Expression analyses of Rac3, a Rho family small GTPase, during mouse brain development

2021 ◽  
Author(s):  
Masashi Nishikawa ◽  
Hidenori Ito ◽  
Mariko Noda ◽  
Nanako Hamada ◽  
Hidenori Tabata ◽  
...  
Keyword(s):  
Brain Development ◽  
Mouse Brain ◽  
Expression Profile ◽  
Neurodevelopmental Disorders ◽  
Small Gtpases ◽  
Similar Distribution ◽  
Dependent Manner ◽  
Brain Anomalies ◽  
Rho Family ◽  
Structural Brain

Rac3 is a member of Rho family small GTPases which regulates cellular signaling and cytoskeletal dynamics. The RAC3 gene abnormalities have been shown to cause neurodevelopmental disorders with structural brain anomalies, including polymicrogyria/dysgyria, callosal abnormalities, brainstem anomalies, and cerebellar dysplasia. Although this evidence indicates that Rac3 is essential in brain development, not only its molecular mechanism but also the expression profile is yet to be elucidated. In this study, we carried out expression analyses of Rac3 with mouse brain tissues. In immunoblotting, Rac3 exhibited a tissue-dependent expression profile in the young adult mouse and was expressed in a developmental stage-dependent manner in brain. In primary cultured hippocampal neurons, while Rac3 was distributed mainly in the cytoplasm, it was visualized in axon and dendrites with partial localization at synapses, in consistent with the observation in biochemical fractionation analyses. In immunofluorescence analyses with brain slices, Rac3 was distributed strongly and moderately in the axon and cytoplasm, respectively, of cerebral cortex at postnatal day (P) 2 and P18. Similar distribution profile was also observed in hippocampus. Taken together, the results obtained strongly suggest that Rac3 plays an important physiological role in neuronal tissues during corticogenesis, and defects in the Rac3 function induce structural brain anomalies leading to pathogenesis of neurodevelopmental disorders.

scholarly journals A chronological expression profile of gene activity during embryonic mouse brain development

2013 ◽  
Vol 24 (11-12) ◽  
pp. 459-472 ◽  
Author(s):  
P. Goggolidou ◽  
S. Soneji ◽  
N. Powles-Glover ◽  
D. Williams ◽  
S. Sethi ◽  
...  
Keyword(s):  
Brain Development ◽  
Mouse Brain ◽  
Expression Profile ◽  
Gene Activity ◽  
Embryonic Mouse ◽  
Mouse Brain Development

scholarly journals Physiological Significance of WDR45, A Responsible Gene for β-propeller Protein Associated Neurodegeneration (BPAN), in Brain Development

2020 ◽  
Author(s):  
Mariko Noda ◽  
Hidenori Ito ◽  
Koh-ichi Nagata
Keyword(s):  
Brain Development ◽  
Mouse Brain ◽  
Cortical Neurons ◽  
De Novo ◽  
Early Stage ◽  
Physiological Role ◽  
Global Developmental Delay ◽  
Dependent Manner ◽  
Biochemical Fractionation ◽  
Biochemical Analyses

Abstract WDR45 plays an essential role in the early stage of autophagy. De novo heterozygous mutations in WDR45 have been known to cause b-propeller protein-associated neurodegeneration (BPAN), a subtype of neurodegeneration with brain iron accumulation (NBIA). Although BPAN patients display global developmental delay including intellectual disability, neurodevelopmental pathophysiology of BPAN remains largely unknown. In the present study, we analyzed the physiological role of Wdr45 and pathophysiological significance of the gene abnormality during mouse brain development. Morphological and biochemical analyses revealed that Wdr45 is expressed in a developmental stage-dependent manner in mouse brain. Wdr45 was also found to be located in the excitatory synapses in biochemical fractionation. Since the WDR45 mutations are thought to cause protein degradation, we conducted acute knockdown experiments by an in utero electroporation method with mice to recapitulate the pathophysiological conditions of BPAN. Silencing of Wdr45 caused abnormal dendritic development and synaptogenesis during corticogenesis, both of which were significantly rescued by co-expression with RNAi-resistant version of Wdr45. In addition, terminal arbors of callosal axons were less developed in Wdr45-deficient cortical neurons of adult mouse when compared to the control cells. These results strongly suggest a pathophysiological significance of WDR45 gene abnormalities in neurodevelopmental aspects of BPAN.

