Biomedical subjects

Y Tillet

Publications and source records attributed to Y Tillet.

At least 19 recordsLinked to original sources

PubMed2006

Kisspeptin immunoreactive cells of the ovine preoptic area and arcuate nucleus co-express estrogen receptor alpha.

Kisspeptins are peptide ligands of the G protein-coupled receptor GPR54, recently shown to be essential to reproductive function. We have raised specific rabbit antisera against a highly conserved 10 amino acid-amidated peptide (kp10) common to all kisspeptin isoforms isolated so far and mapped the distribution of kp10-immunoreactive (ir) cells in the ovine hypothalamus. Kp10-ir cells were predominant in the caudal arcuate nucleus, the dorsomedial nucleus and the medial preoptic area. Numerous varicose kp10-ir fibers were found in the preoptic area where GnRH neurons reside and in the median eminence, seemingly projecting around small capillaries in its external zone. Within the caudal arcuate nucleus, nearly all kp10-ir cells showed an intense estradiol receptor alpha immunofluorescent signal compared with approximately half of kp10-ir cells in the preoptic area. The pattern of distribution of kp10 immunoreactivity in the hypothalamus suggests a role for kisspeptin in the estrogen-dependent regulation of GnRH and LH secretion in the ewe.

Animals↗

PubMed2005

Long-lasting effects of serotonin deficiency on differentiating peptidergic neurons in the rat suprachiasmatic nucleus.

Serotonin (5-HT, 5-hydroxytryptamine) is known to be an inductor of the brain development [Whitaker-Azmitia, P.M., Druse, M., Walker, P., Lauder, J.M., 1996. Serotonin as a developmental signal. Behav. Brain Res. 73, 19-29; Ugrumov, M.V., 1997. Hypothalamic monoaminergic systems in ontogenesis: development and functional significance. Int. J. Dev. Biol. 41, 809-816]. This study was aimed to test whether it provides long-lasting effects on the differentiating vasoactive intestinal polypeptide (VIP) and vasopressin (VP) neurons of the suprachiasmatic nucleus (SCN) in rats. To this aim, 5-HT was depleted in fetal brain by daily injections of p-chlorophenylalanine (pCPA), an inhibitor of 5-HT synthesis, to pregnant rats from the 13th to the 21st day of gestation. Pregnant rats injected with saline served as controls. The offsprings (males) of pCPA-treated and control pregnant rats were maintained after birth for two months under normal laboratory conditions. Then, the SCN was processed for immunocytochemistry of VIP and VP and in situ hybridization of appropriate mRNAs. There were no differences in concentrations of VIP and VP mRNAs in the SCN in adult offsprings of the 5-HT-depleted pregnant rats compared to the controls. Moreover, 5-HT deficiency did not induce any change in size of VIP-immunoreactive (IR) and VP-IR neurons. Conversely, both the numbers of VIP- and VP-immunoreactive neurons and concentrations of the peptides in cell bodies increased significantly. It is concluded that 5-HT provides long-lasting effects on differentiating VIP and VP neurons in the SCN resulting in attenuated release rather than elevated synthesis of both peptides in adulthood.

Analysis of Variance↗

PubMed2005

Oestradiol effect on galanin-immunoreactive neurones in the diencephalon of the ewe.

