WormMine: AnatomyTerm WBbt:0006799

• Definition: A cell which gives rise to spermatids by meiosis.
• Name: spermatocyte
• Primary Identifier: WBbt:0006799

2 Children

Definition	Name	Synonym	Primary Identifier
A diploid immature germ cell which forms in the gonad when a spermatogonial stem cell buds off from the rachis to form a single cell that quickly enters metaphase I and divides to form secondary spermatocytes attached to a residual body.	primary spermatocyte		WBbt:0008591
A haploid immature germ cell which forms in the gonad by the division of a diploid primary spermatocyte. It undergoes a second meiotic division to complete meiosis, at which time the daughter cells detach from the residual body to become spermatids.	secondary spermatocyte		WBbt:0008592

2 Expression Clusters

Regulated By Treatment	Description	Algorithm	Primary Identifier
	Single-cell RNA-Seq cell group 144 expressed in: Syncitial pachytene spermatocytes .	CellRanger, DecontX, Monocle3, Louvain algorithm.	WBPaper00065623:144
	Single-cell RNA-Seq cell group 38 expressed in: Spermatocytes.	CellRanger, DecontX, Monocle3, Louvain algorithm.	WBPaper00065623:38

33 Expression Patterns

• Primary Identifier: Expr4370
• Remark: Note that genes on extrachromosomal arrays in transgenic worms are usually not expressed in gonads because of strong silencing effects.
• Pattern: CDC-48.1::GFP was expressed from embryos through to adult worms and predominantly in spermatocytes at L4 larvae stage, and in spermathecae at adult stage.

• Primary Identifier: Expr11592
• Pattern: UBXN-1, UBXN-2 and UBXN-3 were relatively strongly expressed in proximal gonads where MSP is localized, although UBXN-1, UBXN-2 and UBXN- 3 were ubiquitously localized in the gonads. More precisely, UBXN-1, UBXN-2 and UBXN-3 primarily localized at the periphery of spermatocyte nuclei, but not in mature sperm.

• Primary Identifier: Expr14606
• Pattern: Native GFP::CATP-7 is expressed closely associated with the plasma membrane of the amphid sensory neurons (localized to the sensilla), the gonadal sheath cells, the spermatheca, the hypodermis and the excretory cel. GFP::CATP-7 is expressed in most pharyngeal cells, where it localizes to the plasma membrane, plus internal membranous tubules. GFP::CATP- 7 is also expressed in the intestine, where it is associated with tubular structures in the basolateral domain and with vesicles in the apical domain immediately below the microvilli. GFP::CATP-7 is strongly expressed in spermatocytes and spermatids.

• Primary Identifier: Expr16275
• Pattern: Consistent with the western blot experiments, immunostaining of endogenous GSKL-1-FLAG and GSKL-2-Ollas worms revealed strong fluorescence at multiple stages of spermatogenesis. In males, immunofluorescence was brightest in the proximal gonad, which is where spermatogenesis occurs. Immunofluorescence was also observed in the cytoplasm of primary and secondary spermatocytes, in a punctate pattern for both tagged proteins, but was not detected in similar immunostaining experiments using wild-type untagged control worms.

• Primary Identifier: Expr8679
• Remark: Picture: Fig 3.
• Pattern: Expression in the alimentary canal: Strong and consistent expression in anterior arcades, posterior arcades, pharyngeal epithelium, pm4, pm8, g1, g2, vir, K.a/K' cells. inx-11 is more strongly expressed in the most posterior (int 9) intestinal cell. Weak or rare expression in pm1, pm2, pm3, pm5, pm6, pm7. Expression in the nervous system: CEPsh, DVC, LUA. Expression in the reproductive system: In adult stage, expressed in utse. In developing larva stage, expressed in uterus, sperm (spermatocytes, spermatids). Expression of inx-11 appears in pharyngeal tissue around two-fold stage, and by three-fold stage, strong expression becomes restricted to g1, g2, pm4, and pm8. inx-11 is expressed in the hypodermal cells of the animal in postembryonic stages.

