Immunofluorescently labeling glutamic acid decarboxylase 65 coupled with confocal imaging for identifying GABAergic somata in the rat dentate gyrus—A comparison with labeling glutamic acid decarboxylase 67

Wang, Xiaochen; Gao, Fei; Zhu, Jie; Guo, Enpu; Song, Xuhong; Wang, Shuanglian; Zhan, Ren‐Zhi

doi:10.1016/j.jchemneu.2014.07.002

Cited by 14 publications

(11 citation statements)

References 65 publications

(80 reference statements)

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“…Results indicate that CTR in the LDTg does not co-localize with ChAT, a marker for cholinergic neurons (Figure 7B; n=6). After colchicine treatment (47), 13.7% of CTR neurons in the LDTg, specifically in the caudal LDTg (−8.6mm to −9.1mm from bregma) co-localize with the GABAergic neuronal marker, Gad67 (Figure 7C; n=1). These data suggest that at least a portion of amylin receptor-expressing cells in the LDTg are GABAergic neurons, though we cannot rule out the possibility that colchicine treatment did not result in labeling of all Gad67 cells.…”

Section: Resultsmentioning

confidence: 99%

Amylin Acts in the Lateral Dorsal Tegmental Nucleus to Regulate Energy Balance Through Gamma-Aminobutyric Acid Signaling

Reiner¹,

Mietlicki‐Baase²,

Olivos³

et al. 2017

Biological Psychiatry

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Background The pancreatic- and brain-derived hormone amylin promotes negative energy balance and is receiving increasing attention as a promising obesity therapeutic. However, the neurobiological substrates mediating amylin’s effects are not fully characterized. We postulated that amylin acts in the lateral dorsal tegmental nucleus (LDTg), an understudied neural-processing hub for reward and homeostatic feeding signals. Methods We used immunohistochemical (IHC) and quantitative PCR analyses to examine expression of the amylin receptor complex in rat LDTg tissue. Behavioral experiments were performed to examine the mechanisms underlying the hypophagic effects of amylin receptor activation in the LDTg. Results IHC and quantitative PCR analyses show expression of the amylin receptor complex in the LDTg. Activation of LDTg amylin receptors by the agonist salmon calcitonin dose-dependently reduces body weight, food intake, and motivated feeding behaviors. Acute pharmacological studies and longer-term adeno-associated viral (AAV)-knockdown experiments indicate that LDTg amylin receptor signaling is physiologically and potentially pre-clinically relevant for energy balance control. Finally, IHC data indicate that LDTg amylin receptors are expressed on gamma-aminobutyric acid (GABA)-ergic neurons and behavioral results suggest that local GABA receptor signaling mediates the hypophagia following LDTg amylin receptor activation. Conclusions These findings identify the LDTg as a novel nucleus with therapeutic potential in mediating amylin’s effects on energy balance through GABA receptor signaling.

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Section: Resultsmentioning

confidence: 99%

Amylin Acts in the Lateral Dorsal Tegmental Nucleus to Regulate Energy Balance Through Gamma-Aminobutyric Acid Signaling

Reiner¹,

Mietlicki‐Baase²,

Olivos³

et al. 2017

Biological Psychiatry

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“…Thereafter, the tissue block was sequentially immersed in 10% sucrose in 0.1 M PBS for 4 h, 15% sucrose in 0.1 M PBS for 8 h, and finally 20% sucrose in 0.1 M PBS at 4°C overnight. After the tissue block was firmly embedded with a medium that consisted of 30% (w/v) chicken egg albumin (Sigma, St. Louis, MO, USA), 0.5% (w/v) gelatin and 0.9% (v/v) glutaraldehyde in 0.1 M PBS as described previously (Wang X. et al, 2014 ), it was horizontally cut through the hippocampus into 40 μm-thick-sections with a vibratome (VT1000 S, Leica Biosystems, Wetzlar, Germany) to yield serial hippocampal sections. Hippocampal sections with “C” shaped dentate gyri (mainly representing the ventral part of the hippocampal formation) were orderly collected into 0.1 M PBS and used for detecting the expression of IEGs in GABAergic cells in the above mentioned four time points and the activation of different subtypes of interneurons more than 10 weeks after SE.…”

Section: Methodsmentioning

confidence: 99%

“…In addition to excitatory granule cells and mossy cells, the dentate gyrus also contains GABAergic interneurons, which function to prevent runaway excitation and to control the temporal and spatial dynamics of granule cell firing (Amaral et al, 2007 ; Hosp et al, 2014 ). Like GABAergic interneurons in the cortex, GABAergic interneurons in the dentate gyrus are diverse, varying in morphological appearances, neurochemical identities, electrophysiological properties, and axonal projection domains (Han et al, 1993 ; Mott et al, 1997 ; Houser, 2007 ; Hosp et al, 2014 ; Wang X. et al, 2014 ). Epilepsy is associated with numerous alterations in hippocampal dentate GABAergic interneurons.…”

Section: Introductionmentioning

confidence: 99%

Persistent Hyperactivity of Hippocampal Dentate Interneurons After a Silent Period in the Rat Pilocarpine Model of Epilepsy

Wang

Song

et al. 2016

Front. Cell. Neurosci.

