Viral vectors for opto-electrode recording and photometry-based imaging of oxytocin neurons in anesthetized and socially interacting rats

Tang, Yan; Benusiglio, Diego; Lefèvre, Arthur; Küppers, Stephanie; Łapieś, Olga; Kerspern, Damien; Charlet, Alexandre; Grinevich, Valery

doi:10.1016/j.xpro.2021.101032

Cited by 10 publications

(12 citation statements)

References 8 publications

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“…However, we currently lack the ability to identify oxytocin neurons in vitro but such approaches are available. [51][52][53] Additional experiments using such approaches will be required to determine how BDL effects oxytocin neurons in male and female rats.…”

Section: Discussionmentioning

confidence: 99%

“…It is possible that some of the unlabeled cells that demonstrated impaired inhibition to muscimol were oxytocin cells. However, we currently lack the ability to identify oxytocin neurons in vitro but such approaches are available 51–53 . Additional experiments using such approaches will be required to determine how BDL effects oxytocin neurons in male and female rats.…”

Section: Discussionmentioning

confidence: 99%

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Effects of bile duct ligation on the inhibitory control of supraoptic vasopressin neurons

Aikins

Farmer

Little

et al. 2023

J Neuroendocrinology

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Dilutional hyponatremia due to increased plasma arginine vasopressin (AVP) is associated with liver cirrhosis. However, plasma AVP remains elevated despite progressive hypoosmolality. This study investigated changes to inhibitory control of supraoptic nucleus (SON) AVP neurons during liver cirrhosis. Experiments were conducted with adult male Sprague-Dawley rats. Bile duct ligation was used as a model of chronic liver cirrhosis. An adeno-associated virus containing a construct with an AVP promoter and either green fluorescent protein (GFP) or a ratiometric chloride indicator, ClopHensorN, was bilaterally injected into the SON of rats.After 2 weeks, rats received either BDL or sham surgery, and liver cirrhosis was allowed to develop for 4 weeks. In vitro, loose patch recordings of action potentials were obtained from GFP-labeled and unlabeled SON neurons in response to a brief focal application of the GABA A agonist muscimol (100 μM). Changes to intracellular chloride ([Cl]i) following muscimol application were determined by changes to the fluorescence ratio of ClopHensorN. The contribution of cation chloride cotransporters NKCC1 and KCC2 to changes in intracellular chloride was investigated using their respective antagonists, bumetanide (BU, 10 μM) and VU0240551 (10 μM). Plasma osmolality and hematocrit were measured as a marker of dilutional hyponatremia. The results showed reduced or absent GABA A -mediated inhibition in a greater proportion of AVP neurons from BDL rats as compared to sham rats (100% inhibition in sham vs. 47% in BDL, p = .001).Muscimol application was associated with increased [Cl]i in most cells from BDL as compared to cells from sham rats (χ 2 = 30.24, p < .001). NKCC1 contributed to the impaired inhibition observed in BDL rats (p < .001 BDL À BU vs. BDL + BU).The results show that impaired inhibition of SON AVP neurons and increased intracellular chloride contribute to the sustained dilutional hyponatremia in liver cirrhosis.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Effects of bile duct ligation on the inhibitory control of supraoptic vasopressin neurons

Aikins

Farmer

Little

et al. 2023

J Neuroendocrinology

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“…Adult female Sprague Dawley rats were anesthetized with 2-4% isoflurane and mounted onto the stereotaxic frame. As described previously 97 , an AAV expressing oxytocin promoter (pOT) - ChrimsonR-tdTomato was injected into the PVN, and an AAV expressing hSyn-OT1.0 was injected into the PVN or SON of SD-rat using the following coordinates (PVN, AP: −1.8 mm, ML: 0.4 mm, DV: −8.0 mm relative to Bregma; SON, AP: −1.2 mm, ML: 1.8 mm, DV: −9.2 mm relative to Bregma); 3 weeks after virus injections, an optical fiber (400 um, 0.5NA) was implanted in the PVN or SON to activate oxytocin neuron somas or axons and record oxytocin release. To this purpose it was coupled with a fiber patch cord (200 um, 0.37NA) and linked to an integrated fluorescence minicube (Doric ilFMC5).…”

Section: Methodsmentioning

confidence: 99%

Compartmental Neuropeptide Release Measured Using a New Oxytocin Sensor

Qian

Wang

et al. 2022

Preprint

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As a peptide hormone and neuromodulator, oxytocin (OT) plays a critical role in a variety of physiological and pathophysiological processes in both the central nervous system and the periphery. However, the processes that regulate spatial OT release in the brain remain enigmatic. Here, we developed a genetically encoded GPCR activation-based (GRAB) OT sensor called GRABOT1.0. Using this sensor, we directly visualized stimulation-induced OT release from specific compartments of OT neurons in acute brain slices, and discovered that N-type calcium channels predominantly mediate axonal OT release, while L-type calcium channels mediate somatodendritic OT release. In addition, we found that components in the fusion machinery of OT release differ between axon terminals versus somata and dendrites. Finally, we demonstrated the sensor responses to the activation of OT neurons in various brain regions in vivo and revealed region specific OT release during male courtship behavior. Taken together, these results provide key insights regarding the role of compartmental OT release in the control of physiological and behavioral functions.

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“…All surgeries were performed on rats anesthetized with 2.5% isoflurane and receiving Bupivacaine (s.c., 2mg/kg) or carprofen (i.p., 5mg/kg) and lidocaine applied locally (Tang et al, 2022). rAAVs were injected into the PVN, SON and vlPAG in different combinations, as needed by each experiment, and allowed to express for four weeks.…”

Section: Stereotaxic Injectionsmentioning

confidence: 99%

A novel analgesic pathway from parvocellular oxytocin neurons to the periaqueductal gray

Iwasaki

Lefèvre

Althammer

et al. 2022

Preprint

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The hypothalamic neuropeptide, oxytocin (OT), exerts prominent analgesic effects via central and peripheral action. Here we discovered a novel subset of OT neurons whose projections preferentially terminate on OT receptor (OTR)-expressing neurons in the ventrolateral periaqueductal gray (vlPAG). Using a newly generated line of transgenic rats (OTR-IRES-Cre), we determined that most of the vlPAG OTR expressing cells being targeted by OT projections are GABAergic in nature. Both optogenetically-evoked axonal OT release in the vlPAG as well as chemogenetic activation of OTR vlPAG neurons results in a long-lasting overall increase of vlPAG neuronal activity. This then leads to an indirect suppression of sensory neuron activity in the spinal cord and strong analgesia. Finally, we describe a novel OT-vlPAG-spinal cord circuit that seems critical for analgesia in the context of both inflammatory and neuropathic pain.

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Viral vectors for opto-electrode recording and photometry-based imaging of oxytocin neurons in anesthetized and socially interacting rats

Cited by 10 publications

References 8 publications

Effects of bile duct ligation on the inhibitory control of supraoptic vasopressin neurons

Effects of bile duct ligation on the inhibitory control of supraoptic vasopressin neurons

Compartmental Neuropeptide Release Measured Using a New Oxytocin Sensor

A novel analgesic pathway from parvocellular oxytocin neurons to the periaqueductal gray

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