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Cell–Cell Interactions—Structural

Chapter · January 2018

DOI: 10.1016/B978-0-12-801238-3.64564-6

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 Cell–Cell Interactions—Structural
Nathália de Lima e Martins Lara, Gleide Fernandes de Avelar, Guilherme Mattos Jardim Costa, and Samyra
Maria dos Santos Nassif Lacerda, Federal University of Minas Gerais, Belo Horizonte, Brazil
Rex A Hess, University of Illinois at Urbana-Champaign, Urbana, IL, United States
Luiz Renato de França, Federal University of Minas Gerais, Belo Horizonte, Brazil and National Institute for Amazonian
Research, Manaus, Brazil
© 2017.

Email addresses: nathalialimalara@gmail.com (N. de Lima e Martins Lara); gleideav@yahoo.com.br (G.F. de Avelar); costagmj@gmail.
com (G.M.J. Costa); samyranassif@hotmail.com (S.M. dos Santos Nassif Lacerda); rexhess@illinois.edu (R.A. Hess);
lrfranca@icb.ufmg. br, lrfranca@inpa.gov.br (L.R. de França)
Introduction

Interactions between cells are required for the maintenance or alterations/changes of cellular function, differentiation and growth, being
therefore crucial for the normal physiology of tissues and organs. A great array of cellular interactions are required for the coordination
of reproductive processes, at the structural and molecular levels. As millions or even billions of spermatozoa are daily produced in the
testis, in a very active and complex process named spermatogenesis that occurs in the seminiferous epithelium, this naturally requires a
very coordinated and organized interactions among testicular cells. For instance, since seminiferous cords/tubules formation, extensive
interactions between the germ and Sertoli cells are needed for the development/self-renew and differentiation of germ cells, particularly in
relation to the structural and nutritional aspects. In addition, the formation of the blood-testis or Sertoli cell barrier (SCB) between
adjacent Sertoli cells, which creates a specialized microenvironment, is crucial for the development of more mature germ cells. In order to
provide an optimal androgen milieu, in the intertubular compartment there is also a set of complex interactions and communications
between the steroidogenic Leydig cells and other important interstitial cells. In this article, the main structural cellular interactions present
in the testis (summarized in Table 1) is discussed, focusing particularly on its composition and importance for spermatogenesis.

Tubular Compartment
Sertoli–Sertoli Cell
During spermatogenesis, especially when germ cells are undergoing meiosis they show a different profile of surface antigens that are
usually detected by the immune system and lead to an autoimmune reaction. In order to protect the development of these germ cells,
Sertoli cells form the Sertoli cell barrier (SCB) (the most important component of the so-called blood-testis barrier), which helps to create
an immune privileged environment inside the seminiferous epithelium, where foreign or autoimmunogenic antigens can be tolerated
without inducing a detrimental immune response, leading ultimately to male infertility (França et al., 2012).
Initial evidence for the existence of the SCB came from different sources. Firstly, it was observed that the composition of the fluid
present in the lumen of the seminiferous tubule is different from blood plasma or lymph. This difference would not exist if the tubule
walls were permeable to molecules from the intertubular and basal compartments. Other evidence came from the use of tracers and dyes,
determining the entry rate of different fluids and the relationship of a SCB to the fluid composition and solubility. Therefore, it was
concluded that there must be a mechanism for controlling the entry of substances into the seminiferous tubule.
The SCB is formed by specialized junctional complexes between adjacent Sertoli cells, near the basement membrane, dividing the
seminiferous epithelium into two compartments: basal and adluminal. Spermatogonia and early primary spermatocytes (preleptotene/
leptotene stage) are located in the basal compartment. Spermatocytes must move from basal to the adluminal compartment, through a
transient, intermediate compartment, without damaging the barrier structure. This intermediate compartment is created by a newly formed
barrier, basal to the migrating cell, separating it from the basal compartment. The old junctional complex, luminal to the spermatocyte,
is then dismantled and these cells enter the adluminal compartment. In this way, more advanced spermatocytes (from leptotene/zygotene
onward) and the round and elongated spermatids are located in the protected adluminal compartment (Cheng and Mruk, 2012).
The main junctional constituent of the SCB is the tight junction, but desmosomes, gap junctions, tubulobulbar complexes and basal
ectoplasmic specialization (the last two are testis-specific types of adherens junctions and will be discussed in more detail in the next
section) are also present and collectively maintain this morphological immune barrier of the testis. The proteins involved include
occludins, claudins, cadherins, catenins, and connexins, usually anchored by actin or vimentin filaments. This particular arrangement
gives strength to the SCB, making it one of the tightest barriers in mammalian tissues.
Beyond strength and immunological protection, the barrier also provides for communication between the adjacent Sertoli cells, which
is essential for coordination and signaling during the different stages of the seminiferous epithelial cycle. In addition, the junctional
complexes restrict and control the transit of molecules from blood and interstitial spaces to the seminiferous tubule (and vice-versa),
effectively limiting the immune system's access to the adluminal compartment of the seminiferous epithelium. The specialized

