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Introduction: In the treatment of very low rectal cancer, a distal resection margin of more than 1 cm can be obtained by partial internal sphincteric resection, allowing a sphincter preserving surgery. Thus, intersphincteric resection (ISR) has been proposed as an alternative to abdominoperineal resection for selected low rectal cancer., Objective: The aim of our study was to assess the morbidity, mortality, and the long-term oncologic and functional results of ISR., Methods: Charts of patients who had ISR between 1992 and 2004 were reviewed. Cancer-related survival and locoregional recurrence rates were calculated using the Kaplan-Meier method. Functional outcome was assessed by using a standardized gastrointestinal functional questionnaire. Incontinence was assessed by the continence score of Wexner., Results: Ninety patients (59 males, 31 females) with a tumor at a median distance of 35 mm (range, 22-52) from the anal verge had an ISR. Thirty-seven patients (41%) had preoperative radiotherapy. Histologically complete remission after neoadjuvant radiotherapy (ypT0) was observed in 7 patients (8%), 12 patients (13%) were pT1, 35 patients (39%) pT2, 32 patients (36%) pT3, and 4 patients (4%) pT4. Five patients (5.5%) had synchronous liver metastases. R0 resection was obtained in 85 patients (94.4%). The median distal resection margin on the fixed specimen was 12 mm (range, 5-35) and was positive in 1 case. The circumferential margin was positive (< or =1 mm) in 4 patients (4.4%). There was no mortality. Complication rate was 18.8%: anastomotic leakage occurred in 8 patients (8.8%) and 1 patient had an anovaginal fistula. Five patients (5.6%) underwent secondary abdominoperineal resection: 1 for positive distal margin, 1 for colonic J-pouch necrosis, and 3 for local recurrence. ONCOLOGIC RESULTS: After a median follow-up of 56.2 months (range, 13.3-168.4), local, distant, and combined recurrence occurred in 6 (6.6%), 8 (8.8%), and 2 patients, respectively. Thirteen patients (14.4%) died of cancer recurrence. Five-year overall and disease-free survival was 82% (80-97) and 75% (64-86), respectively. In univariate analysis, overall survival was significantly influenced by pTNM stage and T stage (pT 1-2 vs. 3-4: P = 0.008 and stage I-II vs. III-IV: P = 0.03). In multivariate analysis, we did not find any impact on local recurrence-free survival for the investigated prognostic variables., Functional Results: For a total of 83 patients the mean stool frequency was 2.3 +/- 1.3 per 24 hours. Forty-one percent of patients had stool fragmentation, one-third nocturnal defecation, 19% fecal urgency, and 36% followed low fiber diet. Thirty-four patients (41%) were fully continent, 29 patients (35%) had minor continence problems, and 20 patients (24%) were incontinent. After adjustment for age, gender, tumor level, and pTNM stage, preoperative radiotherapy was the only factor associated with a risk of fecal incontinence [OR (IC 95%) = 3.1 (1.0-9.0), P = 0.04]., Conclusion: In selected patients, ISR is a safe operation with good oncologic results. It achieves good functional results in 76% of patients. Functional results are significantly altered by preoperative radiotherapy.

Objective: To assess in a population of stress incontinent patients without genital prolapse whether urethrovesical junction mobility is associated with global pelvic floor laxity., Methods: Dynamic MRI of 40 patients referred prior to surgery for urinary stress incontinence were reviewed retrospectively. The orientation of the urethrovesical junction was evaluated at rest and at straining in reference to the pubococcygeal line, and defined as the bladder neck pubococcygeal angle. The urethrovesical junction mobility was calculated as the difference between the bladder neck pubococcygeal angles at rest and at straining. Urethrovesical junction mobility and bladder neck pubococcygeal angles at rest and at straining were tested for correlations with the resting and straining position of different pelvic organs, urogenital hiatus size, levator plate angle, and anterior rectal bulging when straining., Results: Urethrovesical junction mobility was correlated with the position of the bladder neck (p<0.0001), bladder base (p<0.0001) and uterine cervix (p<0.0001) at straining, as well as the hiatus length (p=0.0012) and width (p=0.0002), and levator plate angle (p <0.0001). The bladder neck pubococcygeal angle at rest was correlated with the resting position of the bladder neck (p <0.0001), bladder base (p <0.0001), uterine cervix (p=0.02), and the hiatus length (p=0.0004) and width (p=0.045) at rest, whereas the bladder neck pubococcygeal angle at straining was correlated with the straining position of the bladder neck (p <0.0001), bladder base (p=0.0001), uterine cervix (p <0.0001), and hiatus length (p=0.0005) and width (p=0.0004), and levator plate angle (p <0.0001) at straining., Conclusion: In a population of stress incontinent patients, the urethrovesical junction mobility was correlated with global pelvic floor laxity.

