CLF associates with CLC to form a functional heteromeric ligand for the CNTF receptor complex.

Ciliary neurotrophic factor (CNTF) is a cytokine supporting the differentiation and survival of various cell types in the peripheral and central nervous systems. Its receptor complex consists of a non-signaling alpha chain, CNTFR, and two signaling beta chains, gp130 and the leukemia inhibitory factor receptor (LIFR). Striking phenotypic differences between CNTF- and CNTFR-deficient mice suggest that CNTFR serves as a receptor for a second, developmentally important ligand. We have identified this factor as a stable secreted complex of cardiotrophin-like cytokine (CLC) and the soluble receptor cytokine-like factor-1 (CLF). CLF expression was required for CLC secretion, and the complex acted only on cells expressing functional CNTF receptors. The CLF/CLC complex activated gp130, LIFR and signal transducer and activator of transcription 3 (STAT3) and supported motor neuron survival. Our results indicate that the CLF/CLC complex is a second ligand for CNTFR with potentially important implications in nervous system development.

Hydroxocobalamin vs cobalt toxicity on rat cardiac and diaphragmatic muscles.

BACKGROUND: Hydroxocobalamin has been shown to be a rapid and powerful antidote in acute cyanide poisoning and to prevent cyanide poisoning during sodium nitroprusside administration. This cobalt-containing compound has been shown to be devoid of significant immediate side effects during acute administration. However, its potential delayed toxicity related to cobalt accumulation in tissue remains unknown. Therefore, we evaluated the toxicity of hydroxocobalamin as compared with that of cobalt salts on rat cardiac and diaphragmatic muscles. METHODS: For a 21-day period, rats were treated intraperitoneally with either hydroxocobalamin (70 mg kg-1 per day, n = 14), cobalt chloride hexahydrate (12 mg kg-1 per day, n = 14) or saline (n = 10). Hydroxocobalamin and cobalt chloride groups received equimolar doses of cobalt. We studied: (1) the mechanical properties of isolated left ventricular papillary muscles and diaphragmatic strips, (2) the cardiac and diaphragmatic cobalt tissue concentrations, and (3) the myocardial histological aspect. RESULTS: During the study period, no significant increase in body weight was noted in the cobalt-treated group (-4 +/- 1%), which was in contrast to the hydroxocobalamin-treated group (+21 +/- 2%) and the saline-treated group (22 +/- 2%). Compared with controls, the mechanical properties of cardiac and diaphragmatic muscles were unchanged after either hydroxocobalamin or cobalt salt treatments, and myocardial histological characteristics were similar in all groups. Conversely, large amounts of cobalt deposit were observed in the cobalt-treated group in both the diaphragm (41.90 +/- 16.30 vs 0.70 +/- 0.40 mu mol mu g-1 in the control group, P < 0.001) and the myocardium (16.90 +/- 6.40 vs 0.14 +/- 0.01 mu mol mu g-1 in the control group, P < 0.001). After hydroxocobalamin administration, cobalt concentrations were significantly lower in the diaphragm (25.10 +/- 16.50 mu mol mu g-1, P < 0.001 vs cobalt-treated group) and the myocardium (4.50 +/- 1.20 mu mol mu g, P < 0.001 vs cobalt-treated group). CONCLUSION: These results indicate that repeated administration of hydroxocobalamin was devoid of significant diaphragmatic and cardiac muscle toxicity and therefore remains a safe antidote for acute cyanide poisoning.

Mechanics of human quadriceps muscle.

Mechanics of human quadriceps muscle strips (vastus lateralis; n = 10) were investigated over the whole load continuum. Mechanical experiments were performed at 29 degrees C and in both twitch and tetanus modes. For a given level of isotonic total load (P) and over a large part of the contraction phase, instantaneous velocity (V) was shown to be a unique function of instantaneous length (L), regardless of time and initial length. By considering this time- and initial length-independent mechanical property between instantaneous L and instantaneous V over the whole P continuum, a three-dimensional P-V-L relationship was constructed. Any variations in stimulation conditions modified the time-independent P-V-L diagram. Such modifications in the P-V-L relationship were characteristics of changes in contractile performance. Moreover, characteristics of the P-V relationship were investigated in both twitch and tetanus modes. The curvature of the P-V hyperbola was significantly higher in tetanus at 30 Hz than in twitch mode (P < 0.001). In conclusion, our study indicates that, in human quadriceps muscles, contractility can be defined as the time- and initial length-invariant part of a three-dimensional P-V-L relationship. Moreover, our data are consistent with an increase in economy of force generation in tetanus contractions compared with that in twitches.

