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tumor necrosis factor  (TNF) and transplantation of hematopoietic stem Primer

interleukin 1, both of which are cells from allogeneic-matched donors
elevated in FA. remain the best options for treatment
of the FA-associated hematologic
Spermatogenesis
What happens when the pathway manifestations. Unfortunately,
is lost? Biallelic germline mutations androgen therapy is not effective Hitoshi Nishimura1
in the FANCA, FANCB, FANCC, in the long term and has been and Steven W. L’Hernault2
BRCA2/FANCD1, FANCD2, FANCE, associated with liver malignancies.
FANCF, FANCG, FANCI, BRIP1/ Promising recent work is focusing on Most organisms consist of two cell
FANCJ, FANCL, PALB2/FANCN, ex vivo hematopoietic stem cell gene lineages — somatic cells and germ
SLX4/FANCP, XPF/FANCQ and therapy and re-introduction of the cells. The former are required for the
UBE2T/FANCT genes cause bona corrected cells into the FA patient. current generation, and the latter
fide FA, the most frequently inherited Pharmacological interventions also create offspring. Male and female
bone marrow failure syndrome. FA is seem feasible, such as the use of germ cells are usually produced during
characterized by hypersensitivity to metformin or inhibitors of the TGF- spermatogenesis and oogenesis, which
ICL-inducing agents, predisposition pathway. TGF- inhibition has been take place in the testis and the ovary,
to myelodysplastic syndrome/acute shown to restore DNA damage repair respectively.
myeloid leukemia, solid tumors and and facilitate improved survival of FA Spermatogenesis involves the
congenital malformations, including cells. differentiation of spermatogonial stem
absent radii, and cardiac and renal cells into spermatocytes via mitotic
malformations (30% of FA individuals Where can I find out more? cell division and the production
might have subtle or absent physical Ceccaldi, R., Sarangi, P., and D’Andrea, A.D. of haploid spermatids from the
(2016). The Fanconi anaemia pathway: new
anomalies). Mutations in FANCA, players and new functions. Nat. Rev. Mol. Cell tetraploid primary spermatocytes
FANCC and FANCG account for Biol. 17, 337–349. via meiotic cell division. Spermatids
Chen, Y.H., Jones, M.J., Yin, Y., Crist, S.B.,
approximately 90% of all reported FA Colnaghi, L., Sims, R.J., and Huang, T.T. subsequently give rise to spermatozoa
cases. Mutations in BRCA1/FANCS, (2015). ATR-mediated phosphorylation of in the final phase of spermatogenesis,
FANCI regulates dormant origin firing in
RAD51/FANCR, RAD51C/FANCO and response to replication stress. Mol. Cell 58, called spermiogenesis. These
XRCC2/FANCU genes cause a FA- 323–338. fundamental steps, where mitotic
García-Rubio, M.L., Pérez-Calero, C., Barroso, S.I.,
like syndrome, without bone marrow Tumini, E., Herrera-Moyano, E., Rosado, I.V.,
proliferation precedes meiosis during
failure or leukemia predisposition. In and Aguilera, A. (2015). The Fanconi anemia spermatogenesis, are observed in a
addition, an increased risk to breast pathway protects genome integrity from wide variety of organisms. However,
R-loops. PLoS Genet. 11, e1005674.
and ovarian familial cancer has been Rodríguez, A., Sosa, D., Torres, L., Molina, B., developing a comprehensive