Objective: The aim of this study was to determine the frequency of MYH mutations in one large population of polyposis patients without APC mutation identified., Summary Background Data: Familial adenomatous polyposis (FAP) is the most known inherited colorectal cancer syndrome. In 70% to 80% of polyposis patients, an APC mutation is found. Patients with polyposis but no APC mutation are considered as APC-muted patients and followed as their relatives accordingly. Biallelic mutation of MYH has been found to responsible of colorectal polyposis and cancer in an autosomal recessive pattern of inheritance., Methods: Between 1978 and 2004, 433 patients were operated for polyposis. A mutation on APC was identified in 322 patients. Among the remaining patients, 44 were identified as possible MYH-muted patients and contacted, and 31 signed informed consent. Clinical data were obtained from the patients' medical notes. Germline mutation of MYH was searched by sequencing the whole gene. To confirm the deleterious effects of biallelic MYH mutation, transversions on K-ras and APC were searched., Results: There were 9 women and 22 men with a mean age of 53.9 years (range, 22-68 years) at the time of diagnosis. The mean number of polyps was 62.8 (range, 11-266). Eighteen patients (58.1%) had a colorectal cancer. We found biallelic MYH mutation in 6 patients (19.3%; 95% confidence interval, 5.2%-33.5%) and 5 (83.3%) had transversions in K-ras and/or APC., Conclusion: MYH is a new gene responsible for about 1.4% of all adenomatous polyposis and about 20% of adenomatous polyposis without APC mutation identified. Search for MYH biallelic mutation in these patients should be systematic as it changes their and relatives'surveillance.

Transmembrane isoforms of adenylate cyclases (AC) integrate a wide variety of extracellular signals from neurotransmitters to morphogens and can also regulate cAMP production in response to calcium entry. Based on observations in the barrelless mouse strain, the Adcy1 gene (AC1) was involved in the segregation of binocular retinal inputs. To determine the potential role of other AC isoforms we localized the Adcy genes in the visual centres during development, using in situ hybridization. Six different AC subtypes were found in the developing retinal ganglion cell layer (RGC; AC1, AC2, AC3, AC5, AC8, and AC9), and three AC subtypes were expressed in the central brain targets, the dorsal lateral geniculate nucleus (AC1 and AC8), the ventral lateral geniculate nucleus (AC2 and AC8) and the superior colliculus (AC1, AC2, AC8). Using a genetic approach we tested the role of the calcium modulated cyclases AC1, AC5 and AC8 for the segregation retinal fibres. Ipsilateral retinal axons remained exuberant in the AC1(-/-) mice, with overlapping retinal projections from both eyes in the superior colliculus and the visual thalamus. These abnormalities were similar to those of barrelless mouse mutants. No abnormalities were detectable in the AC5(-/-) or the AC8(-/-) mice. Similar abnormalities were noted in the single AC1(-/-) and the AC1/AC8 double-knockout mice (DKO). Thus, only AC1 is required for the maturation of the retinal axon terminals whereas AC5 and AC8 are not needed. The specificity of AC1's action is linked to its cellular localization in the RGCs and to its distinctive functional profile, compared with the other cyclases expressed in the same cells.

Background: Various surgical procedures are currently performed for the treatment of posthysterectomy vesicovaginal fistula. Most of them are carried out via an abdominal incision., Objective: Report of our experience with Latzko's operation, a simple purely transvaginal procedure., Methods: Retrospective study of vesicovaginal fistulas treated by a Latzko procedure between June 1991 and June 2005., Results: Eleven patients were operated on. Two (18%) had a prior failed attempt of vesicovaginal fistula repair. Mean size of the fistulas was 12+/-11 mm (range: 2-40 mm). Mean operative duration was 62+/-33 min (range: 20-110 min). All fistulas healed. No intraoperative complications were observed. There was only one postoperative complication, a lower urinary tract infection in one patient. Mean hospital stay was 6+/-4 days (range: 2-12 days)., Conclusions: The Latzko procedure is an efficient, safe, and simple technique for the management of vault vesicovaginal fistulas, and can therefore be proposed as the first-line surgical treatment.