Galanin is a neuropeptide involved in the regulation of numerous functions such as reproduction. In female rats, this peptide stimulates gonadotropin-releasing hormone (GnRH)/luteinizing hormone release and its synthesis is stimulated by oestradiol. It could therefore be an intermediary between the oestrogenic signal from the ovaries and the GnRH neurones (e.g. during the time course leading to the preovulatory GnRH surge). However, although the involvement of galanin is well-known in rodents, it is poorly understood in ewes. Using immunohistochemistry with a specific antigalanin antiserum, we detected the peptide in neurones of two groups of ovariectomized ewes treated for 6 h with subcutaneous implants, either with oestradiol (experimental group) or empty (control group). The galanin-immunoreactive neurones were counted in three areas, the preoptic area, the bed nucleus of the stria terminalis and the infundibular nucleus, using a computerized image analysis system. There was no change in the mean number of galanin-immunoreactive (GAL-ir) neurones in the infundibular nucleus (37 +/- 12 neurones/section in treated animals and 31 +/- 11 in controls) or in the bed nucleus of the stria terminalis (22 +/- 5 neurones/section in treated animals and 16 +/- 4 in controls), but the number of GAL-ir neurones was higher in the preoptic area in treated than in control ewes (35 +/- 4 versus 14 +/- 10, P < 0.001). To determine whether the neurones of the preoptic area were directly sensitive to oestradiol, we performed double immunohistochemical labelling for oestradiol receptor alpha and galanin. More than 50% of the GAL-ir neurones contained the oestradiol receptor alpha and therefore could be directly regulated by oestradiol. These results indicate that oestradiol might act directly on a GAL-ir neuronal population situated in the preoptic area, without any effect on the GAL-ir neurones of the infundibular nucleus or the bed nucleus of the stria terminalis. Because a 6-h oestradiol treatment can induce a preovulatory GnRH surge in ewes, the GAL-ir neuronal population of the preoptic area might be one of the neuronal systems by which oestradiol activates the GnRH neurones. However, although the morphological relationships between galanin and GnRH neurones have been described in rodents, they remain to be demonstrated in the ewe.

Animals↗

PubMed2005

Nutrition and hypothalamic neuropeptides in sheep: histochemical studies.

The identification and role of neuropeptides in the control of food intake and energy balance have been extensively studied in rodents, and for more than ten years, similar studies have been performed in sheep. As a photoperiodic ruminant, sheep are an interesting alternative animal model to rodents. In this review, we summarize the results obtained in sheep concerning the distribution of peptide-containing neurones in the hypothalamus and their central role in the control of food intake and energy balance, and compared them with relevant data from rodents. Even if the general organization and the role of hypothalamic neuropeptides are similar in sheep and rodents, numerous differences have been observed between these two species. In sheep, the magnocellular neurones of the paraventricular and supraoptic nuclei are characterized by the low density and the lack of galanin- and neuropeptide-Y-containing neurones, respectively. The sheep pituitary stalk presents neurones containing neuropeptides such as neuropeptide-Y or beta-endorphin, which are also found in the deep part of the infundibular nucleus. In this structure, several neuronal populations, including galanin, agouti-gene related peptide, somatostatin, are sensitive to energy balance variations, undernutrition or overfeeding, which may specifically modify neuropeptide levels in discrete neuronal subgroups. This feature is well illustrated by the number of neuropeptide-Y labelled neurones, that increases in the lateral part of the infundibular nucleus of undernourished ewes and decreases in the ventral part of overfed ewes. Conversely, after 24 hours of food deprivation, the number of neuropeptide-Y-immunolabelled neurones is unchanged in the sheep infundibular nucleus, whereas increased levels of this neuropeptide are described, in rats, by radioimmuno-assay. In conclusion, our review shows that peptide-containing neurone systems, involved in the regulation of food intake and energy balance in sheep, are generally similar to those observed in other species, but they present specific differences according to the physiological characteristics of the animal model.

Animals↗

PubMed2003

Sensitivity of galanin- and melanin-concentrating hormone-containing neurones to nutritional status: an immunohistochemical study in the ovariectomized ewe.