observed in the carriers of monoallelic Frías, S., and Mendoza, L. (2012). A Boolean understanding of the cell biology
network model of the FA/BRCA pathway.
mutations in the FANCD1/BRCA2, Bioinformatics 28, 858–866. and genetics of spermatogenesis is
FANCS/BRCA1, FANCJ/BRIP1, Sumpter, R., Sirasanagandla, S., Fernández, difficult for most species because it
Á. F., Wei, Y., Dong, X., Franco, L., and
FANCM, FANCN/PALB2 and FANCO/ Hanenberg, H. (2016). Fanconi anemia proteins occurs within a complex testicular
RAD51C genes. Finally, a sizeable function in mitophagy and immunity. Cell 165, environment characterized by the
867–881.
fraction of high-grade serous ovarian van Twest, S., Murphy, V.J., Hodson, C., Tan,
intimate association of developing
cancer, triple-negative breast cancer, W., Swuec, P., O’Rourke, J.J., and Deans, sperm with accessory cells. In this
and metastatic prostate cancer have A.J. (2016). Mechanism of ubiquitination Primer, we summarize the processes
and deubiquitination in the Fanconi anemia
been shown to have alterations in the pathway. Mol. Cell 65, 247–259. of spermatogenesis occurring in
FA pathway. Vinciguerra, P., Godinho, S.A., Parmar, K., Pellman, two pivotal model animals — mouse
D., and D’Andrea, A.D. (2010). Cytokinesis
There is a consensus that the bone failure occurs in Fanconi anemia pathway– and Caenorhabditis elegans —
marrow failure in FA patients is caused deficient murine and human bone marrow and compare them to consider
hematopoietic cells. J. Clin. Invest. 120,
by attrition of the hematopoietic stem 3834–3842.
which important features might be
and progenitor cell compartment, Zhang, H., Kozono, D.E., O’Connor, K.W., Vidal- evolutionarily conserved.
which has been mainly attributed Cardenas, S., Rousseau, A., Hamilton, A., and
Soulier, J. (2016). TGF- inhibition rescues
to DNA damage accumulation and hematopoietic stem cell defects and bone Mouse spermatogenesis
activation of apoptosis. However, marrow failure in Fanconi anemia. Cell Stem In the mouse testis, germ cells
Cell 18, 668–681.
a pro-inflammatory bone marrow Zhang, Q.S., Tang, W., Deater, M., Phan, N., differentiate into mature spermatozoa
environment is also detected in Marcogliese, A.N., Li, H., and Grompe, M. within seminiferous tubules, which
(2016). Metformin improves defective
FA, which could be due to defects hematopoiesis and delays tumor formation in are highly organized, complicated
in mitophagy mediated by the FA Fanconi anemia mice. Blood 128, 2774–2784. structures. As shown in Figure 1,
pathway. In this scenario, pro- seminiferous tubules contain germ
1
inflammatory TNF may contribute to Department of Radiation Oncology and cells at many different developmental
apoptosis of FA hematopoietic stem Center for DNA Damage and Repair, stages and in intimate association
Dana Farber Cancer Institute, Harvard
cells through activation of the extrinsic Medical School, Boston, MA 02215, USA.
with somatic Sertoli cells. A basement
apoptotic pathway. 2
Laboratorio de Citogenética, Instituto membrane and myoid cells surround
Nacional de Pediatría, Mexico City 04530, the seminiferous tubules, and Leydig
What are the current treatments for Mexico. cells and blood vessels are found in the
FA? Currently, androgen therapy and *E-mail: alan_dandrea@dfci.harvard.edu interstitium.