Background: To describe the static and dynamic MRI features of the levator ani, and evaluate whether they are associated with the MRI evaluation of the severity of genital prolapse., Methods: Static and dynamic MRI of 40 patients, referred for evaluation prior to genital prolapse surgery, were reviewed retrospectively. Prolapse severity was evaluated on MRI at maximal straining by descent of the bladder neck under the pubococcygeal line for the anterior compartment, by descent of the uterine cervix under the pubococcygeal line for the middle compartment, and by anterior bulging of the rectum for the posterior compartment. For evaluation of the levator ani, the following parameters were recorded: (1) at rest: thinning or defects in both puborectalis and iliococcygeus muscles, (2) at rest and at straining: urogenital hiatus length and width, M line, iliococcygeal and levator plate angles. The levator ani features were tested for potential associations with the MRI evaluation of prolapse severity., Results: Bladder neck descent at straining was correlated with the levator plate angle at rest (p=0.001), and with the hiatus length at rest (p=0.02), and at straining (p=0.008). Uterine cervix descent at straining was correlated with the hiatus length (p=0.0005), and width (p=0.014) at straining, M line (p=0.002) and levator plate angle (p=0.007) at straining, whereas anterior rectal bulging at straining was paradoxically inversely correlated with the hiatus width at rest (p = 0.04)., Conclusion: In a population of women with genital prolapse, MRI evaluation of the levator ani was associated with MRI evaluation of the severity of genital prolapse.

Chameleons (Order, Reptilia: Family, Lacertilia) are unique among vertebrates in being able to make independent eye movements. The organisation of their retina, however, closely ressembles that of other diurnal lizards; based on morphological studies, it is typically described as containing only cone photoreceptors. We show here that a subpopulation of the photoreceptors are immunolabelled by an antibody directed against rhodopsin, suggesting the presence of rods. We conclude that in the nonmammalian retina, rods and cones cannot be exclusively distinguished on purely morphological grounds.

Antibodies directed against gamma-aminobutyric acid (GABA) and L-glutamic acid decarboxylases 65 and 67 kDa (GAD65 and -67) were used to study the GABAergic cell populations of the chameleon retina. GABA immunoreactivity was found in the two main types of retinal interneurons, amacrine and horizontal cells. Amacrine, displaced amacrine, and intra- and interplexiform cells displayed the strongest GABA immunoreactivity of all the retinal cell types. Horizontal cells formed a continuous GABA-immunoreactive cell layer lying against the outermost portion of the inner nuclear layer. In contrast to previous studies (Quesada et al. [1996] Cell Biol. Int. 20:395-400; [1999] Eur. J. Anat. 3:13-25), the present results demonstrate that the horizontal cells of the chameleon retina are GABA immunoreactive and that a subpopulation of these is immunolabelled by an antibody against GAD65. These results indicate that GABAergic synaptic transmission plays a key role in the outer plexiform layer of the vertebrate retina., (Copyright 2003 Wiley-Liss, Inc.)

The distribution of delta sleep-inducing peptide immunoreactive cell bodies, fibers, and terminal-like structures was investigated in the normal human hypothalamus during the first postnatal year, using immunohistofluorescence and peroxidase anti-peroxidase techniques. Immunolabeled perikarya were relatively few and were mostly scattered through the anterior (preoptic) and mediobasal regions (infundibular nucleus) of the hypothalamus. DSIP-immunoreactive fibers and terminal-like fibers were observed throughout the entire rostrocaudal extent of the hypothalamus. They exhibit high densities in the preoptic region, the organum vasculosum of lamina terminalis, infundibular nucleus and median eminence. Moderate to low densities of DSIP-immunoreactive fibers were observed in the other hypothalamic structures, located in the anterior and mediobasal regions of hypothalamus, such as periventricular, paraventricular, suprachiasmatic, ventromedial, dorsomedial and parafornical nuclei. In the present study, the analysis of the immunohistochemical pattern of DSIP-immunoreactive neuronal elements in the human infant hypothalamus during the first postnatal year provided evidence of the presence of several differences. We have found qualitative age-related changes in the density of DSIP immunoreactivity in several hypothalamic structures such as the anterior region and the median eminence.

The optic nerve of adult chameleons was investigated with an electron microscope. The total number of retinal ganglion cell axons, the proportion of myelinated axons, the frequency distributions of myelinated and unmyelinated axon diameters were estimated, together with the volume occupied by glial processes. These were distinguished from unmyelinated axons using an antibody directed against glial fibrillary acidic protein, in a post-embedding procedure. The total number of fibers was estimated to be 405,235 +/- 60,000 axons. The proportion of myelinated fibers varied with position between the eyeball and the chiasma; being 22-27% close to the eyeball, rising to 42-47% halfway along the optic nerve and to 56-62% close to the chiasma. Myelinated and unmyelinated fiber diameter distributions were unimodal and positively skewed, with modes of 0.7 microm and 0.2 microm, respectively. There was a significant regional variation in the size of optic nerve axons. Large myelinated axons were observed in the dorsal and ventral periphery, whereas smaller myelinated fibers and a high proportion of unmyelinated fibers were found in the center of the nerve., (Copyright 2001 S. Karger AG, Basel)

