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PubMed1996

Expression of zebrafish connexin43.4 in the notochord and tail bud of wild-type and mutant no tail embryos.

Communication via gap junctions provides a mechanism for the cell-cell transfer and coordination of developmental signals. The spatial restriction of gap junctions may also serve to organize cells into domains of coordinated behavior. To investigate the role of gap junctions during embryogenesis, we have characterized the expression of a member of the gap junction gene family, zebrafish connexin43.4, a homolog of connexin45 in chicken and mammals. Expression of connexin43.4 was induced in the early gastrula, coincident with the first definitive assignments of axial cell fate and the onset of the cell movements comprising convergence and extension in zebrafish. In situ hybridization and immunohistochemistry revealed that during gastrulation connexin43.4 mRNA and protein were progressively enriched in the germ ring and in the notochord primordia on the dorsal side of the embryo. Later in development connexin43.4 expression was detected in the notochord, the paraxial mesoderm, and the tail bud but was not observed after the differentiation of these tissues. In no tail mutant embryos which are defective in tail formation and proper morphogenesis of the notochord, connexin43.4 expression was absent during gastrulation from the caudal embryonic shield and notochord primordia. During somite stages in no tail embryos, connexin43.4 expression remained absent in the notochordal precursor cells and was lost in the tail bud. Thus, the no tail gene product, a transcription factor, was required for the expression of connexin43.4 in both the notochord and tail bud during morphogenesis. By microinjection of mRNA coding for a connexin43.4/green fluorescent protein fusion in the 1-cell zebrafish embryo, we showed that connexin43.4 is capable of assembling into structures reminiscent of gap junctions. The progressively restricted, developmental expression of the zebrafish connexin43.4 gene suggests that this gap junctional protein participates in the coordination of gastrulation and the formation of the notochord and tail.

Animals↗
PubMed2001

Conformational properties of alpha-tubulin tail peptide: implications for tail-body interaction.

The carboxy-terminal segments of the alpha/beta-tubulins are flexible regions rich in acidic amino acid residues. It is generally believed that these regions play crucial roles in tubulin polymerization and interaction with many ligands, including colchicine. Exactly how these effects are exerted are not known at present. One such interesting aspect is the pH dependence of colchicine-tubulin interaction and the influence of the alpha-tail on the binding interaction. We have investigated the location of the colchicine-binding site on tubulin by docking. It has been located on the alpha/beta interface on the N-terminal side, which is also supported by much of the solution data. This location is too far from the tail regions, suggesting that influence of the tail region is transmitted by a pH-dependent conformational change. Two-dimensional NMR studies indicate that at pH 7 a 13-residue peptide corresponding to alpha-tubulin tail shows little NOE constraints, suggesting extended conformation. On the contrary, at pH 5, a relatively compact structure was deduced from the interproton NOE constraints. Pulsed field gradient measurement of diffusion constant indicates that the peptide at pH 5 is substantially faster diffusing than at pH 7. The Perrin factors calculated from diffusion data indicates that the peptide structure at pH is significantly more compact than at pH 7. Temperature coefficients of several amide protons at pH 5 fall below 5 ppb/(o)K, indicating a degree of protection. A difference is also seen in the CD spectra obtained at different pHs, consistent with the NMR data. We have investigated the probable spatial organization of the tail of the alpha-subunit of tubulin, in the high pH extended form and the low pH compact form. On the basis of correlation of pH dependence of many properties of tubulin and the conformation of the alpha-tail peptide, we propose that the intrinsic conformational preference of the tail-region modulate the tail-body interaction, which in turn has important bearing on colchicine binding properties.

Binding Sites↗
PubMed2000

DNA sequence-dependent contributions of core histone tails to nucleosome stability: differential effects of acetylation and proteolytic tail removal.