The sensitivities of galanin and melanin-concentrating hormone (MCH) neuronal systems to nutrition are poorly understood in sheep compared to rodents. The aim of this study was to describe the changes in the numbers of galanin and MCH neurones in ovariectomized ewes submitted to different nutritional levels. In the first experiment, ewes were fed ad libitum or food deprived for 24 h. In the second experiment, two groups of ewes were fed at maintenance level (group 100) or undernourished (group 40) for 167 days, after which one-half of each group was killed or refed ad libitum (group 100R and 40R) for 4 days. The MCH neuronal population located in the lateral hypothalamic area was not affected by these nutritional changes. Long-term undernutrition enhanced the number of galanin neurones located in the infundibular nucleus and the dorsal hypothalamic area (DHA), refeeding resulted in an increase of neurones in the DHA and preoptic area, but short-term starvation had no effect on any galanin subpopulations. Our data suggest that the sensitivity of MCH neuronal populations to nutrition in sheep differs from that of rodents. Various populations of galanin-containing neurones differ in sensitivity in ewes subjected to long undernutrition and refeeding but not to short starvation.

Animals↗

PubMed2003

Prolonged neurogenesis during early development of gonadotropin-releasing hormone neurones in sheep (Ovis Aries): in vivo and in vitro studies.

Gonadotropin-releasing hormone (GnRH) neurons involved in controlling the reproductive function in vertebrates are derived from the olfactory placode. However, in the sheep and the rat species, GnRH-immunoreactive (GnRH-IR) neurons could not be detected in the olfactory region during the earliest phase of GnRH system development. Using in situ hybridization (ISH) and immunohistochemistry (IHC) in sheep embryos ranging from 26 to 53 days' gestational age (G26-G53), the present work confirmed that GnRH expression could not be detected during the earliest steps of migration. The first ISH+ cells were detected in the nasal septum and at the entrance of the telencephalon at G33 stage. [(3)H]-thymidine pulses applied to in vitro olfactory explant cultures showed that GnRH neuron precursor cells have an extended multiplication period corresponding to G26-G36 before entering the neuronal differentiation process. Therefore, the lack of GnRH neuron detection during the early phase of development in the sheep compared to the mouse and other vertebrates represents a major difference in the early development of GnRH neurons. In the mouse, GnRH neuron precursors have a limited multiplication period in the vomeronasal pit and only postmitotic neurons start migration, whereas in the sheep embryo, the multiplication period is extended to about 10 days as demonstrated in olfactory explant cultures.

Animals↗

PubMed2003

Bioavailability of oral vs intramuscular iodinated oil (Lipiodol UF) in healthy subjects.

BACKGROUND: In order to fight against iodine deficiency, the essential cause of endemic goiter and cretinism, several health organizations promoted campaigns of iodinated oil (Lipiodol UF) administration using iodinated oil administered intramuscularly. However, it seems preferable to administer iodinated oil orally, as this is more appropriate and since the efficacy of this route has been demonstrated as well as for intramuscular route by controlled clinical trials. OBJECTIVE: To assess the bioavailability of iodinated oil (Lipiodol UF) administered via two different administration routes and the safety profile of this agent. DESIGN: A randomized bioavailability study was performed comparing a single oral dose of 3 capsules (570 mg of iodine) vs a single intramuscular injection of 1 ml of Lipiodol UF (480 mg of iodine) in 36 healthy subjects followed for 9 months. RESULTS: The results show that, at these dosages, the 24 h urinary iodine values are above baseline for both oral and intramuscular administrations (im: >12 months/oral: 6 months) for prolonged period of time. In terms of safety, Lipiodol, administered by im injection or orally, did not induce any undesirable effects or any alteration of thyroid function tests in this study. CONCLUSIONS: In conclusion, this study shows that im or oral administration of a single dose of Lipiodol provides a significant and prolonged iodine supplement. The results obtained confirm the possibility of protection of exposed populations after annual administration of an appropriate single oral dose, without inducing any clinical or laboratory adverse effects. The product, by either route of administration, has a prolonged efficacy in iodine-deficient subjects (im: 2-3 years/oral: 1 year).

Administration, Oral↗

PubMed2002

Long-term undernutrition followed by short-term refeeding effects on the corticotropin-releasing hormone containing neurones in the paraventricular nucleus: an immunohistochemical study in sheep.

