RATIONALE: The crustacean stomatogastric nervous system (STNS) is a classic experimental model to derive basic knowledge about neuronal functions and how they coordinate with each other to generate neural circuits. To investigate the components of the neuromodulators and how they are distributed in such a system is essential to understand the underlying mechanism. In this study, in situ mass spectrometry based techniques were employed to fulfill this goal. METHODS: Offline high-performance liquid chromatography (HPLC) separation was coupled with matrix-assisted laser desorption/ionization time-of-flight/time-of-flight (MALDI-TOF/TOF) to analyze the neuropeptides in the stomatogastric ganglion (STG) tissue extract from the Jonah crab Cancer borealis. Direct tissue analysis was employed to investigate the neuropeptides present in the STNS. MALDI imaging was also applied to map the localization of multiple neuropeptide families in the STG and the upstream nerve fibers. RESULTS: Fifty-seven neuropeptides were detected from a single desheathed STG using direct tissue analysis, and they were from eleven different neuropeptide families, including FaRP, AST-A, AST-B, etc. Differential neuropeptide profiles from three different types of ganglia and two types of nerve fiber tissues from the STNS were documented. The direct tissue analysis was shown better for studying neuropeptides from small neural organs like the STG as compared to the large-scale HPLC/MALDI analysis. MALDI images were also acquired to study the distribution of neuropeptides in the STG. CONCLUSIONS: In this study, the components and distribution of neuropeptides have been analyzed in the STNS from C. borealis using direct tissue profiling and MALDI imaging. The results show that the direct tissue analysis of desheathed neural tissues can provide higher sensitivity for neuropeptide study compared to large-scale HPLC/MALDI analysis of pooled tissues. The results are valuable for understanding the functions of neuropeptides in neural network generation.
RATIONALE: The crustacean stomatogastric nervous system (STNS) is a classic experimental model to derive basic knowledge about neuronal functions and how they coordinate with each other to generate neural circuits. To investigate the components of the neuromodulators and how they are distributed in such a system is essential to understand the underlying mechanism. In this study, in situ mass spectrometry based techniques were employed to fulfill this goal. METHODS: Offline high-performance liquid chromatography (HPLC) separation was coupled with matrix-assisted laser desorption/ionization time-of-flight/time-of-flight (MALDI-TOF/TOF) to analyze the neuropeptides in the stomatogastric ganglion (STG) tissue extract from the Jonah crabCancer borealis. Direct tissue analysis was employed to investigate the neuropeptides present in the STNS. MALDI imaging was also applied to map the localization of multiple neuropeptide families in the STG and the upstream nerve fibers. RESULTS: Fifty-seven neuropeptides were detected from a single desheathed STG using direct tissue analysis, and they were from eleven different neuropeptide families, including FaRP, AST-A, AST-B, etc. Differential neuropeptide profiles from three different types of ganglia and two types of nerve fiber tissues from the STNS were documented. The direct tissue analysis was shown better for studying neuropeptides from small neural organs like the STG as compared to the large-scale HPLC/MALDI analysis. MALDI images were also acquired to study the distribution of neuropeptides in the STG. CONCLUSIONS: In this study, the components and distribution of neuropeptides have been analyzed in the STNS from C. borealis using direct tissue profiling and MALDI imaging. The results show that the direct tissue analysis of desheathed neural tissues can provide higher sensitivity for neuropeptide study compared to large-scale HPLC/MALDI analysis of pooled tissues. The results are valuable for understanding the functions of neuropeptides in neural network generation.
Authors: A F Maarten Altelaar; Ivo Klinkert; Kees Jalink; Robert P J de Lange; Roger A H Adan; Ron M A Heeren; Sander R Piersma Journal: Anal Chem Date: 2006-02-01 Impact factor: 6.986
Authors: Elizabeth A Stemmler; Christopher R Cashman; Daniel I Messinger; Noah P Gardner; Patsy S Dickinson; Andrew E Christie Journal: Peptides Date: 2007-08-25 Impact factor: 3.750
Authors: Emily G Tillmaand; Ning Yang; Callie A C Kindt; Elena V Romanova; Stanislav S Rubakhin; Jonathan V Sweedler Journal: J Am Soc Mass Spectrom Date: 2015-09-21 Impact factor: 3.109
Authors: Christopher J Konop; Jennifer J Knickelbine; Molly S Sygulla; Colin D Wruck; Martha M Vestling; Antony O W Stretton Journal: J Am Soc Mass Spectrom Date: 2015-07-15 Impact factor: 3.109