• Primary Identifier: Expr8680
• Remark: Picture: N.A.
• Pattern: Expression in the alimentary canal: Strong and consistent expression in M2, rectal epithelial cells. Weak or rare expression in anterior arcades, posterior arcades. Expression in the nervous system: Amsh, CEPso, ILso, OLso, PDEso, AVH (early larva), AVJ (early larva), AVK, CAN, IL1 (early larva), PDE, PVD, SIB (early larva), URB, VAn, VBn, M2. Expression in the reproductive system: In adult stage, expressed in vulva, spermatheca, sperm(spermatocytes, spermatids). In developing larva stage, expressed in vulva. inx-12 expressionstarts in seam precursors around 1.5-fold stage, followed by expression in arcade cells by 2-fold and head neurons by 3-fold stage. inx-12 is expressed in the hypodermal cells of the animal in postembryonic stages.

• Primary Identifier: Expr15826
• Pattern: During spermatogenesis in L4 hermaphrodites and in males, both MIPs form puncta in spermatocytes, are eliminated into residual bodies, and are undetectable in mature sperm.

• Primary Identifier: Expr9634
• Pattern: peel-1 is expressed exclusively in sperm. In both males and hermaphrodites, a GFP reporter driven by the peel-1 promoter was expressed strongly in spermatocytes but not in any other tissue.

• Primary Identifier: Expr9635
• Pattern: peel-1 transcripts were observed in spermatocytes but not in mature sperm.

• Primary Identifier: Expr11861
• Pattern: Expression of snf-10 was visible in the male and hermaphrodite germline in cells undergoing differentiation into spermatocytes as well as at later stages of spermatogenesis. No fluorescence was visible in somatic tissues,in the mitotic or meiotic-prophase regions of the germline, or in germ-line cells developing into oocytes.

• Primary Identifier: Expr11465
• Remark: Temporal description
• Pattern: CSR-1 is abundant in mature sperm. CSR-1 was associated with P-granules throughout the syncytial male germline and into differentiating spermatocytes, where P-granules disperse and disappear. In developing gametes, CSR-1 localized to large cytoplasmic foci and in discrete chromatin domains of spermatocytes undergoing nuclear condensationas well as in haploid spermatids.

• Primary Identifier: Expr16424
• Pattern: We found a similar pulse of NHR-23::mScarlet in L4.3 vulval precursor cells as that reported by Kinney et al.. nhr-23::mScarlet was also detectable in seam and hypodermal cells at this stage, epithelial cells that synthesize cuticular components. NHR-23::mScarlet was detected in pachytene nuclei in L4 animals and its zone of expression became restricted in young adult animals. This expression pattern is again similar to NHR-23::GFP::AID*::3xFLAG (Ragle et al. 2020). However, we also observed some interesting differences compared to NHR-23::GFP::AID*::3xFLAG. In our previous work, we observed that NHR::23::GFP became undetectable in ovulating adults (Ragle et al. 2020). In contrast, NHR-23::mScarlet appears to be diffusely expressed in young adult residual bodies and spermatocytes. This diffuse expression persists into ovulating adults, and appears restricted to the spermatheca. The NHR-23::mScarlet signal is specific to nhr-23::mScarlet::3xMyc animals as no diffuse expression in the germline is observed in wild-type animals. One can observe the diffuse expression pattern in L4.5 germlines several rows of cells after NHR-23::mScarlet::3xMyc localization is lost from nuclei.

• Primary Identifier: Expr1116
• Pattern: immunohistochemistry shows that L1 larvae have high HSP12 levels throughout the body. Although overall levels of HSP12s are much lower in L4 larvae and in adult hermaphrodites, these smHSPs are locally abundant in the spermatheca and in specific vulval cells under unstressed conditions. In the spermatheca of hermaphrodites, HSP12s are localized in sperm. At higher magnification, the distribution of HSP12 cytoplasmic staining in sperm cells is seen to be spherically symmetrical. In males, spermatids are stored in the seminal vesicle, and most sperm cells mature following ejaculation; the extensive labelling of HSP12 in male germ cells therefore indicates localization of this smHSP to spermatids and perhaps spermatocytes. Cells in the mitotic region of the male gonad, however, are not labelled. In hermaphrodites, HSP12s are also expressed in a subset of vulval muscle cells (A9A12). The specificity of the antibody for HSP12s is demonstrated by the loss of signal when the antibody is pre-incubated with excess recombinant HSP12.3.
• Subcellular Localization: At higher magnification, the distribution of HSP12 cytoplasmic staining in sperm cells is seen to be spherically symmetrical.

