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C Abbadie

Publications and source records attributed to C Abbadie.

35 records · Page 2Linked to original sources
PubMed1997

Inflammation increases the distribution of dorsal horn neurons that internalize the neurokinin-1 receptor in response to noxious and non-noxious stimulation.

Although the neurokinin-1 (NK-1)/substance P (SP) receptor is expressed by neurons throughout the spinal dorsal horn, noxious chemical stimulation in the normal rat only induces internalization of the receptor in cell bodies and dendrites of lamina I. Here we compared the effects of mechanical and thermal stimulation in normal rats and in rats with persistent hindpaw inflammation. Electron microscopic analysis confirmed the upregulation of receptor that occurs with inflammation and demonstrated that in the absence of superimposed stimulation, the increased receptor was, as in normal rats, concentrated on the plasma membrane. In general, noxious mechanical was more effective than noxious thermal stimulation in inducing NK-1 receptor internalization, and this was increased in the setting of inflammation. Although a 5 sec noxious mechanical stimulus only induced internalization in 22% of lamina I neurons in normal rats, after inflammation, it evoked near-maximal (98%) internalization in lamina I, produced significant changes in laminae III-VI, and expanded the rostrocaudal distribution of neurons with internalized receptor. Even non-noxious (brush) stimulation of the inflamed hindpaw induced internalization in large numbers of superficial and deep neurons. For thermal stimulation, the percentage of cells with internalized receptor increased linearly at >45 degrees C, but in normal rats, these were restricted to lamina I. After inflammation, however, the 52 degrees C stimulus also induced internalization in 25% of laminae III-IV cells. These studies provide a new perspective on the reorganization of dorsal horn circuits in the setting of persistent injury and demonstrate a critical contribution of SP.

Animals↗
PubMed1997

The differential contribution of capsaicin-sensitive afferents to behavioral and cardiovascular measures of brief and persistent nociception and to Fos expression in the formalin test.

Intraplantar injection of dilute formalin evokes brief (Phase 1) and persistent (Phase 2) increases in primary afferent activity, pain behavior, and cardiovascular responses, and induces spinal cord Fos-like immunoreactivity (Fos-LI). Although previous studies demonstrated that the destruction of small diameter primary afferents with neonatal capsaicin treatment decrease formalin-evoked nociception, these studies only evaluated behavioral responses, and did not distinguish between Phase 1 and 2. To address these questions, we simultaneously evaluated formalin-evoked pain behavior (flinching of the afflicted paw), cardiovascular responses (heart rate and mean arterial pressure), and lumbar spinal cord Fos expression in control rats and in rats treated with capsaicin (100 mg/kg) one day postpartum. We found that neonatal capsaicin-treated rats, compared to controls, exhibited similar cardiovascular responses and slightly less flinching behavior during Phase 1. During Phase 2, however, capsaicin-treated rats exhibited 59% less flinching and 45% smaller heart rate responses. Also, in capsaicin-treated rats, we counted 59% fewer Fos-labeled neurons in the spinal cord. These results indicate that capsaicin-sensitive afferents contribute to formalin-evoked behavioral and cardiovascular responses and to spinal cord neuronal responses. The differential effect of neonatal capsaicin on nociception during Phase 1 and Phase 2 suggests that sensitization mechanisms during Phase 1 do not contribute to the magnitude of nociceptive responses during Phase 2.

Afferent Pathways↗
PubMed1997

Contribution of sacral spinal cord neurons to the autonomic and somatic consequences of withdrawal from morphine in the rat.

