Optimization of whole-brain rabies virus tracing technology for small cell populations

Roelofs, Theresia J. M.; Menting-Henry, Shanice; Gol, Lieke M.; Speel, Annelijn M.; Wielenga, Vera H.; Garner, Keith M.; Luijendijk, Mieneke C. M.; Hennrich, Alexandru A; Conzelmann, Karl-Klaus; Adan, Roger A.H.

doi:10.1038/s41598-021-89862-5

Cited by 5 publications

(4 citation statements)

References 55 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To figure out which of the brain regions host cholinergic cells innervating GnRH neurons, a monosynaptic, rabies virus retrograde tract tracing technique was used (Wall et al, 2010; Roelofs et al, 2021) in combination with immunocytochemistry. GnRH-Cre neurons ( Fig.…”

Section: Resultsmentioning

confidence: 99%

Cholinergic control of GnRH neuron physiology and luteinizing hormone secretion: involvement of ACh/GABA co-transmission

Vastagh,

Farkas,

Csillag

et al. 2023

Preprint

View full text Add to dashboard Cite

Gonadotropin-releasing hormone (GnRH)-synthesizing neurons orchestrate reproduction centrally. They receive commands from various neuronal networks for regulation of the hypothalamo-pituitary-gonadal (HPG)-axis. Here we report acetylcholine (ACh) as a critical regulator of GnRH neurons and reproduction in mice. In acute brain slice preparation, the acetylcholine receptor (AChR) agonists, ACh and carbachol, evoked a biphasic effect on the firing rate in GnRH neurons, first increasing afterwards diminishing it. In the presence of tetrodotoxin, carbachol induced an inward current followed by a decline in frequency of mPSCs. RT-PCR and whole cell patch-clamp studies revealed that GnRH neurons expressed both muscarinic (M1-M5) and nicotinic (α4β2, α3β4 and α7) ACh receptors. Muscarine via M1 and M3 receptors increased, while via M2 and M4 reduced the frequency of both miniature postsynaptic currents (mPSCs) and firing. The identified nicotinic AChRs contributed to the nicotine-elicited inward current and the rise of firing rate. Optogenetic activation of channelrhodopsin-2-tagged cholinergic axons modified GnRH neuronal activity and evoked co-transmission of ACh and GABA from a subpopulation of boutons. 3DISCO immunocytochemistry and electron microscopy revealed innervation of GnRH neurons by cholinergic axons. Rabies retrograde viral labelling started from GnRH-Cre neurons identified the medial septum and the diagonal band of Broca as exclusive sites of origin for specific cholinergic afferents.In vivopharmacogenetic activation of cholinergic system resulted in an increased secretion of luteinizing hormone (LH) in gonadectomized mice. Our findings collectively corroborate that the central cholinergic system exerts an important regulatory influence upon GnRH neurons and activates the HPG-axis via ACh and ACh/GABA neurotransmissions.Graphical abstract

show abstract

Section: Resultsmentioning

confidence: 99%

Cholinergic control of GnRH neuron physiology and luteinizing hormone secretion: involvement of ACh/GABA co-transmission

Vastagh,

Farkas,

Csillag

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

“…It is also important to mention that the transfection efficiency can vary significantly as the result of numerous factors (promoter, viral titer, injection protocols, injection efficacy, the cell type targeted, and transgenic mouse lines used); therefore, the number of starter cells can vary between different studies as well as between animals within a single study (Callaway and Luo, 2015;Roelofs et al, 2021). For this reason, it is convention to view the results as ratios instead of absolute numbers.…”

Section: Discussionmentioning

confidence: 99%

Whole-brain mapping of long-range inputs to the VIP-expressing inhibitory neurons in the primary motor cortex

Lee,

Côté,

Raman

et al. 2023

Front. Neural Circuits

View full text Add to dashboard Cite

The primary motor cortex (MOp) is an important site for motor skill learning. Interestingly, neurons in MOp possess reward-related activity, presumably to facilitate reward-based motor learning. While pyramidal neurons (PNs) and different subtypes of GABAergic inhibitory interneurons (INs) in MOp all undergo cell-type specific plastic changes during motor learning, the vasoactive intestinal peptide-expressing inhibitory interneurons (VIP-INs) in MOp have been shown to preferentially respond to reward and play a critical role in the early phases of motor learning by triggering local circuit plasticity. To understand how VIP-INs might integrate various streams of information, such as sensory, pre-motor, and reward-related inputs, to regulate local plasticity in MOp, we performed monosynaptic rabies tracing experiments and employed an automated cell counting pipeline to generate a comprehensive map of brain-wide inputs to VIP-INs in MOp. We then compared this input profile to the brain-wide inputs to somatostatin-expressing inhibitory interneurons (SST-INs) and parvalbumin-expressing inhibitory interneurons (PV-INs) in MOp. We found that while all cell types received major inputs from sensory, motor, and prefrontal cortical regions, as well as from various thalamic nuclei, VIP-INs received more inputs from the orbital frontal cortex (ORB) – a region associated with reinforcement learning and value predictions. Our findings provide insight on how the brain leverages microcircuit motifs by both integrating and partitioning different streams of long-range input to modulate local circuit activity and plasticity.