Modulation of nucleosome stability in chromatin plays an important role in eukaryotic gene expression. The core histone N-terminal tail domains are believed to modulate the stability of wrapping nucleosomal DNA and the stability of the chromatin filament. We analyzed the contribution of the tail domains to the stability of nucleosomes containing selected DNA sequences that are intrinsically straight, curved, flexible, or inflexible. We find that the presence of the histone tail domains stabilizes nucleosomes containing DNA sequences that are intrinsically straight or curved. However, the tails do not significantly contribute to the free energy of nucleosome formation with flexible DNA. Interestingly, hyperacetylation of the core histone tail domains does not recapitulate the effect of tail removal by limited proteolysis with regard to nucleosome stability. We find that acetylation of the tails has the same minor effect on nucleosome stability for all the selected DNA sequences. A comparison of histone partitioning between long donor chromatin, acceptor DNA, and free histones in solution shows that the core histone tails mediate internucleosomal interactions within an H1-depleted chromatin fiber amounting to an average free energy of about 1 kcal/mol. Thus, such interactions would be significant with regard to the free energies of sequence-dependent nucleosome positioning. Last, we analyzed the contribution of the H2A/H2B dimers to nucleosome stability. We find that the intact nucleosome is stabilized by 900 cal/mol by the presence of the dimers regardless of sequence. The biological implications of these observations are discussed.

Acetylation↗
PubMed1990

Identification of functional regions on the tail of Acanthamoeba myosin-II using recombinant fusion proteins. II. Assembly properties of tails with NH2- and COOH-terminal deletions.

We used purified fusion proteins containing parts of the Acanthamoeba myosin-II tail to localize those regions of the tail responsible for each of the three steps in the successive dimerization mechanism (Sinard, J. H., W. F. Stafford, and T. D. Pollard. 1989. J. Cell Biol. 107:1537-1547) for Acanthamoeba myosin-II minifiliment assembly. Fusion proteins containing the terminal approximately 90% of the myosin-II tail assemble normally, but deletions within the last 100 amino acids of the tail sequence alter or prevent assembly. The first step in minifilament assembly, formation of antiparallel dimers, requires the COOH-terminal approximately 30 amino acids that are thought to form a nonhelical domain at the end of the coiled-coil. The second step, formation of antiparallel tetramers, requires the last approximately 40 residues in the coiled-coil. The final step, the association of two antiparallel tetramers to form the completed octameric minifilament, requires residues approximately 40-70 from the end of the coiled-coil. A region of the tail near the junction with the heads is important for tight packing of the tails in the minifilaments. Divalent cations induce the lateral aggregation of minifilaments formed from native myosin-II or fusion proteins containing a nonmyosin "head," but under the same conditions fusion proteins composed essentially only of myosin tail sequences with very little nonmyosin sequences form paracrystals. The region of the tail necessary for this paracrystal formation lies NH2-terminal to amino acid residue 1,468 in the native myosin-II sequence.

Acanthamoeba↗
PubMed2001

DNA inversion in the tail fiber gene alters the host range specificity of carotovoricin Er, a phage-tail-like bacteriocin of phytopathogenic Erwinia carotovora subsp. carotovora Er.

Carotovoricin Er is a phage-tail-like bacteriocin produced by Erwinia carotovora subsp. carotovora strain Er, a causative agent for soft rot disease in plants. Here we studied binding and killing spectra of carotovoricin Er preparations for various strains of the bacterium (strains 645Ar, EC-2, N786, and P7) and found that the preparations contain two types of carotovoricin Er with different host specificities; carotovoricin Era possessing a tail fiber protein of 68 kDa killed strains 645Ar and EC-2, while carotovoricin Erb with a tail fiber protein of 76 kDa killed strains N786 and P7. The tail fiber proteins of 68 and 76 kDa had identical N-terminal amino acid sequences for at least 11 residues. A search of the carotovoricin Er region in the chromosome of strain Er indicated the occurrence of a DNA inversion system for the tail fiber protein consisting of (i) two 26-bp inverted repeats inside and downstream of the tail fiber gene that flank a 790-bp fragment and (ii) a putative DNA invertase gene with a 90-bp recombinational enhancer sequence. In fact, when a 1,400-bp region containing the 790-bp fragment was amplified by a PCR using the chromosomal DNA of strain Er as the template, both the forward and the reverse nucleotide sequences of the 790-bp fragment were detected. DNA inversion of the 790-bp fragment also occurred in Escherichia coli DH5alpha when two compatible plasmids carrying either the 790-bp fragment or the invertase gene were cotransformed into the bacterium. Furthermore, hybrid carotovoricin CGE possessing the tail fiber protein of 68 or 76 kDa exhibited a host range specificity corresponding to that of carotovoricin Era or Erb, respectively. Thus, a DNA inversion altered the C-terminal part of the tail fiber protein of carotovoricin Er, altering the host range specificity of the bacteriocin.

