Expression and regulation of the immediate-early gene product Arc in the accessory olfactory bulb after mating in male rat

doi:10.1016/S0306-4522(01)00620-0

Neuroscience

Volume 111, Issue 2, 10 May 2002, Pages 251-258

https://doi.org/10.1016/S0306-4522(01)00620-0 Get rights and content

Abstract

Recent studies of the accessory olfactory bulb have shown that the expression of immediate-early genes, e.g., c-fos, c-jun and egr-1, can be used as a marker of neuronal activity in response to pheromonal cues. In this study, we analyzed the expression pattern, in response to mating, of the novel immediate-early gene product Arc (an activity-regulated cytoskeleton-associated protein). Arc is hypothesized to play a role in activity-dependent neuronal plasticity in the hippocampus. In a control group of male rats, only a small number of Arc-immunoreactive cells were observed in the accessory olfactory bulb. In a mating group, however, a marked increase in the number of Arc-immunoreactive cells was observed only in the granule cell layer of the accessory olfactory bulb. The increase in the number of Arc-immunoreactive cells after mating was similar to that observed for other immediate-early genes. However, for the mating group, the increase in Arc-positive cells was limited to the granule cell layer. Granule cells have been shown to exhibit a strong synaptic plasticity in response to pheromonal stimulation.

From these findings we suggest that Arc plays an important role in neuronal plasticity in the accessory olfactory bulb.

Section snippets

Animal treatment and tissue preparation

Sexually inexperienced male Sprague–Dawley (SD) rats (Clea Japan, Japan), 7–8 weeks old, were used for immunoblot and immunocytochemical analyses. Animals were housed in standard laboratory cages (40×30×20 cm, one per cage) with food and water available ad libitum under 12-h reversed light cycles (lights on 8.00 h). The animals were randomly divided into two groups: (1) rats placed in contact with an estrous female in a cage with the soiled bedding of females which had not been cleaned for more

Immunoblot analysis

To investigate whether the Arc protein is induced in the AOB after mating, the anti-Arc antibody was used for western blot analysis. Consequently, single protein bands with an apparent molecular weight of 55 kDa were identified by western blot analysis of AOB of male rats. The Arc protein was observed in the AOB of control group, but it was strongly induced in that of the mating group (Fig. 1). The c-Fos protein was also induced in the AOB of the males of the mating group. In addition, we

Localization of Arc protein in the AOB after mating

The vomeronasal system has an important role in the mammalian reproductive function (Wysocki, 1979, Halpern, 1987). The AOB is the first relay station in the vomeronasal system. The aim of the present study was to investigate the changes of the novel IEG product, Arc, in the AOB of male rats after mating. Firstly, we performed western blot analysis of AOB of male rats. A single protein band with an apparent molecular weight of 55 kDa was identified, and induction of the protein in the AOB of

Acknowledgements

We wish to thank Dr. R.M. Costanzo for helpful discussions and assistance. This work was supported in part by CREST of JST (M.M. and M.I.), a Grant-in-Aid for Scientific Research (13760192) from the Japan Society for the Promotion of Science and the Ministry of Education, Science, Sports and Culture of Japan (M.M.), grants from the Japan Society for the Promotion of Science and the Ministry of Education, Science, Sports and Culture of Japan (K.Y.) and a Grant-in-Aid for Scientific Research (C)

