Volume 4 • Issue 1 • 1000139
J Cell Sci Ther
ISSN: 2157-7013 JCEST, an open access journal
Open Access Research Article
Alhad Ashok Ketkar and KVR Reddy, J Cell Sci Ther 2013, 4:1
DOI: 10.4172/2157-7013.1000139
Keywords: Spermatogenesis; Spermatogonial stem cells (SSCs);
Microarray; days post partum (dpp)
Introduction
Spermatogenesis is a complex and well-orchestrated process, in
which Spermatogonial Stem Cells (SSCs) divide and diferentiate to
produce unlimited numbers of mature spermatozoa. Tis process
is continuous throughout the adult life in most mammals. Te
seminiferous epithelium consists of only one somatic cell type, sertoli
cell [1] and many diferent germ cell types [2]. Sertoli cells play a
nurturing role in coordinating and supporting important events of
spermatogenesis which occurs in mitotic, meiotic and post meiotic
phases. Spermatogenesis is initiated by the division of stem cells
(primitive type A spermatogonia) to form type B spermatogonia. Tese
diferentiated spermatogonial cells enter a meiotic prophase as pre-
leptotene spermatocytes. Te spermatocytes continue to mature as they
go through the leptotene, zygotene and pachytene phases of meiosis.
At the end of meiotic prophase, they undergo the fnal meiotic division
to form haploid spermatids. Tese spermatids undergo spermiogenesis
to form mature spermatozoa [3]. Tis complex process is orchestrated
through expression of thousands of genes encoding proteins which
are developmentally regulated during spermatogenesis and play
essential roles during specifc phases of germ cell development. Both
transcriptional and translational control mechanisms are responsible
for temporal and stage-specifc expression pattern of genes [4,5].
Each cell contains the same genome, but not all the genes are
used in each cell. Some genes are expressed when needed and many
genes are used to specify features unique to each type of cells. In
order to understand how genes are controlled and expressed at
specific time and which genes function uniquely in special cells, an
important step is defining gene expression profiles i.e. comparing
patterns of gene expression in different tissues and at different
developmental stages, in normal versus treated or diseased states.
This can be accomplished by RT-PCR, RNase protection assays or
Northern blot analysis, but these methods focus on only a few genes
at a time. A more promising approach for analyzing multiple genes
simultaneously is the hybridization of entire cDNA populations to
nucleic acid arrays such as microarray, a method adopted for high-
throughput analysis of gene expression [6-9].
In a similar vein, microarray technology can be applied to gain a
comprehensive view of expression pattern of genes involved in early
stages of spermatogenesis with the purpose of studying the mechanisms
and regulation of spermatogenesis at the genetic level. In rodents, frst
wave of spermatogenesis sets the basis for spermatogenesis process in
adult animals. In mouse, at 0 days post partum (dpp), gonocytes are in
the quiescent stage, around 3 dpp gonocytes are converted into SSCs
which then migrate to the basement membrane of seminiferous tubules,
at 5 dpp proliferation of SSCs starts, at 10 and 20 dpp spermatocytes
and spermatids appear, and at 35 dpp frst appearance of spermatozoa
thereby marking the end of the frst wave of spermatogenesis [10,11].
In our earlier studies, we deduced the expression patterns of Oct-4 as
well as Plzf and functional signifcance of Oct-4 during the early stages
of spermatogenesis in mice [12,13]. Both Oct-4 and Plzf are known to
be expressed in undiferentiated SSCs and considered to be important
for SSC self-renewal. Te results of study showed Oct-4 and Plzf
expression to be highest in the testes of 10 dpp mice both at the mRNA
and protein level [12]. Apart from Oct-4 and Plzf, the SSC self-renewal
and proliferation process involves many other genes which are crucial
to maintain SSCs in the undiferentiated state. Terefore, the main aim
of the present study is to elucidate the expression pattern of all those
*Corresponding authors: KVR Reddy, Division of Molecular Immunology and
Microbiology (MIM), National Institute for Research in Reproductive Health
(NIRRH), JM Street, Parel, Mumbai-400012, India, Tel: +91-22-24192016;
Fax: +91-22-24139412; E-Mail: reddyk@nirrh.res.in
Received December 19, 2012; Accepted March 19, 2013; Published March 21,
2013
Citation: Alhad Ashok Ketkar, KVR Reddy (2013) Differential Expression of Genes
Involved in Early Events of Spermatogenesis in Mice. J Cell Sci Ther 4: 139.
doi:10.4172/2157-7013.1000139
Copyright: © 2013 Ketkar AA, et al. This is an open-access article distributed under
the terms of the Creative Commons Attribution License, which permits unrestricted
use, distribution, and reproduction in any medium, provided the original author and
source are credited.
Abstract
Spermatogenesis is a highly complex process involved in the transmission of genetic information from one
generation to next generation. The process takes place in the seminiferous tubules of the testis and is regulated
by genes that control cell division, cell-cell interaction and morphogenetic changes in both the somatic and germ
cell lineages in a highly orchestrated pattern. The frst wave of spermatogenesis in mouse is typically characterized
by the differential expression of genes involved in spermatogenesis. The purpose of this study was to analyze
the differential expression of genes involved in Spermatogonial Stem Cells (SSCs) self-renewal, proliferation
and differentiation using microarray approach in the testes of 35 vs. 5 Days Post Partum (dpp) mice. Our results
demonstrate that genes involved in SSC self-renewal and proliferation were signifcantly down-regulated while
genes involved in SSC differentiation were signifcantly up-regulated in the testes of 35 vs. 5 dpp mice. There was
up-regulation in the expression of genes involved in cell cycle regulation. Pro-apoptotic genes were found to be up-
regulated on the contrary anti-apoptotic genes were down-regulated in the testes of 35 vs. 5 dpp mice. Thus, our
study helps in understanding the differential expression profle of genes involved in early stages of spermatogenesis
in mice.
Differential Expression of Genes Involved in Early Events of
Spermatogenesis in Mice
Alhad Ashok Ketkar and KVR Reddy*
Division of Molecular Immunology and Microbiology (MIM), National Institute for Research in Reproductive Health (NIRRH), JM Street, Parel, Mumbai-400012, India
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