Bacteriocins↗
PubMed1984

The tail flattening reflex in Lumbricus: reconstitution after tail amputation and modifications in segmental nerve roots.

On stimulation of the head, the caudal 20-30 segments of intact earthworms exhibit a marked dorso-ventral flattening reflex which is an element of the escape response. When the tail is removed this reflex is reconstituted gradually in segments anterior to the amputation level without regeneration of new tail segments. It takes longer to reestablish the flattening if larger parts of the body are cut off, but after 60 days p.o. all animals regained the full response independent of the number of segments amputated. Extirpation of three consecutive ganglia from the ventral nerve cord also suffices to develop flattening anterior to the operated segments, but the reaction is less pronounced and appears later than after tail amputation. Moreover, it vanishes at the same time the normal tail flattening is observed again, presumably after regeneration across the gap in the cord. In different regions of the body, area and perimeter distributions of transversely sectioned axons in the three pairs of segmental nerves (SN I-III) of the cord were measured. Characteristic changes along the length of the animal were found for SN I and SN III. In tail segments the axonal size distributions are more variable than "typical" segments. The flattening reflex is mediated by axons in SN III (Pallas and Drewes, 1981). In body segments having reconstituted behavioral characteristics of the lost tail segments, the area of the largest axons and the general size distribution in SN III approximate those found in normal tail segments.

Animals↗
PubMed1988

Response latencies in the tail-flick test depend on tail skin temperature.

Tail skin temperatures and tail-flick latencies were simultaneously recorded in male Sprague-Dawley rats exposed to various ambient temperatures (22-30 degrees C). There was a positive correlation between tail skin temperature and ambient temperature and a negative correlation between tail-flick latency and ambient temperature. Importantly, a highly significant negative correlation was present between tail-flick latency and skin temperature, even at constant ambient temperature (22.1 or 23.3 degrees C). Thus, the results of tail-flick testing are highly affected by skin temperature and factors altering the skin temperature must be considered when tail-flick latencies are interpreted in terms of nociception.

Animals↗
PubMed2005

Formation of detached tail and coiled tail of sperm in a beagle dog.

The right testis and epididymis were excised from a Beagle dog that ejaculated high percentages of sperm with detached tails and with coiled tails. Cross sections of the organs were stamped on glass slides and histological examination of the organs was performed to find the portion where sperm with the abnormal tails appear. Many sperm with tails whose axoneme was exposed near the neck region were observed in the testis and they decreased in order from the caput, to the corpus, and the cauda epididymis. Sperm with detached tails and sperm with coiled tails gradually increased in the epididymis. These findings indicate that the tails of sperm with an exposed axoneme detached in the epididymis.

Animals↗
PubMed2005

An improved technique for tail-cuff blood pressure measurements with dark-tailed mice.

Study of the genetics of hypertension has been facilitated greatly by the use of mice with modified genes that affect blood pressure. A current successful method for measuring blood pressure in mice relies on detection of light passing through the tail to determine the pressure in a tail-cuff necessary to stop pulsed flow. Success in obtaining reliable blood pressure measurements in light-tailed strains of mice (e.g., C57BL/6J) has been excellent. However, in our and others' experience, mice having highly pigmented tails (e.g., 129S6/SvEvTac) have yielded less consistent measurements. We report here that simple modifications to the channel containing the pulse detection sensor can greatly improve the pulse detection of dark-tailed mice. The first modification--lining the sensor channel with four layers of clear plastic wrap--increased the frequency of successful blood pressure measurements of 129S6/SvEvTac mice twofold and reduced variability by one-third. The second modification--lining the sides of the channel with reflective foil--also improved the success rate with dark-tailed mice. Mean blood pressures were unaffected by these modifications, which enhance detection of the pulse wave and likely will be helpful in diverse applications in which blood pressure is measured in rodent strains with pigmented tails.