Reference Module in Biomedical Research, Volume ■ https://doi.org/10.1016/B978-0-12-801238-3.64564-6 1

2 Cell–Cell Interactions—

Table 1 Summary of the main junctions present in the testis and the cell types involved

Junction Cells involved

Tight junctions Sertoli–Sertoli cell (SCBa)

Adherens junctions Sertoli cell-spermatogonia
Leydig–Leydig cell
Gap junctions Sertoli–Sertoli cell (SCB)
Sertoli–germ cells
Leydig–Leydig cell
Desmosomes Sertoli–Sertoli cell (SCB)
Sertoli cell-spermatocytes/early spermatids
Hemidesmosomes Sertoli cell-basement membrane
Tubulobulbar complex Sertoli–Sertoli cell (SCB)
Sertoli cell-elongated spermatid (around spermiation)
Ectoplasmic specialization (ES) Sertoli–Sertoli cell (SCB; basal ES)
Sertoli cell-elongated spermatid (apical ES)
Intercellular bridge Germ–germ cell
Leydig–Leydig cell (Fetal)
Digitations Leydig cell-macrophage
a
SCB, Sertoli cell barrier.

microenvironment created is enhanced by immunomodulatory factors produced locally that cause an immunosuppressive response,
instead of a possible destructive proinflammatory reaction. Sertoli cell polarity is also conferred by the presence of the barrier,
illustrated by the usual basal location of the nucleus and an uneven distribution of cytoplasmic organelles within the Sertoli cell. This
asymmetric organization is very important for the support of spermatogenesis, as physiological and cellular events take place in the
corresponding cellular compartments in a stage-specific manner (Fig. 1).
In mammals, the establishment of the SCB occurs when the Sertoli cells stop proliferating and terminally differentiate, coinciding with
the production of primary spermatocytes, tubular fluid secretion and lumen formation. As observed in the SCARKO mice model (mice
with a selective ablation of the androgen receptor in Sertoli cells), germ cells are also important for the establishment and effectiveness of
the barrier, as the meiotic phase of spermatogenesis can only be initiated in the presence of a competent SCB. Permeability and structure
of the barrier is highly dynamic. As spermatogenesis proceeds, different molecules are transported to and from the interior of the tubule,
depending on the spermatogenic cycle phase and the needs of the developing germ cells. Due to the presence of the barrier, hormones,
such as FSH and LH for example, may act indirectly in germ cells, via Sertoli cells, which allows for a higher level of control by these
supporting somatic cells over the spermatogenic process (França et al., 2016).

Sertoli–Germ Cell
The interactions between the Sertoli and germ cells in the seminiferous epithelium must be tightly regulated, because Sertoli cells provide
the main control over spermatogenesis, providing nutritional and structural support for all developing germ cells. This Sertoli/germ cell
interaction occurs through different types of junctions, such as adherens junctions, desmosomes, and gap junctions, depending on the
stage of the seminiferous epithelium cycle and germ cell-specific requirements. For example, adherens junctions are usually detected
between Sertoli and spermatogonia located in the stem cell niche, being important for stem cell homing and colonization. Desmosomes,
in turn, are the major anchoring junction between spermatocytes/early spermatids and the Sertoli cells. Additionally, gap junctions
provide crucial communications between adjacent germ cells, which is essential for the coordination of germ cell development throughout
the different stages of differentiation (Fig. 2).
The testis forms a unique type of adherens junction, called ectoplasmic specialization (ES), which is not found in any other epithelium
and is usually considered as one of the strongest anchoring junctions in the mammalian body ( Cheng and Mruk, 2015). Despite
participating in the SCB (basal ES), apical ectoplasmic specialization is also found at the Sertoli cell-elongated spermatid interface,
conferring spermatid adhesion and polarity. Spermatid polarity involves the elongating heads pointing perpendicularly toward the
basement membrane of the seminiferous tubule, allowing for maximum packing of developing spermatids in the seminiferous epithelium.
This polarity and the very well-coordinated spermatogenic process allows the usually very high sperm output observed during
spermatogenesis. Loss of spermatid adhesion and early release of the spermatids to the lumen of the seminiferous tubule is often caused
by the disruption of cell polarity. In this context, it has also been shown that DICER (involved in the miRNA biosynthesis pathway) is an
important player in the regulation of these Sertoli/germ cell junctions (Korhonen et al., 2015).
Structurally, the ectoplasmic specializations consist on bundles of actin microfilaments sandwiched between flattened cisternae of
the Sertoli cell endoplasmic reticulum, and are present in the Sertoli cell membrane that faces the spermatid. This structure is closely
associated with the microtubule network of the Sertoli cell, and is probably responsible for the movement of elongating spermatids across
the seminiferous epithelium during the stages of the cycle. Injection of a microtubule stabilizer (taxol) lead to the retention of spermatids
deep within the Sertoli cell crypts, while disruption of both ES and microtubules trapped the spermatids inside the epithelium, blocking
 Cell–Cell Interactions— 3