In this study, we monitored Fos-like immunoreactivity in the sacral spinal cord to identify neurons that are likely to contribute to the autonomic manifestations of opioid antagonist-precipitated withdrawal in morphine-tolerant rats. Injection of systemic antagonist increased the Fos-like immunoreactivity throughout the first sacral segment, particularly in laminae I/II, X, and in the sacral parasympathetic nucleus (SPN). Selective peripheral withdrawal, with a hydrophilic antagonist that does not cross the blood-brain barrier (BBB), induced diarrhea, but no other withdrawal signs were evident. Compared to rats that withdrew systemically, peripherally withdrawal evoked significantly less Fos-like immunoreactivity in laminae V/VI, X and the SPN. By contrast, selective spinal withdrawal, by intrathecal injection of an opioid antagonist that does not cross the BBB, provoked hyperactivity of the hindlimbs and tail, but no diarrhea. These animals demonstrated significantly increased Fos-like immunoreactivity in laminae I/II, V/VI, the SPN, and the ventral horn compared to rats that withdrew systemically. Animals treated neonatally with capsaicin, to eliminate C-fiber input, demonstrated withdrawal behavior similar to intact withdrawing rats, except that the capsaicin-pretreated rats had significantly greater weight loss. However, this group had less Fos-like immunoreactivity in laminae V/VI, X and SPN compared to the intact withdrawing rats. These data suggest that withdrawal from morphine evokes hyperactivity of sacral neurons, particularly those involved in regions that process nociceptive and autonomic information. Peripheral withdrawal is sufficient to induce diarrhea, but it does not fully explain the associated weight loss. Unmyelinated primary afferents may contribute a tonic peripheral inhibition of circuits that regulate gut motility and intestinal fluid transport. Taken together, these data suggest that chronic exposure to opioids induces a latent sensitization in sacral cord neurons that can be manifested as neuronal hyperactivity during withdrawal; this mechanism may underlie withdrawal-induced hyperalgesia and gut hypermotility.

Animals↗
PubMed1997

Differential contribution of the two phases of the formalin test to the pattern of c-fos expression in the rat spinal cord: studies with remifentanil and lidocaine.

Injection of formalin in the rat hindpaw produces two phases of nociceptive behavior. Although it is generally agreed that the first phase results from direct chemical activation of nociceptive primary afferent fibers, the factors that contribute to the second phase are not established. In the present study, we monitored the expression of the c-fos protein to evaluate whether the pattern of activity of dorsal horn neurons differs as a result of ongoing afferent activity during the two phases. To selectively block the first or second phase, we respectively used remifentanil, a potent and short acting opiate agonist, and QX-314, a quaternary derivative of lidocaine, which does not cross the blood brain barrier. We also evaluated the effect of eliminating nociceptive behavior in both phases using both remifentanil and lidocaine or a combination of local anesthetics, bupivicaine and quaternary lidocaine. In all groups, formalin (5%, 50 microliters) was injected subcutaneously into the plantar surface of the hindpaw. To assess the nociceptive behavior produced by formalin, we monitored the number of flinches. Injection of remifentanil during the first phase completely blocked the first phase formalin-evoked nociceptive behavior, and had no effect on the second phase. Injection of lidocaine during the interphase completely blocked second phase nociceptive behavior. As expected, when remifentanil was administered during the first phase and lidocaine during the second phase, all formalin-evoked nociceptive behavior was blocked. The same was true for rats that received injections of bupivicaine and lidocaine during phases 1 and 2, respectively. In laminae I-II of the L4-L5 segment, the magnitude of the decrease in Fos expression was comparable for remifentanil (26.5%) and lidocaine (27.3%); the decrease was greater when both remifentanil and lidocaine were administered (50.5%), and even greater when bupivicaine and lidocaine were used (74.2%). In laminae V-VI, remifentanil, by itself, decreased c-fos expression by 39.4%; for lidocaine alone, the decrease was 58.4%. We did not observe further significant decreases when both remifentanil and lidocaine, or bupivacaine and lidocaine were injected (69.7% and 74.6%, respectively). Our results not only provide strong evidence that activity during the second phase is necessary for maintaining the maximal expression of c-fos in the spinal cord, but also reveal significant regional differences in the central patterns of activity generated during the two phases. These results also confirm our previous reports that c-fos expression is not eliminated when the behavioral manifestation of the noxious stimulus is completely blocked.