Animals↗
PubMed1994

The actin binding site in the tail domain of Dictyostelium myosin IC (myoC) resides within the glycine- and proline-rich sequence (tail homology region 2).

The majority of protozoan myosins I possess tail domains composed of three distinct and conserved regions of sequence, referred to as tail homology regions 1, 2 and 3 (TH.1, TH.2 and TH.3). While the N-terminal approximately half of the tail (corresponding to TH.1) has been implicated in membrane binding, all or some portion of the C-terminal approximately half of the tail (corresponding to TH.2 plus TH.3) has been implicated in binding to F-actin in a nucleotide-insensitive fashion. Here we show, using fusion proteins containing portions of the Dictyostelium myosin IC (myoC) tail domain and F-actin sedimentation assays, that the ability of the myoC tail to bind to actin resides entirely within the glycine- and proline-rich TH.2 domain. The src-like TH.3 domain does not bind to actin, nor does it augment the binding properties of the TH.2 domain. In addition to defining more precisely the location of the actin binding site in the tail domain of a protozoan myosin I, these results have implications for the function of the src-like TH.3 domain in myosins I and other proteins.

Actins↗
PubMed2005

Temperature-dependent tail-tail dynamics of pyrene-labeled poly(dimethylsiloxane) oligomers dissolved in ethyl acetate.

We have determined the tail-tail dynamics and energetics of linear poly(dimethylsiloxane) (PDMS) polymers (M(n) = 3100; M(w)/M(n) = 1.07) that have been terminally labeled with pyrene (Py-PDMS-Py) in ethyl acetate between 255 and 323 K. The upper critical solution temperature (theta(u)) for the PDMS/ethyl acetate system is 278 K. The Py-PDMS-Py photophysics follow the Birks model at all of the temperatures studied. However, there is evidence for an increase in the local composition of PDMS surrounding the Py- residues in Py-PDMS-Py below the theta temperature. The recovered activation energy for Py-PDMS-Py tail-tail cyclization is the same as the activation energy for diffusion in ethyl acetate. The activation energy for cyclization is about fourfold larger in comparison to the barrier for PDMS internal backbone rotations. The estimated internal activation barrier for Py-PDMS-Py in ethyl acetate is in line with the internal barrier rotation for PDMS backbone rotation. The intramolecular excimer binding energy for Py-PDMS-Py in ethyl acetate is similar to the intermolecular pyrene excimer. In the ground state, the mean distance between the Py-PDMS-Py tail segments does not change over the temperature range studied. A comparison of these temperature-dependent Py-PDMS-Py/ethyl acetate results with previous results on Py-PDMS-Py in pure supercritical CO2 and CO2-dilated liquids shows that the addition of CO2 to the liquid phase offers the way to tune to Py-PDMS-Py tail-tail dynamics over the greatest range.

Journal Article↗
PubMed1997

The isolated comet tail pseudopodium of Listeria monocytogenes: a tail of two actin filament populations, long and axial and short and random.

Listeria monocytogenes is driven through infected host cytoplasm by a comet tail of actin filaments that serves to project the bacterium out of the cell surface, in pseudopodia, to invade neighboring cells. The characteristics of pseudopodia differ according to the infected cell type. In PtK2 cells, they reach a maximum length of approximately 15 microm and can gyrate actively for several minutes before reentering the same or an adjacent cell. In contrast, the pseudopodia of the macrophage cell line DMBM5 can extend to >100 microm in length, with the bacteria at their tips moving at the same speed as when at the head of comet tails in bulk cytoplasm. We have now isolated the pseudopodia from PtK2 cells and macrophages and determined the organization of actin filaments within them. It is shown that they possess a major component of long actin filaments that are more or less splayed out in the region proximal to the bacterium and form a bundle along the remainder of the tail. This axial component of filaments is traversed by variable numbers of short, randomly arranged filaments whose number decays along the length of the pseudopodium. The tapering of the tail is attributed to a grading in length of the long, axial filaments. The exit of a comet tail from bulk cytoplasm into a pseudopodium is associated with a reduction in total F-actin, as judged by phalloidin staining, the shedding of alpha-actinin, and the accumulation of ezrin. We propose that this transition reflects the loss of a major complement of short, random filaments from the comet, and that these filaments are mainly required to maintain the bundled form of the tail when its borders are not restrained by an enveloping pseudopodium membrane. A simple model is put forward to explain the origin of the axial and randomly oriented filaments in the comet tail.

