Identification and Characterization of Myosin from Rat Testicular Peritubular Myoid Cells1

Fernández, Darío; Bertoldi, María Victoria; Gómez, Laura Collados; Morales, Alfonsina; Callegari, Eduardo; López, Leonardo

doi:10.1095/biolreprod.107.066472

Cited by 12 publications

(4 citation statements)

References 40 publications

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“…The levels of Acta2 messenger RNA (mRNA) were low in these cultures even when compared to that in adult testicular tissue where peritubular myoid cell mRNA was greatly diluted with germ cell mRNA (Figure 3d). The same was true (Figure 3d) for Des and Myh11, which are also known to express in peritubular myoid cells (Fernández et al, 2008;Virtanen et al, 1986).…”

Section: Facs With Pdgfra Antibody Effectively Removes Peritubular My...supporting

confidence: 70%

Establishment of a pure culture of immature Sertoli cells by PDGFRA staining and cell sorting

Малолина

Galiakberova

Дашинимаев

et al. 2022

Molecular Reproduction Devel

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Sertoli cells are key somatic cells in the testis that form seminiferous tubules and support spermatogenesis. The isolation of pure Sertoli cells is important for their study. However, it is a difficult effort because of the close association of Sertoli cells with peritubular myoid cells surrounding seminiferous tubules. Here, we propose a novel approach to the establishment of a pure Sertoli cell culture from immature mouse testes. It is based on the staining of testicular cells for platelet-derived growth factor receptor alpha (PDGFRA), followed by fluorescence-activated cell sorting and culturing of a PDGFRA-negative cell population. Cells positive for a Sertoli cell marker WT1 accounted for more than 96% of cells in cultures from 6 to 12 days postpartum (dpp) mice. The numbers of peritubular myoid cells identified by ACTA2 staining did not exceed 4%. Cells in the cultures were also positive for Sertoli cell proteins SOX9 and DMRT1. Amh and Hsd17b3 expression decreased and Ar and Gata1 expression increased in 12 dpp cultures compared to 6 dpp cultures, which suggests that cultured Sertoli cells at least partially retained their differentiation status. This method can be employed in various applications including the analysis of differential gene expression and functional studies.

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Section: Facs With Pdgfra Antibody Effectively Removes Peritubular My...supporting

confidence: 70%

Establishment of a pure culture of immature Sertoli cells by PDGFRA staining and cell sorting

Малолина

Galiakberova

Дашинимаев

et al. 2022

Molecular Reproduction Devel

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“…PMCs were isolated from 3-week-old wild-type and Lgr4 −/− mice and plated in plastic culture dishes in MEM according to Fernández et al (Fernández et al, 2008). The cells were treated with or without recombinant R-spondin 1 (cat.…”

Section: Primary Pmc Culturementioning

confidence: 99%

Lgr4-mediated Wnt/β-catenin signaling in peritubular myoid cells is essential for spermatogenesis

Liu

Guan

et al. 2013

Development

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SUMMARYPeritubular myoid cells (PMCs) are myofibroblast-like cells that surround the seminiferous tubules and play essential roles in male fertility. How these cells modulate spermatogenesis and the signaling pathways that are involved are largely unknown. Here we report that Lgr4 is selectively expressed in mouse PMCs in the testes, and loss of Lgr4 leads to germ cells arresting at meiosis I and then undergoing apoptosis. In PMCs of Lgr4 mutant mice, the expression of androgen receptor, alpha-smooth muscle actin and extracellular matrix proteins was dramatically reduced. Malfunctioning PMCs further affected Sertoli cell nuclear localization and functional protein expression in Lgr4 −/− mice. In addition, Wnt/β-catenin signaling was activated in wild-type PMCs but attenuated in those of Lgr4 −/− mice. When Wnt/β-catenin signaling was reactivated by crossing with Apc min/+ mice or by Gsk3β inhibitor treatment, the Lgr4 deficiency phenotype in testis was partially rescued. Together, these data demonstrate that Lgr4 signaling through Wnt/β-catenin regulates PMCs and is essential for spermatogenesis.

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“…However, a causal relationship between contractility (or the lack thereof) and male (in)fertility has never been established. In fact, seminiferous tubule contractions had, so far, never been observed in vivo and most in vitro reports were based on indirect and non-quantitative evidence, for example from post-hoc fluorescence or scanning electron microscopy ( Barone et al, 2002 ; Fernández et al, 2008 ; Losinno et al, 2016 ; Losinno et al, 2012 ; Tripiciano et al, 1999 ; Tripiciano et al, 1997 ; Tripiciano et al, 1996 ), morphometry of single cells in culture ( Rossi et al, 2002 ; Santiemma et al, 2001 ; Santiemma et al, 1996 ; Tripiciano et al, 1996 ), or intraluminal pressure analysis ( Miyake et al, 1986 ; Yamamoto et al, 1989 ). The fact that expression of TPC contractility proteins initiates with puberty under androgen control and that selective androgen receptor knock-out in TPCs renders mice infertile ( Welsh et al, 2009 ) underscores a potential role of TPC contractions in male fertility.…”

Section: Discussionmentioning

confidence: 99%

ATP activation of peritubular cells drives testicular sperm transport

Fleck

Kenzler

Mundt

et al. 2021

eLife

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Spermatogenesis, the complex process of male germ cell proliferation, differentiation, and maturation, is the basis of male fertility. In the seminiferous tubules of the testes, spermatozoa are constantly generated from spermatogonial stem cells through a stereotyped sequence of mitotic and meiotic divisions. The basic physiological principles, however, that control both maturation and luminal transport of the still immotile spermatozoa within the seminiferous tubules remain poorly, if at all, defined. Here, we show that coordinated contractions of smooth muscle-like testicular peritubular cells provide the propulsive force for luminal sperm transport toward the rete testis. Using a mouse model for in vivo imaging, we describe and quantify spontaneous tubular contractions and show a causal relationship between peritubular Ca2+ waves and peristaltic transport. Moreover, we identify P2 receptor-dependent purinergic signaling pathways as physiological triggers of tubular contractions both in vitro and in vivo. When challenged with extracellular ATP, transport of luminal content inside the seminiferous tubules displays stage-dependent directionality. We thus suggest that paracrine purinergic signaling coordinates peristaltic recurrent contractions of the mouse seminiferous tubules to propel immotile spermatozoa to the rete testis.

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Identification and Characterization of Myosin from Rat Testicular Peritubular Myoid Cells1

Cited by 12 publications

References 40 publications

Establishment of a pure culture of immature Sertoli cells by PDGFRA staining and cell sorting

Establishment of a pure culture of immature Sertoli cells by PDGFRA staining and cell sorting

Lgr4-mediated Wnt/β-catenin signaling in peritubular myoid cells is essential for spermatogenesis

ATP activation of peritubular cells drives testicular sperm transport

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