Sarcomere relaxation in hamster diaphragm muscle.

We characterized instantaneous sarcomere relaxation over the load continuum in isolated hamster diaphragm muscles by means of laser diffraction. In afterloaded twitches, sarcomere relaxation displayed two consecutive phases. The bulk of sarcomere lengthening occurred during the first phase and corresponded in time to muscle lengthening. The second phase of sarcomere relaxation was slower and corresponded in time to tension decay. At initial muscle length, the peak velocity of sarcomere lengthening (SVL) was linearly related to both the maximum extent of sarcomere shortening (delta SL) and sarcomere length at peak shortening (SLmin; each P < 0.01). Varying preload modified the SVL vs. SLmin relationship but not the SVL vs. delta SL relationship. At a given preload, muscle tension decay began at a similar sarcomere length, regardless of the afterload level. In conclusion, our results support the role played by sarcomere length in regulating the diaphragm muscle-lengthening rate but not the rate of tension decline.

Mechanical determinants of isotonic relaxation in isolated diaphragm muscle.

Determinants of lengthening velocity have not been investigated in the diaphragm muscle. This study was undertaken to define the mechanical determinants of isotonic relaxation rate over the entire load continuum in isolated rat diaphragm (n = 30). We tested the hypothesis that the determinants of lengthening could include loading conditions, namely, preload and afterload; abrupt changes of load during the contraction phase; end-shortening muscle length (ESL); extent of shortening (delta L); time; stimulation mode; and stimulation frequency. In afterloaded contractions preloaded at optimal initial length and stimulated in tetanus at 30 Hz, peak lengthening velocity (+dL/dtmax) was linearly related to delta L, ESL, and/or total load. Varying initial muscle length, ESL, afterload, or the load imposed on the muscle during the isotonic lengthening process did not modify +dL/dtmax vs. delta L relationship, whereas +dL/dtmax vs. load and +dL/dtmax vs. ESL relationships were modified by these procedures. For a given delta L, +dL/dtmax could be modified when lengthening was delayed by reversing the relaxation sequence and when twitch and tetanus modes were compared. In conclusion, our results demonstrate that in isolated diaphragm muscle, delta L is the main determinant of +dL/dtmax over a wide range of loads and under various experimental conditions, independent of ESL and initial muscle length and independent of the load imposed on the muscle during the lengthening process. Time and stimulation mode were also shown to modulate the lengthening rate in diaphragm muscle.

Isometric relaxation of isolated diaphragm muscle: influence of load, length, time, and stimulation.

Determinants of the isometric relaxation rate were investigated in isolated rat diaphragm (n = 30). We tested the hypothesis that these determinants could include loading conditions, namely preload and afterload; abrupt changes in load during the contraction phase; stimulation conditions; and time. Two relaxation sequences were studied. When isometric relaxation occurred at initial muscle length (isotonic-isometric sequence), an increase in total load (P) accelerated the negative peak rate of tension decline (-dP/dtmax). Variations in initial length, stimulation, and onset of relaxation did not modify the -dP/dtmax vs. afterload relationship. When isometric relaxation was analyzed after -dP/dtmax, for a given afterload level the instantaneous rate of tension decline (-dP/dt) was a unique function of instantaneous tension, regardless of previous loading conditions, stimulation mode, or time. When the isometric relaxation occurred at end-shortening muscle length (isometric-isotonic sequence), the -dP/dtmax vs. P relationship was flat. The rate of tension decay, as attested by either -dP/dtmax or instantaneous -dP/dt vs. instantaneous tension phase plane, differed markedly depending on stimulation conditions. Thus the regulation of isometric relaxation rate differed according to the relaxation sequence. In muscle isometrically relaxing at initial muscle length, peak isometric relaxation rate was mainly determined by afterload. Conversely, in muscle isometrically relaxing at end-shortening length, isometric relaxation rate was highly dependent on the level of activation and was independent of preload and afterload.

Disappearance of xenogenic astrocytes transplanted into newborn mice is associated with a T-cell response.

Following transplantation of fragments of embryonic rabbit brain into the brains of newborn mice, the proportion of mice bearing detectable xenogenic astrocytes increases to over 80% in the first 3-4 weeks. Recent studies have demonstrated that the host response at this time was dominated by non-specific elements of host defense: macrophages, microglia and astrocytes. In the second phase, the proportion of mice bearing xenogenic astrocytes declines rapidly after 4 weeks and reaches zero by 10 weeks. In the present experiments, designed to characterize the host defense during this period, a dramatic increase in the proportion of mice displaying T-cells in the brain in the fourth and fifth weeks after transplantation was found. This corresponded with a marked decline of xenogenic astrocytes. Both subsets of T-cell, helper-inducer (L3T4) and cytotoxic-suppressor (Lyt2), were found, with L3T4 more numerous in many samples. T-cells were found at the site of transplantation and at sites of migration. The division of the host-defense response in this model into a phase of antigen non-specific cells followed by a period when T-cells appear and transplanted astrocytes disappear, should facilitate kinetic studies into the mechanisms of brain-graft rejection.

