Analysis of various physiological salines for heart rate, CNS function, and synaptic transmission at neuromuscular junctions in Drosophila melanogaster larvae

Castro, Clara de; Titlow, Josh; Majeed, Zana R.; Cooper, Robin L.

doi:10.1007/s00359-013-0864-0

Cited by 45 publications

(23 citation statements)

References 56 publications

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“…After collection, early third instar larvae were pinned ventral side up on a glass plate and dissected in a droplet of saline (Cooper et al 2009). The Drosophila heart is very sensitive to pH (Gu and Singh 1995); therefore, the saline is adjusted to pH 7.1 and maintained with the high concentration of buffer as described in de Castro et al (2014). The larval dissection is described in detail by Gu and Singh (1995) and in video by Cooper et al (2009).…”

Section: Heart Rate Assaymentioning

confidence: 99%

“…Fly saline, modified Hemolymph-like 3 (HL3) (Stewart et al 1994) containing: (in mmol/L) 70 NaCl, 5 KCl, 20 MgCl 2 , 10 NaHCO 3 , 1 CaCl 2 , 5 trehalose, 115 sucrose, 25N,N-Bis-(2-hydroxyethyl)-2-aminoethane sulfonic acid (BES) was used. The following modifications were made to the HL3 saline: pH was decreased from 7.2 to 7.1 and BES buffer was increased from 5.0 to 25.0 mmol/L to maintain a stable pH (de Castro et al 2014).…”

Section: Pharmacologymentioning

confidence: 99%

“…In addition, the adult heart is modulated by neuronal inputs, which complicates addressing the function of the intrinsic cardiac pacemaker and ionic regulation in an intact heart Levine 2003, 2005). The larval heart is easily exposed, myogenic, and its activity can be maintained for hours with a newly developed physiological saline (de Castro et al 2014). Whereas previous research has utilized intact pupa or larvae with drug administration via injection, we directly expose an open preparation with pharmacological agents at known concentrations.…”

Section: Introductionmentioning

confidence: 99%

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Pharmacological identification of cholinergic receptor subtypes on Drosophila melanogaster larval heart

et al. 2015

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The Drosophila melanogaster heart is a popular model in which to study cardiac physiology and development. Progress has been made in understanding the role of endogenous compounds in regulating cardiac function in this model. It is well characterized that common neurotransmitters act on many peripheral and non-neuronal tissues as they flow through the hemolymph of insects. Many of these neuromodulators, including acetylcholine (ACh), have been shown to act directly on the D. melanogaster larval heart. ACh is a primary neurotransmitter in the central nervous system (CNS) of vertebrates and at the neuromuscular junctions on skeletal and cardiac tissue. In insects, ACh is the primary excitatory neurotransmitter of sensory neurons and is also prominent in the CNS. A full understanding regarding the regulation of the Drosophila cardiac physiology by the cholinergic system remains poorly understood. Here we use semi-intact D. melanogaster larvae to study the pharmacological profile of cholinergic receptor subtypes, nicotinic acetylcholine receptors (nAChRs) and muscarinic acetylcholine receptors (mAChRs), in modulating heart rate (HR). Cholinergic receptor agonists, nicotine and muscarine both increase HR, while nAChR agonist clothianidin exhibits no significant effect when exposed to an open preparation at concentrations as low as 100 nM. In addition, both nAChR and mAChR antagonists increase HR as well but also display capabilities of blocking agonist actions. These results provide evidence that both of these receptor subtypes display functional significance in regulating the larval heart's pacemaker activity.