Analgesics, Opioid↗
PubMed1997

The contribution of supraspinal, peripheral and intrinsic spinal circuits to the pattern and magnitude of Fos-like immunoreactivity in the lumbar spinal cord of the rat withdrawing from morphine.

Withdrawal from morphine evokes increases in Fos-like immunoreactivity in the spinal cord, particularly in the superficial dorsal horn, laminae I/II. To determine the origin of the increased Fos-like immunoreactivity, we selectively targeted central or peripheral opioid receptors with naloxone-methiodide, an antagonist that does not cross the blood-brain barrier, or induced withdrawal after eliminating possible sources of input to the superficial dorsal horn. To induce tolerance, we implanted rats with morphine or placebo pellets (75 mg, six pellets over three days). On day 4, withdrawal was precipitated and after 1 h, the rats were killed, their spinal cords removed and 50 microm transverse sections of the spinal cord immunoreacted with a rabbit polyclonal antiserum directed against the Fos protein. In placebo-pelleted rats, none of the different procedures, viz. spinal transection, unilateral dorsal rhizotomy (L4-S2), neonatal capsaicin treatment or direct intrathecal opioid antagonist injection, induced expression of the Fos protein. However, both spinally transected and rhizotomized withdrawing animals showed significant increases in Fos-like immunoreactivity in laminae I/II, compared to intact withdrawing rats. Neonatal treatment with capsaicin, which eliminates C-fibres, did not alter Fos-like-immunoreactivity. Selective withdrawal of morphine from peripheral opioid receptors by naloxone-methiodide did not induce Fos-like immunoreactivity in the lumbar spinal cord greater than that recorded in nonwithdrawing rats. However, intrathecal injection of naloxone-methiodide increased Fos-like immunoreactivity in laminae I/II and the ventral horn to a greater extent than did subcutaneous injection of naloxone. We hypothesize that the increased Fos expression after systemic withdrawal in spinally-transected rats results from a loss of descending inhibitory control that is activated during withdrawal. The increase in withdrawal-induced Fos-like immunoreactivity after rhizotomy may be secondary to loss of inhibitory controls exerted by large diameter primary afferents or to deafferentation-induced reorganization in the dorsal horn. Since capsaicin did not alter the magnitude of Fos-like immunoreactivity in withdrawing rats, we conclude that hyperactivity of opioid receptor-laden C-fibres is not a necessary contributor to the withdrawal-induced increase in Fos-like immunoreactivity in laminae I and II. Taken together with the results recorded after intrathecal injection of naloxone-methiodide in tolerant rats, we conclude that the pattern of lumbar spinal cord Fos expression following systemic withdrawal is primarily a consequence of increased activity in opioid receptor-containing circuits intrinsic to the dorsal horn and that the magnitude of Fos expression is normally dampened by supraspinal and primary afferent-derived inhibitory inputs.

Animals↗
PubMed1997

The avian transcription factor c-Rel is induced and translocates into the nucleus of thymocytes undergoing apoptosis.

This study investigates the involvement of the avian transcription factor c-Rel in thymocyte apoptosis occurring either in vivo or in organotypic culture. In vivo, only a few cortical thymocytes express the c-Rel protein. Their number, localization and morphology resemble that of apoptotic cells evidenced by TUNEL staining. In organotypic culture, the expression of c-Rel is induced in medullary thymocytes as apoptosis is triggered. This induction would be post-transcriptional since no increase in the c-rel gene expression is detected. Moreover, c-Rel translocates into the nucleus of medullary thymocytes during the time course of apoptosis. This translocation is preceded by a decrease in ikba expression, the gene which encodes the avian homologue of IkappaBalpha. Altogether these results suggest that the proto-oncogene c-rel could take an active part in apoptosis of cortical thymocytes occurring in vivo during T-cell selection as well as in experimentally-induced apoptosis of medullary thymocytes.