In situ transformation of striatal glia into cerebellar-like glia after brain transplantation.

Transplants of striatum from rabbit embryo were implanted into the colliculus posterior of newborn mice. After 4 weeks, astroglial cells derived from the transplant had migrated into the cerebellum of the host. Whenever they had settled in the cerebellum they presented forms similar to local glia. Some migrated glial cells were found to transform into forms of glia, such as radial-like glia, which are not present in the striatum. This observation confirms that glial precursor cells are highly plastic. It is an in vivo demonstration that local conditions alone define the morphology of glial cells. After grafting in an heterotopic location they take on forms that they were not destined to express in the region of origin.

[Role of sarcoplasmic reticulum in the regulation of myocardial relaxation]. / Rôle du réticulum sarcoplasmique dans la régulation de la relaxation myocardique.

In order to investigate cardiac muscle behavior after inhibition of either sarcoplasmic reticulum (SR) Ca2+ release or SR Ca2+ uptake, 35 papillary muscles of adult Wistar rats were studied after a 60 minutes exposure to ryanodine 10(-7) M (n = 11) or to cyclopiazonic acid (CPA) 10(-5) M (n = 14) and compared with a control group containing the solvent alone (n = 10). We measured the maximum extent of muscle shortening of the preloaded twitch (DLp) and the normalized total force of the fully isometric twitch (FTi). The peak lengthening velocity of the preloaded twitch (VRp) and the normalized negative peak force derivative of the fully isometric twitch (-DFi) tested the lusitropic state. Intrinsic changes in the relaxation phase, independent of the contractile state, i.e., the relaxant effects, were analysed using 1) two ratios; the VRp/DLp ratio of the preloaded twitch and the -DFi/FTi ratio of the fully isometric twitch and 2) the slopes of the VR versus DL and of the -DF versus FT relationship over the whole continuum of load. Ryanodine induced a marked negative inotropic effect associated with a decrease in VRp from 2.7 +/- 0.2 to 1.4 +/- 0.2 Lmax/s (p < 0.001). The VRp/DLp ratio and the slope of the VR versus DL relationship remained unchanged, indicating that ryanodine was devoid of intrinsic relaxant effect under isotonic conditions. At a 10/min stimulation frequency, inhibition of Ca(2+)-uptake function of the SR with CPA had no inotropic effect but decreased VRp from 2.9 +/- 0.1 to 2.2 +/- 0.1 Lmax/s (p < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

[Efficacy of Spirolept vaccine against human leptospirosis as estimated by passive protection of laboratory rodents]. / Etude de l'efficacité du vaccin Spirolept contre la leptospirose par la protection passive de rongeurs de laboratoire.

OBJECTIVE: This study had for aim to assess the serological response induced by the Spirolept vaccine against human leptospirosis. METHOD: A serological follow-up was made on 31 patients at a risk of occupational exposure. The antibody titers of vaccinated patients were assessed by MAT and ELISA. In a second step, vaccinal protection was studied in vivo by checking the seroprotective effect of the human sera injected in an animal model (Meriones unguiculatus) naturally susceptible to the disease. The passive protection was studied by comparing the death rate on five batches of animals to which the bacterium was inoculated. Thus, four batches of animals were injected subcutaneously with a pooled sera of vaccinated people sampled at D0, D15, D135, and D320 after Spirolept vaccination. One control batch was given PBS. One day after injection, the latter batch was inoculated with the homologous strain Verdun of Leptospira interrogans ss icterohemorrhagiae (serogroup Icterohemorrhagiae) used to make the vaccine. RESULTS: The death rate was significantly decreased as soon as D15 after the first injection, even with pooled sera of vaccinated people negative for the MAT. COMMENTS: The Spirolept vaccine induces a protective response against icterohemorrhagiae, which can be transmitted to the animal model and thus is linked to a humoral response.

[Mechanical determinants of isotonic relaxation of the isolated diaphragmatic muscle]. / Déterminants mécaniques de la relaxation isotonique du muscle diaphragmatique isolé.