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Spermatogonia and Sertoli cells both

adhere to the basement membrane.
While the type A spermatogonia
undergo mitotic cell divisions to
differentiate from type A1 to A4 cells,
some of the type A4 cells differentiate Spermatozoa
into intermediate spermatogonia,
which then divide once to form type
B spermatogonia. The tetraploid
Lumen
primary spermatocytes are generated Sertoli cell
Elongating
from the type B cells via one mitotic spermatids
division and subsequently undergo
meiosis I to produce diploid secondary Apical
spermatocytes. After completion of Apical ES protein ES
meiosis II, two haploid spermatids complex
Cytoplasmic * Round
are formed from each of the two
Tubulobulbar droplet spermatids
secondary spermatocytes. During complex *
the cell divisions and maturation AdC
* Secondary
associated with the transition of type A1 Adherens junction *
* spermatocytes
spermatogonia into spermatids, these
*
types of germ cells migrate from the * Desmosome-like Spermatogonia **
junction
basement membrane to the luminal Type A Type B Primary
* spermatocyte
side of the seminiferous tubules. Gap junction *
Subsequently, spermatids undergo *
Basal ES protein *
transformation into spermatozoa Basal ES BTB
complex
during spermiogenesis, which involves **
condensation and elongation of the Tight junction *
BsC
nucleus, biogenesis of the acrosome (a
Golgi-derived, specialized lysosome- * *
BM
like organelle that later plays a role
during interactions with the egg during Myoid cells
fertilization), and formation of the
Leydig cells
flagellum. Spermiogenesis involves
the selective placement of certain
cellular constituents of the spermatid
that are not required for its subsequent
differentiation into a functional Blood vessel
Current Biology
spermatozoon (such as ribosomes)
into a cytoplasmic droplet termed the
residual body at the time when the Figure 1. Spermatogenesis in the mouse testis.
Mouse spermatogenesis occurs in thousands of seminiferous tubules within each testis; only a small
spermatids align at the luminal surface portion of a cross-section is shown here schematically. Male germ cells differentiate into spermatozoa as
of the seminiferous tubule. The resulting a result of close nutritional and structural communications with Sertoli cells. For simplicity, not all protein
mature spermatozoa are then released complexes and junctions that are required for physical interactions between Sertoli cells and between
into the lumen (spermiation). Sertoli cells and germ cells are shown. ES, ectoplasmic specialization; AdC, adluminal compartment;
BTB, blood–testis barrier; BsC, basal compartment; BM, basement membrane.
Communication between germ cells
and Sertoli cells that plays an essential role during and between germ cells and Sertoli
To complete spermatogenesis, mitotic differentiation and meiotic entry cells are also crucial to maintain
Sertoli cells must maintain intimate of spermatogonia) via heterodimers spermatogenesis. Sertoli cells form
structural and signaling contact with of an ATRA receptor (RAR) and a the blood–testis barrier (BTB), a highly
developing germ cells throughout the rexinoid receptor (RXR), leading to the selective barrier that distinguishes the
maturation of these cells (Figure 1). For differentiation of type Aaligned, which basal and adluminal compartments
example, vitamin A is required for the is the final phase of undifferentiated of the seminiferous tubules. The BTB
differentiation of spermatogonia and type A spermatogonia. In contrast, contains tight junctions (TJs) and
their subsequent entry into meiosis. All- the meiotic entry of spermatocytes ectoplasmic specializations (ESs),
trans retinoic acid (ATRA) is the active probably requires a paracrine as well as desmosome-like junctions
metabolite of vitamin A, and rexinoids molecule(s) that is produced by ATRA and gap junctions. These types of
are synthetic agonists of retinoic acids. signaling in Sertoli cells, but not in the cell-to-cell interactions promote the
In spermatogonia, ATRA promotes the germ cells. migration of developing germ cells from
expression of genes such as Kit (which Physical interactions between the basement membrane towards the
encodes a receptor tyrosine kinase germ cells, between Sertoli cells tubule lumen.

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Transition Mitotic

Plasma Distal tip cell

membrane

Seminal
Nucleus Rachis vesicles Vas deferens
Pachytene

Spermatogenesis

Residual body

Rachis

Gonad wall Primary Secondary Terminal Round

spermatocyte spermatocytes spermatocytes spermatids

Spermiogenesis in uterus

TRY-5?
Pseudopod
Ejaculation of spermatids

with seminal fluid

Round
spermatids Spermatozoa
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Figure 2. Spermatogenesis in the C. elegans male gonad.

C. elegans spermatogenesis occurs in the male gonad, which is composed of a single tube closed at the distal tip. Differentiation of male germline
stem cells into spermatocytes progresses proximally from the mitotic to the pachytene regions. Therefore, germ cells near the distal tip are at the
earliest developmental stages. These germ cells in the distal gonad arm share a common core cytoplasm (the rachis) because their nuclei are in-
completely separated by membranes from adjacent nuclei. In the spermatogenesis region of the male gonad, primary spermatocytes bud off from
the rachis as individual cells and undergo meiosis I. Unlike mouse spermatogenesis, germ cells released from the C. elegans male gonad are round
spermatids, not spermatozoa. The male-derived round spermatids transform into spermatozoa in the uterus of hermaphrodites after ejaculation with
seminal fluids. TRY-5, a worm serine protease, is a candidate inducer of spermiogenesis. The number of sperm in a C. elegans ejaculate varies from
10 to 100, while a mouse ejaculate has ~5 x 107.

