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Coexpression analysis of nine neuropeptides in the neurosecretory preoptic area of larval zebrafish.

Herget U, Ryu S - Front Neuroanat (2015)

Bottom Line: To identify distinct cell types present in the larval NPO, we also generated a comprehensive 3D map of 9 zebrafish homologs of typical neuropeptides found in the mammalian PVN (arginine vasopressin (AVP), corticotropin-releasing hormone (CRH), proenkephalin a (penka)/b (penkb), neurotensin (NTS), oxytocin (OXT), vasoactive intestinal peptide (VIP), cholecystokinin (CCK), and somatostatin (SST)).Our results allowed the subclassification of NPO cell types, and differences in variability of coexpression profiles suggest potential targets of biochemical plasticity.Thus, this work provides an important basis for the analysis of the development, function, and plasticity of the primary neuroendocrine brain region in larval zebrafish.

View Article: PubMed Central - PubMed

Affiliation: Developmental Genetics of the Nervous System, Max Planck Institute for Medical Research Heidelberg, Germany ; The Hartmut Hoffmann-Berling International Graduate School of Molecular and Cellular Biology, University of Heidelberg Heidelberg, Germany.

ABSTRACT
The paraventricular nucleus (PVN) of the hypothalamus in mammals coordinates neuroendocrine, autonomic and behavioral responses pivotal for homeostasis and the stress response. A large amount of studies in rodents has documented that the PVN contains diverse neuronal cell types which can be identified by the expression of distinct secretory neuropeptides. Interestingly, PVN cell types often coexpress multiple neuropeptides whose relative coexpression levels are subject to environment-induced plasticity. Due to their small size and transparency, zebrafish larvae offer the possibility to comprehensively study the development and plasticity of the PVN in large groups of intact animals, yet important anatomical information about the larval zebrafish PVN-homologous region has been missing. Therefore we recently defined the location and borders of the larval neurosecretory preoptic area (NPO) as the PVN-homologous region in larval zebrafish based on transcription factor expression and cell type clustering. To identify distinct cell types present in the larval NPO, we also generated a comprehensive 3D map of 9 zebrafish homologs of typical neuropeptides found in the mammalian PVN (arginine vasopressin (AVP), corticotropin-releasing hormone (CRH), proenkephalin a (penka)/b (penkb), neurotensin (NTS), oxytocin (OXT), vasoactive intestinal peptide (VIP), cholecystokinin (CCK), and somatostatin (SST)). Here we extend this chemoarchitectural map to include the degrees of coexpression of two neuropeptides in the same cell by performing systematic pairwise comparisons. Our results allowed the subclassification of NPO cell types, and differences in variability of coexpression profiles suggest potential targets of biochemical plasticity. Thus, this work provides an important basis for the analysis of the development, function, and plasticity of the primary neuroendocrine brain region in larval zebrafish.

No MeSH data available.


Schematic lateral view of a 5 dpf larval zebrafish brain showing the location of the NPO (dashed line) within the otpa-positive part (dark gray line) of the preoptic area, and the spatial distribution of nine cell types expressing the indicated neuropeptides. Cells clustering within the NPO are opaque. For more details, the reader is referred to our previously reported chemoarchitectural map (Herget et al., 2014). Abbreviations: ac, anterior commissure; d, dorsal; H, hypothalamus; Ha, habenula; NPO, neurosecretory preoptic area; oc, optic chiasm; PO, preoptic area; poc, postoptic commissure; PT, posterior tuberculum; PTh, prethalamus; r, rostral; Tel, telencephalon; TeO, optic tectum; Th, thalamus.
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Figure 1: Schematic lateral view of a 5 dpf larval zebrafish brain showing the location of the NPO (dashed line) within the otpa-positive part (dark gray line) of the preoptic area, and the spatial distribution of nine cell types expressing the indicated neuropeptides. Cells clustering within the NPO are opaque. For more details, the reader is referred to our previously reported chemoarchitectural map (Herget et al., 2014). Abbreviations: ac, anterior commissure; d, dorsal; H, hypothalamus; Ha, habenula; NPO, neurosecretory preoptic area; oc, optic chiasm; PO, preoptic area; poc, postoptic commissure; PT, posterior tuberculum; PTh, prethalamus; r, rostral; Tel, telencephalon; TeO, optic tectum; Th, thalamus.

Mentions: To comprehensively analyze the degree of coexpression of two peptides in the same cell, we performed cell by cell comparisons of pairwise combinatorial ISH staining of nine peptide markers that we had previously identified to be expressed in the 5 dpf larval NPO. The NPO is defined by the dense clustering of cells expressing these peptides within the transcription factor orthopedia a (otpa)-positive preoptic area (Figure 1). Altogether we examined 36 pairwise combinations of neuropeptides, analyzing a minimum of 5 animals per pair, but the sample size varied and was larger for some peptide combinations. All data was obtained from 5 dpf larvae, since the original NPO cell type map was generated for that stage. We found broadly three different categories of coexpression extent: (1) Absence of coexpression in all animals; (2) Occurance of coexpression in a single, few or several cells only in some animals analyzed (“variable coexpression”); (3) Coexpression in several cells in all animals analyzed (“consistent coexpression”).