The effect of nutritional level on the immunoreactivity of corticotropin-releasing hormone (CRH) in neurones of the hypothalamic paraventricular nucleus was described in sheep, a ruminant, whose feeding strategy differs from that of monogastric species. Two groups of ewes were underfed (40%), or fed at maintenance (100%) for 167 days, after which one-half of each group was killed or ad libitum refed (at least 150% of maintenance) for 4 days before killing. The presence of CRH in the paraventricular nucleus was examined by immunohistochemistry. The number of CRH immunoreactive neurones was increased in underfed ewes, but without modification of the plasma concentration of cortisol, indicating that the rise of CRH was not released in the portal blood nor linked to the pituitary-adrenal axis. Refeeding did not modify significantly the number of CRH immunoreactive neurones in the nucleus although these neurones were increased, only in refed ewes that were previously underfed. These data differ from those for rats and mice where CRH expression is decreased or not modified by underfeeding which could reflect different effects of undernutrition on CRH immunoreactive neurones in monogastric compared to ruminants species.

Animal Nutritional Physiological Phenomena↗

PubMed2001

Distribution and co-localization of choline acetyltransferase and p75 neurotrophin receptors in the sheep basal forebrain: implications for the use of a specific cholinergic immunotoxin.

The basal forebrain cholinergic system is involved in different forms of memory. To study its role in social memory in sheep, an immunotoxin, ME20.4 immunoglobulin G (IgG)-saporin, was developed that is specific to basal forebrain cholinergic neurons bearing the p75 neurotrophin receptor. The distribution of sheep cholinergic neurons was mapped with an antibody against choline acetyltransferase. To assess the localization of the p75 receptor on basal forebrain cholinergic neurons, the distribution of p75 receptor-immunoreactive neurons with ME20.4 IgG was examined, and a double-labeling study with antibodies against choline acetyltransferase and p75 receptor was undertaken. The loss of basal forebrain cholinergic neurons and acetylcholinesterase fibers in basal forebrain projection areas was assessed in ewes that had received intracerebroventricular injections of the immunotoxin (50, 100 or 150 microg) alone, as well as, in some of the ewes treated with the highest dose, with bilateral immunotoxin injections in the nucleus basalis (11 microg/side). Results indicated that choline acetyltransferase- and p75 receptor-immunoreactive cells had similar distributions in the medial septum, the vertical and horizontal limbs of the band of Broca, and the nucleus basalis. The double-labeling procedure revealed that 100% of the cholinergic neurons are also p75 receptor positive in the medial septum and in the vertical and horizontal limbs of the band of Broca, and 82% in the nucleus basalis. Moreover, 100% of the p75 receptor-immunoreactive cells of these four nuclei were cholinergic. Combined immunotoxin injections into ventricles and the nucleus basalis produced a near complete loss (80-95%) of basal forebrain cholinergic neurons and acetylcholinesterase-positive fibers in the hippocampus, olfactory bulb and entorhinal cortex. This study provides the first anatomical data concerning the basal forebrain cholinergic system in ungulates. The availability of a selective cholinergic immunotoxin effective in sheep provides a new tool to probe the involvement of basal forebrain cholinergic neurons in cognitive processes in this species.

Animals↗

PubMed2001

Dopaminergic input to the ventromedial hypothalamus facilitates the oestrogen-induced luteinizing hormone surge in ewes.