The TJs are formed between Sertoli compartment to complete their transit the BTB (the basal ES) and between
cells to restrict solutes from crossing across the BTB. During this process, spermatids and Sertoli cells in the
the extracellular space, creating a claudin 3 (CLDN3), a TJ protein, is adluminal compartment (the apical
specialized environment within the transiently incorporated into new TJs ES). Cadherins are Ca2+-dependent,
seminiferous tubules that protects and subsequently replaced by CLDN11. adhesive transmembrane proteins
germ cells from attack by the host Migration of germ cells across the BTB found in both the basal and apical ESs
immune system. Interestingly, germ is therefore facilitated by internalization of the seminiferous epithelium and
cells can travel across the TJs without of TJ-related proteins, so that the BTB form protein complexes with catenins
disrupting junctional integrity. Sertoli is transiently dissolved. Testosterone in the cytoplasm. For example, CDH1
cells engulf germ cells within the then promotes BTB re-assembly after (E-cadherin) and CDH2 (N-cadherin)
transient compartments that are germ cells cross the barrier to enter the are present on the surfaces of both
formed between the old and new TJs; adluminal compartment. germ cells and Sertoli cells, and their
dissolution of the old TJs releases the The ESs are anchoring junctions status is regulated by cytokines and
moving germ cells into the adluminal between Sertoli cells at the site of testosterone. It is generally known

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A B
Incompetent to fuse Competent to fuse Incompetent to fuse Competent to fuse
with the oocyte with the oocyte with the oocyte with the oocyte

Spermatid Sperm
Inner acrosomal
IZUMO1 membrane (IAM) Cell body
Outer acrosomal
membrane (OAM) PM/OAM

Pseudopod
Acrosome

Membranous organelle
(MO)
Plasma
membrane (PM) PM
Equatorial
segment
Some SPE-9 class proteins
Nucleus

Midpiece

Collar

Acrosome- Acrosome- Some relocate to the pseudopod,

intact sperm reacted sperm some stay on the cell body surface
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Figure 3. Comparison of the mechanism of fertilization competence of mouse and C. elegans sperm.
(A) The acrosome reaction of mouse spermatozoa. The plasma membrane (PM) fuses with the outer acrosomal membrane (OAM) during the acro-
some reaction, so that acrosomal components (gray-filled circles) are released extracellularly and the equatorial segment is newly formed. IZUMO1
(green-filled circle), an essential protein for gamete fusion, relocates from the OAM onto the surface of the equatorial segment during the acrosome
reaction, where spermatozoa bind to and fuse with the oocyte PM. (B) Spermiogenesis of C. elegans round spermatids. A C. elegans round sperma-
tid contains acrosome-like organelles called the membranous organelles (MOs), which localize beneath the spermatid PM. Like the mouse sperm
acrosome, soluble components (red-filled circles) are contained within the MOs and released into the extracellular space during spermiogenesis.
Moreover, some SPE-9 class proteins localize within the MOs and translocate to the cell surface when MOs fuse with the PM. Some of these proteins
are found on the pseudopod surface, some on the cell body surface and some in both places.

that the turnover of the ESs is partly of elongated spermatids within the hermaphrodites of this organism during
controlled by the endocytosis of these seminiferous tubules. the fourth larval stage, but it completely
cadherins, followed by their recycling As described above, mammalian switches to oogenesis after entering
back to the cell surface. The addition spermatogenesis is a complex the young adult stage. This indicates
of testosterone to cultured Sertoli process that requires a variety of that C. elegans adult hermaphrodites
cells results in an increase in the rates physical interactions between germ are somatically females that carry self-
at which adhesion proteins including cells and Sertoli cells, in addition sperm. For simplicity, here we focus on
cadherins are endocytosed and then to communication between these spermatogenesis occurring in the male
recycled to the surface of Sertoli cells, cell types, mediated by humoral gonads (Figure 2) and will not discuss
implying that testosterone might assist (both endocrine and paracrine) the few aspects that differ between
the cyclical reformation of the BTB. factors. While all of these complex males and hermaphrodites.
Maintenance of the ESs requires interactions are required for successful There is only limited information
members of the nectin and afadin spermatogenesis, they make it very about how germline stem cells are
families. While all four members of the challenging to establish distinctive roles controlled to produce male germ cells
nectin family (NECTIN1 to NECTIN4) for each cell type. throughout the life cycle. The mitotic
are present in the testis, only NECTIN2 region of the male gonad contains
and NECTIN3 are found at the apical C. elegans spermatogenesis mitotically dividing germ cells. This
ES, where they localize to the surface Unlike most animals, C. elegans is region seems to contain germline stem
of Sertoli cells and spermatids, principally a self-fertile hermaphrodite, cells, like the mitotic region in the
respectively. Previous studies showed although it also produces males. hermaphrodite gonad. Two male distal
that the interaction between NECTIN2 As in most animals, C. elegans tip cells generate a niche to maintain
and NECTIN3 is indispensable for males undergo spematogenesis germline stem cells, and Notch
apical ES functions, in addition to in their gonads. The male-specific signaling regulates self-renewal of
the development and positioning event also takes place in self-fertile these stem cells. Since hermaphrodite