The distribution of DSIP-IR cell bodies and fibers was investigated in the normal human hypothalamus during the first postnatal year using the indirect immunofluorescence technique. The analysis of the immunohistochemical patterns obtained in the seven cases analyzed showed regional differences in the localization of cell bodies and fibers. Immunoreactive perikarya were relatively few, and were mostly scattered throughout the anterior and the mediobasal hypothalamus. DSIP-IR fibers and terminal-like structures were observed throughout the rostro-caudal extent of the hypothalamic region. In the present study, we noticed qualitative changes in the density of DSIP immunoreactivity in several hypothalamic structures such as the preoptic area and the median eminence with respect to age. These postnatal differences observed for DSIP could be related to neuronal maturation processes occurring at this period in the central nervous system as well as other physiological processes controlling the evolution of DSIP concentrations. These data are compatible with the proposed role of the neuropeptide in the regulation of many postnatal physiological functions.

The topography and localization of motoneurons innervating the six extraocular muscles in the chameleon (Chamaeleo chameleon) was studied following HRP injection in each of these individual muscles. Four muscles were innervated ipsilaterally: medial rectus, inferior rectus, inferior oblique and lateral rectus. The medial rectus muscle was innervated by the dorsomedial part of the oculomotor nucleus. The innervation to the inferior rectus muscle arose from the lateral part of the intermediate oculomotor subnucleus, which extended to the lateral part of the dorsal subdivision. The lateral rectus muscle was innervated by the abducens nucleus, which was composed by two subgroups of labeled cells, respectively observed in the principal and accessory abducens subnuclei, whereas efferents to the inferior oblique muscle originated from both the ventral and intermediate oculomotor subnuclei. The contralateral pattern consisted of motoneurons innervating the superior rectus and the superior oblique that were located respectively in the caudal portion of the ventral oculomotor nucleus and in the trochlear nucleus. These results confirmed data reported in most vertebrate species, and were discussed from a comparative and functional point of view.

Using in vitro labelling and autoradiographic techniques, we have analyzed the fine and the detailed distribution of benzodiazepine binding sites in the post-mortem human hypothalamus. Binding sites were labelled in mounted tissue sections from adult brains, using the selective high affinity ligand [3H]-Flunitrazepam. A heterogeneous distribution of benzodiazepine binding sites was found throughout the rostrocaudal extent of human hypothalamus. The autoradiographic labelling was shown in the three hypothalamic parts, i.e., anterior, mediobasal and posterior levels. At the anterior level, the highest densities were present in the diagonal band of Broca, the preoptic area (medial and lateral parts) and the septohypothalamic nucleus. At the mediobasal hypothalamic level, the highest densities were mainly localized in the ventromedial nucleus, whereas the other structures were moderately labelled with [3H]-Flunitrazepam. The mammillary complex as well as the posterior hypothalamic area represented the most heavily labelled structures in the posterior hypothalamus. The results obtained in this study, indicate the presence of a large and heterogeneous distribution of benzodiazepine binding sites in human adult hypothalamus. This could support their implication in the control of distinct neural functions (like neuroendocrine role).

The cervico-ocular reflex (COR) was investigated in the chameleon. Two kinds of responses were observed by oscillating the body (sine-wave stimuli) in the fixed-head animal: a "smooth response" of very low gain (around 0.08) and a saccadic response composed of 1-12 saccades per cycle of stimulation (depending on the stimulation frequency). Both responses were elicited in the compensatory direction (same direction as the stimulation) and exhibited a frequency dependence with low-pass properties. The saccadic response was especially developed and displayed a higher gain (up to 0.4) than the smooth response. In darkness, the saccades were triggered near the zero point (head-body alignment), whereas in the presence of a fixed visual surround they were elicited more regularly throughout the stimulation cycle. The amplitude of saccades was increased in the light. Consequently, the gain and the phase lag of the saccadic response were enhanced by the visual input. No visuo-cervical interaction was observed for the smooth response. Oscillating the body at a constant velocity (seesaw or ramp stimuli) revealed a frequency effect on the number of saccades (during a cycle of stimulation), but not on the gain of the response. Increasing the amplitude of oscillations augmented only very slightly the amplitude of saccades and consequently decreased the gain. Hence, the best working range of the saccadic response corresponds to body or head movements of low amplitude (up to +/- 20 deg) and low frequency (up to 0.25 Hz), and is improved by a visual input. These properties are discussed on a comparative point of view. It is proposed that, in chameleons, the saccadic response could contribute to gaze stabilization and add to the vestibulo-ocular and the optokinetic responses.

