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Published on: November 7, 2017
Locustatachykinin immunoreactivity in the blowfly central nervous system and intestine
C T Lundquist1, F L Clottens, G M Holman
1Department of Zoology, Stockholm University, Sweden.
Insights
An antiserum detected locustatachykinin-like immunoreactive neurons and endocrine cells in the blowfly nervous system and intestine. This suggests these peptides act as neurotransmitters or modulators in the central nervous system and regulate the gut.
Area of Science:
- Neuroscience
- Insect Physiology
- Immunocytochemistry
Background:
- Locustatachykinins (LomTKs) are myotropic peptides found in locusts.
- Understanding the distribution and function of LomTKs in other insects is crucial.
Purpose of the Study:
- To characterize an antiserum against locustatachykinin I.
- To detect and map locustatachykinin-like immunoreactive (LomTK-LI) neurons and endocrine cells in the blowfly Calliphora vomitoria.
- To investigate the potential role of LomTKs in the blowfly nervous system and intestine.
Main Methods:
- Enzyme-linked immunosorbent assay (ELISA) for antiserum characterization and detection of LomTK-LI material in blowfly extracts.
- High-performance liquid chromatography (HPLC) to analyze LomTK-LI material.
- Immunocytochemistry to visualize LomTK-LI neurons and cells in adult and larval blowflies.
Main Results:
- The antiserum specifically recognized locustatachykinins I-III, with minimal cross-reactivity to LomTK-IV and no cross-reactivity to vertebrate tachykinins.
- Extensive networks of LomTK-LI neurons were found in the blowfly brain (approx. 160 neurons) and thoracicoabdominal ganglion (46 neurons).
- LomTK-LI material was present in interneurons throughout the central nervous system and in endocrine cells of the midgut, but not in efferent/afferent axons or neurosecretory cells.
Conclusions:
- The widespread distribution of LomTK-LI material in interneurons suggests a role as a neurotransmitter or neuromodulator in the blowfly central nervous system.
- The presence of LomTK-LI in midgut endocrine cells indicates a potential regulatory function in the blowfly intestine.
Abstract:
An antiserum raised against locustatachykinin I, one of four myotropic peptides that have been isolated from the locust brain and corpora cardiaca, was characterized by enzyme-linked immunosorbent assay (ELISA) and used for immunocytochemical detection of neurons and endocrine cells in the nervous system and intestine of the blowfly Calliphora vomitoria. The ELISA characterization indicated that the antiserum recognizes the common C-terminus sequence of the locustatachykinins I-III. Hence, the cross reaction with locustatachykinin IV is less, and in competitive ELISAs no cross reaction was detected with a series of vertebrate tachykinins tested. It was also shown that the antiserum recognized material in extracts of blowfly heads, as measured in ELISA. In high-performance liquid chromatography the extracted locustatachykinin-like immunoreactive (LomTK-LI) material eluted in two different ranges. A fairly large number of LomTK-LI neurons was detected in the blowfly brain and thoracicoabdominal ganglion. A total of about 160 LomTK-LI neurons was seen in the proto-, deuto-, and tritocerebrum and subesophageal ganglion. Immunoreactive processes from these neurons could be traced in many neuropil regions of the brain: superior and dorsomedian protocerebrum, optic tubercle, fan-shaped body and ventral bodies of the central complex, all the glomeruli of the antennal lobes, and tritocerebral and subesophageal neuropil. No immunoreactivity was seen in the mushroom bodies or the optic lobes. In the fused thoracicoabdominal ganglion, 46 LomTK-LI neurons could be resolved. The less evolved larval nervous system was also investigated to obtain additional information on the morphology and projections of immunoreactive neurons. In neither the larval nor the adult nervous systems could we identify any efferent or afferent immunoreactive axons or neurosecretory cells. The widespread distribution of LomTK-LI material in interneurons suggests an important role of the native peptide(s) as a neurotransmitter or neuromodulator within the central nervous system. Additionally a regulatory function in the intestine is indicated by the presence of immunoreactivity in endocrine cells of the midgut.
