Anat Cell Biol.  2012 Jun;45(2):97-102. 10.5115/acb.2012.45.2.97.

Differential expression levels of synaptophysin through developmental stages in hippocampal region of mouse brain

• Pyeon HJ
• Lee YI

• KMID: 2005872
• DOI: http://doi.org/10.5115/acb.2012.45.2.97

Abstract

The formation of neural synapses according to the development and growth of neurite were usually studied with various markers. Of these markers, synaptophysin is a kind of synaptic protein located in the synaptic vesicle of neuron or neuroendocrine cell known to be distributed consistently in all neural synapses. The purpose of this study was to investigate differential expression levels and patterns of synaptic marker (synaptophysin) in the mouse hippocampal region according to the developmental stages of embryonic, neonatal, and adulthood respectively. In the embryonic and neonatal groups, synaptophysin immunofluorescence was almost defined to cornu ammonis subfields (CA1 and CA3) of hippocampus and subiculum proper in the hippocampal region. However in dentate gyrus, synaptophysin immunoreactivities were insignificant or absent in all developmental stages. In embryonic and neonatal hippocampus, the intensities of immunofluorescence were significantly different between molecular and oriens layers. Furthermore, those intensities were decreased considerably in both layers of neonatal group compared to embryonic. The results from this study will contribute to characterizing synaptogenic activities in the central nervous system through developmental stages.

Keyword

Synaptophysin; Hippocampus; Cornu ammonis; Dentate gyrus; Developmental stages

MeSH Terms

Animals
Brain
Central Nervous System
Dentate Gyrus
Fluorescent Antibody Technique
Growth and Development
Hippocampus
Mice
Neurites
Neuroendocrine Cells
Neurons
Synapses
Synaptic Vesicles
Synaptophysin
Synaptophysin

Figure

• 

  Fig. 1 Confocal densitometric analysis methods for the morphological quantification of synaptophysin in the mice hippocampal region (hippocampus and subiculum proper). (A) Diagram of coronal thin section (10 µm) of the mouse brain. Hippocampal areas in red rectangular region were used for confocal microscopic analysis. (B) Confocal image of coronal sectioned hippocampal region. Synaptophysin volume data were obtained by analyzing this image using the system calculating penetration ratio of light intensity into optical density values. Optical density measurement was obtained by manually positioning the area (white dashed square, superficial molecular layer [Mol] and lacunosum molecular layer [LMol]; white straight square, deep oriens layer [Or]) of immunofluorescence-stained section. (C) ZEN 2009 (ver. 5,5,0,375, Carl Zeiss) software used for image acquisition and analysis. The synaptophysin densities were assayed in relation to the areas outlined, and results were appeared on the analysis software (C). Py, pyramidal cell layer; Rad, radiatum layer.

• 

  Fig. 2 Immunofluorescence of synaptophysin in the hippocampal region of mouse brain in each developmental stage of embryonic (A), neonatal (B), and adult (C). (D) Magnified picture of (A). Note synaptophysin immunoreactivities connecting pyramidal cell layer (Py) and lacunosum molecular layer (LMol) through radiatum layer (Rad) (arrow). (E) Differential synaptophysin immunoreactivites between molecular layer (Mol) and oriens layers (Or) of hippocampal region in each developmental stages. *P<0.05. n=6-10. Nuclei are counterstained with DAPI. Scale bars=100 µm.

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