Immunohistochemical techniques were used to determine the distribution of cholecystokinin-8 (CCK8) immunoreactivity in the brain of the chameleon. In the telencephalon, CCK8-immunopositive somata were sparse and observed principally in the olfactory tubercle at the ventromedial edge of the rostral telencephalon and in the medial septum. Immunopositive fibers were observed mainly in the medial septal region and the ventral telencephalon. In the diencephalon, numerous CCK8-reactive fibers were densely concentrated in the periventricular region, the dorsolateral hypothalamus and the external zone of the median eminence. In the thalamus, labelled fibers were restricted to the peri-rotundal nuclei and the lateral part of the habenula. Immunoreactive cell bodies were observed in the medial part of the dorsal lateral geniculate nucleus, in the periventricular, ventral and lateral regions of the hypothalamus. In the mesencephalon, the densest accumulations of immunopositive fibers were observed in the area pretectalis, the periventricular gray matter, the medial tegmentum and the isthmus. Labelled neurons were observed in the deep, and occasionally intermediate, tectal layers and in the laminar nucleus of the torus semicircularis. In the rhombencephalon, labelled fibers were observed at the highest density in the central gray matter and the locus coeruleus; labelled somata were observed only in the nucleus of the tractus solitarius.

Intraocular injections of rhodamine and horseradish peroxidase in chameleon, labelled retrogradely neurons in the ventromedial tegmental region of the mesencephalon and the ventrolateral thalamus of the diencephalon. In both areas, staining was observed contralaterally to the injected eye. Labelling was occasionally observed in some rhombencephalic motor nuclei. These results indicate that chameleons, unlike other reptilian species, have two retinopetal nuclei.

Chameleons display a number of well-described physiological peculiarities of their visual system, but there is no information on the topography of the retinal ganglion cell layer. In the present study, ganglion cell density of the chameleon retina was constructed from whole mounts of the retina stained with cresyl violet. For the identification of ganglion cells, these latter cells were labelled retrogradely with horseradish peroxidase applied to the optic nerve. Using this criterion, the proportion of ganglion cells was estimated to represent 80% of retinal cells, while glial cells and amacrine cells represented 14 and 6%, respectively, of the total cell population of the retina. As for the main features of the retinal map, first, ganglion cells were distributed inhomogeneously within the ganglion cell layer, and revealed the existence of a putative area centralis. Second, a horizontal visual streak, which showed two peak density areas, was identified. These features point out the degree of specialisation of the chameleon retina and the complexity of its visual system.

The NADPH-diaphorase histochemical technique, which reflects the activity of nitric oxide synthase was used in order to map the corresponding chameleon brain structures. In the forebrain, stained neurons were found particularly in the dorsal ventricular ridge and the ventral telencephalon, but no stained somata were observed in the cortical areas nor in the septum. In the midbrain, few NADPH-stained structures were seen. The stained cells were confined in the nucleus dorsomedialis, the nucleus geniculatus lateralis dorsalis, and the ventromedial hypothalamus. More caudally, some stained neurons were observed in the nucleus opticus mediodorsalis hypothalami posterior. In the brainstem, NADPH-stained cells were located in the optic tectum, the torus semicircularis, the substantia nigra, the ventral tegmental area, and all motor nuclei of the cranial nerves. The present results showed some striking differences and similarities with birds and mammals. Moreover, the presence of NADPH-staining in most of retinofugal, retinopetal and oculomotor nuclei suggests that nitric oxide system plays a prominent role in the visual and oculomotor functions.

There is little evidence allowing the hypothesis of the existence of direct pathways from the sensorimotor cortex (SMC) to main cardiovascular medullary nuclei: the dorsal motor nucleus of the vagus (DMV), the nucleus of the solitary tract (NTS) and the rostral ventrolateral medulla (RVLM). The purpose of this study was to identify in the rat direct SMC-NTS/DMV and-RVLM projections descending through the pyramidal tract (PT) and corticospinal neurones projecting to spinal somatic centers and sending collaterals to the NTS/DMV and the RVLM. The first group of animals (N = 15) received injections of anterograde tracers into the SMC: wheat germ agglutinin conjugated to horseradish peroxydase (WGA-HRP) or rhodamine-conjugated dextran (DR). In the second group (n = 35), retrograde tracers were injected: fluorogold (FG) into the NTS/DMV or into the RVLM and DR into the lateral thoracic cord (Th2-Th4). Anterograde transport of WGA-HRP and DR allowed corticofugal fibers to be followed inside the PT ipsilaterally to the site of cortical injection and showed bilateral labeled projections to the NTS/DMV and RVLM. After retrograde transport, bilateral FG or DR labeled cells were distributed in the SMC, mainly in the medial (AGm) and lateral (AGl) agranular cortex. After spinal and bulbar injections, double-labeled cells were distributed in same cortical areas. After injections in RVLM, 49% of labeled cells showed a double-labeling in the frontal cortex (rostral AGm and premotor cortex) while only 24% were observed in the posterior SMC (caudal AGl). On the contrary, when injections were done in NTS/DMV, double-labeled neurons were respectively of 11% in the frontal cortex and 4% in the posterior SMC. In the present work it was shown that the SMC sent direct projections to bulbar cardiovascular nuclei by means of fibers descending through the PT and corticospinal collaterals. The hypothesis which may be drawn from this study is that cortical motor areas probably program cardiovascular adjustments, preparatory or concomitant to the control of striate muscles.