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Profile of GABAergic interneuron activity after pilocarpine-induced status epilepticus (SE) was examined in the rat hippocampal dentate gyrus by analyzing immediate early gene expression and recording spontaneous firing at near resting membrane potential (REM). SE for exact 2 h or more than 2 h was induced in the male Sprague-Dawley rats by an intraperitoneal injection of pilocarpine. Expression of immediate early genes (IEGs) was examined at 1 h, 1 week, 2 weeks or more than 10 weeks after SE. For animals to be examined at 1 h after SE, SE lasted for exact 2 h was terminated by an intraperitoneal injection of diazepam. Spontaneous firing at near the REM was recorded in interneurons located along the border between the granule cell layer and the hilus more than 10 weeks after SE. Results showed that both c-fos and activity-regulated cytoskeleton associated protein (Arc) in hilar GABAergic interneurons were up-regulated after SE in a biphasic manner; they were increased at 1 h and more than 2 weeks, but not at 1 week after SE. Ten weeks after SE, nearly 60% of hilar GABAergic cells expressed c-fos. With the exception of calretinin (CR)-positive cells, percentages of hilar neuronal nitric oxide synthase (nNOS)-, neuropeptide Y (NPY)-, parvalbumin (PV)-, and somatostatin (SOM)-positive cells with c-fos expression are significantly higher than those of controls more than 10 weeks after SE. Without the REM to be more depolarizing and changed threshold potential level in SE-induced rats, cell-attached recording revealed that nearly 90% of hilar interneurons fired spontaneously at near the REM while only 22% of the same cell population did so in the controls. In conclusion, pilocarpine-induced SE eventually leads to a state in which surviving dentate GABAergic interneurons become hyperactive with a subtype-dependent manner; this implies that a fragile balance between excitation and inhibition exists in the dentate gyrus and in addition, the activity-dependent up-regulation of IEGs may underlie plastic changes seen in some types of GABAergic cells in the pilocarpine model of epilepsy.

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“…The glutamate decarboxylase 67 kDa isoform (GAD67) antibody was raised against recombinant GAD67 protein in mouse (manufacturer's technical information). This antibody recognized GAD67 with no cross-reactivity with the 65 kDa isoform (GAD65) by Western blot analysis on the samples isolated from rat cerebellar cortex (Fong, Stornetta, Foley, & Potts, 2005;Wang et al, 2014). GAD67 immunoreactivity can be visualized in cell bodies, fibers and punctate structures in cortical cultures (King, Chung, Vickers, & Dickson, 2006), layer 5 somatosensory cortex (Haberl et al, 2015), and dentate gyrus (Wang et al, 2014).…”

Section: Antibody Characterizationmentioning

confidence: 99%

Laminar specific gene expression reveals differences in postnatal laminar maturation in mouse auditory, visual, and somatosensory cortex

Chang¹,

Suzuki²,

Kawai³

2018

J of Comparative Neurology

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Proper formation of laminar structures in sensory cortexes is critical for sensory information processing. Previous studies suggested that the timing of neuronal migration and the laminar position of cortical neurons differ among sensory cortexes. How they differ during postnatal development has not been systematically investigated. Here, identifying laminas using transcription factors, we examined postnatal changes in neuronal density and distribution in presumptive primary auditory (ACx), visual (VCx), and somatosensory cortexes (SCx) in a strain of mice using immunofluorescence techniques. Development of laminar thickness and its cortical proportion differed among the sensory cortexes. Layers 2-4 defined by Cut-like homeobox 1 (Cux1)-expressing neurons were narrower, and layer 5 was wider in ACx compared to those in VCx or SCx, while Forkhead-box protein P2 (Foxp2)-defined layer 6 was wider in SCx than the other two sensory cortexes throughout postnatal development. Meanwhile, thalamocortical input layers identified by Cux1-expressing neurons formed later in ACx than in the other two cortical regions. The cell densities of ETS-related protein 81-expressing neurons increased in both lower and upper layers but at distinct timing, while those of COUP-TF-interacting protein 2 expressing neurons in the lower layers changed bidirectionally (i.e., increased or decreased) both in layer- and cortical region-specific manners. Foxp2-expressing cells in layer 6 distributed differently and declined at different timing among the sensory cortexes. Overall, we demonstrate that the maturational timing of lamina differs among the sensory cortexes and that postnatal age-dependent changes in neuronal distribution are unique to each of the sensory cortexes.

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Immunofluorescently labeling glutamic acid decarboxylase 65 coupled with confocal imaging for identifying GABAergic somata in the rat dentate gyrus—A comparison with labeling glutamic acid decarboxylase 67

Cited by 14 publications

References 65 publications

Amylin Acts in the Lateral Dorsal Tegmental Nucleus to Regulate Energy Balance Through Gamma-Aminobutyric Acid Signaling

Amylin Acts in the Lateral Dorsal Tegmental Nucleus to Regulate Energy Balance Through Gamma-Aminobutyric Acid Signaling

Persistent Hyperactivity of Hippocampal Dentate Interneurons After a Silent Period in the Rat Pilocarpine Model of Epilepsy

Laminar specific gene expression reveals differences in postnatal laminar maturation in mouse auditory, visual, and somatosensory cortex

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