scholarly journals Dependence of Dbl and Dbs Transformation on MEK and NF-κB Activation

1999 ◽  
Vol 19 (11) ◽  
pp. 7759-7770 ◽  
Author(s):  
Ian P. Whitehead ◽  
Que T. Lambert ◽  
Judith A. Glaven ◽  
Karon Abe ◽  
Kent L. Rossman ◽  
...  
Keyword(s):  
Cyclin D1 ◽  
Structural Integrity ◽  
Small Gtpases ◽  
Dependent Manner ◽  
Nucleotide Exchange ◽  
Ph Domain ◽  
Transforming Activity ◽  
Rho Family ◽  
Altered Gene

ABSTRACT Dbs was identified initially as a transforming protein and is a member of the Dbl family of proteins (>20 mammalian members). Here we show that Dbs, like its rat homolog Ost and the closely related Dbl, exhibited guanine nucleotide exchange activity for the Rho family members RhoA and Cdc42, but not Rac1, in vitro. Dbs transforming activity was blocked by specific inhibitors of RhoA and Cdc42 function, demonstrating the importance of these small GTPases in Dbs-mediated growth deregulation. Although Dbs transformation was dependent upon the structural integrity of its pleckstrin homology (PH) domain, replacement of the PH domain with a membrane localization signal restored transforming activity. Thus, the PH domain of Dbs (but not Dbl) may be important in modulating association with the plasma membrane, where its GTPase substrates reside. Both Dbs and Dbl activate multiple signaling pathways that include activation of the Elk-1, Jun, and NF-κB transcription factors and stimulation of transcription from the cyclin D1 promoter. We found that Elk-1 and NF-κB, but not Jun, activation was necessary for Dbl and Dbs transformation. Finally, we have observed that Dbl and Dbs regulated transcription from the cyclin D1 promoter in a NF-κB-dependent manner. Previous studies have dissociated actin cytoskeletal activity from the transforming potential of RhoA and Cdc42. These observations, when taken together with those of the present study, suggest that altered gene expression, and not actin reorganization, is the critical mediator of Dbl and Rho family protein transformation.

scholarly journals Physiological significance of WDR45, a responsible gene for β-propeller protein associated neurodegeneration (BPAN), in brain development

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Mariko Noda ◽  
Hidenori Ito ◽  
Koh-ichi Nagata
Keyword(s):  
Brain Development ◽  
Mouse Brain ◽  
Cortical Neurons ◽  
De Novo ◽  
Early Stage ◽  
Physiological Role ◽  
Global Developmental Delay ◽  
Dependent Manner ◽  
Biochemical Fractionation ◽  
Biochemical Analyses

AbstractWDR45 plays an essential role in the early stage of autophagy. De novo heterozygous mutations in WDR45 have been known to cause β-propeller protein-associated neurodegeneration (BPAN), a subtype of neurodegeneration with brain iron accumulation (NBIA). Although BPAN patients display global developmental delay with intellectual disability, the neurodevelopmental pathophysiology of BPAN remains largely unknown. In the present study, we analyzed the physiological role of Wdr45 and pathophysiological significance of the gene abnormality during mouse brain development. Morphological and biochemical analyses revealed that Wdr45 is expressed in a developmental stage-dependent manner in mouse brain. Wdr45 was also found to be located in excitatory synapses by biochemical fractionation. Since WDR45 mutations are thought to cause protein degradation, we conducted acute knockdown experiments by in utero electroporation in mice to recapitulate the pathophysiological conditions of BPAN. Knockdown of Wdr45 caused abnormal dendritic development and synaptogenesis during corticogenesis, both of which were significantly rescued by co-expression with RNAi-resistant version of Wdr45. In addition, terminal arbors of callosal axons were less developed in Wdr45-deficient cortical neurons of adult mouse when compared to control cells. These results strongly suggest a pathophysiological significance of WDR45 gene abnormalities in neurodevelopmental aspects of BPAN.