In this study we examined the release of dopamine and noradrenaline in the ventromedial hypothalamus (VMH) of ovariectomized ewes during the oestrogen-induced luteinizing hormone (LH) surge by measuring their respective metabolites, 3,4-dihydroxyphenylacetic acid (DOPAC) and methoxyhydroxyphenylglycol (MHPG) using microdialysis. Further we investigated whether inhibition of catecholamine synthesis in the VMH by bilateral reverse dialysis of alpha-methyl-p-tyrosine (alpha-MPT) would block the oestrogen-induced LH and/or prolactin surges. Oestradiol treatment (50 microg oestradiol benzoate) of ovariectomized ewes resulted in a biphasic LH response, significantly (p < 0.05) decreasing LH concentrations from 2.5 to 10.5 h after injection, followed by an LH surge beginning at 16 h. Prolactin concentrations were also significantly (p < 0.05) increased in oestradiol-treated ewes from 13 h. VMH DOPAC concentrations in oil-vehicle-treated animals were at the level of detection (0.02 ng/ml) in most samples over the 24-hour sampling period. In oestradiol-treated ewes, VMH DOPAC levels were initially low before and up to 8 h after oestradiol injection but then increased significantly (p < 0.05) at 10-12 h and remained elevated up to 20 h after injection. In contrast, oestradiol injection had no effect on MHPG concentrations in the VMH. Bilateral reverse microdialysis of alpha-MPT into the VMH significantly (p < 0.05) delayed the time from oestradiol injection to the onset of the LH surge, the time to peak LH concentration and attenuated the LH surge compared with reverse dialysis of Ringer solution alone. In contrast, alpha-MPT treatment had no effect upon the oestradiol-induced increase in prolactin concentrations. This study provides evidence that the VMH is an important hypothalamic site in the neuro-endocrine control of the LH surge in ewes. The results suggest that dopaminergic neurons with terminals in the VMH are part of a neuronal pathway mediating the positive feedback effects of oestradiol on gonadotropin-releasing hormone secretion and the LH surge.

3,4-Dihydroxyphenylacetic Acid↗

PubMed2000

Neuronal projections to the lateral retrochiasmatic area of sheep with special reference to catecholaminergic afferents: immunohistochemical and retrograde tract-tracing studies.

The retrochiasmatic area contains the A15 catecholaminergic group and numerous monoaminergic afferents whose discrete cell origins are unknown in sheep. Using tract-tracing methods with a specific retrograde fluorescent tracer, fluorogold, we examined the cells of origin of afferents to the retrochiasmatic area in sheep. The retrogradely labeled cells were seen by observation of the tracer by direct fluorescence or by immunohistochemistry with specific antibodies raised in rabbits or horses. Among the retrogradely labeled neurons, double immunohistochemistry for tyrosine hydroxylase, dopamine-beta-hydroxylase, and serotonin were used to characterize catecholamine and serotonin FG labeled neurons. The retrochiasmatic area, which included the A15 dopaminergic group and the accessory supraoptic nucleus (SON), received major inputs from the lateral septum (LS), the bed nucleus of the stria terminalis (BNST), the thalamic paraventricular nucleus, hypothalamic paraventricular and supraoptic nuclei, the perimamillary area, the amygdala, the ventral part of the hippocampus and the parabrachial nucleus (PBN). Further, numerous scattered retrogradely labeled neurons were observed in the preoptic area, the ventromedial part of the hypothalamus. the periventricular area, the periaqueductal central gray (CG), the ventrolateral medulla and the dorsal vagal complex. Most of the noradrenergic afferents came from the ventro-lateral medulla (Al group), and only a few from the locus coeruleus complex (A6/A7 groups). A few dopaminergic neurons retrogradely labeled with flurogold were observed in the periventricular area of the hypothalamus. Rare serotoninergic fluorogold labeled neurons belonged to the dorsal raphe nucleus. Most of these afferents came from both sides of the brain, except for hypothalamic supraoptic and paraventricular nuclei. In the light of these anatomical data, we compared our results with data obtained from rats, and we discussed the putative role of these afferents in sheep in the regulation of several specific functions in which the retrochiasmatic area may be involved, such as reproduction.

Afferent Pathways↗

PubMed2000

Effect of feeding on Fos protein expression in sheep hypothalamus with special reference to the supraoptic and paraventricular nuclei: an immunohistochemical study.

