System-Wide Immunohistochemical Analysis of Protein Co-Localization

Background: The analysis of co-localized protein expression in a tissue section is often conducted with immunofluorescence histochemical staining which is typically visualized in localized regions. On the other hand, chromogenic immunohistochemical staining, in general, is not suitable for the detection of protein co-localization. Here, we developed a new protocol, based on chromogenic immunohistochemical stain, for system-wide detection of protein co-localization and differential expression. Methodology/Principal Findings: In combination with a removable chromogenic stain, an efficient antibody stripping method was developed to enable sequential immunostaining with different primary antibodies regardless of antibody’s host species. Sections were scanned after each staining, and the images were superimposed together for the detection of protein co-localization and differential expression. As a proof of principle, differential expression and co-localization of glutamic acid decarboxylase67 (GAD67) and parvalbumin proteins was examined in mouse cortex. Conclusions/Significance: All parvalbumin-containing neurons express GAD67 protein, and GAD67-positive neurons that do not express parvalbumin were readily visualized from thousands of other neurons across mouse cortex. The method provided a global view of protein co-localization as well as differential expression across an entire tissue section. Repeate

Mapping of neurokinin-like immunoreactivity in the human brainstem

BACKGROUND: Using an indirect immunoperoxidase technique, we have studied the distribution of immunoreactive fibers and cell bodies containing neurokinin in the adult human brainstem with no prior history of neurological or psychiatric disease. RESULTS: Clusters of immunoreactive cell bodies and high densities of neurokinin-immunoreactive fibers were located in the periaqueductal gray, the dorsal motor nucleus of the vagus and in the reticular formation of the medulla, pons and mesencephalon. Moreover, immunoreactive cell bodies were found in the inferior colliculus, the raphe obscurus, the nucleus prepositus hypoglossi, and in the midline of the anterior medulla oblongata. In general, immunoreactive fibers containing neurokinin were observed throughout the whole brainstem. In addition to the nuclei mentioned above, the highest densities of such immunoreactive fibers were located in the spinal trigeminal nucleus, the lateral reticular nucleus, the nucleus of the solitary tract, the superior colliculus, the substantia nigra, the nucleus ambiguus, the gracile nucleus, the cuneate nucleus, the motor hypoglossal nucleus, the medial and superior vestibular nuclei, the nucleus prepositus hypoglossi and the interpeduncular nucleus. CONCLUSION: The widespread distribution of immunoreactive structures containing neurokinin in the human brainstem indicates that neurokinin might be involved in several physiological mechanisms, acting as a neurotransmitter and/or neuromodulator

Substrats neuroanatomiques et cellulaires de l'anorexie associée à une inflammation périphérique chez le rat

BORDEAUX1-BU Sciences-Talence (335222101) / SudocSudocFranceF

Etude de l'induction de la production des ARNm codant la neurotensine dans le noyau paraventriculaire de l'hypothalamus de rat lors de l'inflammation provoquée par le lipopolysaccharide

Lors d'une inflammation, les cellules immunitaires libèrent des cytokines qui agissent sur le système nerveux central pour déclencher une réponse au niveau végétatif, neuroendocrinien et comportemental. Plus particulièrement, l'inflammation stiumle le neurone à corticolibérine (CHR) du noyau paraventriculaire (NPV). Une manière d'aborder le problème de l'impact de l'infection sur la production peptidique de ce neurone est de mobiliser le système immunitaire par administration du lipopolysaccharide (LPS). chez le rat, quelques heures après son administration il se produit une augmentation de la production d'ARNm de la CHR dans le NPV, une augmentation de la production d'ARNm de la cholécystikinine dans la majorité de ces neurones et une induction d'ARNm de la neurotensine dans envrion 1/3 de ces mêmes neurones qui n'en expriment pas à l'état basal. Puisqu'elle résulte d'une induction, la neurotensine produite dans le NPV après traitement au LPS a probablement un rôle spécifique dans la réponse à l'inflammation. L'influence qu'à l'activation du système immunitaire sur la production d'ARNm de la neurotensine semble être spécifique au neurone à CHR du NPV. ces données sont en faveur d'un rôle de la neurotensine dans la réponse corticotrope à l'inflammation. De plus, nous montrons que les neurones exprimant l'ARN de la neurotensine durant l'inflammation sont des neurones qui se projettent dans l'éminence médiane. La neurotensine est alors probablement déversée dans le sang porte-hypophysaire....BORDEAUX1-BU Sciences-Talence (335222101) / SudocSudocFranceF

Distribution of galanin immunoreactivity in the sheep diencephalon

International audienceAlthough 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 awes, 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

Hypotension orthostatique asympathicotonique révélatrice d une dysautonomie associée à une hépatite C chronique

L'hypotension orthostatique est un problème fréquent, notamment dans la population âgée. Elle est dite asympathicotonique quand l'accélération cardiaque lors de la chute tensionnelle est absente ou modeste, et signe alors une atteinte organique du système nerveux autonome (SNA). Le but de cet exposé est tout d'abord de rappeler les grands principes de la prise en charge clinique de l'hypotension orthostatique puis des dysautonomies. La physiopathologie, le diagnostic étiologique et les traitements y seront ainsi détaillés. Ce propos sera ensuite illustré par deux cas cliniques présentant des dysautonomies sévères à priori primitives. Dans ces deux observations, est déouverte fortuitement une hépatite C chronique. Sur la base de cette constatation et au travers d'une revue de la littérature, nous discuterons de la relation de cause à effet éventuelle entre l'hépatite C et la dysautonomie. L'insuffisance hépatocellulaire s'accompagne fréquemment de dysfonctionnements du SNA dont l'importance est grosso modo corrélée au degré d'insuffisance hépatique. Dans les hépatites C, les études ne mettent en évidence que des désordres minimes, infracliniques.BORDEAUX2-BU Santé (330632101) / SudocSudocFranceF

Influence du niveau nutritionnel sur les neurones à corticolibérine (CRF) du noyau paraventriculaire. Etude immunohistochimique chez la brebis

National audienc

Presence of galanin in dopaminergic neurons of the sheep infundibular nucleus: a double staining immunohistochemical study

International audienc

Immunohistochemical characterization of the sheep suprachiasmatic nucleus

International audienc