fg-9041 and 5-carboxamidotryptamine

Trauma during early life is a major risk factor for the development of anxiety disorders and suggests that the developing brain may be particularly sensitive to perturbation. Increased vulnerability most likely involves altering neural circuits involved in emotional regulation. The role of serotonin in emotional regulation is well established, but little is known about the postnatal development of the raphe where serotonin is made. Using whole-cell patch-clamp recording and immunohistochemistry, we tested whether serotonin circuitry in the dorsal and median raphe was functionally mature during the first 3 postnatal weeks in mice. Serotonin neurons at postnatal day 4 (P4) were hyperexcitable. The increased excitability was due to depolarized resting membrane potential, increased resistance, increased firing rate, lack of 5-HT1A autoreceptor response, and lack of GABA synaptic activity. Over the next 2 weeks, membrane resistance decreased and resting membrane potential hyperpolarized due in part to potassium current activation. The 5-HT1A autoreceptor-mediated inhibition did not develop until P21. The frequency of spontaneous inhibitory and excitatory events increased as neurons extended and refined their dendritic arbor. Serotonin colocalized with vGlut3 at P4 as in adulthood, suggesting enhanced release of glutamate alongside enhanced serotonin release. Because serotonin affects circuit development in other brain regions, altering the developmental trajectory of serotonin neuron excitability and release could have many downstream consequences. We conclude that serotonin neuron structure and function change substantially during the first 3 weeks of life during which external stressors could potentially alter circuit formation.

Topics: Age Factors; Animals; Animals, Newborn; Bacterial Proteins; Bicuculline; Environment; Excitatory Amino Acid Antagonists; Excitatory Postsynaptic Potentials; GABA-A Receptor Antagonists; Guanosine 5'-O-(3-Thiotriphosphate); In Vitro Techniques; Luminescent Proteins; Male; Mice; Mice, Transgenic; Quinoxalines; Raphe Nuclei; Serotonergic Neurons; Serotonin; Serotonin Receptor Agonists; Transcription Factors

The olfactory tubercle (OT) is found in the brains of mammals that are highly dependent on their sense of smell. Its human analogue is the poorly understood anterior perforated substance. Previous work on rat brain slices identified two types of field potential responses from the OT. The association fibre (AF) pathway was sensitive to muscarinic modulation, whereas the lateral olfactory tract (LOT) fibre pathway was not. Here, we establish that serotonin (5-hydroxytryptamine; 5-HT) also inhibits field potential excitatory postsynaptic potentials (EPSPs) in the AF, but not in the LOT fibre, pathway. Parallel experiments with adenosine (ADO) excluded ADO mediation of the 5-HT effect. Exogenous 5-HT at 30 microm caused a long-lasting approximately 40% reduction in the amplitude of AF postsynaptic responses, without affecting the time-course of EPSP decline, indicating a fairly restricted disposition of the 5-HT receptors responsible. The 5-HT(1)-preferring, 5-HT(5)-preferring and 5-HT(7)-preferring agonist 5-carboxamidotryptamine caused similar inhibition at approximately 100 nm. The 5-HT(1A)-preferring ligand 8-hydroxy-di-n-propylamino-tetralin at 10 microm, and the 5-HT uptake inhibitor citalopram at 3 microm, caused inhibition of AF-stimulated field potential responses in the 5-10% range. Order-of-potency information suggested a receptor of the 5-HT(1B) or 5-HT(1D) subtype. The 5-HT(1D) agonist L-694,247 (1 microm) suppressed the AF response by approximately 10% when used on its own. After washing out of L-694,427, inhibition by 30 microm 5-HT was reduced to negligible levels. Allowing for a partial agonist action of L-694,427 and complex interactions of 5-HT receptors within the OT, these results support the presence of active 5-HT(1D)-type receptors in the principal cell layer of the OT.

Topics: 8-Hydroxy-2-(di-n-propylamino)tetralin; Animals; Citalopram; Excitatory Amino Acid Antagonists; Excitatory Postsynaptic Potentials; In Vitro Techniques; Male; Olfactory Pathways; Oxadiazoles; Quinoxalines; Rats; Rats, Sprague-Dawley; Selective Serotonin Reuptake Inhibitors; Serotonin; Serotonin Receptor Agonists; Synaptic Transmission; Tryptamines