A method for immunolabeling neurons in intact cuticularized insect appendages
Autor: | George Boyan, Tatjana Kleele, Erica Ehrhardt |
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Rok vydání: | 2015 |
Předmět: |
Arthropod Antennae
Nervous system animal structures media_common.quotation_subject Sensory system Grasshoppers Insect Nervous System Immunolabeling Genetics medicine Animals Axon Grasshopper media_common Neurons Appendage biology fungi Extremities Anatomy biology.organism_classification Immunohistochemistry Axons medicine.anatomical_structure Peripheral nervous system Developmental Biology |
Zdroj: | Development Genes and Evolution. 225:187-194 |
ISSN: | 1432-041X 0949-944X |
Popis: | The antennae of the grasshopper Schistocerca gregaria possess a pair of nerve pathways which are established by so-called pioneer neurons early in embryonic development. Subsequently, sensory cell clusters mediating olfaction, flight, optomotor responses, and phase changes differentiate from the antennal epithelium at stereotypic locations and direct their axons onto those of the pioneers to then project to the brain. Early in embryonic development, before the antennae become cuticularized, immunolabeling can be used to follow axogenesis in these pioneers and sensory cells. At later stages, immunolabeling becomes problematical as the cuticle is laid down and masks internal antigen sites. In order to immunolabel the nervous system of cuticularized late embryonic and first instar grasshopper antennae, we modified a procedure known as sonication in which the appendage is exposed to ultrasound thereby rendering it porous to antibodies. Comparisons of the immunolabeled nervous system of sectioned and sonicated antennae show that the cellular organization of sensory clusters and their axon projections is intact. The expression patterns of neuron-specific, microtubule-specific, and proliferative cell-specific labels in treated antennae are consistent with those reported for earlier developmental stages where sonication is not necessary, suggesting that these molecular epitopes are also conserved. The method ensures reliable immunolabeling in intact, cuticularized appendages so that the peripheral nervous system can be reconstructed directly via confocal microscopy throughout development. |
Databáze: | OpenAIRE |
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