Neurons containing catecholamine, indoleamine, and gamma-aminobutyric acid (GABA) were identified by immunohistochemistry in the chameleon retina. Tyrosine hydroxylase (TH) and serotonin (5HT) were observed mostly in two subtypes of orthotopic amacrine cells differing in their soma size and process distribution within the IPL. Some labelled cells were displaced either to the IPL (5HT) or the GCL (TH and 5HT). A multiplicity of retinal cell types contained GABA including cones, horizontal, amacrine, and ganglion cells. Our results confirmed those obtained in the retinas of other lizards except for the presence of interstitial and displaced amacrine cells containing TH or 5HT of which this is the first report.

We opted for a symphysiotomy approach in the management of cervico-urethro-vaginal fistulae in two patients. This approach offers an excellent exposure of the region of the bladder neck. After section of the symphysis pubis with a Gigli's wire saw, the anterior wall of the bladder was taken down to the fistulae and we excised the fistulous tract. Longitudinal closure of the fistulae and the bladder followed. Results were satisfactory and there were no urinary incontinence.

The distribution of vasopressin (VP)-like and oxytocin (OX)-like immunoreactivities was investigated in the chameleon brain with antisera specific for either peptide. VP-like and OX-like neurons are found only in two nuclei: in the supraoptic and in the paraventricular nuclei, whereas both peptidergic innervations are restricted to some ventral areas, especially to the hypothalamus. However, in these areas, OX-like innervation is less dense than the VP-like innervation. Even though, no sex differences were noted for the two systems, differences are present between the normal female and the preoviposit one concerning VP-like system. In the latter female, numerous VPergic-like cell bodies occurred in the ependymal layer, making likely direct contact with the cerebrospinal fluid (CSF) of the third ventricle. In addition, some VP-like nerve fibers seem to be in supraependymal position at the septal wall, probably also in contact with CSF.

The distribution of substance P-like immunoreactivity in the chameleon brain and spinal cord was studied with immunohistochemical methods using polyclonal antibodies against substance P. In the telencephalon, immunoreactive cell bodies and fibers were located primarily in the striatum and in the globus pallidus. In addition, few substance P-like fibers were observed in the cortical areas, in the septum, and in the amygdala. In the diencephalon, a high density of immunostained neurons and fibers were seen in the periventricular and ventrolateral hypothalamus. Another group of cell bodies was located in the optic tectum and particularly in the stratum griseum central. A large number of immunoreactive fibers were also detected in the thalamic nuclei and in the median eminence. In the mesencephalon, few immunoreactive neurons were observed in the ventral tegmental area, in the substantia nigra, and in the nucleus reticularis isthmi. These latter nuclei, the periventricular area, the posterior commissure, the nucleus lentiformis mesencephali, the oculomotor nucleus, and the raphe nuclei contained a dense plexus of substance P immunoreactive fibers. No immunoreactive cell bodies were observed in raphe nuclei. In the spinal cord, no substance P-like immunoreactive neurons were observed, but a large number of substance P immunostained fibers were seen in the dorsal and lateral part of the dorsal horn and surrounding the dorsal parts of the central canal. The results of the present study are discussed with respect to those obtained in other species of reptiles, the main differences concerning the lateral septum, the habenula, the area of the paraventricular organ, and the raphe nuclei.

The retinofugal projections of the chameleon were investigated by means of autoradiography, horseradish peroxidase and fluorescent techniques after intraocular injection of tracers. An ipsilateral contingent of visual fibers and projections is absent. The retinal fibers decussate completely in alternating fascicles at the optic chiasma and course to terminate in two hypothalamic nuclei (nucleus suprachiasmaticus and nucleus opticus periventricularis hypothalami posterior), six thalamic nuclei (nucleus ovalis, nucleus geniculatus lateralis dorsalis partes lateralis and medialis, nucleus geniculatus lateralis ventralis, lateral part of nucleus dorsolateralis anterior, and nucleus ventrobasalis), four pretectal nuclei (nucleus griseus tectalis, nucleus lentiformis mesencephali, nucleus geniculatus pretectalis and nucleus posterodorsalis), the optic tectum (stratum griseum et fibrosum superficiale) and the tegmental nucleus opticus tegmenti. Our findings are, in general, compatible with previous descriptions of the primary visual system in other species of lizards. However, they indicate three features particular to chameleons: first, the total absence of an ipsilateral retinofugal projection; second, the existence of an additional hypothalamic visual center located in the posterior mediodorsal hypothalamus; and third, the large size of nucleus opticus tegmenti. These features are discussed in terms of the taxonomic position of chameleons with respect to other lizards.

