Jean Gascuel

Specific expression of olfactory binding protein in the aerial olfactory cavity of adult and developing Xenopus

Olfactory binding proteins (OBP), commonly associated with aerial olfaction, are found in the olfactory mucus of mammals but have never been identified in fish. It is still not clear whether the presence of OBP in aerial olfactory systems is due to phylogenetic or to functional differences linked to the adaptation of the olfactory system to an aerial environment. To test this alternative, the olfactory system of Xenopus offers a unique opportunity because it includes two olfactory cavities, one of which is thought to be devoted to aquatic olfaction and the other to aerial olfaction. We therefore purified and cloned OBPs in two Xenopus species. Xenopus laevis OBP (XlaeOBP) and Xenopus tropicalis OBP (XtroOBP) exhibit 158 and 160 amino acids, respectively, sharing 89 residues. cRNA probes allowed us to demonstrate that XlaeOBP and XtroOBP are expressed at the level of Bowmans gland specifically in the aerial olfactory cavity, as confirmed using anti‐XlaeOBP antiserum. OBP mRNA transcription occurs early during metamorphosis, as early as stage 57. This is the first study to demonstrate that OBPs are exclusively present in the aerial chamber and are only expressed as the tadpole becomes an adult in species which possess both aquatic and aerial olfactory organs.

Hypothalamus-Olfactory System Crosstalk: Orexin A Immunostaining in Mice

It is well known that olfaction influences food intake, and conversely, that an individual’s nutritional status modulates olfactory sensitivity. However, what is still poorly understood is the neuronal correlate of this relationship, as well as the connections between the olfactory bulb and the hypothalamus. The goal of this report is to analyze the relationship between the olfactory bulb and hypothalamus, focusing on orexin A immunostaining, a hypothalamic neuropeptide that is thought to play a role in states of sleep/wakefulness. Interestingly, orexin A has also been described as a food intake stimulator. Such an effect may be due in part to the stimulation of the olfactory bulbar pathway. In rats, orexin positive cells are concentrated strictly in the lateral hypothalamus, while their projections invade nearly the entire brain including the olfactory system. Therefore, orexin appears to be a good candidate to play a pivotal role in connecting olfactory and hypothalamic pathways. So far, orexin has been described in rats, however, there is still a lack of information concerning its expression in the brains of adult and developing mice. In this context, we revisited the orexin A pattern in adult and developing mice using immunohistological methods and confocal microscopy. Besides minor differences, orexin A immunostaining in mice shares many features with those observed in rats. In the olfactory bulb, even though there are few orexin projections, they reach all the different layers of the olfactory bulb. In contrast to the presence of orexin projections in the main olfactory bulb, almost none have been found in the accessory olfactory bulb. The developmental expression of orexin A supports the hypothesis that orexin expression only appears post-natally.

A morphometric classification of pupal honeybee antennal lobe neurones in culture

BOTH intrinsic (programmed), and extrinsic (non-programmed) factors are thought to play a role in the morphogenesis of neurones in the honeybee antennal lobe (the first relay station in the olfactory pathway) during development. We present here a morphometric, and statistical analysis of a large population of pupal honeybee antennal lobe neurones grown in primary culture. Quantitative parameters were used to characterize neuronal shapes. On the basis of such morphometric measurements, an automatic classification allowed three morphological types to be distinguished. Their main characteristics are presented, and the role of intrinsic, and extrinsic factors in their determination is discussed.

Glial fibrillary acidic protein and vimentin expression in the frog olfactory system during metamorphosis

In the present study, we investigated glial cell organization in the olfactory system of adult and tadpole Xenopus laevis using glial fibrillary acidic protein and vimentin antibodies. Our results showed for the first time that glial fibrillary acidic protein was strongly expressed at the level of the olfactory nerve from tadpole to adult and was likely to be expressed by ensheathing glia. In the olfactory bulb, the nerve layer was stained, and no staining was observed in glomeruli. By contrast, vimentin decorated radial glia in the bulb but faintly stained the olfactory nerve. Interestingly, glial fibrillary acidic protein and vimentin presented complementary staining patterns, with glial fibrillary acidic protein being expressed in the peripheral olfactory system and vimentin being expressed in the central part of the olfactory system.

Brain FOS-like expression in developing and adult honeybees.

This study was undertaken to examine the expression of the protooncogene c-fos in an invertebrate, the honeybee. We investigated first the expression of Fos-like proteins, testing different methodologies (immunohistochemistry, immunocytochemistry, immunoblotting). These studies were carried out at the embryonic, nymphal and adult stages. We found immunoreactive bands at approximately 30 kD and 57 kD. The first ones are revealed at the embryonic, nymphal and adult stages. The approximately 57 kD proteins are present at the adult stage. Beside this qualitative variation, intensive modification occurs with a burst of protein expression a few days after the insect emergence. The results suggest that Fos-like proteins are found in the developing and maturing nervous system. Basal level of expression is found in the olfactory pathway, convincing to explore the protooncogene c-fos expression in the cellular and molecular analyses of olfactory plasticity.