Journal Article↗
PubMed1996

Spinal cord substance P receptor immunoreactivity increases in both inflammatory and nerve injury models of persistent pain.

Numerous studies have implicated the primary afferent derived neuropeptide, substance P, which exerts its effects via the neurokinin-1/substance P receptor, in the transmission of nociceptive messages at the level of the spinal cord. Immunocytochemical studies demonstrate that the substance P receptor is concentrated in neurons of lamina I of the superficial dorsal horn. Since alterations in the number and distribution of the receptor may underlie persistent pain conditions, we have used immunocytochemistry to study the distribution of the receptor in two very different rat models of persistent pain: chronic inflammation, which is associated with increased levels of substance P, and sciatic nerve section, which is associated with decreased levels of substance P in the dorsal horn. Inflammation was produced by unilateral hindpaw injection of complete Freund's adjuvant. We report that there is an up-regulation of substance P receptor immunoreactivity in the superficial laminae of the dorsal horn in both injury models. The increase was found at all time points studied (up to one week after induction of inflammation and up to two weeks after sciatic nerve section). The increase in substance P receptor immunoreactivity was not only present in the medial part of the dorsal horn at segment L4, which is the region of input of the afferents from the hindpaw, but also in the lateral parts of the dorsal horn, and at segments rostral (L1) and caudal (S1) to the afferent input from the hindpaw. These results indicate that the up-regulation of the receptor is not predictable merely by the change in the concentration of substance P in the dorsal horn. Furthermore, the non-topographic up-regulation of substance P receptor in these different conditions may contribute to the central sensitization of dorsal horn nociceptors under conditions of persistent pain.

Animals↗
PubMed1994

Effects of opioids and non-opioids on c-Fos-like immunoreactivity induced in rat lumbar spinal cord neurons by noxious heat stimulation.

This study evaluated Fos-like immunoreactivity in rat lumbar spinal cord neurons following peripheral noxious heat stimulation and the modifications induced by pharmacological agents. Under urethane anaesthesia, the hindpaw was stimulated by dipping it in a regulated temperature bath at various temperatures (44-65 degrees C) and for various durations (5 s to 2 min). There was no Fos-like immunoreactivity in lumbar spinal cord neurons when the paw was stimulated at 44 degrees C for 15 s. From 46 to 52 degrees C, the number of Fos-like immunoreactivity neurons increased with increasing stimulation temperature, but was decreased at 65 degrees C as compared to 52 degrees C. At 52 degrees C, the number of Fos-like immunoreactivity neurons increased with the duration of stimulation. Fos-like immunoreactive neurons in the L4 segment were almost exclusively located in laminae I-II. On the basis of the results of the latter experiments, we chose a stimulation of 52 degrees C for 15 s to perform pharmacological investigations. The number of Fos-like immunoreactive neurons induced by the heat stimulation was significantly decreased by pretreatment with morphine (42, 64 and 75% decrease as compared to control values after 2.5, 5 and 7.5 mg/kg i.v. respectively), and these effects were blocked by naloxone. When various stimulation intensities (46-52 degrees C) were used, the effects of morphine (5 mg/kg i.v.) were most marked when the temperature was highest. In morphine-tolerant rats, morphine (5 mg/kg i.v.) was half as potent in decreasing Fos-like immunoreactivity induced by the heat stimulation than in non-tolerant rats. RB 101, a systemically active mixed inhibitor of enkephalin-metabolising enzymes, significantly decreased Fos-like immunoreactivity induced by heat stimulation (19, 29 and 48% decreases as compared to control values at 10, 20 and 40 mg/kg i.v. respectively) and these effects were blocked by naloxone. Aspirin (150 mg/kg i.v.), proacetaminophen (300 mg/kg i.v.) and tizanidine, a centrally acting myorelaxant (0.25-1 mg/kg i.v.), had no effect on the number of Fos-like immunoreactivity neurons induced by heat stimulation. The use of immunochemistry of the c-Fos protein as a pharmacological test in order to gauge antinociceptive effects at the dorsal horn level is discussed.

