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Cellular and Molecular Neurobiology

, Volume 38, Issue 4, pp 919–927 | Cite as

Co-Localization and Interaction of Pax5 with Iba1 in Brain of Mice

  • Shashank Kumar Maurya
  • Rajnikant Mishra
Original Research

Abstract

The Pax5, a B-cell-Specific Activator Protein (BSAP) and redox-sensitive transcription factor, is expressed in the immune-privileged brain, B-lymphocytes, lymph nodes and spleen. PAX5-mediated immune pathway has also been described in the progression of Glioblastoma multiforme. However, the status of Pax5 and its role in brain immunity are not yet elucidated. In silico analysis of Pax5 interacting proteins predicts its interaction with proteins of cell proliferation, differentiation of hematopoietic cells, neurogenesis and several cell signalling pathways. Promoter analysis shows multiple binding sites for Pax5 in promoter of ionized calcium-binding adapter molecule 1 (Iba1). Like Iba1, Pax5 is also associated with inflammatory and immune response, activation of leukocyte and remodelling of actin cytoskeleton. Therefore, localization and interaction of Pax5 with Iba1 in brain of mice were studied using Chromatin Immunoprecipitation (ChIP), Co-Immunoprecipitation (Co-IP) and Immuno-fluorescence assay. The Pax5- and Iba1-positive cells were observed in cerebral cortex, cerebellum, olfactory bulb, hippocampus, and ventricles of brain. The co-localization of Pax5 and Iba1 was evident in microglia in almost all evaluated regions of brain. In some regions, Pax5- and Iba1-positive were distinctly compartmentalized. The Pax5a/b interacts with Iba1 and binds to its regulatory sequences. Results indicate Pax5-associated activities of Iba1 in microglia in brain of mice.

Keywords

Pax5 Iba1 Microglia Brain immunity 

Abbreviations

Pax5

Paired box 5

BSAP

B-cell-Specific Activator Protein

Iba1

Ionized calcium-binding adapter molecule 1

IFN-γ

Interferon-γ

CSF

Cerebrospinal fluid

IHC

Immunohistochemistry

ChIP

Chromatin Immunoprecipitation

Co-IP

Co-Immunoprecipitation

Cc

Cerebral cortex

C

Cerebellum

ChP

Choroid plexus

OB

Olfactory bulb

H

Hippocampus

TV

Third ventricle

LV

Lateral ventricles

CNS

Central nervous system

Whm

White matter

Gm

Grey matter

ML

Molecular layer

OlGl

Olfactory glomerular layer

PGC

Peri-glomerular cells

BV

Blood vessels

IML

Inner molecular layer

GCL

Granular cell layer

SGZ

Sub-granular zone

AGC

Adult granule cells

Rm

Ramified microglia

Am

Amoeboid microglia

rm

Round microglia

Notes

Acknowledgements

RM acknowledges partial financial supports from DST-PURSE, UGC-CAS & UPE-II, and BHU. We are thankful to DBT-ISLS-BHU for extending confocal microscopy and Prof. A.K. Mishra, Department of Botany, BHU for Real-time PCR facilities.

Compliance with Ethical Standards

Conflict of interest

Dr. Rajnikant Mishra and Shashank Kumar Maurya declare that they have no conflict of interest.

Ethical Approval

All the experiments were approved by the Animal Ethical Committee of Institute of Science, Banaras Hindu University, Varanasi 221005, India.

Supplementary material

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Supplementary material 1 (TIFF 279161 kb)
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Supplementary material 2 (TIFF 281690 kb)
10571_2017_566_MOESM3_ESM.tif (61.9 mb)
Supplementary material 3 (TIFF 63385 kb)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2017

Authors and Affiliations

  1. 1.Department of Zoology, Biochemistry and Molecular Biology LaboratoryInstitute of Science, Banaras Hindu UniversityVaranasiIndia

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