Fig. 1 Sertoli cell barrier (SCB). (A) Schematic representation of the junctions that form the Sertoli cell barrier junctional complex. (B)
Immunohistochemistry for Connexin-43 (Cx43; arrows), which is the main component of the gap junctions in the SCB. (C) The SCB delimited by
the rectangle area and viewed by electron microscopy. (D) Higher magnification of the area depicted in C. (E) Tubulobulbar complex observed in
the SCB. SC, Sertoli cell; GC, Germ cell; Spg, Spermatogonia; Spt, Spermatocyte; PMC, Peritubular myoid cell; Red arrowheads, Sertoli cell
barrier (SCB); Black arrowheads, Tubulobulbar complex.

their release at spermiation, until they were phagocytosed by the Sertoli cells. The molecular composition of the ES is constituted by
nectin-, cadherin- and integrin-based protein complexes. A polarity protein complex, composed of partitioning-defective3/partitioning-
defective6/ atypical protein kinase C (Par3/Par6/aPKC), is also an integrated component of the apical ectoplasmic specialization.
The apical ectoplasmic specialization appears in step 8 spermatids, replacing gap junctions and desmosomes as the only anchoring
device between the spermatid and the Sertoli cell. Around the time of spermiation (steps 18–19 of spermiogenesis in rats, and steps 15–16
in mice), the apical ES undergoes degeneration, through the formation of structures similar to endocytic vesicles, which initiates the
elimination of cytoplasmic debris resulting from the release of the spermatids. These vesicular structures are called tubulobulbar
complexes. They form narrow tubular projections of the plasma membrane of the elongating spermatid head, penetrating an invagination
of the Sertoli cell plasma membrane and forming a bulbous end. The tubular portion of the complex is surrounded by actin filaments,
while the bulb is more associated with cisternae of the endoplasmic reticulum. The tubulobulbar complex facilitates the internalization of
the disassembled apical ES junctions, in preparation for the release of sperm cells during spermiation (Vogl et al., 2013). Steroids,
especially androgens, may be involved with the regulation of tubulobulbar complex formation, as it occurs concomitant with the
occurrence of androgen-dependent stages of the seminiferous epithelium cycle (Fig. 3).
4 Cell–Cell Interactions—

Fig. 2 (A, B) Desmosome-like junctions between Sertoli cell (SC) and germ cell (GC). The insert represents a higher magnification of the area
depicted by the small box. Opposing red arrowheads, Sertoli cell barrier; TP, Tunica propria.