After small pressure injections of Fluorogold (FG), and Dextran-tetramethylrodamine (DR) into the dorsal motor nucleus of the vagus/nucleus of the solitary tract (DMV/NTS) and the rostral ventrolateral medulla (RVLM), respectively, retrograde FG-labelled cells were found mainly in the sensorimotor cortex; retrograde DR-labelled cells were located in the same cortical areas and in the prefrontal cortex. Double-labelled cells were also found in the sensorimotor cortical areas. These results provide evidence of direct projections from the sensorimotor cortex to the DMV/NTS and RVLM and suggest that somatic cortical areas directly control cardiovascular output during sensory and somatic processes.

Spontaneous eye movements as well as visual, vestibular, and proprioceptive cervical reflexes which contribute to gaze stabilization were investigated in the chameleon using the magnetic search-coil technique. The oculomotor range of each eye was very large (180 deg horizontally x 80 deg vertically). Spontaneous ocular saccades were independent in the two eyes and could have very large amplitudes. The fast phases of nystagmus during the stabilization reflexes were also independent in the eyes. In the head-restrained condition, optokinetic nystagmus (OKN) had a low gain in both horizontal and vertical planes (0.35 at 5 deg/s) and showed little binocular interaction. The vestibulo-ocular reflex (VOR) exhibited a low gain (0.2-0.3 from 0.05-1 Hz) and a high-phase lead at low frequency (140 deg at 0.05 Hz). Rotation of the animal in the presence of a visible surround increased the overall gain of gaze stabilization to 0.4-0.5 (P < 0.01) and considerably reduced the phase lead (38 deg at 0.05 Hz). In the head-free condition, head and eye reflexes were active simultaneously during both optokinetic and vestibular stimulation, but nystagmic head movements appeared only occasionally with a rather loose eye-head coordination. During optokinetic stimulation, eye movements contributed more than head movements to gaze stabilization, whereas, during vestibular or visuo-vestibular stimulation, the relative contribution of eye and head responses varied with stimulus frequency. When the head was freed, overall gain for gaze stabilization increased from 0.35 to 0.45 (P < 0.05) for optokinetic stimulation at 5 deg/s and from 0.2-0.3 to 0.4-0.75 (P < 0.001) for vestibular stimulation at 0.05-1 Hz.(ABSTRACT TRUNCATED AT 250 WORDS)

An immunocytochemical method, using glutaraldehyde fixation and an antiserum developed against a GABA-glutaraldehyde protein conjugate, permitted direct visualization of GABAergic structures in the brain of a reptile (chameleon). GABA immunoreactive cell bodies and nerve terminals were observed to be evenly distributed throughout the forebrain and midbrain. In the forebrain, GABA-positive perikarya were shown in all cortical areas, the septal area, the striatum, the dorsal ventricular ridge, and in the nucleus accumbens. In the midbrain, the optic tectum contained a dense and laminar distribution of GABA neurons. These neurons were also observed in the lateral geniculate nucleus, nucleus profundus mesencephali, nucleus opticus tegmenti and substantia nigra. Immunoreactive nerve fibers and terminals were observed in the same structures and, additionally, in the tractus septo-hypothalamicus, habenula complex, median eminence, intermediate lobe of the pituitary, basal part of the subcommissural organ, torus semicircularis and nucleus reticularis isthmi. These results provide a framework for a further electron microscopic analysis of the GABAergic innervation of some encephalic areas involved in physiological regulations particular to this species especially the visual system.

The distribution of serotonin immunoreactive cell bodies and fibers was studied in the chameleon brain by using the immunohistochemical technique with antisera against serotonin coupled to a carrier with glutaraldehyde. Serotonin perikarya were found in the caudal midbrain tegmentum, in the lateral part of the nucleus reticularis isthmi, the lateral part of the nucleus interpeduncularis and along the midline in the raphe superior. More caudally, the serotonin immunoreactive cell bodies were located along the nucleus raphe inferior and ventrolaterally in the vicinity of the olivary complex. No immunoreactive cell bodies were found in the spinal cord nor in the paraventricular organ (PVO) of the hypothalamus. Immunoreactive fibers were observed in the entire brain. Prominent concentrations were found in the dorsal cortex, lateral septum, lateral geniculate nucleus, median eminence, pretectal nucleus, nucleus interpeduncularis, vestibular nucleus and olivary complex. Descending serotonin immunoreactive fibers were found in particular in the ventral motoneuron area in the spinal cord. One of the most interesting findings in this study was the lack of immunoreactive CSF contacting neurons in the PVO and the observation of an extensive plexus of supraependymal fibers, a feature reported so far only in mammals.