Actins↗
PubMed2002

How do birds' tails work? Delta-wing theory fails to predict tail shape during flight.

Birds appear to use their tails during flight, but until recently the aerodynamic role that tails fulfil was largely unknown. In recent years delta-wing theory, devised to predict the aerodynamics of high-performance aircraft, has been applied to the tails of birds and has been successful in providing a model for the aerodynamics of a bird's tail. This theory now provides the conventional explanation for how birds' tails work. A delta-wing theory (slender-wing theory) has been used, as part of a variable-geometry model to predict how tail and wing shape should vary during flight at different airspeeds. We tested these predictions using barn swallows flying in a wind tunnel. We show that the predictions are not quantitatively well supported. This suggests that a new theory or a modified version of delta-wing theory is needed to adequately explain the way in which morphology varies during flight.

Animals↗
PubMed1986

[Tail necrosis and tail biting in veal calves in group housing].

Two cases of lesions of the tail due to tail-biting in veal calves kept in a loose housing system are reported. On one farm, necrosis of the tail accompanied by abscess formation occurred. In fattening bulls necrosis of the tail may appear after traumatic lesions due to treading the tails, but the exact pathogenesis is obscure. The reason for tail-biting in veal calves is also not known. The therapeutic and preventive measures described therefore are rather non-specific.

Animals↗
PubMed1986

Centrifugal modulation of the rat tail flick reflex evoked by graded noxious heating of the tail.

Centrifugal modulation from the midbrain, pons and medulla of the spinal nociceptive tail flick (TF) reflex evoked by graded noxious heating of the tail was studied in lightly pentobarbital-anaesthetized rats. In initial experiments, the relationship between the intensity of the noxious thermal stimulus and the TF latency was characterized. The thermal stimulus was provided by a lamp focused on the ventral surface of a rat's tail. Five different rates of heating of the tail were varied systematically by altering the voltage supplied to the lamp and were characterized using a thermocouple to measure the temperature of tissue exposed to the radiant heat. A linear stimulus-response function relating the inverse of the latency of the TF to the rate of heating of the tail was established. However, the mean cutaneous tissue temperature of the exposed tail at the time of the TF was found to be invariant and independent of the rate of heating. Focal electrical stimulation in the midbrain, pons and medulla modulated the TF reflex in two different ways, analogous to modulations of stimulus-response functions of single-cell recordings of spinal dorsal horn neurons. A Type I modulation, analogous to the parallel shift in response threshold seen in spinal dorsal horn neurons, was an absolute increase in the thermal threshold of the TF reflex. A Type II modulation, analogous to a change in slope or gain seen in spinal dorsal horn neurons, was a linear increase in the thermal threshold of the TF reflex as a function of the rate of heating. Type I modulations were produced by electrical stimulation in the ventromedial medulla (n. raphe magnus and n. reticularis gigantocellularis) and lateral periaqueductal gray of the midbrain. Type II modulations were produced by electrical stimulation in the dorsolateral pons, locus coeruleus-subcoeruleus and in the medial periaqueductal gray. This experimental approach has shown itself to be useful in the characterization of descending inhibition of nociception. Much simpler and less invasive than analogous spinal dorsal horn single cell electrophysiologic studies, it can be used to study the mechanisms of centrifugal modulation of nociceptive flexion reflexes and further establishes the utility of the lightly anaesthetized rat preparation for studies of nociception-antinociception.

Animals↗
PubMed2000

Intrathecal endomorphin-1 produces antinociceptive activities modulated by alpha 2-adrenoceptors in the rat tail flick, tail pressure and formalin tests.