Analgesia↗
PubMed1994

Intense cold noxious stimulation of the rat hindpaw induces c-fos expression in lumbar spinal cord neurons.

This study evaluated Fos-like immunoreactivity in lumbar spinal cord neurons following intense cold stimulation and then the modifications induced by opioid administration. Under urethane anaesthesia, the rat's right foot was stimulated by holding it in a regulated temperature bath at 15, 10, 0, -10, -15, -17.5 or -20 degrees C. There was no or little Fos-like immunoreactivity in lumbar spinal cord neurons when the paw was at temperatures between 15 and -10 degrees C (0-5 Fos-like immunoreactive neurons/section). The threshold to induce consistent c-fos expression was -15 degrees C. From -15 to -20 degrees C, the number of Fos-like immunoreactive neurons increased with decreases in temperature. At -20 degrees C, Fos-like immunoreactive neurons were numerous in L3 and L4 segments, in laminae I-II (approximately 60 Fos-like immunoreactive neurons/section) and to a lesser extent in laminae V-VI (approximately 20). Almost no Fos-like immunoreactivity was present in laminae III-IV (< 5). At -20 degrees C, the number of Fos-like immunoreactive neurons increased with the duration of the stimulation. The number of Fos-like immunoreactive neurons induced by the cold stimulation temperatures was significantly decreased by pretreatment with 10 mg/kg s.c. morphine and moderately decreased by 5 mg/kg s.c. This effect was antagonized by the combined administration of morphine (10 mg/kg s.c.) and naloxone (2 mg/kg s.c.). Naloxone (2 mg/kg s.c.) significantly increased the number of Fos-like immunoreactive neurons induced by -20 degrees C as compared to saline-injected rats. This study showed that Fos-like immunoreactivity distribution is in good agreement with the location of neurons receiving noxious inputs and that the threshold to induce c-fos expression with cold was unexpectedly low at -15 degrees C. Taking into account, on the one hand, previous investigations using the same technique using noxious heat stimulation and, on the other hand, electrophysiological and psychophysiological studies using cold stimulation in animals and humans, our results suggest that Fos-like immunoreactivity induced by extremely cold stimulation, which seems to reproduce frostbite, may reflect activation of nociceptors due to vasoconstriction.

Animals↗
PubMed1994

c-Fos expression in the spinal cord and pain-related symptoms induced by chronic arthritis in the rat are prevented by pretreatment with Freund adjuvant.

We previously showed that pathological adjuvant-induced arthritis (AIA), a chronic pain model, is associated with an increase in the number of Fos-like immunoreactivity (Fos-LI) in lumbar spinal cord neurons; maximal Fos-LI corresponds to the peak of pathological and behavioral signs of the disease. On the other hand, AIA is prevented in rats that have previously been injected with diluted complete Freund adjuvant (CFA). Here, the effects of pretreatment with CFA were studied in parallel on both pathological parameters and pain-related tests, and on Fos-LI, 3 weeks after the inoculation of the arthritogenic solution during the acute phase of the disease, that is, when hyperalgesia is maximal. One group of rats (vaccinated group) was pretreated with diluted CFA, 3 weeks and 1 week before inoculation (AIA) (with the concentrated arthritogenic solution), while a second group (arthritic group) was pretreated with the vehicle for the CFA solution and then inoculated with the concentrated arthritogenic solution. Control groups consisted of rats treated with various combinations of either saline or vehicle solutions during the pretreatment periods or at the time of inoculation. The arthritic group developed all pathological symptoms of AIA (i.e., loss of weight, difficulties in walking, increases in paw diameter, and decreases in the vocalization thresholds to mechanical pressure of the hindpaw, and finally an increase in the vocalization response to flexion and extension). In contrast, the vaccinated group did not develop AIA symptoms.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

High levels of c-rel expression are associated with programmed cell death in the developing avian embryo and in bone marrow cells in vitro.