The hypothalamus plays an important role in the control of food intake in different species, but there is little relevant information for ruminants like sheep. In order to study the putative role of several hypothalamic nuclei in food intake in sheep, Fos expression, a marker of cellular activity, was compared by immunohistochemistry between fed and unfed ewes. The expression of Fos protein was stimulated in the supraoptic nucleus of fed ewes, whereas it was increased in the paraventricular nucleus of unfed animals. In the latter nucleus, Fos immunoreactivity was mainly localized close to the third ventricle, an area corresponding to the parvocellular system of the nucleus, but never in the magnocellular system. In the paraventricular nucleus, the number of corticotrophin releasing factor-immunoreactive neurons and the number of Fos/corticotrophin releasing factor double-labelled neurons were not affected by feeding or lack of feeding. The number of Fos-immunoreactive neurons was higher in the lateral septum, the infundibular, the ventromedial and in the dorsomedial nuclei of unfed ewes than in those of fed ewes. Our results show for the first time that the dorsomedial and ventromedial nuclei are involved in the control of feeding in sheep as in rodents. The supraoptic nucleus of sheep is activated by the same conditions as in rodents but, conversely, the paraventricular nucleus is activated in unfed sheep, whereas in rodents and primates, this nucleus is activated by satiety as well as by fasting. In sheep, unlike in rodents, corticotrophin releasing factor did not appear to be involved in short-term regulation of food intake.

Animals↗

PubMed1999

Small intensely fluorescent cells of the rat paracervical ganglion synthesize adrenaline, receive afferent innervation from postganglionic cholinergic neurones, and contain muscarinic receptors.

In the paracervical ganglion (PCG) of the rat, double-labelling immunofluorescence for catecholamine-synthesizing enzymes and HPLC measurement of catecholamine contents were first performed to evaluate whether intraganglionic small intensely fluorescent (SIF) cells are capable of synthesizing adrenaline. Immunolabelling for tyrosine hydroxylase (TH), dopamine beta-hydroxylase and phenylethanolamine-N-methyl transferase (PNMT) occurred in all SIF cells of the PCG, thus demonstrating the presence of all the enzymes required for adrenaline biosynthesis. Adrenaline levels were undetectable in the PCG but to test the hypothesis that PNMT is active in SIF cells, catecholamines were measured in ganglia of rats pretreated with pargyline, an inhibitor of the monoamine oxidase, the major enzyme involved in the catecholamine degradation. Pargyline treatment increased adrenaline levels in the PCG, thus demonstrating that SIF cells are capable of adrenaline synthesis. The undetectable levels of adrenaline in the PCG of untreated rats suggested a slow rate of biosynthesis of adrenaline in the ganglion. Furthermore, the use of double-labelling showed that SIF cells of the PCG were stained for muscarinic receptors and were approached by varicose ChAT-immunoreactive nerve fibres. Nerve fibres immunoreactive for ChAT were also observed associated with nerve cell bodies of ganglion neurones. Following deafferentation of the PCG, the ChAT-immunoreactive nerve fibres surrounding nerve cell bodies totally disappeared indicating their preganglionic origin, while those associated with SIF cells did not degenerate, which demonstrate that they derived from intraganglionic cholinergic neurones. Taken together, the results show that adrenaline may be a transmitter for SIF cells in the PCG and suggest that cholinergic neurones of the parasympathetic division of the PCG can modulate the SIF cell activity through the activation of muscarinic receptors.

Afferent Pathways↗

PubMed1999

Afferents to the rostral olfactory bulb in sheep with special emphasis on the cholinergic, noradrenergic and serotonergic connections.