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Table 1. Mouse genes required for acrosome biogenesis/acrosome reaction and C. elegans genes required for MO formation/spermiogenesis1
Encoded protein

Gene Structure, type, domain Function

Mouse genes
Acr Acrosin, Ser protease Dispersal of acrosomal components during the acrosome
reaction
Cd46 CD46, Complement regulatory protein Increasing the spontaneous acrosome reaction
Csnk2a2 Casein kinase 2, alpha prime polypeptide Biogenesis of the acrosome
Eqtn Equatorin, sperm acrosome associated protein Fusion of the outer acrosomal and plasma membranes
during the acrosome reaction
Spaca1 Sperm acrosome associated protein Biogenesis of the acrosome
Spesp1 Sperm equatorial segment protein Stability of the equatorial segment membrane
Tssk6 Testis-specific Ser/Thr kinase Relocation of IZUMO1 onto the equatorial segment
Zpbp1 Zona pellucida binding protein Biogenesis of the acrosome
Zpbp2 Zona pellucida binding protein Biogenesis of the acrosome
C. elegans genes
fer-1 Protein of the Ferlin family Fusion of the membranous organelles (MOs) and plasma
membrane (PM) during spermiogenesis
snf-10 Solute carrier 6 (SLC6) transporter Acts in the male-dependent spermiogenesis pathway
spe-4 Presenilin Formation of the MOs, inhibition of ectopic initiation of
spermiogenesis
spe-5 B subunit of V-ATPase Formation of the MOs
spe-6 Casein kinase 1-like Ser/Thr kinase Inhibition of ectopic initiation of spermiogenesis
spe-8 Non-receptor type Tyr kinase with an SH2 domain Acting in the hermaphrodite-dependent spermiogenesis
pathway
spe-10 Four-pass transmembrane protein with a DHHC-CRD Fusion of the MOs and PM during spermiogenesis
zinc-finger motif and palmitoyl transferase activity
spe-15 Heavy chain of myosin VI Formation of the MOs
spe-39 Protein associated with the HOPS complex in vitro Formation of the MOs
spe-46 Six-pass transmembrane protein Inhibition of ectopic initiation of spermiogenesis
swm-1 Trypsin inhibitor in seminal fluids Inhibition of TRY-5 activity to prevent ectopic initiation of
spermiogenesis
try-5 Ser protease in seminal fluids Induction of spermiogenesis

1
All data shown in this table are based on mutant phenotypes and were obtained from Mouse Genome Informatics (www.informatics.jax.org) or
WormBase (www.wormbase.org).

distal tip cells express the LAG-2 and C. elegans spermatogenesis is During spermatogenesis, the
APX-1 (members of the Delta/Serrate/ completed without any intimate cytokinesis that accompanies meiosis
LAG-2 (DSL) family) ligands for the support from accessory cells: there I produces either two completely or
Notch receptor GLP-1, these two DSL is no equivalent to the Sertoli cell two incompletely divided secondary
proteins possibly participate in the found in mammals. C. elegans primary spermatocytes from each primary
Notch signaling induced by male distal spermatocytes differentiate into round spermatocyte (Figure 2; note that
tip cells. spermatids in chemically defined only the two incompletely divided
As germ cells migrate in a proximal medium and, following the addition of spermatocytes are depicted). Each
direction away from the controlling appropriate chemicals or proteases as secondary spermatocyte divides
influence of the male distal tip cells, activators, the round spermatids can to yield two haploid spermatids
they enter meiosis I. The spermatocytes transform into spermatozoa in vitro. In after completion of meiosis II. The
initially form a syncytium with a addition to these properties, thus far ~60 spermatids bud from an anucleate
cytoplasmic core called the rachis spe (sperm-defective) mutants have been residual body that contains many
and, after the pachytene stage, isolated; the affected genes in many of cellular constituents present in the
they bud off from the rachis as these mutants have been identified and secondary spermatocytes, such as
primary spermatocytes to initiate are helping to clarify the molecular basis most voltage-gated ion channels,
spermatogenesis. of C. elegans spermatogenesis. tubulin, actin and all ribosomes.

