Immunolocalization of synaptotagmin for the study of synapses in the developing antennal lobe of Manduca sexta

Suzanne H. Dubuque, Joachim Schachtner, Alan J Nighorn, Kaushiki P. Menon, Kai Zinn, Leslie P Tolbert

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

In the mature olfactory systems of most organisms that possess a sense of smell, synapses between olfactory receptor neurons and central neurons occur in specialized neuropil structures called glomeruli. The development of olfactory glomeruli has been studied particularly heavily in the antennal lobe of the moth Manduca sexta. In the current study, we address the development of synapses within the antennal lobe of M. sexta by reporting on the localization of synaptotagmin, a ubiquitous synaptic vesicle protein, throughout development. A cDNA clone coding for M. sexta synaptotagmin was characterized and found to encode a protein that shares 67% amino acid identity with Drosophila synaptotagmin and 56% amino acid identity with human synaptotagmin I. Conservation was especially high in the C2 domains near the C-terminus and very low near the N-terminus. A polyclonal antiserum (MSYT) was raised against the unique N-terminus of M. sexta synaptotagmin, and a monoclonal antibody (DSYT) was raised against the highly conserved C-terminus of D. melanogaster synaptotagmin. In Western blot analyses, both antibodies labeled a 60 kD protein, which very likely corresponds to synaptotagmin. On sections, both antibodies labeled known synaptic neuropils in M. sexta and yielded similar labeling patterns in the developing antennal lobe. In addition, DSYT detected synaptotagmin-like protein in three other insect species examined. Analysis of synaptotagmin labeling at the light microscopic level during development of the antennal lobe of M. sexta confirmed and extended previous electron microscopic studies. Additional synapses in the coarse neuropil and a refinement of synaptic densities in the glomeruli during the last one-third of metamorphic development were revealed.

Original languageEnglish (US)
Pages (from-to)277-287
Number of pages11
JournalJournal of Comparative Neurology
Volume441
Issue number4
DOIs
StatePublished - Dec 24 2001

Fingerprint

Synaptotagmins
Manduca
Synapses
Neuropil
Proteins
Synaptotagmin I
Olfactory Receptor Neurons
Amino Acids
Moths
Antibodies
Smell
Synaptic Vesicles
Olfactory Bulb
Drosophila
Insects
Immune Sera
Complementary DNA
Clone Cells
Western Blotting
Monoclonal Antibodies

Keywords

  • Development
  • Glomeruli
  • Olfactory system
  • Synapses
  • Synaptotagmin

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Immunolocalization of synaptotagmin for the study of synapses in the developing antennal lobe of Manduca sexta. / Dubuque, Suzanne H.; Schachtner, Joachim; Nighorn, Alan J; Menon, Kaushiki P.; Zinn, Kai; Tolbert, Leslie P.

In: Journal of Comparative Neurology, Vol. 441, No. 4, 24.12.2001, p. 277-287.

Research output: Contribution to journalArticle

Dubuque, Suzanne H. ; Schachtner, Joachim ; Nighorn, Alan J ; Menon, Kaushiki P. ; Zinn, Kai ; Tolbert, Leslie P. / Immunolocalization of synaptotagmin for the study of synapses in the developing antennal lobe of Manduca sexta. In: Journal of Comparative Neurology. 2001 ; Vol. 441, No. 4. pp. 277-287.
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