To determine the physiological processes in which the transcription factor c-Rel may act, we have examined its pattern of expression in the avian embryo by in situ hybridization. These studies showed that c-rel is expressed ubiquitously at low levels and at high levels in isolated cells undergoing programmed cell death by apoptosis or autophagocytosis. To further establish a functional link between expression of c-rel and cell death, we examined the biological consequences of c-rel overexpression in vitro. In primary avian fibroblasts, overexpression of c-rel leads to transformation and dramatic life span extension. In contrast, bone marrow cells expressing high levels of c-rel undergo a process of programmed cell death displaying features of both apoptosis and autophagocytic cell death. Thus, these experiments suggest a critical role for c-rel not only in the control of cell proliferation, but also in the induction of cell death.

Age Factors↗
PubMed1993

C-fos expression in rat lumbar spinal cord following peripheral stimulation in adjuvant-induced arthritic and normal rats.

Our previous data reported a maximal expression of the c-fos immediate-early gene in the lumbar spinal cord of the non-stimulated polyarthritic rat neurons, three weeks after Freund's adjuvant injection. The present study utilises c-fos expression to judge the reactivity of spinal neurons to calibrated mechanical pressure applied to the ankle joint, in both normal and arthritic rats under ketamine anesthesia. The results indicate that the number of Fos-like immunoreactive neurons (1) is slightly decreased in ketamine-anesthetized non-stimulated arthritic rats as compared to the non-anesthetized non-stimulated ones, (2) is significantly higher in both stimulated normal and arthritic animals as compared to non-stimulated animals, particularly in laminae I, II, V and VI of L3 and L4, and (3) is significantly increased in stimulated arthritic as compared to stimulated normal rats, in all laminae of lumbar spinal segments. The appearance of 'basal' Fos labeling during the adjuvant-induced arthritic disease and the increased number of Fos-like immunoreactive neurons in stimulated arthritic rats compared to stimulated normal animals indirectly suggests that these neurons are abnormally active and thus involved in the hyperalgesia of arthritic disease. Therefore the use of Fos-like immunoreactivity in the chronic pain model seems to be an appropriate tool to study possible effects of various pharmacological compounds, such as analgesics and anti-inflammatory drugs.

Anesthesia↗
PubMed1992

Fos-like immunoreactivity in the rat superficial dorsal horn induced by formalin injection in the forepaw: effects of dorsal rhizotomies.

As previously described at the lumbar spinal level, we found that 2 h after subcutaneous formalin injection in the distal part of the fore-limb, Fos-like immunoreactivity (FLI) was induced in the ipsilateral cervical enlargement. Not surprisingly, as the injection site corresponds to the distal part of the C6-C8 dorsal root dermatomes, maximal labelling which predominated in the superficial laminae, was observed in the C6-C8 segments and to a lesser extent in C5. Similar experiments were performed on rats which underwent various types of unilateral dorsal rhizotomies (DRh) 7 days before formalin injection. In animals with C4, C5, T1 and T2 DRh sparing C6-C8 the rostrocaudal distribution was similar to the intact one. But, in animals having C4-T2 DRh sparing one single root, C7, the segmental FLI distribution was modified: it was slightly increased in C7, decreased in C6 and significantly decreased in C8. As expected, no FLI was found in animals with C4 to T2 DRh. The spared root model provides information about the segmental distribution in the cervical spinal cord of the input brought by a single root following stimulation of the distal forelimb, i.e., maximal distribution in the entry segment, but also in the two rostral and one caudal segments.

Animals↗
PubMed1992

c-fos expression in rat lumbar spinal cord during the development of adjuvant-induced arthritis.

