Establishment of two cell lines from embryonic tissue of the tobacco hornworm,Manduca sexta (L.)

Eide, Paul E.; Caldwell, J. D.; Marks, Edwin P.

doi:10.1007/bf02616376

Cited by 53 publications

(23 citation statements)

References 11 publications

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“…The culture medium consisted of two parts of L-15 medium (Life Technologies) supplemented with 5% fetal bovine serum (FBS; Hyclone International, Logan, UT) and one part of Grace medium (Life Technologies). The Grace medium was conditioned with Manduca embryonic cell line MRRL-CH1 (Eide et al, 1975) that was supplemented with 3 g/L TC-yeastolate (Beckton, Dickinson & Co, Sparks, MD), 3 g/L lactalbumin hydrosylate (Sigma, Oakville, ON, Canada), and 10% FBS. Neurons differentiated and grew long processes within a few hours after plating and a variety of non-neural cells also appeared rapidly.…”

Section: Cell Culturementioning

confidence: 99%

Expression of muscarinic acetylcholine receptors and choline acetyltransferase enzyme in cultured antennal sensory neurons and non-neural cells of the developing mothManduca sexta

2005

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Antennal sensory neurons of Manduca sexta emerge from epidermal cells that also give rise to sheath cells surrounding the peripheral parts of the neurons and to glial cells that enwrap the sensory axons in the antennal nerve. Reciprocal interactions between sensory neurons and glial cells are believed to aid in axon growth and guidance, but the exact nature of these interactions is not known. We investigated the possibility of cholinergic interactions in this process by locating muscarinic acetylcholine receptors (mAChRs) and choline acetyltransferase (ChAT) enzyme in cultured antennal sensory neurons and non-neural cells. ChAT and mAChRs were present in the sensory neurons from the first day in culture. Therefore, the sensory neurons are probably cholinergic, as previously suggested, but they may also be controlled by ACh. In 7-day-old cultures a subgroup of small non-neural cells with processes expressed ChAT activity, and in 14-day-old cultures non-neural cells that formed lamellipodia and scaffoldlike structures on the culture substrate were labeled with ChAT antibody. mAChR activity was detected in similar non-neural cells but only in areas surrounding the nuclei. In addition, mAChRs were found in flat lamellipodia and filopodia forming cells that were present in 1-day-old cultures and grew in size during the 2 week investigation period. These findings suggest muscarinic cholinergic interactions between the neural and non-neural cells during the development of Manduca antenna.

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Section: Cell Culturementioning

confidence: 99%

Expression of muscarinic acetylcholine receptors and choline acetyltransferase enzyme in cultured antennal sensory neurons and non-neural cells of the developing mothManduca sexta

2005

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“…Cells were mechanically and enzymatically dissociated. Dispersed cells were plated onto uncoated Falcon Petri dishes in three parts of Leibovitz L15 medium, two parts of Grace medium conditioned on the embryonic cell line MRRL-CH1 (Eide et al, 1975), and 5% of fetal bovine serum (Invitrogen). Cultures were maintained in an incubator at 20°C before use.…”

Section: Methodsmentioning

confidence: 99%

Calcium Activates a Chloride Conductance Likely Involved in Olfactory Receptor Neuron Repolarization in the MothSpodoptera littoralis

Pezier¹,

Grauso²,

Acquistapace³

et al. 2010

J. Neurosci.

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“…Dispersed cells were plated on glass-bottom culture dishes, which were coated with concanavalin A and poly-L-lysine, and allowed to settle for 30 min. Then, 1 ml of a 2:1 cell culture medium [two parts of Leibovitz L-15 medium supplemented with 10% fetal bovine serum and one part of Manduca embryonic cell line conditioned medium MRLL-CH1 (Eide et al 1975) was added. The medium was completely replaced within 24 h after dispersion.…”

Section: Primary Cell Culturesmentioning

confidence: 99%

Cyclic Nucleotide-Activated Currents in Cultured Olfactory Receptor Neurons of the HawkmothManduca sexta

Krannich

Stengl

2008

Journal of Neurophysiology

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Krannich S, Stengl M. Cyclic nucleotide-activated currents in cultured olfactory receptor neurons of the hawkmoth Manduca sexta. J Neurophysiol 100: 2866 -2877, 2008. First published August 6, 2008 doi:10.1152/jn.01400.2007. Moth pheromones cause rises in intracellular Ca 2ϩ concentrations that activate Ca 2ϩ -dependent cation channels in antennal olfactory receptor neurons. In addition, mechanisms of adaptation and sensitization depend on changes in cyclic nucleotide concentrations. Here, cyclic nucleotide-activated currents in cultured olfactory receptor neurons of the moth Manduca sexta are described, which share properties with currents through vertebrate cyclic nucleotide-gated channels. The cyclic nucleotide-activated currents of M. sexta carried Ca 2ϩ and monovalent cations. They were directly activated by cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP), modulated by Ca 2ϩ /calmodulin, and inhibited by lanthanum. M. sexta cyclic nucleotide-activated currents developed in an all-or-none manner, which suggests that the underlying channels are coupled and act coordinately. At least one cAMP-and two cGMP-activated nonselective cation currents could be distinguished. Compared with the cAMP-activated current, both cGMP-activated currents appeared to conduct more Ca 2ϩ and showed a stronger down-regulation by Ca 2ϩ /calmodulin-dependent negative feedback. Furthermore, both cGMP-activated currents differed in their Ca 2ϩ -dependent inhibition. Thus M. sexta olfactory receptor neurons, like vertebrate sensory neurons, appear to express nonselective cyclic nucleotide-activated cation channels with different subunit compositions. Besides the nonselective cyclic nucleotide-activated cation currents, olfactory receptor neurons express a cAMP-dependent current. This current resembled a protein kinase-modulated low-voltageactivated Ca 2ϩ current.

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Establishment of two cell lines from embryonic tissue of the tobacco hornworm,Manduca sexta (L.)

Cited by 53 publications

References 11 publications

Expression of muscarinic acetylcholine receptors and choline acetyltransferase enzyme in cultured antennal sensory neurons and non-neural cells of the developing mothManduca sexta

Expression of muscarinic acetylcholine receptors and choline acetyltransferase enzyme in cultured antennal sensory neurons and non-neural cells of the developing mothManduca sexta

Calcium Activates a Chloride Conductance Likely Involved in Olfactory Receptor Neuron Repolarization in the MothSpodoptera littoralis

Cyclic Nucleotide-Activated Currents in Cultured Olfactory Receptor Neurons of the HawkmothManduca sexta

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