The aim of this study was to define the mechanical determinants of isotonic relaxation of isolated diaphragm muscle in the rat over a load range (n = 30). We tested several hypotheses to determine the effect of i) load conditions (preload, post-load), ii) sudden changes in load during contraction, iii) length of the muscle at peak shortening, iv) maximal amplitude of shortening (delta L) and v) stimulation conditions on peak rate of isotonic reelongation (+dL/dmax). At tetanus at 30 Hz, +dL/dmax was linearly correlated to delta L peak shortening and total load. Variations in preload, peak shortening or postload did not modify the +dL/dmax vs delta L relationship but such variations did affected the relationships +dL/dmax vs total load or +dL/dmax vs peak shortening. For a given value of L, +dL/dmax was weaker for twitch than for tetanus. In conclusion, four findings show that over a wide lad range the maximal amplitude of shortening the main mechanical determinant of the rate of isotonic reelongation of the isolated diaphragm muscle, independently of the length of the muscle at peak shortening, the initial length of the muscle and independently of the load during reelongation.

Migration patterns of donor astrocytes after reciprocal striatum-cerebellum transplantation into newborn hosts.

Fragments of striatum or cerebellum from E 25 rabbit embryo were implanted into either the striatum or the mesencephalon of newborn mice. Implanted rabbit astrocytes were selectively identified by monoclonal antibodies to the GFAP which are unable to combine with mouse GFAP. Previous investigations had shown that xenogenic astrocytes have the capacity to migrate in host CNS. The purpose of this study was to compare the patterns of migration of transplant-derived astroglial cells according to the topographic origin of the transplant and location of the grafting site. We found that the migration pattern of the grafted cells from any of both selected sites of implantation was independent from the topographic origin of the transplant. The routes as well as the distances of migration were similar after homo- or heterotopic transplantation. We conclude that astroglial cells or their precursors do not express information which would direct them to move specifically toward a defined region in the host brain according to the region of origin in the donor.

Short-term post-grafting morphological alterations of glia from an adult brain transplant.

Fragments of corpus callosum from adult rabbit have been implanted into the brain of newborn mice. Previous studies had shown that under such conditions transplant-derived astroglial cells differentiate in the host and survive for at least 2 months. The present study was devised to clarify the fate of the differentiated astrocytes present in the adult transplant by using combined ultrastructural and immunohistochemical approaches. These mature cells are shown to degenerate and die within 2 days after the implantation. Therefore, we suggest that stem cells present in adult tissue would account for the surviving population of transplant-derived glial cells.

Comparative migration and development of astroglial and oligodendroglial cell populations from a brain xenograft.

In previous studies of brain transplantation, the fate of the implanted glial cells has been investigated separately; that is, the interest has been focused either on the astroglia or on the oligodendroglia. However, the two populations of implanted glial cells may interact with each other, for example by secreting species-specific factors or by inducing reactions by the host. We have used two different models of brain transplantation: one that allows the identification of the implanted astrocytes, and another that allows the identification of the implanted oligodendroglia. The present model is a combination of both; it consists of the grafting of embryonic rabbit brain fragments into the brains of neonatal Shiverer mice. The myelin made by the implanted oligodendrocytes is identified by anti-myelin basic protein immunohistochemistry. The implanted astrocytes are identified by a monoclonal antibody that combines with rabbit but not with mouse glial fibrillary acidic protein. This study shows that although they use the same major routes of migration, both populations of glial cells tend to move differently. They demonstrate areas of common settlement but also areas where only one population of implanted glia is present. From the site of implantation in the dorsal striatum, the major routes of migration are the corpus callosum, the white matter fascicles in the striatum, and the internal capsule. After a delay of 6 weeks, no significant prevalence of one population of implanted glial cells over the other was observed.

A comparison of cyclopiazonic acid and ryanodine effects on cardiac muscle relaxation.