A parallel clinical and behavioral study of adjuvant-induced arthritis in the rat showed four stages in the time-course of the disease: preclinical (first week), acute (weeks 2-4), post-acute (weeks 5-8) and recovery weeks 9-11) [Calvino et al. (1987) Behav. Brain Res. 24, 11-29]. As several studies have reported the expression of the proto-oncogene c-fos in spinal cord neurons following acute noxious peripheral stimuli, the aim of this study was to quantitatively assess Fos-like immunoreactivity in lumbar spinal cord neurons at various times of adjuvant-induced arthritis development, i.e. one, two, three, 11 and 22 weeks post-inoculation. The total number of Fos-like immunoreactive neurons in the lumbar enlargement correlated with the observed development of adjuvant-induced arthritis, i.e. Fos-like immunoreactivity was absent at one week, moderate at two weeks, greatly increased at three weeks, decreased at 11 weeks and returned to control values at 22 weeks. At three weeks, at the peak of Fos-like immunoreactivity distribution and acute stage of hyperalgesia, maximal labeling was observed in L3 and L4 spinal segments. In these segments, the most densely labeled region was the neck (laminae V and VI) of the dorsal horn (55%) and the ventral horn (35%) as compared to the superficial laminae (laminae I and II; 5%) and the nucleus proprius (laminae III and IV; 5%). These data indicate that c-fos expression induced by chronic inflammation is better expressed in deeper laminae than in the superficial ones, and that the number of Fos-positive cells correlates with behavioral studies.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

The acentriolar state of the Drosophila cell lines 1182.

A Drosophila melanogaster cell line devoid of centrioles has been recently described. In order to achieve an easier characterization of these acentriolar cells, we used the monoclonal antibody Bx 63 of M. Frasch which recognizes the Drosophila centrosome. Although centrosomes are detected at every mitotic pole in Drosophila cells with centrioles, no such structure has been observed in 1182-4 acentriolar cells. The antigenic material is, however, present in these cells. Moreover, we noticed a certain proportion of acentriolar cells in 4 other 1182 lines. The lack of centrioles previously found only in the 1182-4 cells seems therefore not accidental and should be linked to their particular origin.

Animals↗
PubMed1985

Monoclonal antibodies to lampbrush chromosome antigens of Pleurodeles waltlii.

Germinal vesicles of oocytes from Pleurodeles waltlii were used for immunization of BALB/c mice to obtain hybridomas secreting monoclonal antibodies. The hybridomas were screened for reactivity of their antibodies against lampbrush chromosomes of oocytes, as revealed by indirect immunostaining. Antibodies labelling the lampbrush chromosomes were also tested on histological sections of oocytes, embryos, and larvae of Pleurodeles. Characterization of the antigens was accomplished through immunoblotting of two-dimensional electrophoretic gels of germinal vesicle proteins. The ten monoclonal antibodies giving a positive reaction were classed into five groups. Group 1, exemplified by antibody A33, recognizes all the lampbrush chromosome transcribing sites (loops). Moreover, it differentially labels the cell nuclei during embryonic and larval development. Group 2, antibody B71, also stains all the loops of the lampbrush chromosomes, but does not react with cell nuclei of embryos and larvae. Group 3, antibody A1, labels specific loops, some of which are heterozygous in the strain of P. waltlii used. These heterozygosities have allowed us to localize and to characterize a chromosomal segment on bivalent IV which is heteromorphic in the two partners of the bivalent. We suggest that this heteromorphism represents a morphological distinction between Z and W heterochromosomes. Moreover, this antibody reacts with only one transcription unit along a loop that contains several units. Group 4, antibody B24, stains the only two structures in the lampbrush chromosomes of P. waltlii that do not have a loop organization, the mass "M" and the spheres. Group 5, antibody A35, reacts with the chromomeres. The antigens corresponding to antibodies A33 and B24 have been identified as proteins, which have apparent molecular weights of 80 and 104 kilodaltons, respectively. They correspond to proteins abundant in the germinal vesicles. All the antibodies described here cross-react with the lampbrush chromosomes of five other species of Urodeles.

Animals↗