It is known that spinal morphine produces antinociception that is modulated by alpha 2-adrenoceptors. Endomorphin-1, a newly-isolated endogenous opioid ligand, shows the greatest selectivity and affinity for the mu-opiate receptor of any endogenous substance found to date and may serve as a natural ligand for the mu-opiate receptor. We examined the antinociceptive effects of endomorphin-1 administered intrathecally (i.t.) in the rat tail flick, tail pressure and formalin tests. Intrathecal endomorphin-1 produced dose-dependent antinociceptive effects in the three tests. ED50 (CI95) values for antinociception of i.t. endomorphin-1 in the tail flick test and tail pressure test were 1.9 (0.96-3.76) nmol and 1.8 (0.8-4.2) nmol, respectively. ED50 (CI95) values for phase 1 and phase 2 in the formalin test were 12.5 (7.9-19.8) nmol and 17.5 (10.2-30) nmol, respectively. Pretreatment with i.t. beta-funaltrexamine (a mu-opioid receptor selective antagonist) significantly antagonized the antinociceptive effects of endomorphin-1 in the three tests. Beta-funaltrexamine alone had not effects on the three tests. The antinociceptive effects of endomorphin-1 were also antagonized by i.t. yohimbine (an alpha 2-adrenoceptor selective antagonist). The combination of ineffective doses of i.t. clonidine (an alpha 2-adrenoceptor agonist) and endomorphin-1 produced a significant antinociception in the three tests. The results showed that intrathecal endomorphin-1 produced antinociception in a dose-dependent manner in the rat tail flick, tail pressure and formalin tests, which was mediated by spinal mu-opioid receptors and modulated by alpha 2-adrenoceptors.

Adrenergic alpha-Antagonists↗
PubMed2000

Myosin-II tails confer unique functions in Schizosaccharomyces pombe: characterization of a novel myosin-II tail.

Schizosaccharomyces pombe has two myosin-IIs, Myo2p and Myp2p, which both concentrate in the cleavage furrow during cytokinesis. We studied the phenotype of mutant myosin-II strains to examine whether these myosins have overlapping functions in the cell. myo2(+) is essential. myp2(+) cannot rescue loss of myo2(+) even at elevated levels of expression. myp2(+) is required under specific nutritional conditions; thus myo2(+) cannot rescue under these conditions. Studies with chimeras show that the tails rather than the structurally similar heads determine the gene-specific functions of myp2(+) and myo2(+). The Myo2p tail is a rod-shaped coiled-coil dimer that aggregates in low salt like other myosin-II tails. The Myp2p tail is monomeric in high salt and is insoluble in low salt. Biophysical properties of the full-length Myp2p tail and smaller subdomains indicate that two predicted coiled-coil regions fold back on themselves to form a rod-shaped antiparallel coiled coil. This suggests that Myp2p is the first type II myosin with only one head. The C-terminal two-thirds of Myp2p tail are essential for function in vivo and may interact with components of the salt response pathway.

Animals↗
PubMed2001

A new G-tailing method for the determination of the poly(A) tail length applied to hepatitis A virus RNA.

To study the role of the poly(A) tail length during the replication of poly(A)-containing plus-strand RNA virus, we have developed a simple reverse transcription polymerase chain reaction (RT-PCR)-based method that substantially improves the previously reported PAT [poly(A) test] assay. In contrast to the PAT assay, the new method is based on the enzymatic 3' elongation of mRNA with guanosine residues, thus immediately preserving the 3' end of the RNA and creating a unique poly(A)-oligo(G) junction. The oligo(G)-protected full-length poly(A) tail is reverse transcribed using the universal anti-sense primer oligo(dC(9)T(6)) and amplified by PCR with a gene-specific sense primer. After sequencing the resulting RT-PCR product the length of the poly(A) tail was unequivocally deduced from the number of adenosine residues between the oligo(G) stretch and the sequence upstream of the poly(A) tail. The efficiency and specificity of the newly developed assay was demonstrated by analysing the poly(A) tail length of the hepatitis A virus (HAV) RNA. We show here that the poly(A) tail of HAV RNA rescued after transfection of in vitro transcripts was elongated in the course of HAV replication.

Adenosine↗