Insect olfactory neurons in vitro: morphological and immunocytochemical characterization of male-specific antennal receptor cells from developing antennae of male Manduca sexta

Stengl, Monika; Hildebrand, John G.

doi:10.1523/jneurosci.10-03-00837.1990

Cited by 43 publications

(55 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This can be done by eluting the antibody itself (Tramu et al, 1978) or by completely masking the antibody by deposition ofa permanent reaction product such as silver granules (Roth et al, 1990). Alternatively, primary antibodies can be directly conjugated to fluorophores (Stengl and Hildebrand, 1990), enzymes (Boorsma, 1984), or other molecules (Hartig et al, 1995; Wurden and Homberg, 1993) so that the conjugate itself can then be detected instead of detecting the immunoglobulin portion of the primary antibody. These detection systems require additional steps to modlfy the primary antisera and often are time consuming and less sensitive than those using conventional fluorophore-conjugated secondary antisera.…”

Section: Introductionmentioning

confidence: 99%

Double immunofluorescent staining using two unconjugated primary antisera raised in the same species.

Shindler

Roth²

1996

J Histochem Cytochem.

206

169

View full text Add to dashboard Cite

Monoclonal antibodies (MAbs) capable of recognizing developmental stage-specific neuronal epitopes are becoming increasingly available. Because most of these MAbs are raised in a single species (mouse), simultaneous immunofluorescent detection of multiple epitopes has been diflticult. We have taken advantage of the high sensitivity of tyramide signal amplification to develop a protocol that permits simultaneous detection of two antibodies raised in the same species. One primary antibody was applied at a concentration below the detection limit of fluorescently labeled secondary antibodies, yet sufftcient for detection with the tyramide system. This first primary antibody was then efkively neglected during application of a second primary antibody that was detected by conventional fluorescently labeled sec-

show abstract

Section: Introductionmentioning

confidence: 99%

Double immunofluorescent staining using two unconjugated primary antisera raised in the same species.

Shindler

Roth²

1996

J Histochem Cytochem.

206

169

View full text Add to dashboard Cite

show abstract

“…The yield of cells for the primary culture was still much better than in other studies on dissociated scolopidial sensory neurons (5-30 out of 2000 cells [14]). Nevertheless, mechanosensory neurons had been investigated physiologically in primary cell culture [27].…”

Section: Methodsmentioning

confidence: 99%

“…On mechanosensory neurons of cockroaches patch clamp experiments on chloride channels have been performed [14]. Mechanosensitive channels could be detected in cultured neurons of Manduca sexta [27]. So far it could only be speculated why the auditory sensory cells of the locust did not show any action potentials.…”

Section: Physiologymentioning

confidence: 99%

Thoracic Interneurons, Motorneurons and Sensory Neurons of Locusta Migratoria (Insecta: Orthoptera) in Primary Cell Culture

Pfahlert¹,

Lakes‐Harlan²

2008

TOENTOJ

View full text Add to dashboard Cite

Abstract:The aim of this study was to establish a cell culture system with identified classes of locust neurons (interneurons, motorneurons and sensory neurons). The cells belonging to the different classes were distinguished in cell culture by vital dyes, which had been applied to the neurons in situ. From the various dyes tested fluorescent marked dextrans (10.000MW) gave the best results. The cells survived for up to 28 days in culture and approx. half of the cells grew processes, except of the sensory neurons, which never formed any processes. The different neurons were comparatively investigated, e.g. with immunhistochemistry: 86% of motorneuron were glutamate immunoreactive and 50% of the interneurons exhibited GABA-like immunoreactivity. The cells had resting potentials between -20 and -60mV and did not show spontanous action potentials. Action potentials could be elicited by current injection in 8% of interneurons and 26% of motorneurons, but not in sensory neurons. The vital marking of cells allowed to study distinct neurons in cell culture and to compare their morphology and physiology.

show abstract

“…The cell culture protocol was modified after Stengl and Hildebrand (1990). Briefly, antennae of male M. sexta pupae were dissected in Hanks' balanced salt solution containing 1% penicillin-streptomycin solution (HBSS/PS).…”

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

View full text Add to dashboard Cite

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.

show abstract

Insect olfactory neurons in vitro: morphological and immunocytochemical characterization of male-specific antennal receptor cells from developing antennae of male Manduca sexta

Cited by 43 publications

References 26 publications

Double immunofluorescent staining using two unconjugated primary antisera raised in the same species.

Double immunofluorescent staining using two unconjugated primary antisera raised in the same species.

Thoracic Interneurons, Motorneurons and Sensory Neurons of Locusta Migratoria (Insecta: Orthoptera) in Primary Cell Culture

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

Contact Info

Product

Resources

About