Expression Analyses of POGZ, A Responsible Gene for Neurodevelopmental Disorders, during Mouse Brain Development

2019 ◽  
Vol 41 (1-2) ◽  
pp. 139-148 ◽  
Author(s):  
Kyoko Ibaraki ◽  
Nanako Hamada ◽  
Ikuko Iwamoto ◽  
Hidenori Ito ◽  
Noriko Kawamura ◽  
...  
Keyword(s):  
Gene Expression ◽  
Brain Development ◽  
Mouse Brain ◽  
Neurodevelopmental Disorders ◽  
Hippocampal Neurons ◽  
Morphological Characterization ◽  
Synaptic Function ◽  
Early Developmental Stage ◽  
Heterochromatin Protein 1 ◽  
Biochemical Fractionation

POGZ is a heterochromatin protein 1 α-binding protein and regulates gene expression. On the other hand, accumulating pieces of evidence indicate that the POGZ gene abnormalities are involved in various neurodevelopmental disorders. In this study, we prepared a specific antibody against POGZ, anti-POGZ, and carried out biochemical and morphological characterization with mouse brain tissues. Western blotting analyses revealed that POGZ is expressed strongly at embryonic day 13 and then gradually decreased throughout the brain development process. In immunohistochemical analyses, POGZ was found to be enriched in cerebrocortical and hippocampal neurons in the early developmental stage. The nuclear expression was also detected in Purkinje cells in cerebellum at postnatal day (P)7 and P15 but disappeared at P30. In primary cultured hippocampal neurons, while POGZ was distributed mainly in the nucleus, it was also visualized in axon and dendrites with partial localization at synapses in consistency with the results obtained in biochemical fractionation analyses. The obtained results suggest that POGZ takes part in the regulation of synaptic function as well as gene expression during brain development.

Control of neutrophil pseudopods by fluid shear: role of Rho family GTPases

2005 ◽  
Vol 288 (4) ◽  
pp. C863-C871 ◽  
Author(s):  
Ayako Makino ◽  
Michael Glogauer ◽  
Gary M. Bokoch ◽  
Shu Chien ◽  
Geert W. Schmid-Schönbein
Keyword(s):  
Signal Transduction ◽  
Shear Stress ◽  
Fluid Shear Stress ◽  
Small Gtpases ◽  
Dependent Manner ◽  
Fluid Shear ◽  
Rho Family

Blood vessels and blood cells are under continuous fluid shear. Studies on vascular endothelium and smooth muscle cells have shown the importance of this mechanical stress in cell signal transduction, gene expression, vascular remodeling, and cell survival. However, in circulating leukocytes, shear-induced signal transduction has not been investigated. Here we examine in vivo and in vitro the control of pseudopods in leukocytes under the influence of fluid shear stress and the role of the Rho family small GTPases. We used a combination of HL-60 cells differentiated into neutrophils (1.4% dimethyl sulfoxide for 5 days) and fresh leukocytes from Rac knockout mice. The cells responded to shear stress (5 dyn/cm2) with retraction of pseudopods and reduction of their projected cell area. The Rac1 and Rac2 activities were decreased by fluid shear in a time- and magnitude-dependent manner, whereas the Cdc42 activity remained unchanged (up to 5 dyn/cm2). The Rho activity was transiently increased and recovered to static levels after 10 min of shear exposure (5 dyn/cm2). Inhibition of either Rac1 or Rac2 slightly but significantly diminished the fluid shear response. Transfection with Rac1-positive mutant enhanced the pseudopod formation during shear. Leukocytes from Rac1-null and Rac2-null mice had an ability to form pseudopods in response to platelet-activating factor but did not respond to fluid shear in vitro. Leukocytes in wild-type mice retracted pseudopods after physiological shear exposure, whereas cells in Rac1-null mice showed no retraction during equal shear. On leukocytes from Rac2-null mice, however, fluid shear exerted a biphasic effect. Leukocytes with extended pseudopods slightly decreased in length, whereas initially round cells increased in length after shear application. The disruption of Rac activity made leukocytes nonresponsive to fluid shear, induced cell adhesion and microvascular stasis, and decreased microvascular density. These results suggest that deactivation of Rac activity by fluid shear plays an important role in stable circulation of leukocytes.

