoeuvrability in streamerless hirundines, Hirundinidae.

Recent studies have suggested that the proximal part of the swallow (Hirundo rustica) tail streamer appears to aid turning flight, as expected if streamers evolved initially purely through natural selection for enhanced manoeuvrability. However, the evolution of slender aerodynamically advantageous streamers is also predicted by an alternative hypothesis, which suggests that such a trait could develop primarily to ameliorate the aerodynamic cost of a long size-dimorphic tail. To distinguish between these hypotheses, we have investigated for the effect on manoeuvrability of trimming the tips of the outer tail feathers into short streamers, without lengthening these feathers, in two streamerless hirundine species--the house martin (Delichon urbica) and the sand martin (Riparia riparia). This allowed us to examine the aerodynamic costs and benefits of streamers at an early evolutionary stage that predates elongation of the outermost tail feathers through female choice. We showed that such initial streamers enhance manoeuvrability in streamerless hirundines, confirming the findings of recent studies. However, in contrast to these studies, we showed that improved manoeuvrability resulting from streamers could arise before the outermost tail feathers have become elongated (e.g. owing to female choice). The occurrence of such an aerodynamic advantage depends on the ancestral shape of a forked tail. This provides support for the hypothesis that streamers, like those in the barn swallow, might evolve initially purely through natural selection for enhanced manoeuvrability.

Analysis of Variance↗

Temporal variation in cellular proliferation during recornification of mouse tail skin.

The influence of the time of injury on subsequent epidermal regeneration is unknown. Epidermal cell proliferation of tail skin in C57BL/6J mice in response to tape stripping was followed for 7 days by radiolabelled thymidine incorporation and autoradiography. The homeostatic labelling index (LI) of the basal epidermis of unmanipulated, unwounded (control) animals was 7.6% and did not vary depending on the time of day. Tape stripping increased the LI of epidermal basal cells 110% above control values 24 h after injury. Labelling indexes of epidermal basal cells in the skin adjacent to the wounded area were 7.0%. Basal cell DNA synthesis stimulated by wounding exhibited a distinct temporal variation at 24 h postinjury, with tail skin wounded at 12.00 h found to be 275% greater than control values and elevated 78% from LIs recorded at any other time point. This temporal spike was due to the time of day at which wounding occurred rather than the time point when the LI was determined. Mice wounded at 12.00 h and terminated 27 h later (15.00 h) had LIs that were 52% greater than wounds created at 09.00 h and examined at 12.00 h the following day. Higher levels of DNA synthesis in tail skin injured at 12.00 h compared to wounding at 09.00 h was detected 12-48 h after injury. Furthermore, DNA synthesis in wounds created at 12.00 h returned to baseline levels 1-2 days earlier than tail skin wounded at 09.00 h. Investigation of other strains of mice detected differences in radiolabelling of epidermal basal cells 24 h after tape stripping at 12.00 h or 09.00 h in CD-1 and BALB/cJ mice, but not in the C3H/HeJ strain. These results indicate: (a) there is no diurnal variation in the LI of mouse tail skin under normal homeostatic conditions (b) tape stripping is a potent stimulator of basal cell turnover in the epidermis (c) the time of wounding determines the magnitude of the increase in the LI of basal cells following injury, and (d) the proliferative response to wounding of the tail is dependent on the strain of mouse.

Animals↗

Effects of extracellular matrix components on the differentiation of chick embryo tail bud mesenchyme in culture.

The mesenchymal cells of the chick tail bud comprise the remains of Hensen's node and the primitive streak after gastrulation. This mass of cells, situated at the caudal limit of the chick embryo, is morphologically homogeneous but pluripotent, with the ability to differentiate into a variety of tissues that are both ectoderm- and mesoderm-derived elsewhere in the embryo. These tissues include neuroectoderm, neurons, myoblasts and chondrocytes. As the factors regulating the differentiation of tail bud mesenchyme into so many cell types are unclear, and because the extracellular matrix (ECM) is known to have a profound effect on cellular differentiation in many embryonic systems, we studied the differentiation of tail bud mesenchyme explanted onto a variety of different ECM components as substrata. We report that the histogenetic potential of isolated tail buds in culture compares favourably with that in situ. Using various antibody markers, we have demonstrated that tail bud mesenchyme cultured upon different ECM components as substrata is able to differentiate into neurons, neuroepithelium, melanocytes, muscle and cartilage. Laminin and laminin-containing substrata (Matrigel) were found to promote the differentiation of neural crest derivatives (neurons and melanocytes) and neuroepithelial cells; type I collagen promoted both myogenesis and chondrogenesis; while type IV collagen promoted myogenesis only. We have therefore demonstrated that differentiation of tail bud mesenchyme in vitro is substratum-dependent.

Animals↗

Immunological cross-reactivity between electric-eel acetylcholinesterase and rat-tail-tendon collagen.

Immunological cross-reactivity between acetylcholinesterase from the electric organ of the electric eel and rat tail tendon collagen was examined both on the cellular and humoral levels. 1. Guinea pigs immunized with rat tail tendon collagen displayed a strong delayed-type skin reaction when tested with the elongated acetylcholinesterase preparation (i.e. 14-S + 18-S molecular forms). However, when the glubular 11-S enzyme was tested, almost no cross-reactivity was obtained. Similarly, guinea pigs immunized with 14-S + 18-S preparation exhibited skin sensitization to rat tail tendon collagen. 2. Using a radioimmunoassay, it was observed that 125I-labeled 14-S + 18-S acetylcholinesterase binds efficiently to rabbit antiserum elicited against rat tail tendon collagen, whereas 125I-labeled 11-S enzyme does not bind at all to this antiserum. Similar results were obtained by passive hemagglutination assay. The experiments suggest that 14-S + 18-S acetylcholinesterase, but not 11-S enzyme, which is devoid of the tail structure, has antigenic determinants in common with collagen from rat tail tendon.

Acetylcholinesterase↗

Lipoic acid prevents collagen abnormalities in tail tendon of high-fructose-fed rats.

BACKGROUND: Non-enzymatic glycation of proteins, leading to chemical modification and cross-linking are of importance in the pathology of diabetic complications. We studied the effect of alpha-lipoic acid (LA) on the glycation and cross-linking of collagen from tail tendon of high-fructose-fed rats. METHODS: The rats were divided into four groups of six each. Two groups of rats were fed with a high-fructose diet (60/100 g diet) and administered with either LA (35 mg/kg body weight, intraperitoneally) (FRU + LA) or saline (FRU) for 45 days. The other two groups were fed control diet containing starch (60/100 g diet) and administered with either saline (CON) or LA (CON + LA). The rats were maintained for 45 days and then sacrificed. Collagen was isolated from tail tendon and the extent of glycation, fluorescence, aldehyde and peroxidation levels were measured. The tail tendons were separated, and shrinkage temperature was also measured. Acid-soluble collagen was extracted from tail tendon and subjected to sodium dodecyl sulphate polyacrylamide gel electrophoresis. RESULTS: Fructose administration caused accumulation of collagen in tail tendon. Glycation, collagen-linked fluorescence, aldehyde and peroxide contents were elevated. The gel pattern of acid-soluble collagen from the fructose-fed rat showed elevated beta-component. These changes were alleviated by the simultaneous administration of LA. CONCLUSION: Administration of LA has a positive influence on tail tendon collagen glycation and other variables in high-fructose-fed rats.

Animals↗