Sertoli Cell–Tunica Propria

The exterior wall of the seminiferous tubule—the tunica propria—is composed mainly by the peritubular myoid cells, separated from the
Sertoli cells by a basement membrane. This cell layer is responsible for structural integrity and contraction of the seminiferous tubule
and regulatory interactions between the interstitial and tubular compartments. The peritubular myoid cells are in constant interaction with
the nearby Sertoli cells and participate actively in the SCB (Skinner, 1993) and establishment of the stem cell niche. Their interaction is
mainly mediated by the extracellular matrix of the basement membrane present between them, produced cooperatively by both peritubular
and Sertoli cells. The peritubular myoid cells are capable of influencing the Sertoli cell function and vice-versa, changing the cellular
synthetic profile, proliferation, differentiation and hormone responsiveness. Some paracrine factors have been shown to be involved in
this regulation, such as activin, PModS, and growth factors (TGFα, TGFβ, IGF-1), for example, but no structural interaction is described
directly between these cells. Therefore, although not in direct contact, the Sertoli and peritubular myoid cells have a close association that
involves a number of cell–cell interactions, mainly paracrine, in order to maintain Sertoli cell function, retention of peritubular myoid cell
phenotype/activity, and spermatogenesis (Verhoeven et al., 2000).
Additionally, recent reports discuss the existence of a local apical ES-blood-testis barrier-basement membrane functional axis, mainly
mediated by polarity proteins that could coordinate the events of junction restructuring in the seminiferous epithelium during
spermatogenesis. This can be illustrated by proteins at the Sertoli cell–extracellular matrix interface, such as integrins, which can provide
a feedback regulatory mechanism to modulate barrier integrity. In addition, proteins released during spermiation (e.g., fragments of
laminins from the apical ES) can collaborate with polarity proteins to regulate SCB remodeling, directly or indirectly, via
hemidesmosomes in the basement membrane. Experimental manipulation of hemidesmosomes integrins can disrupt the barrier function.
These findings are supported by the fact that three cellular events that involve cellular interactions but at opposite ends of the Sertoli cell
epithelium, namely
(1) spermiation; (2) transit of primary spermatocytes and SCB restructuring; and (3) germ cell-cycle progression through meiosis that
occurs concomitant in time. Therefore, it seems logical to assume that a regulatory loop would coordinate these cellular events (Wong
and Cheng, 2009).

Germ–Germ Cell
During spermatogenesis, the germ cells go through successive cell divisions but cytokinesis is usually incomplete, resulting in daughter
cells that are connected by cytoplasmic bridges. This process forms a syncytium of clonal cells derived from one spermatogonial stem
cell. These bridges are maintained throughout spermatogenesis, even when germ cells migrate from the basal to the adluminal
compartment through the SCB, and are eliminated only at the final stage of spermatid development, along with the residual cytoplasmic
bodies. The presence of this syncytium is responsible for the synchronous development of germ cells by sharing essential signals,
proteins, mRNA and organelles, which helps to coordinate mitosis, meiosis, differentiation and apoptosis of these cells (Smith and
Braun, 2012).
The intercellular bridges are large and stable cytoplasmic channels, with an electron dense wall composed mainly of actin. Some
proteins were shown to participate in these intercellular bridges, with Tex14 (Testis-Expressed Gene 14) being the most studied. This
protein forms the mitotic spindle matrix and its absence in mice leads to spermatogenesis interruption at the meiotic phase and no
formation of intercellular bridges. Tex14 blocks the complex centralspindlin that is formed by the proteins MKLP1 (Mitotic Kinesin-
Like Protein 1) and MgcRacGAP (Male Germ Cell Rac GTPase-activating protein). This complex recruits abscission effector proteins,
such as Cep55 (Centrosomal Protein 55-KDa), that are responsible for the completion of cytokinesis (Greenbaum et al., 2011).
In summary, the presence of cytoplasmic bridges between germ cells is a unique mode of intercellular communication, essential
for fertility. However, the specific roles of the intercellular bridges for stem cell self-renewal, proliferation and differentiation of more
 Cell–Cell Interactions— 5

Fig. 3 Ectoplasmic specialization (ES) and tubulobulbar complex in elongating spermatids. (A) Schematic representation of the ectoplasmic
specialization (left) and the tubulobulbar complex (right) and the relationship between the elongating spermatids and the Sertoli cell. (B) Electron
microscopy of an elongating spermatid showing the ES structure, composed by actin filaments (yellow asterisks) sandwiched between smooth
endoplasmic reticulum cisternae (arrows) and the apposing plasma membranes of the Sertoli and the germ cell (opposing white arrowheads). (C,
D) Cross-section of mouse seminiferous tubules illustrating the intimate relationship between the Sertoli cell and the elongating spermatids
delimited by the rectangle. (C) Stage I tubule showing the elongating spermatids deep within the cripts of the Sertoli cell. (D) Stage VII tubule,
showing elongated spermatids at the edge of the seminiferous epithelium, close to the lumen, in preparation for spermiation. ES, Ectoplasmic
specialization; SC, Sertoli cell; Ac, Acrosome; M, Mitochondria; PL, Preleptotene spermatocytes; P, Pachytene spermatocytes; RS, Round
spermatids; E, Elongating spermatids; RB, Residual body; PMC, Peritubular myoid cell.

advanced germ cells still need to be elucidated. The study of the germ cell's ability to inactivate abscission during cytokinesis could
provide relevant information leading to the ability to regulate this cell division event in abnormal cells found in cancers (Fig. 4).