Expression Analyses of Mediator Complex Subunit 13-Like: A Responsible Gene for Neurodevelopmental Disorders during Mouse Brain Development

2021 ◽  
pp. 1-10
Author(s):  
Nanako Hamada ◽  
Ikuko Iwamoto ◽  
Masashi Nishikawa ◽  
Koh-ichi Nagata
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Cerebral Cortex ◽  
Brain Development ◽  
Developmental Stage ◽  
Expression Profile ◽  
Neurodevelopmental Disorders ◽  
Mediator Complex ◽  
Cortical Plate ◽  
The Central Nervous System

MED13L (mediator complex subunit 13-like) is a component of the mediator complex, which functions as a regulator for gene transcription. Since gene abnormalities in MED13L are responsible for neurodevelopmental disorders, MED13L is presumed to play an essential role in brain development. In this study, we prepared a specific antibody against MED13L, anti-MED13L, and analyzed its expression profile in mouse tissues with focusing on the central nervous system. In Western blotting, MED13L exhibited a tissue-dependent expression profile in the adult mouse and was expressed in a developmental stage-dependent manner in brain. In immunofluorescence analyses, MED13L was at least partially colocalized with pre- and post-synaptic markers, synaptophysin, and PSD95, in primary cultured hippocampal neurons. Immunohistochemical analyses revealed that MED13L was relatively highly expressed in ventricular zone surface of cerebral cortex, and was also located both in the cytoplasm and nucleus of neurons in the cortical plate at embryonic day 14. Then, MED13L showed diffuse cytoplasmic distribution throughout the cerebral cortex at the postnatal day (P) 30. In addition, MED13L appeared to be localized in cell type- and developmental stage-specific manners in the hippocampus and cerebellum. These results suggest that MED13L is involved in the development of the central nervous system and synaptic function.

scholarly journals Immunolocalization of Estrogen Receptor β in the Mouse Brain: Comparison with Estrogen Receptor α

Endocrinology ◽  
2003 ◽  
Vol 144 (5) ◽  
pp. 2055-2067 ◽  
Author(s):  
Sudha Warrier Mitra ◽  
Elena Hoskin ◽  
Joel Yudkovitz ◽  
Lisset Pear ◽  
Hilary A. Wilkinson ◽  
...  
Keyword(s):  
Estrogen Receptor ◽  
Cerebral Cortex ◽  
Olfactory Bulb ◽  
Mouse Brain ◽  
Preoptic Area ◽  
Estrogen Receptor Α ◽  
Hypothalamic Nucleus ◽  
Similar Distribution ◽  
Dependent Manner ◽  
The Brain

Estrogen receptor α (ERα) and ERβ are members of the steroid nuclear receptor family that modulate gene transcription in an estrogen-dependent manner. ER mRNA and protein have been detected both peripherally and in the central nervous system, with most data having come from the rat. Here we report the development of an ERβ-selective antibody that cross-reacts with mouse, rat, and human ERβ protein and its use to determine the distribution of ERβ in the murine brain. Further, a previously characterized polyclonal antibody to ERα was used to compare the distribution of the two receptors in the first comprehensive description of ER distribution specifically in the mouse brain. ERβ immunoreactivity (ir) was primarily localized to cell nuclei within select regions of the brain, including the olfactory bulb, cerebral cortex, septum, preoptic area, bed nucleus of the stria terminalis, amygdala, paraventricular hypothalamic nucleus, thalamus, ventral tegmental area, substantia nigra, dorsal raphe, locus coeruleus, and cerebellum. Extranuclear-ir was detected in several areas, including fibers of the olfactory bulb, CA3 stratum lucidum, and CA1 stratum radiatum of the hippocampus and cerebellum. Although both receptors were generally expressed in a similar distribution through the brain, nuclear ERα-ir was the predominant subtype in the hippocampus, preoptic area, and most of the hypothalamus, whereas it was sparse or absent from the cerebral cortex and cerebellum. Collectively, these findings demonstrate the region-selective expression of ERβ and ERα in the adult ovariectomized mouse brain. These data provide an anatomical framework for understanding the mechanisms by which estrogen regulates specific neural systems in the mouse.

Sign in / Sign up

Export Citation Format

Share Document