The olfactory bulb (OB) is involved in the processing of olfactory information particularly through the activation of its afferents. To localize their cell origin in sheep, a specific retrograde fluorescent tracer, Fluoro-Gold, was injected into the olfactory bulb of seven ewes. By using immunocytochemical techniques, retrogradely labeled neurons were colocalized with choline acetyltransferase, tyrosine hydroxylase, dopamine-beta-hydroxylase and serotonin to characterize cholinergic, noradrenergic and serotonergic Fluoro-Gold-labeled neurons. Most afferents originated from the ipsilateral side of the injection site. The OB received major inputs from the anterior olfactory nucleus (AON), the piriform cortex (PC), the olfactory tubercle, the diagonal band of Broca (DBB) and the amygdala. Other retrogradely labeled neurons were observed in the taenia tecta, the septum, the nucleus of the lateral olfactory tract, the preoptic area, the lateral hypothalamic area, the mediobasal hypothalamus, the lateral part of the premammillary nucleus, the paraventricular nucleus of the hypothalamus, the paraventricular thalamic nucleus, the central grey, the substantia nigra (SN), the ventral tegmental area (VTA), the lateral nucleus to the interpeduncular nucleus (IIP), the raphe and the locus coeruleus (LC). Contralateral labeling was also found in the AON, the PC, the SN compacta, the VTA, the IIP and the LC. Cholinergic Fluoro-Gold-labeled neurons belonged to the horizontal and vertical branch of the DBB. Noradrenergic afferents came from the LC and serotoninergic afferents came from the medial raphe nuclei and the 1IP. These data are discussed in relation with olfactory learning in the context of maternal behavior in sheep.

Animals↗

PubMed1999

Distribution of galanin immunoreactivity in the sheep diencephalon.

Although the physiological role of galanin has been demonstrated in several endocrine regulations in sheep, the anatomical characteristics of this neuronal system has never been studied. The distribution of galanin-containing neurones was described by immunohistochemistry using galanin antiserum in the diencephalon of adult ewes, both ovariectomized or treated with colchicine. Galanin-immunoreactivity was found throughout the diencephalon. In the ovariectomized ewes, galanin-immunoreactive neurones were mainly observed in the medial preoptic area and the infundibular nucleus. The highest density of immunoreactive fibres was found in the external layer of the median eminence. Numerous galanin-immunoreactive fibres were also observed in the preoptic area, the mediobasal hypothalamus, the periphery of the supraoptic and the paraventricular nuclei. With colchicine treatment, the number of labelled neurones increased, and additional galanin-immunoreactive perikarya were observed in the bed nucleus of the stria terminalis, the lateral septum, the supraoptic, the paraventricular and the periventricular nuclei and the paraventricular nucleus of the thalamus. In the caudal part of the diencephalon, the density of labelled neurones was lower in both groups of animals than in other species studied. Regardless of treatment, labelling was not seen in the suprachiasmatic nucleus and only rarely in the ventromedial nucleus. These results describe, for the first time, the distribution of galanin-immunoreactive neurones in the sheep diencephalon. Compared to other species studied, distribution in the sheep diencephalon has several distinct differences. In ovariectomized animals, the medial preoptic area presents more labelled neurones in sheep than in monkeys, whereas in the supraoptic nucleus the density of labelled neurones is lower in sheep than in humans or opossums. After colchicine treatment only very few differences were observed between sheep and rats, but in contrast to other species, the suprachiasmatic nucleus of the sheep does not contain labelled neurones.

Animals↗

PubMed1999

Localization of estrogen-receptive neurons projecting to the GnRH neuron-containing rostral preoptic area of the ewe.