Intertubular Compartment

Leydig cells are the main component of the intertubular (interstitial) compartment of the testis. Among other important functions, these
steroidogenic cells are responsible for synthesizing and secreting androgens, which are essential hormones for spermatogenesis and
development of male sex characteristics. Other important components of the interstitial compartment include macrophages, fibroblasts,
lymphocytes, mast cells, blood, and lymphatic vessels. The interaction between these components and the Leydig cells is very important
6 Cell–Cell Interactions—

Fig. 4 Intercellular bridges between germ cells. (A) Electron microscopy section of the seminiferous epithelium, showing three pachytene
spermatocytes (Sp) connected by intercellular bridges (opposing arrowheads). (B) Semithin cross-section of an intercellular bridge between
pachytene spermatocytes. (C, E) Higher magnification of the intercellular bridges observed by electron microscopy. Note that, due to the large size
of the bridge, the cells form a syncytium, where signals, proteins, mRNA and even organelles can be shared. SC, Sertoli cell; Spt, Spermatocyte; L,
Lipid droplet; TP, Tunica propria; *, Mitochondria; G, Golgi complex.

for their development and differentiation and for the correct function of the testis. In this context, the most important and studied cell to
cell structural interaction is between Leydig cells and macrophages, while less is known about the interactions with the other interstitial
cells. It is worth mentioning that, although anatomically separated, Sertoli and Leydig cells highly modulate each other's development and
function through paracrine interactions and secreted factors, such as androgens, inhibin, interleukin-1, grown factors, and estrogen.

Leydig–Leydig Cell
Each Leydig cell is coupled to another Leydig cell through adherens and gap junctions and these connections are present in both fetal and
adult populations. The presence of these junctions allows the exchange of ions and small molecules, besides controlling androgen
secretion. Various proteins are involved in this communication, such as connexins (especially connexin-43, which abundance is dependent
on LH levels) and cadherins, cell adhesion molecules (You et al., 2000; Pointis et al., 2010). Intercellular bridges with continuous
cytoplasm can also be observed between some fetal Leydig cells (FLC).
In the adult population of Leydig cells (ALC), the gap junctions form before the increase in secretory activity of these cells, which is
associated with puberty, when this population is established. The structural diversity and presence of these junctions increase along with
the maturity of the adult Leydig cell and, in general, the ALC tend to form more gap junctions in comparison to the FLCs (Haider et al.,
2007). Some of the gap junctions have a distinctive annular appearance, as they are formed between a cell process and the main
cytoplasmic region of the other cell. However, the reason for this characteristic is still unknown. The presence of gap junctions between a
group of Leydig cells strongly suggests a coordinated activity, which could be related to the variation in androgenic activity of Leydig
cells depending on the stage of the spermatogenic cycle, as well as proximity of these Leydig cell groups to the tunica propria and the
seminiferous tubule itself (Fig. 5).

Leydig-Immune Cells
In most species, Leydig cells are the most prevalent cell type in the testicular interestitium, while macrophages are the second most
frequent cell. Testicular macrophages and Leydig cell numbers increase proportionately throughout development, as changes in their
 Cell–Cell Interactions— 7

Fig. 5 Gap junctions between Leydig cells. (A, B) Electron microscopy images of the contact between two Leydig cells. The insert shows the
gap junctions in higher magnification. (C, D) Immunohistochemistry for Connexin-43 in the mouse testis, between fetal Leydig cells (FLC;
embryonic day 18.5) (C) and adult Leydig cells (ALC; 70 postnatal day) (D). Observe that the staining is also present between the Sertoli cells in
the seminiferous cord/tubule. LC, Leydig cell; SC, Sertoli cell; BV, Blood vessel; Arrows, Gap junctions.