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The AOB subdivisions in rats respond differently to social stimulation
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The animal's ability to recognise and differentiate between familiar and unfamiliar conspecifics, plays a crucial role in their social behaviour. To study the neural mechanisms underlying individual recognition, the expression patterns of the Immediate Early Genes c-Fos, Egr.1, and Arc were investigated immunocytochemically in the brains of socially stimulated rats. Juvenile stimulated rats showed a significantly increased expression of c-Fos, Egr.1, and Arc in the accessory olfactory bulb in comparison with the control and carvone stimulated groups. Also, in the anterior AOB, more cells were activated than in the posterior AOB.
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The brain-specific immediate early gene Arc/Arg3.1 is induced in response to a variety of stimuli, including sensory and behavior-linked neural activity. Here we report the generation of transgenic mice, termed TgArc/Arg3.1-d4EGFP, expressing a 4-h half-life form of enhanced green fluorescent protein (d4EGFP) under the control of the Arc/Arg3.1 promoter. We show that d4EGFP-mediated fluorescence faithfully reports Arc/Arg3.1 induction in response to physiological, pathological and pharmacological stimuli, and that this fluorescence permits electrical recording from activated neurons in the live mouse. Moreover, the fluorescent Arc/Arg3.1 indicator revealed activity changes in circumscribed brain areas in distinct modes of stress and in a mouse model of Alzheimer's disease. These findings identify the TgArc/Arg3.1-d4EGFP mouse as a versatile tool to monitor Arc/Arg3.1 induction in neural circuits, both in vitro and in vivo.
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Axon guidance molecules and related proteins such as semaphorin 3A, neuropilin-1, plexin-1, netrin-1, growth-associated protein, olfactory marker protein, cypin and collapsin response mediator proteins guide the development of neural circuits in the olfactory bulb. In this study, transcriptions of these genes were examined in the olfactory bulb of female, male and neonatal testosterone propionate-treated female rats at the ages of 2, 5, 10, 15, 20, 25, 30 and 45 days. The semaphorin 3A, neuropilin-1, growth-associated protein and collapsin response mediator protein 1–5 genes were expressed significantly higher during the early development stages than in adulthood while the opposite is true for the olfactory marker protein. The expression profile of cypin and netrin-1 was relatively constant through development. A late effect of the neonatal testosterone propionate treatment on netrin-1, growth-associated protein, olfactory marker protein, collapsin response mediator proteins 1, 3, 4 and cypin gene expression was observed. The expression profiles of collapsin response mediator proteins and their related genes in the developing olfactory bulb confirmed most studies on the relationship between collapsin response mediator proteins and development in the brain. Sex differences of semaphorin 3A, neuropilin-1 as well as collapsin response mediator protein 3 at the early development stage and the late effect of neonatal testosterone propionate treatment on the expressions of netrin-1, growth-associated marker protein, cypin and collapsin response mediator proteins 1, 3 and 5 genes may indicate a possible role of these molecules on sexual differentiation of the olfactory bulb.
Rapid induction of Arc is observed in the granule cell dendrites in the accessory olfactory bulb after mating
2003, Brain Research
Citation Excerpt :
Similarly, Arc induction can be used as a marker for neuronal activity in response to chemosensory cues, and indeed, it has been shown that various chemosensory cues induce the synthesis of Arc protein [5,25]. Recently, we reported that increases in Arc-immunoreactive (Arc-ir) cells in the AOB of male rats were observed after mating [22]. The induction of Arc after mating localized in the granule cell layer of the AOB, a few Arc-ir cells were observed in the main olfactory bulb [24].
The activity-regulated cytoskeleton-associated protein (Arc), encoded by the immediate early gene arc, is enriched in the brain and is hypothesized to play a role in the activity-dependent neuronal plasticity in the hippocampus. In the present study, the time course of Arc expression during the post-mating period was determined immunocytochemically, and the localization of Arc in the neurons in the accessory olfactory bulb (AOB) of female mice after mating was analyzed using immunocytochemical electron microscopy. Transient increases in the number of Arc-immunoreactive cells were observed in the glomerular, mitral/tufted cell and granule cell layers of the AOB after mating. In particular, the increase in the granule cell layer was remarkable, and larger than the increases in the other layers. In addition, electron microscopic observation revealed that Arc immunoreactivity was in the dendrites of the granule cells 1.5 h after mating. These results indicate that expression of Arc protein is induced rapidly and transiently in granule cell dendrites after mating. It is postulated that Arc protein has a role in the neuronal plasticity of the AOB after mating.
Structure and function of the vomeronasal system: An update
2003, Progress in Neurobiology
Several developments during the past 15 years have profoundly affected our understanding of the vomeronasal system (VNS) of vertebrates. In the mid 1990s, the vomeronasal epithelium of mammals was found to contain two populations of receptor cells, based on their expression of G-proteins. These two populations of neurons were subsequently found to project their axons to different parts of the accessory olfactory bulb (AOB), forming the basis of segregated pathways with possibly heterogeneous functions. A related discovery was the cloning of members of at least two gene families of putative vomeronasal G-protein-coupled receptors (GPRs) in the vomeronasal epithelium. Ligand binding to these receptors was found to activate a phospholipase C (PLC)-dependent signal transduction pathway that primarily involves an increase in intracellular inositol-tris-phosphate and intracellular calcium. In contrast to what was previously believed, neuron replacement in the vomeronasal epithelium appears to occur through a process of vertical migration in most mammals. New anatomical studies of the central pathways of the olfactory and vomeronasal systems indicated that these two systems converge on neurons in the telencephalon, providing an anatomical substrate for functional interactions. Combined anatomical, physiological and behavioral studies in mice provided new information that furthered our understanding of one of the most striking pheromonal phenomena, the Bruce effect. Finally, contrary to prior observations, new anatomical studies indicated that a vomeronasal organ (VNO) was present in human adults and reports were published indicating that this system might be functional. These latter observations are still controversial and require confirmation from independent laboratories.
Mating behavior induces differential Arc expression in the main and accessory olfactory bulbs of adult rats
2002, Neuroscience Letters
The expression of activity-related cytoskeleton-associated protein, Arc, could be useful as a marker for neuronal activity. We investigated Arc-immunoreactivity in both the accessory olfactory bulb (AOB) and the main olfactory bulb (MOB) of adult male rats in response to mating or exposure to female pheromones. Mating behavior strongly enhanced the Arc-immunoreactivity in the granule cell layer of the AOB. However, the enhancement of Arc-immunoreactivity by mating behavior was not observed in the MOB. These results showed that Arc-immunoreactivity was enhanced when the AOB received both afferent and efferent information during mating behavior. Hence, the expression of Arc in the AOB directly associates the pheromonal information with mating behavior. The AOB will provide a useful model to investigate the function of Arc protein.

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Expression and regulation of the immediate-early gene product Arc in the accessory olfactory bulb after mating in male rat

Abstract

Section snippets

Animal treatment and tissue preparation

Immunoblot analysis

Localization of Arc protein in the AOB after mating

Acknowledgements

Neuroscience

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Neuroscience

Neuroscience

Neuroscience

Brain Res.

Behav. Biol.

Neuron

Neurosci. Res.

Neurosci. Res.

Neuroscience

Brain Res. Bull.

Cell

Neuroscience

Brain Res.

Anim. Behav.

Neuron

Neurosci. Biobehav. Rev.

Olfactory recognition: a simple memory system

Science

An exteroceptive block to pregnancy in the mouse

Nature