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Section: Heart Rate Assaymentioning

confidence: 99%

Section: Pharmacologymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Pharmacological identification of cholinergic receptor subtypes on Drosophila melanogaster larval heart

et al. 2015

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“…49,50 However, it appears a physiological saline with a lower pH of 7.1 is substantially better for maintaining the heart function for exposed preparations. 51 In addition, heart rates increased in response to serotonin with a bathing saline that was buffered and maintained at 7.1. 51 In relation to mammals, we have not found any reports on the influence of pHo and alteration in the effects of neuromodulation by serotonin or dopamine on neuronal function in humans.…”

Section: Discussionmentioning

confidence: 99%

The Effect of CO2, Intracellular pH and Extracellular pH on Mechanosensory Proprioceptor Responses in Crayfish and Crab

Dayaram¹,

Malloy²,

Martha³

et al. 2017

AJUR

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Proprioceptive neurons monitor the movements of limbs and joints to transduce the movements into electrical signals. These neurons function similarly in species from arthropods to humans. These neurons can be compromised in disease states and in adverse environmental conditions such as with changes in external and internal pH. We used two model preparations (the crayfish muscle receptor organ and a chordotonal organ in the limb of a crab) to characterize the responses of these proprioceptors to external and internal pH changes as well as raised CO2. The results demonstrate the proprioceptive organs are not highly sensitive to changes in extracellular pH, when reduced to 5.0 from 7.4. However, if intracellular pH is decreased by exposure to propionic acid or saline containing CO2, there is a rapid decrease in firing rate in response to joint movements. The responses recover quickly upon reintroduction of normal pH (7.4) or saline not tainted with CO2. These basic understandings may help to address the mechanistic properties of mechanosensitive receptors in other organisms, such as muscle spindles in skeletal muscles of mammals and tactile as well as pressure (i.e., blood pressure) sensory receptors. KEYWORDS: Proprioception; Sensory; Invertebrate; Carbon Dioxide; Protons; Mechanosensory; Intracellular pH; Extracellular pH

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“…For dual-color and single-color calcium imaging in fictive preps, freshly dissected brains were mounted on 12mm round Poly-D-Lysine Coverslips (Corning® BioCoat™) in HL3.1 saline (de Castro et al 2014), which were then were placed on 25 mm × 75 mm glass slides to be imaged with a 40× objective on an upright Zeiss LSM-800 confocal microscopy. To simultaneously image two different neurons expressing GCaMP6m we imaged neuron-specific regions of interest (ROI).…”

Section: Methodsmentioning

confidence: 99%

ADrosophilalarval premotor/motor neuron connectome generating two behaviors via distinct spatio-temporal muscle activity

Zarin

Mark

Cardona

et al. 2019

Preprint

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Animals generate diverse motor behaviors, yet how the same motor neurons generate distinct behaviors remains an open question.Drosophilalarvae have multiple behaviors – e.g. forward crawling, backward crawling, self-righting and escape – and all of the body wall motor neurons (MNs) driving these behaviors have been identified. Despite impressive progress in mapping larval motor circuits, the role of most motor neurons in locomotion remains untested, the majority of premotor neurons (PMNs) remain to be identified, and a full understanding of proprioceptor-PMN-MN connectivity is missing. Here we report a comprehensive larval proprioceptor-PMN-MN connectome; describe individual muscle/MN phase activity during both forward and backward locomotor behaviors; identify PMN-MN connectivity motifs that could generate muscle activity phase relationships, plus selected experimental validation; identify proprioceptor-PMN connectivity that provides an anatomical explanation for the role of proprioception in promoting locomotor velocity; and identify a new candidate escape motor circuit. Finally, we generate a recurrent network model that produces the observed sequence of motor activity, showing that the identified pool of premotor neurons is sufficient to generate two distinct larval behaviors. We conclude that different locomotor behaviors can be generated by a specific group of premotor neurons generating behavior-specific motor rhythms.

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Analysis of various physiological salines for heart rate, CNS function, and synaptic transmission at neuromuscular junctions in Drosophila melanogaster larvae

Cited by 45 publications

References 56 publications

Pharmacological identification of cholinergic receptor subtypes on Drosophila melanogaster larval heart

Pharmacological identification of cholinergic receptor subtypes on Drosophila melanogaster larval heart

The Effect of CO2, Intracellular pH and Extracellular pH on Mechanosensory Proprioceptor Responses in Crayfish and Crab

ADrosophilalarval premotor/motor neuron connectome generating two behaviors via distinct spatio-temporal muscle activity

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