morphology are coordinated. This association suggests a functional relationship between both cells, but much remains to be learned. From
studies mainly based on rats, it is estimated that there is approximately one macrophage for every three to five Leydig cells in the testis.
The macrophages are usually in direct contact with adjacent Leydig cells through membrane associations or “digitations.” These
interactions appear as long microvillus-like Leydig cell processes inserted within coated membrane invaginations into the macrophage
cytoplasm. The digitations probably function as anchoring sites and for the exchange of signaling factors between both cells, as multiple
coated vesicles are seen near these structures (Hutson, 2006). This interaction is a remarkable characteristic of testicular macrophages, as
there are no other examples of specialized areas of contact between macrophages and other cell types outside of the immune system.
Another interesting aspect is that aging has a negative effect on steroidogenesis and one of the reasons could be the loss of these
digitations between macrophages and Leydig cells, interrupting their communication and the exchange of molecules, including 25-
hydroxycholesterol. This type of cholesterol is transferred from macrophages to Leydig cells under normal conditions and enter the
steroidogenic pathway (Fig. 6).
The macrophages are capable of influencing Leydig cell proliferation, differentiation, and testosterone secretion. It was shown that,
after Leydig cell destruction by a cytotoxic drug (EDS; ethane 1,2-dimethane sulphonate), testicular macrophages were responsible for the
phagocytosis of the apoptotic Leydig cells. There is a wave of proliferative activity when new Leydig cells repopulate the interstitium and
a concomitant increase in the number of resident macrophages. In addition, the lack of macrophages during the proliferation wave disrupts
Leydig cell recovery. A mouse model with reduced numbers of macrophages (osteopetrotic mouse; op/op) presents poor fertility, reduced
testosterone levels and abnormal Leydig cells. Regarding testosterone production, the macrophages may have positive or negative effects,
depending on the stimulus and the mediators produced. It has also been suggested that the testicular macrophages could metabolize
Leydig cell secreted steroids, as peritoneal macrophages are able to convert androstenedione to testosterone and dihydrotestosterone
(Hales, 2007).
It has also been suggested that Leydig cells could mediate the local regulation of the testicular leukocyte populations and thus provide
an additional source of immunoregulatory polypeptides. Leydig cells have specific receptors and recognition molecules encoded on their
surface and thus have the ability to interact with lymphocytes. As such, Leydig cell are the only highly differentiated endocrine cells
that spontaneously form rosettes with lymphocytes, macrophages or eosinophils. This complex relationship is probably involved in the
maintenance of the testis as an immunologically protected site, where immune cells appear to have restricted proinflammatory activity and
immune responses are usually limited, possibly due to immunomodulatory factors secreted by Leydig cells. Androgens are also known to
be immunosuppressive; therefore, it is also possible that immunological suppression is related to the high levels of intratesticular
testosterone (Diemer et al., 2003).

Leydig-Endothelial Cells
Vascular and lymphatic endothelial cells are abundant in the well vascularized interstitial/intertubular compartment of the testis and
their proximity to Leydig cells suggests their functional interaction. The diffusion of testosterone to blood circulation requires the
communication between Leydig cells and the microvasculature, although the mechanism for this hormone transport is not completely
known and may occur through passive diffusion. In this regard, cell contacts between Leydig and closely associated vascular endothelium
have been described. A paracrine interdependence between Leydig and endothelial cells has been reported to possibly involve hormone
transcytosis and vascular endothelial growth factors produced by the Leydig cells (Haider et al., 2007). Vascular endothelial cells
produce the peptide endothelin, which was shown to stimulate testosterone production in the rat. It is also noted that the lymphatic
endothelium associates with the peritubular wall and Leydig cell clusters. Endothelial cells of the testis are connected to each other by
tight junctions and, in general, the capillaries are unfenestrated. These characteristics provide a supplemental mechanism of
immunoprotection by controlling and protecting the testicular microenvironment, in addition to the SCB. However, further investigation is
needed to clarify the cell interactions involved in this relationship (Martin, 2016).

Fig. 6 Relationship between Leydig cells and testicular macrophages. (A) Electron microscopy image of the Leydig cell contacting a
macrophage in the adult mouse testis. (B) Schematic representation of the micrograph in (A), including the digitations (dotted rectangle), the
structural interaction that exists between Leydig cells and macrophages. LC, Leydig cell; TM, Testicular macrophage; G, Golgi complex; RER,
Rough endoplasmic reticulum; SER, Smooth endoplasmic reticulum; LD, Lipid droplet; black arrowheads, Leydig cell mitochondria (tubular
cristae); red arrowheads, macrophage mitochondria (lamellar cristae); arrows, lysosomes.
8 Cell–Cell Interactions—

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