Estrogen exerts important feedback effects upon the biosynthetic and secretory behavior of gonadotropin-releasing hormone (GnRH) neurons to control reproductive functioning. The mechanism of estrogen action upon these neurons is unclear and seems likely to involve the transsynaptic regulation of GnRH neurons. The objective of the present study was to identify the estrogen-receptive neural populations which project to the general vicinity of the GnRH perikarya in the rostral preoptic area and diagonal band of Broca (rPOA/DBB) of the ewe. Intact breeding-season ewes received an injection of the retrograde tracer fluorogold (FG) into the rPOA/DBB, and their hypothalami and brainstems examined for the presence of FG and estrogen receptor alpha (ERalpha) immunocytochemistry. Retrogradely labeled neurons were identified principally within the lateral septum (LS), lamina terminalis, bed nucleus of the stria terminalis, POA, arcuate nucleus (ARN), ventromedial nucleus (VMN) and median eminence. Smaller numbers of FG-immonoreactive cells were found in the caudal brainstem where they resided mostly in the ventrolateral medulla (VLM). Dual-labeled cells exhibiting both FG and ERalpha staining were prominent in the POA, LS and at all rostrocaudal levels of the VMN and ARN. Small numbers of dual-labeled cells were found in the VLM. These observations indicate that a number of distinct ERalpha-expressing neural populations project to the rPOA/DBB where the majority of the GnRH perikarya are found in the ewe. Although it is not possible to determine the direct connectivity of these projections with GnRH neurons, the findings provide an initial neuroanatomical framework through which the transsynaptic actions of estrogen on ovine GnRH neurons may be tested.

Animals↗

PubMed1998

Histaminergic neurons in the sheep diencephalon.

The distribution of histaminergic neurons in the sheep brain was studied by immunohistochemistry by using antibodies raised against histamine. For the first time in this species, the presence of histamine-immunoreactive neurons was described in the caudal diencephalon, around the mammillary bodies, and in the tuberomammillary area. The general pattern of distribution of these neurons was similar to that described previously in other species, i.e., rodents and humans. The distribution in the five neuronal groups described in rodents was not easy to demonstrate in sheep, because the boundaries between each group were not clear. The labeled neurons appeared to form a continuous cell system, as in humans. Numerous histamine-immunoreactive mast cells were found in the habenula and the thalamus. Histamine-immunoreactive fibers were found in almost all of the structures studied. The highest density of fibers was seen in the tuberomammillary area, from which dense bundles of fibers ran rostrally and dorsally along the third ventricle in a parasagittal plane. Numerous immunostained fibers were found close to the wall of the ventricles; some of them appeared to reach the cerebrospinal fluid through the ependymal cell layer. Some fibers were also observed in the optic tract, and the lowest density was found in the supraoptic and paraventricular nuclei. These results should be useful for developing further physiological studies on the role of histaminergic neuronal systems in sheep.

Animals↗

PubMed1998

Immunolocalization of aromatic L-amino acid decarboxylase, tyrosine hydroxylase, dopamine, and serotonin in the forebrain of Ambystoma mexicanum.

To improve basic knowledge about the neurochemical organization of the urodele brain, and to study discrepancies in the localization of monoaminergic markers, we immunohistochemically charted the distribution of four such markers (tyrosine hydroxylase, aromatic L-amino acid decarboxylase, dopamine, and serotonin) in the axolotl (Ambystoma mexicanum) forebrain. Catecholaminergic and serotoninergic systems were found in similar locations to those seen in other Urodela. As seen in other vertebrates, the localization of the different monoaminergic markers reveals some inconsistencies. Cells that are exclusively tyrosine hydroxylase-immunoreactive are observed in the olfactory bulb, anterior olfactory nucleus/nucleus accumbens region, the epichiasmatic portion of the preoptic nucleus, and in the pars intercalaris thalami, whereas cells that are only labelled by aromatic L-amino acid decarboxylase are seen in the anterior olfactory nucleus/nucleus accumbens region, the bed nuclei of the anterior commissure, the posterior portion of the preoptic nucleus, the ventral hypothalamus, and the pars intercalaris thalami. The presence of cells solely serotonin (5-HT)-immunoreactive is suggested for the nucleus infundibularis dorsalis. Conversely, there were no areas that appeared to be exclusively immunoreactive for dopamine. Double-labelling for aromatic L-amino acid decarboxylase/tyrosine hydroxylase and aromatic L-amino acid decarboxylase/serotonin, together with cell counting, confirmed the existence of neurons that express only one monoaminergic marker in amphibian, supporting the hypothesis that these cells are universally present in the central nervous system of vertebrates.

Ambystoma↗