An immunohistochemical method, using glutaraldehyde fixation and a highly specific monoclonal antibody recently synthetized against dopamine (DA)-glutaraldehyde protein conjugate, permitted direct visualization of DA structures in the brainstem and spinal cord of a reptile (Chameleon). DA-immunoreactive cell bodies occurred in some contiguous areas of the midbrain tegmentum. The first one was located in the ventral tegmental area. Some somata intermingled with the oculomotor nucleus. The second group was the large round or oval DA-Immunostained neurons located in the substantia nigra. More caudally, a third group of round or fusiform DA-cell bodies was seen in an homologous area of so called mammalian A8 and were continuous with the substantia nigra group. In the medulla oblongata, the DA-containing cells were shown in the nucleus of solitary tract and in the dorsal lateral part of the dorsal motor nucleus of the vagus. The density of this DA-Immunoreactive neurons decreased more caudally. At the medullo-spinal level and upper cervical spinal cord, a few labelled cells were distinguished near the central canal. In the spinal cord DA-immunopositive cell bodies were observed in the vicinity of the central canal and formed a continuous column that extended throughout the rostral spinal cord. The apical processes of these neurons seemed to be in contact with the lumen of the central canal. This study constitute the first visualization of the immunoreactive DA-cell bodies at the medullo-spinal level which were already described, as TH immunoreactive in other species of reptiles.

Some visual, vestibular and proprioceptive reflexes which contribute to gaze (head + eye) stabilization were quantified in the chameleon. All the reflexes were analysed in the horizontal plane, and the visual reflexes were also studied in the vertical plane. In restrained-head animals, both the optokinetic nystagmus (OKN) and the vestibulo-ocular reflex (VOR) had low gains. In free-head animals, the head (opto-collic or vestibulo-collic reflex) and eye (OKN or VOR) responses added their effects, thus improving gaze stabilization, especially during vestibular stimulation. Cervical stimulation provoked both a cervico-ocular reflex (COR) in the compensatory direction and a large number of saccades. The saccadic response was especially marked in the presence of patterned visual surroundings.

The effects of stimulation of cervical vagal afferents on the electromyographic activity (e.m.g.) of the gastric antrum and pylorus have been studied both in conscious and decorticate, curarized rabbits. In the latter group, stimulation at 10-15 Hz (0.3-0.4 mA; 5-7 ms duration) inhibited both the antral and pyloric e.m.g. in 21 rabbits, elicited an excitatory response in two more but produced no change in the remaining 13 animals. The inhibitory response was not affected by guanethidine (1 mg/kg, i.v.), phentolamine (1 mg/kg, i.v.) or propranolol (0.1 mg/kg, i.v.) and persisted in rabbits in which the contralateral cervical vagus and both splanchnic nerves had been cut, the coeliac ganglia and both adrenal glands had been extirpated. It was only abolished by combined section of the spinal cord at the level of C1, C2 and the contralateral vagus in the neck. In conscious rabbits, vagal stimulation at 2-5 Hz (0.4 mA; 0.5-1.0 ms duration) consistently inhibited antral and pyloric e.m.g. This inhibitory response persisted in the presence of phentolamine, propranolol and naloxone (0.25-1.00 mg/kg), and in adrenalectomized rabbits with cut splanchnic nerves and extirpated coeliac ganglia. However, it was completely abolished by section of both vagi above the diaphragm. It is concluded that (1) the vagal afferents responsible for the inhibition of the gastric e.m.g. are already included in the abdominal vagus nerves, (2) this inhibition of gastric e.m.g. is mediated by non-adrenergic fibers, one group of them emerging from the thoracic spinal cord to join the thoracic vagi via the stellate ganglia.

The GnRH system of the chameleon brain was studied at light microscopic and ultrastructural levels by use of an immunohistochemical technique with antibodies directed against salmon gonadotrophin-releasing hormone. Immunoreactive (IR) perikarya were found in the anterior midbrain tegmentum. At this level numerous IR cell bodies were detected around the fasciculus longitudinalis medialis (FLM). The more rostral neurons were observed dorsal to the FLM and progressively tended to be lateral to it along the midline. More caudally, they were found ventral to the FLM. At the electron microscope level, these cells were seen to contain large granular vesicles and to receive numerous synaptic inputs. A prominent pathway was traced from these cell bodies along the medulla oblongata to the spinal cord. A second IR pathway ascended rostrally to the habenular complex. No IR perikarya were located in the anterior brain including the olfactory bulbs.