Transient Receptor Potential Canonical 3 (TRPC3) Channels Are Required for Hypothalamic Glucose Detection and Energy Homeostasis

The mediobasal hypothalamus (MBH) contains neurons capable of directly detecting metabolic signals such as glucose to control energy homeostasis. Among them, glucose-excited (GE) neurons increase their electrical activity when glucose rises. In view of previous work, we hypothesized that transient receptor potential canonical type 3 (TRPC3) channels are involved in hypothalamic glucose detection and the control of energy homeostasis. To investigate the role of TRPC3, we used constitutive and conditional TRPC3-deficient mouse models. Hypothalamic glucose detection was studied in vivo by measuring food intake and insulin secretion in response to increased brain glucose level. The role of TRPC3 in GE neuron response to glucose was studied by using in vitro calcium imaging on freshly dissociated MBH neurons. We found that whole-body and MBH TRPC3-deficient mice have increased body weight and food intake. The anorectic effect of intracerebroventricular glucose and the insulin secretory response to intracarotid glucose injection are blunted in TRPC3-deficient mice. TRPC3 loss of function or pharmacological inhibition blunts calcium responses to glucose in MBH neurons in vitro. Together, the results demonstrate that TRPC3 channels are required for the response to glucose of MBH GE neurons and the central effect of glucose on insulin secretion and food intake.

Exotic models may offer unique opportunities to decipher specific scientific question: the case of Xenopus olfactory system.

The fact that olfactory systems are highly conserved in all animal species from insects to mammals allow the generalization of findings from one species to another. Most of our knowledge about the anatomy and physiology of the olfactory system comes from data obtained in a very limited number of biological models such as rodents, Zebrafish, Drosophila, and a worm, Caenorhabditis elegans. These models have proved useful to answer most questions in the field of olfaction, and thus concentrating on these few models appear to be a pragmatic strategy. However, the diversity of the organization and physiology of the olfactory system amongst phyla appear to be greater than generally assumed and the four models alone may not be sufficient to address all the questions arising from the study of olfaction. In this article, we will illustrate the idea that we should take advantage of biological diversity to address specific scientific questions and will show that the Xenopus olfactory system is a very good model to investigate: first, olfaction in aerial versus aquatic conditions and second, mechanisms underlying postnatal reorganization of the olfactory system especially those controlled by tyroxine hormone. Anat Rec, 296:1453‐1461, 2013.

Expression of Odorant Receptor Family, Type 2 OR in the Aquatic Olfactory Cavity of Amphibian Frog Xenopus tropicalis

Recent genome wide in silico analyses discovered a new family (type 2 or family H) of odorant receptors (ORs) in teleost fish and frogs. However, since there is no evidence of the expression of these novel OR genes in olfactory sensory neurons (OSN), it remains unknown if type 2 ORs (OR2) function as odorant receptors. In this study, we examined expression of OR2 genes in the frog Xenopus tropicalis. The overall gene expression pattern is highly complex and differs depending on the gene and developmental stage. RT-PCR analysis in larvae showed that all of the OR2η genes we identified were expressed in the peripheral olfactory system and some were detected in the brain and skin. Whole mount in situ hybridization of the larval olfactory cavity confirmed that at least two OR2η genes so far tested are expressed in the OSN. Because tadpoles are aquatic animals, OR2η genes are probably involved in aquatic olfaction. In adults, OR2η genes are expressed in the nose, brain, and testes to different degrees depending on the genes. OR2η expression in the olfactory system is restricted to the medium cavity, which participates in the detection of water-soluble odorants, suggesting that OR2ηs function as receptors for water-soluble odorants. Moreover, the fact that several OR2ηs are significantly expressed in non-olfactory organs suggests unknown roles in a range of biological processes other than putative odorant receptor functions.

Organizing metadata into models and ontologies for lowering annotation costs of a biological image database

In this chapter we present the initial phase towards lowering annotation costs in a specialized image database application. The proposed coupling of model-based description with a domain-specific ontology allows to use metadata during the annotation process. This makes creating annotations feasible even for domain technicians while guaranteeing consistency of annotations from an expert’s point of view.

Expression of the surface antigen A2B7 in adult and developing honeybee olfactory pathway

In order to identify molecules involved in the development of the honeybee olfactory pathway, hybridoma technology has been used. Among different cell lines, A2B7 has been selected. It produces a specific antibody for a surface glycoprotein of 91 kDa. This protein is mainly expressed by both the antennal receptor cells and mushroom body neurons. Based on (i) the spatio‐temporal pattern of expression during pupal development; (ii) the cell surface location of the antigen; and (iii) the partial molecular characterization of the antigen, a putative role for this protein in axonal fasciculation and guidance is discussed.