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Like the cytoplasmic droplet formed and the female reproductive tract, with the ability to fuse with oocytes in
during mammalian spermatogenesis, respectively. both mice and C. elegans. Therefore,
cellular constituents placed into the On the other hand, one of the there are probably orthologs among
residual body are no longer needed important similarities in the male germ mouse genes required for the acrosome
by the spermatid for it to complete cells of these two species is how reaction and C. elegans genes
differentiation into a functional spermatozoa become fertilization required for spermiogenesis. Table 1
spermatozoon. competent (Figure 3). In the mouse, shows a partial list of mouse and C.
The form of germ cell released testicular spermatozoa acquire motility elegans genes that are required for the
from the C. elegans gonad is a round during their transit through the caput, acrosome formation/acrosome reaction
spermatid, not a spermatozoon. corpus and cauda regions of the and MO formation/spermiogenesis,
This indicates that C. elegans epididymis. After mating, ejaculated respectively. Aside from the fact that
spermiogenesis does not take place spermatozoa from the vas deferens the strongly conserved V-ATPase
in the male gonads. The spermatids undergo biochemical and physiological enzyme is likely involved during both
are stored in a seminal vesicle and, modifications during migration from the the acrosome reaction and the fusion
after mating with hermaphrodites, are cervix of the uterus into the oviduct, between the MO and the plasma
ejaculated with seminal fluids through collectively called capacitation. membrane, obvious orthologs have
the vas deferens to enter the uterus. Moreover, the acrosome reaction is a not been currently found in mice and
The C. elegans serine protease TRY-5 prerequisite for sperm–oocyte fusion C. elegans that act in these events. In
is contained in the seminal fluids and in mammals. While soluble acrosomal particular, mouse genes implicated in
induces spermiogenesis in the uterus of contents, presumably required for triggering the acrosome reaction are
hermaphrodites (described in Table 1). regulation of fertilization, are released currently unclear, so it remains possible
This protease forms a protein complex into the extracelluar space during the that they might prove to be related
with SWM-1 (described in Table 1), acrosome reaction, the principle result to the already identified C. elegans
which contains two trypsin inhibitor- of this reaction is the creation of the spermiogenesis-related genes. Perhaps
like domains, that prevents ectopic equatorial segment (Figure 3A), which the complicated cell–cell interactions
spermiogenesis in the male gonad and is the region of the spermatozoon required for successful mouse
seminal vesicle. TRY-5 dissociates that binds to and fuses with the spermatogenesis make it difficult to
from the inhibitor and becomes oocyte plasma membrane. Indeed, identify such genes. As described
proteolytically active in the uterus. certain proteins including IZUMO1, an below, an in vitro spermatogenesis
During spermiogenesis in the uterus, immunoglobulin-like transmembrane system for the mouse would greatly
each sessile, round spermatid extends protein required for gamete fusion, are facilitate a better understanding of the
a pseudopod, which is the motile known to relocate onto the surface molecular basis for spermatogenesis.
machinery that is analogous to the of the equatorial segment during the
flagellum of the mouse spermatozoon. acrosome reaction. Perspectives
The resulting spermatozoa crawl into In C. elegans, spermiogenesis Reproductive biology is characterized
the spermatheca, the chamber in which includes both spermatid transformation by incredible diversity at the
fertilization occurs. into spermatozoa and sperm activation behavioral, anatomical, physiological
(Figure 3B). Each C. elegans spermatid and molecular levels. It is well
Comparison of spermatogenesis in contains ~15 membranous organelles known that genes encoding proteins
mice and C. elegans (MOs), which have certain features involved in reproduction are among
Since fertilization is generally species that suggest they could be considered the most rapidly evolving in a
specific, in order to prevent the to be a distributed acrosome. Like eukaryotic genome, which can make
generation of cross-hybrid organisms, the mouse sperm acrosome, soluble it a challenge to identify orthologs.
gametogenesis likely includes species- components are contained within the It has been the hope that intensive
specific aspects. If so, what are MOs and released to the extracellular study of a small number of model
substantial differences between mouse space during spermiogenesis, although systems, where combined genetic
and C. elegans spermatogenesis? functional roles of the released and molecular approaches are
First, mouse spermatogenesis molecules have yet to be identified. C. possible, would reveal common
requires interaction with accessory elegans spe-9 class genes are specific principles of spermatogenesis and
cells, including Sertoli cells, for to the male germline and encode help to define conserved processes.
development of germ cells, but the transmembrane proteins essential for The development of CRISPR/Cas9-
C. elegans primary spermatocyte can fertilization (i.e. sperm–oocyte fusion). mediated genome editing approaches
differentiate into round spermatids Several of these proteins localize should allow for the production of
cell-autonomously in vitro. Second, on the MO membranes and, during mutants from a more diverse collection
the residual bodies are formed during spermiogenesis, relocate onto the of species, which should facilitate the
spermiogenesis in the mouse testis, cell surface where spermatozoa bind identification of orthologous genes that
whereas C. elegans round spermatids to and fuse with the oocyte plasma have evolutionarily conserved roles in
bud off from the residual body during membrane. spermatogenesis.
meiosis II. Third, transformations of The acrosome and the MO are The complex cell–cell, paracrine and
mouse and C. elegans spermatids into physiologically similar and share a endocrine regulation of mammalian
spermatozoa take place in the testis common role in providing spermatozoa spermatogenesis presents a serious