Coexpression analysis of nine neuropeptides in the neurosecretory preoptic area of larval zebrafish.

Herget U, Ryu S - Front Neuroanat (2015)

Schematic lateral view of a 5 dpf larval zebrafish brain showing the location of the NPO (dashed line) within the otpa-positive part (dark gray line) of the preoptic area, and the spatial distribution of nine cell types expressing the indicated neuropeptides. Cells clustering within the NPO are opaque. For more details, the reader is referred to our previously reported chemoarchitectural map (Herget et al., 2014). Abbreviations: ac, anterior commissure; d, dorsal; H, hypothalamus; Ha, habenula; NPO, neurosecretory preoptic area; oc, optic chiasm; PO, preoptic area; poc, postoptic commissure; PT, posterior tuberculum; PTh, prethalamus; r, rostral; Tel, telencephalon; TeO, optic tectum; Th, thalamus.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4325906&req=5

Figure 1: Schematic lateral view of a 5 dpf larval zebrafish brain showing the location of the NPO (dashed line) within the otpa-positive part (dark gray line) of the preoptic area, and the spatial distribution of nine cell types expressing the indicated neuropeptides. Cells clustering within the NPO are opaque. For more details, the reader is referred to our previously reported chemoarchitectural map (Herget et al., 2014). Abbreviations: ac, anterior commissure; d, dorsal; H, hypothalamus; Ha, habenula; NPO, neurosecretory preoptic area; oc, optic chiasm; PO, preoptic area; poc, postoptic commissure; PT, posterior tuberculum; PTh, prethalamus; r, rostral; Tel, telencephalon; TeO, optic tectum; Th, thalamus.
Mentions: To comprehensively analyze the degree of coexpression of two peptides in the same cell, we performed cell by cell comparisons of pairwise combinatorial ISH staining of nine peptide markers that we had previously identified to be expressed in the 5 dpf larval NPO. The NPO is defined by the dense clustering of cells expressing these peptides within the transcription factor orthopedia a (otpa)-positive preoptic area (Figure 1). Altogether we examined 36 pairwise combinations of neuropeptides, analyzing a minimum of 5 animals per pair, but the sample size varied and was larger for some peptide combinations. All data was obtained from 5 dpf larvae, since the original NPO cell type map was generated for that stage. We found broadly three different categories of coexpression extent: (1) Absence of coexpression in all animals; (2) Occurance of coexpression in a single, few or several cells only in some animals analyzed (“variable coexpression”); (3) Coexpression in several cells in all animals analyzed (“consistent coexpression”).

Bottom Line: To identify distinct cell types present in the larval NPO, we also generated a comprehensive 3D map of 9 zebrafish homologs of typical neuropeptides found in the mammalian PVN (arginine vasopressin (AVP), corticotropin-releasing hormone (CRH), proenkephalin a (penka)/b (penkb), neurotensin (NTS), oxytocin (OXT), vasoactive intestinal peptide (VIP), cholecystokinin (CCK), and somatostatin (SST)).Our results allowed the subclassification of NPO cell types, and differences in variability of coexpression profiles suggest potential targets of biochemical plasticity.Thus, this work provides an important basis for the analysis of the development, function, and plasticity of the primary neuroendocrine brain region in larval zebrafish.

View Article: PubMed Central - PubMed

Affiliation: Developmental Genetics of the Nervous System, Max Planck Institute for Medical Research Heidelberg, Germany ; The Hartmut Hoffmann-Berling International Graduate School of Molecular and Cellular Biology, University of Heidelberg Heidelberg, Germany.

ABSTRACT
The paraventricular nucleus (PVN) of the hypothalamus in mammals coordinates neuroendocrine, autonomic and behavioral responses pivotal for homeostasis and the stress response. A large amount of studies in rodents has documented that the PVN contains diverse neuronal cell types which can be identified by the expression of distinct secretory neuropeptides. Interestingly, PVN cell types often coexpress multiple neuropeptides whose relative coexpression levels are subject to environment-induced plasticity. Due to their small size and transparency, zebrafish larvae offer the possibility to comprehensively study the development and plasticity of the PVN in large groups of intact animals, yet important anatomical information about the larval zebrafish PVN-homologous region has been missing. Therefore we recently defined the location and borders of the larval neurosecretory preoptic area (NPO) as the PVN-homologous region in larval zebrafish based on transcription factor expression and cell type clustering. To identify distinct cell types present in the larval NPO, we also generated a comprehensive 3D map of 9 zebrafish homologs of typical neuropeptides found in the mammalian PVN (arginine vasopressin (AVP), corticotropin-releasing hormone (CRH), proenkephalin a (penka)/b (penkb), neurotensin (NTS), oxytocin (OXT), vasoactive intestinal peptide (VIP), cholecystokinin (CCK), and somatostatin (SST)). Here we extend this chemoarchitectural map to include the degrees of coexpression of two neuropeptides in the same cell by performing systematic pairwise comparisons. Our results allowed the subclassification of NPO cell types, and differences in variability of coexpression profiles suggest potential targets of biochemical plasticity. Thus, this work provides an important basis for the analysis of the development, function, and plasticity of the primary neuroendocrine brain region in larval zebrafish.

No MeSH data available.