We investigated cardiac muscle behavior after inhibition of either sarcoplasmic reticulum (SR) Ca2+ release or SR Ca2+ uptake. Mechanics of 35 rat papillary muscles were studied after either ryanodine 10(-7) M (n = 11) or cyclopiazonic acid (CPA) 10(-5) M (n = 14) and compared with a control group containing the solvent alone (n = 10). We measured the maximum extent of shortening (delta L) of the preloaded twitch (delta Lp), and the normalized total force (TF) of the full isometric twitch (TFi). The peak lengthening velocity (Vl) of the preloaded twitch (Vlp) and the normalized negative peak force derivative of the fully isometric twitch (-DFi) tested the lusitropic state. With the influence of shortening and/or load on relaxation taken into account, analysis of relaxation was performed using 1) Vlp-to-delta Lp and magnitude of -DFi-to-TFi ratios and 2) slopes of the Vl-delta L and magnitude of -DF-TF relationships over the entire continuum of load. Ca(2+)-release inhibition with ryanodine induced a negative inotropic effect and a decrease in Vlp from 2.7 +/- 0.2 to 1.4 +/- 0.2 Lmax/S, where Lmax is the initial length at the peak of the length-active tension curve (P < 0.001). The Vlp-to-delta Lp ratio and the slope of the Vl-delta L relationship were preserved, indicating that ryanodine was devoid of intrinsic relaxant effect under isotonic conditions. Ca(2+)-uptake inhibition with CPA had no inotropic effect but decreased Vlp from 2.9 +/- 0.1 to 2.2 +/- 0.1 Lmax/s (P < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Interspecies identification of astrocytes after intracerebral transplantation.

Species-specificity of the Tp-GFAP 1 (glial fibrillary acidic protein) monoclonal antibodies raised against calf GFAP was established by means of immunochemical techniques. Since it was shown to combine with rabbit GFAP but not with mouse GFAP it allows the characterization of a new experimental model potentially useful in the study of the fate of implanted astrocytes after intracerebral graft of CNS fragments. Preliminary observations indicate that embryonic and newborn rabbit astrocytes are able to survive, express GFAP and migrate when implanted into newborn mouse brain.

Relaxant effects of isoproterenol in isolated cardiac muscle: influence of loading patterns.

The purpose of this study was to test the hypothesis that loading patterns (i.e., loading sequence, total load, and preload) modulate the relaxant effects of isoproterenol. The effects of isoproterenol (10(-6) M) on peak rate of force decline (-dF/dt) were studied in rat left ventricular papillary muscle (n = 24) with respect to two sequences of relaxation: the classical, isotonic-isometric sequence, in which tension fall occurs at initial muscle length, and the physiological, isometric-isotonic sequence, in which tension fall occurs at end-systolic muscle length. The influences of muscle load and initial length were accounted for in the evaluation of relaxation rate by plotting -dF/dt against the entire range of loads both at preload = maximum length (Lmax) and 90% Lmax. The main results are the following: 1) in the classical, isotonic-isometric sequence of relaxation, and whatever the preload, the magnitude of the relaxant effect of isoproterenol increased with load; 2) after reversal into the physiological, isometric-isotonic sequence of relaxation, the relaxant effect of isoproterenol behaved independently of load level in muscle preloaded at Lmax; 3) conversely, in muscle preloaded at 90% Lmax and relaxing according to the physiological sequence, the relaxant effect of isoproterenol increased with load; and 4) the peak relaxant effect of isoproterenol was proportionally higher in the physiological sequence of relaxation than in the classical one and occurred at a similar level of load, whatever the loading sequence and whatever the preload level. Our results indicate that loading patterns finely modulated the relaxant effects of isoproterenol and that muscle length, both before the contraction phase and at the onset of relaxation phase, influenced the effects of isoproterenol on myocardial relaxation rate.

Implantation of rabbit embryo brain fragments into newborn mice: integration and survival of xenogeneic astrocytes.

Brain fragments containing embryonic rabbit glia were implanted into the brains of newborn mice. The hosts developed an astroglial reaction around the transplants and along the needle tracks. Transplant-derived astrocytes were identified in the operated brain by their expression of rabbit GFAP. During the first few days post-implantation (PI) glial cells were exchanged between the transplant and the host. Less than 3 to 5 days PI, the transplant was extensively invaded by host astrocytes. Xenogeneic astroglial cells were first detected 10 days PI in the immediate proximity of the transplant. At 2 to 11 weeks, they could be detected either close to or at distance from the point of implantation. Most often, transplant-derived astrocytes presented a morphology similar to that of neighboring host astrocytes. Xenogeneic glial cells were found to participate in various types of astroglial features: sub-pial, pericapillary, fibrous, and protoplasmic. This morphological integration suggests that they are physiologically integrated, at least to a certain degree, in the host tissue. In spite of their integration into the host, xenogeneic astrocytes disappear after 3 months without signs of an inflammatory reaction.

Survival of astroglial cell lineage from adult brain transplant.

We previously described a transplantation model which enables the selective identification of rabbit astroglial cells in sections of mouse brain tissue. In the present paper, we report that cells of the astroglial lineage from an adult transplant are able to survive after implantation into the brain of an immature host. Surviving xenogenic cells are found outside the graft, forming colonies in the host brain parenchyma. However, after 3 months, transplant-derived astrocytes are no longer detected.