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challenge to our understanding of this Correspondence and afferents distributed throughout

process. Development of a robust the central nervous system, either
in vitro spermatogenesis system for
mammals would provide useful insights.
Restoring monosynaptically or via the nucleus
of the solitary tract (NTS) [5]. The
So far, numerous approaches using the consciousness vagus directly modulates activity in the
entire tissue or separated single cells brainstem and via the NTS it reaches
from the testis in 2D or 3D cell culture with vagus nerve the dorsal raphe nuclei, the thalamus,
systems have been explored, which is
encouraging. Additionally, a robust in
stimulation the amygdala, and the hippocampus
[5]. In humans, vagus nerve stimulation
vitro spermatogenesis system might help increases metabolism in the forebrain,
define the causes of infertility in humans. Martina Corazzol1,2,4, Guillaume Lio1,2,4, thalamus and reticular formation
Arthur Lefevre1,2, Gianluca Deiana1,2, [6]. It also enhances neuronal firing
FURTHER READING Laurence Tell3, in the locus coeruleus which leads
Nathalie André-Obadia3, to massive release of norepinephrine
Ashrafzadeh, A., Karsani, S.A., and Nathan, S. Pierre Bourdillon3, Marc Guenot2,3,
(2013). Mammalian sperm fertility related in the thalamus and hippocampus, a
proteins. Int. J. Med. Sci. 10, 1649–1657. Michel Desmurget1,2, noradrenergic pathway important for
Ellis, R.E., Stanfield, G.M. (2014). The regulation Jacques Luauté2,3,4, arousal, alertness and the fight-or-flight
of spermatogenesis and sperm function in
nematodes. Semin. Cell Dev. Biol. 29, 17–30.
and Angela Sirigu1,2,4,* response [7].
Foster, J.A., and Gerton, G.L. (2016). The acrosomal Following the hypothesis that vagus
matrix. Adv. Anat. Embryol. Cell Biol. 220,
15–33. Patients lying in a vegetative state nerve stimulation functionally reorganizes
Gleason, E.J., Hartley, P.D., Henderson, M., Hill- present severe impairments of the thalamo-cortical network, we tested
Harfe, K.L., Price, P.W., Weimer, R.M., Kroft, T.L.,
Zhu, G.D., Cordovado, S., and L’Hernault, S.W.
consciousness [1] caused by lesions in its effects on the cortical activity of a
(2012). Developmental genetics of secretory the cortex, the brainstem, the thalamus patient lying in a vegetative state for 15
vesicle acidification during Caenorhabditis and the white matter [2]. There is years following traumatic brain injury.
elegans spermatogenesis. Genetics 191,
477–491. agreement that this condition may Behavioral, electroencephalographic
Griswold, M.D. (2016). Spermatogenesis: the involve disconnections in long-range (EEG) and 18F-FDG PET recordings
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1
to the stimulation period [4]. Here analysis identified eight sources
Department of Life Science, Setsunan
we propose to activate the thalamo- responsible for the stimulation-induced
University, Neyagawa, Osaka 572-8508,
Japan. 2Department of Biology, Emory cortical network based on vagus power increase, distributed over the
University, Atlanta, GA 30322, USA. nerve stimulation. The vagus nerve occipito-parietal, inferior temporal and
*E-mail: bioslh@emory.edu carries somatic and visceral efferents fronto-central regions in addition to a

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