Dataset Information

Screening of Integrin Heterodimers Expressed Functionally on the Undifferentiated Spermatogonial Stem Cells in the Outbred ICR Mice.

ABSTRACT: Background and objectives: Outbred mice are widely used in toxicology, pharmacology, and fundamental biomedical research. However, there have been no reports of in vitro culture systems for spermatogonial stem cells (SSCs) derived from these mice.

Methods: As a step towards constructing a non-cellular niche supporting the in vitro maintenance of outbred mouse SSC self-renewal, we systematically investigated the types of integrin heterodimers that are expressed transcriptionally, translationally, and functionally in SSCs derived from Imprinting Control Region (ICR) mice.

Results: Among the genes encoding 25 integrin subunits, integrin ?₁, ?₅, ?₆, ?₉, ?_V, and ?_E, and integrin ?₁ and ?₅ had significantly higher transcriptional levels than the other subunits. Furthermore, at the translational level, integrin ?₅, ?₆, ?₉, ?_V, ?_E, and ?₁ were localized on the surface of SSCs, but integrin ?₁ and ?₅ not. Moreover, significantly stronger translational expression than integrin ?₉ and ?_E was observed in integrin ?₅, ?₆, ?_V, and ?₁. SSCs showed significantly increased adhesion to fibronectin, laminin, tenascin C and vitronectin, and functional blocking of integrin ?₅?₁, ?₆?₁, ?₉?₁ or ?_V?₁ significantly inhibited adhesion to these molecules.

Conclusions: We confirmed that integrin ?₅?₁, ?₆?₁, ?₉?₁ and ?_V?₁ actively function on the surface of undifferentiated SSCs derived from outbred ICR mice.

SUBMITTER: Park HJ

PROVIDER: S-EPMC7691863 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature

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Publications

Screening of Integrin Heterodimers Expressed Functionally on the Undifferentiated Spermatogonial Stem Cells in the Outbred ICR Mice.

Park Hye Jin HJ Yun Jung Im JI Kim Minseok M Choi Kimyung K Lee Eunsong E Lee Seung Tae ST

International journal of stem cells 20201101 3

<h4>Background and objectives</h4>Outbred mice are widely used in toxicology, pharmacology, and fundamental biomedical research. However, there have been no reports of <i>in vitro</i> culture systems for spermatogonial stem cells (SSCs) derived from these mice.<h4>Methods</h4>As a step towards constructing a non-cellular niche supporting the <i>in vitro</i> maintenance of outbred mouse SSC self-renewal, we systematically investigated the types of integrin heterodimers that are expressed transcri ...[more]

PMID: 32840227

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Project description:Integrins are transmembrane cell receptors involved in two crucial mechanisms for successful fertilization, namely, mammalian intracellular signaling and cell adhesion. Integrins α6β4, α3β1 and α6β1 are three major laminin receptors expressed on the surface of mammalian cells including gametes, and the presence of individual integrin subunits α3, α6, β1 and β4 has been previously detected in mammalian sperm. However, to date, proof of the existence of individual heterodimer pairs in sperm and their detailed localization is missing. The major conclusion of this study is evidence that the β4 integrin subunit is expressed in mouse sperm and that it pairs with subunit α6; additionally, there is a detailed identification of integrin heterodimer pairs across individual membranes in an intact mouse sperm head. We also demonstrate the existence of β4 integrin mRNAs in round spermatids and spermatogonia by q-RT-PCR, which was further supported by sequencing the PCR products. Using super-resolution microscopy accompanied by colocalization analysis, we located integrin subunits as follows: α6/β4-inner apical acrosomal membrane and equatorial segment; α3, α6/β1, β4-plasma membrane overlaying the apical acrosome; and α3/β1-outer acrosomal membrane. The existence of α6β4, α3β1 and α6β1 heterodimers was further confirmed by proximity ligation assay (PLA). In conclusion, we delivered detailed characterization of α3, α6, β1 and β4 integrin subunits, showing their presence in distinct compartments of the intact mouse sperm head. Moreover, we identified sperm-specific localization for heterodimers α6β4, α3β1 and α6β1, and their membrane compartmentalization and the presented data show a complexity of membranes overlaying specialized microdomain structures in the sperm head. Their different protein compositions of these individual membrane rafts may play a specialized role, based on their involvement in sperm-epithelium and sperm-egg interaction.

Dataset Information

Screening of Integrin Heterodimers Expressed Functionally on the Undifferentiated Spermatogonial Stem Cells in the Outbred ICR Mice.

Publications

Screening of Integrin Heterodimers Expressed Functionally on the Undifferentiated Spermatogonial Stem Cells in the Outbred ICR Mice.

Similar Datasets

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