Neuroanatomical distribution and function of the vasopressin V1B receptor in the rat brain deciphered using specific fluorescent ligands

2018 ◽  
Vol 258 ◽  
pp. 15-32 ◽  
Author(s):  
Maithé Corbani ◽  
Rafik Marir ◽  
Miguel Trueba ◽  
Magda Chafai ◽  
Anne Vincent ◽  
...  
Keyword(s):  
Rat Brain ◽  
V1b Receptor ◽  
Fluorescent Ligands ◽  
And Function

scholarly journals Phosphorylation-dependent interaction of the synaptic vesicle proteins cysteine string protein and synaptotagmin I

2002 ◽  
Vol 364 (2) ◽  
pp. 343-347 ◽  
Author(s):  
Gareth J.O. EVANS ◽  
Alan MORGAN
Keyword(s):  
Rat Brain ◽  
Direct Interaction ◽  
Secretory Vesicle ◽  
Brain Membrane ◽  
Synaptotagmin I ◽  
Phosphorylated Form ◽  
Order Of Magnitude ◽  
And Function

The secretory vesicle cysteine string proteins (CSPs) are members of the DnaJ family of chaperones, and function at late stages of Ca2+-regulated exocytosis by an unknown mechanism. To determine novel binding partners of CSPs, we employed a pull-down strategy from purified rat brain membrane or cytosolic proteins using recombinant hexahistidine-tagged (His6-)CSP. Western blotting of the CSP-binding proteins identified synaptotagmin I to be a putative binding partner. Furthermore, pull-down assays using cAMP-dependent protein kinase (PKA)-phosphorylated CSP recovered significantly less synaptotagmin. Complexes containing CSP and synaptotagmin were immunoprecipitated from rat brain membranes, further suggesting that these proteins interact in vivo. Binding assays in vitro using recombinant proteins confirmed a direct interaction between the two proteins and demonstrated that the PKA-phosphorylated form of CSP binds synaptotagmin with approximately an order of magnitude lower affinity than the non-phosphorylated form. Genetic studies have implicated each of these proteins in the Ca2+-dependency of exocytosis and, since CSP does not bind Ca2+, this novel interaction might explain the Ca2+-dependent actions of CSP.

scholarly journals Development of a CRISPR-SaCas9 system for projection- and function-specific gene editing in the rat brain

2020 ◽  
Vol 6 (12) ◽  
pp. eaay6687 ◽  
Author(s):  
Haojie Sun ◽  
Su Fu ◽  
Shuang Cui ◽  
Xiangsha Yin ◽  
Xiaoyan Sun ◽  
...  
Keyword(s):  
Rat Brain ◽  
Gene Editing ◽  
High Efficiency ◽  
Protein A ◽  
Cell Types ◽  
Cell Labeling ◽  
Specific Gene ◽  
Creb Binding Protein ◽  
A Genome ◽  
And Function

A genome editing technique based on the clustered regularly interspaced short palindromic repeats (CRISPR)–associated endonuclease Cas9 enables efficient modification of genes in various cell types, including neurons. However, neuronal ensembles even in the same brain region are not anatomically or functionally uniform but divide into distinct subpopulations. Such heterogeneity requires gene editing in specific neuronal populations. We developed a CRISPR-SaCas9 system–based technique, and its combined application with anterograde/retrograde AAV vectors and activity-dependent cell-labeling techniques achieved projection- and function-specific gene editing in the rat brain. As a proof-of-principle application, we knocked down the cbp (CREB-binding protein), a sample target gene, in specific neuronal subpopulations in the medial prefrontal cortex, and demonstrated the significance of the projection- and function-specific CRISPR-SaCas9 system in revealing neuronal and circuit basis of memory. The high efficiency and specificity of our projection- and function-specific CRISPR-SaCas9 system could be widely applied in neural circuitry studies.

scholarly journals Central distribution and function of 5-HTe and 5-HT7 receptor subtypes in the rat brain

1997 ◽  
Vol 73 ◽  
pp. 13
Author(s):  
Mitsuhiro Yoshioka ◽  
Machiko Matsumoto ◽  
Hiroko Togashi ◽  
Kjyoshi Mori ◽  
Hideya Saito
Keyword(s):  
Rat Brain ◽  
Receptor Subtypes ◽  
And Function

Transplantation of cultured sympathetic ganglionic neurons into Parkinsonian rat brain: Survival and function of graft

1995 ◽  
Vol 133 (1-2) ◽  
pp. 61-67 ◽  
Author(s):  
N. Nakao ◽  
T. Itakura ◽  
Y. Uematsu ◽  
N. Komai
Keyword(s):  
Rat Brain ◽  
And Function

The effects of donor stage on the survival and function of embryonic striatal grafts in the adult rat brain

Neuroscience ◽  
1997 ◽  
Vol 79 (3) ◽  
pp. 711-721 ◽  
Author(s):  
R.A Fricker ◽  
E.M Torres ◽  
S.P Hume ◽  
R Myers ◽  
J Opacka-Juffrey ◽  
...  
Keyword(s):  
Rat Brain ◽  
Adult Rat ◽  
Striatal Grafts ◽  
Adult Rat Brain ◽  
And Function

scholarly journals Development and Characterization of Monoclonal Antibodies Specific to the Serotonin 5-HT2A Receptor

1998 ◽  
Vol 46 (7) ◽  
pp. 811-824 ◽  
Author(s):  
Chun Wu ◽  
Elizabeth J. Yoder ◽  
Jean Shih ◽  
Kevin Chen ◽  
Peter Dias ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Rat Brain ◽  
Pyramidal Neurons ◽  
Expression System ◽  
Molecular Size ◽  
Baculovirus Expression System ◽  
Differential Staining ◽  
Receptor Subtypes ◽  
And Function

Serotonin (5-hydroxytryptamine, 5-HT) mediates many functions of the central and peripheral nervous systems by its interaction with specific neuronal and glial receptors. Fourteen serotonin receptors belonging to seven families have been identified through physiological, pharmacological, and molecular cloning studies. Monoclonal antibodies (MAbs) specific for each of these receptor subtypes are needed to characterize their expression, distribution, and function in embryonic, adult, and pathological tissues. In this article we report the development and characterization of MAbs specific to the serotonin 5-HT2A receptor. To generate MAbs against 5-HT2AR, mice were immunized with the N-terminal domain of the receptor. The antigens were produced as glutathionine S-transferase (GST) fusion proteins in insect cells using a Baculovirus expression system. The hybridomas were initially screened by ELISA against the GST-5-HT2AR recombinant proteins and subsequently against GST control proteins to eliminate clones with unwanted reactivity. They were further tested by Western blotting against recombinant GST-5-HT2AR, rat and human brain lysate, and lysate from cell lines transfected with 5-HT2AR cDNA. One of the MAbs G186-1117, which recognizes a portion of the 5-HT2AR N-terminus, was selected for further characterization. G186-1117 reacted with a band of molecular size 55 kD corresponding to the predicted size of 5-HT2AR in lysates from rat brain and a 5-HT2AR-transfected cell line. Its specificity was further confirmed by adsorption of immunoreactivity with recombinant 5-HT2AR but not with recombinant 5-HT2BR and 5-HT2CR. Rat brain sections and Schwann cell cultures were immunohistochemically labeled with this MAb. G186-1117 showed differential staining in various regions of the rat brain, varying from regions with no staining to regions of intense reactivity. In particular, staining of cell bodies and dendrites of the pyramidal neurons in the cortex was observed, which is in agreement with observations of electrophysiological studies.

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