show abstract

“…To identify the location/s/ of ChAT-IR neurons that monosynaptically innervate GnRH neurons, the septo-preoptic area of GnRH-Cre mice was infected in two steps with genetically altered helper and rabies viruses ( Wall et al, 2010 ; Roelofs et al, 2021 ). Helper virus-infected GnRH-Cre neurons expressing avian tumor virus receptor A (TVA) receptor and G-proteins became targets of the modified, envelope protein A (EnvA)-coated, G-deleted rabies virus (RVdG) which then labeled the monosynaptically connected afferent neurons with their fluorescent transgene.…”

Section: Methodsmentioning

confidence: 99%

Cholinergic control of GnRH neuron physiology and luteinizing hormone secretion in male mice: involvement of ACh/GABA co-transmission

Vastagh,

Farkas,

Csillag

et al. 2024

J. Neurosci.

View full text Add to dashboard Cite

GnRH-synthesizing neurons orchestrate reproduction centrally. Early studies have proposed the contribution of acetylcholine (ACh) to hypothalamic control of reproduction, although the causal mechanisms haven't been clarified. Here, we report thatin vivopharmacogenetic activation of the cholinergic system increased the secretion of luteinizing hormone (LH) in orchidectomized mice. 3DISCO immunocytochemistry and electron microscopy revealed the innervation of GnRH neurons by cholinergic axons. Retrograde viral labeling initiated from GnRH-Cre neurons identified the medial septum and the diagonal band of Broca as exclusive sites of origin for cholinergic afferents of GnRH neurons. In acute brain slices, ACh and the ACh receptor (AChR) agonist carbachol evoked a biphasic effect on the firing rate in GnRH neurons, first increasing and then diminishing it. In the presence of tetrodotoxin, carbachol induced an inward current, followed by a decline in the frequency of mPSCs, indicating a direct influence on GnRH cells. RT-PCR and whole-cell patch-clamp studies revealed that GnRH neurons expressed both nicotinic (α4β2, α3β4, and α7) and muscarinic (M1-M5) ACh receptors. The nicotinic AChRs contributed to the nicotine-elicited inward current and the rise in firing rate. Muscarine via M1 and M3 receptors increased, while via M2 and M4 reduced the frequency of both miniature postsynaptic currents (mPSCs) and firing. Optogenetic activation of channelrhodopsin-2-tagged cholinergic axons modified GnRH neuronal activity and evoked co-transmission of ACh and GABA from a subpopulation of boutons. These findings confirm that the central cholinergic system immensely regulates GnRH neurons and activates the HPG-axis via ACh and ACh/GABA neurotransmissions in male mice.Significance statementCholinergic drugs influence reproduction centrally, although the exact neuronal targets and regulatory mechanisms remain unsettled. We found that pharmacogenetic activation of the cholinergic systemin vivoevoked an augmented LH release in male mice. The study also identified cholinergic cell groups in the mouse forebrain that innervate gonadotropin-releasing hormone (GnRH) neurons, the main hypothalamic regulators of reproduction. We also determined the subtypes of nicotinic and muscarinic receptors involved in neuronal information transfer and explored how their ligands affect the electrophysiological activity of GnRH neurons. A subset of cholinergic neurons was found to co-transmit GABA which excites GnRH cells via GABA-A receptors. The findings suggest a novel, cholinergic regulation of the GnRH system activating the pituitary-gonadal axis in adult male mice.

show abstract

Optimization of whole-brain rabies virus tracing technology for small cell populations

Cited by 5 publications

References 55 publications

Cholinergic control of GnRH neuron physiology and luteinizing hormone secretion: involvement of ACh/GABA co-transmission

Cholinergic control of GnRH neuron physiology and luteinizing hormone secretion: involvement of ACh/GABA co-transmission

Whole-brain mapping of long-range inputs to the VIP-expressing inhibitory neurons in the primary motor cortex

Cholinergic control of GnRH neuron physiology and luteinizing hormone secretion in male mice: involvement of ACh/GABA co-transmission

Contact Info

Product

Resources

About