• Primary Identifier: Expr11974
• Pattern: Expression of INX-8, -9, -14, -21 and -22 overlapped closely throughout the gonad in wild-type adult hermaphrodites. In distal arms clusters of fine puncta were associated with each germ cell, as previously reported for INX-14 and INX-22 expression (Govindan et al. 2009). INX-8/9 (antibodies raised against INX-8 shown to cross-react with INX-9) INX-8::GFP and INX-9::GFP were expressed in the distal tip cell (DTC) in addition to the somatic sheath. In the DTC, INX-8/9, INX-8::GFP and INX-9::GFP associated with long processes that extended from the DTC proximally, especially those running between germ cells and intercalating among them. At the distal limit of sheath cell coverage of germ cells, longer formations sometimes appeared associated with the apparent edge of the sheath cells. At the loop region of the gonad arm, individual puncta appeared in larger, higher-density aggregates. In the proximal arm puncta size was more variable, with many appearing to be considerably larger than the fine puncta seen in the distal arm. The expression levels of INX-8, INX-9 appeared higher in the proximal gonad. Throughout the gonad the localization patterns of INX-8::GFP and INX-9::GFP were indistinguishable. In addition to gonadal expression, antibody staining of whole mounts suggested that INX-8/9 may be expressed in some pharyngeal and a few other head neurons (T. Starich, unpublished results), but because the antibody reacted primarily with processes and not cell bodies we did not attempt to identify these neurons. Male gonads were examined and found to express INX-8, -9, -14, -21 and -22 as well. Presumptive gap junctions forming between both male distal tip cells and germ cells were detected with antibodies specific to INX-8/9, INX-14, INX-21, and INX-22. INX-8::GFP and INX-9::GFP were both expressed in the DTCs. No expression was detected in the transition zone, but innexin expression appeared to outline individual germ cells in the pachytene region. All 5 innexins are also expressed in the regions occupied by differentiated spermatocytes and sperm, with evidence of puncta formation. Expression of INX-9::GFP visualized somatic coverage of spermatids, probably by cells of the seminal vesicle, but somatic coverage of germ cells in the pachytene region of the male gonad has not been described. Examination of earlier developmental stages showed that all five innexins are expressed in the primordial gonad, consisting of somatic gonadal precursors Z1 and Z4 and the primordial germ cells Z2 and Z3. Early larval expression patterns differ from adults in that distinct, well-defined puncta potentially corresponding to gap junctions are less clearly discernible. In L2-L4 larval stages, germ cell innexins appear to be continually expressed, and somatic innexins are expressed predominantly in the DTC. In late L2 and early L3 stages, as the gonad arm lengthens, the migrating DTC seems to trail a process behind it that maintains contact with the germ cell compartment. The DTC appears to form gap junctions with germ cells at both long external processes and processes that intercalate between germ cells. At this time a second, more proximal focus of somatic innexin expression was sometimes detected. Though this second focus might represent extensions from the DTC, expression of inx-9::gfp suggested that other somatic cells besides the DTC might be involved. By the late L3 stage, the DTC no longer appears to be in contact with all of the germ cells. To better understand the contacts between germ cells and the DTC, we examined early L4-stage animals by TEM. We observed several processes trailing behind the DTC cell body on the outer edge of the germ line, as well as extensions that dig deeply between germ cells. The immunofluorescence results suggest that gap junctions form between the DTC and germ cells at both the outer and inner DTC arms. The inx-8 promoter was fused to mCherry, and inx-8p::mCherry was expressed during larval development in most or all of the somatic gonad cells derived from Z1 and Z4, including the DTC, sheath/spermathecal precursors and uterine cell precursors. We conclude that although expression of INX-8/9 appears to be strongest in the DTC during larval development, other somatic gonad cells may also express INX-8/9 at this time.

• Primary Identifier: Expr15540
• Pattern: The anti-NSPH-2 antibody labelled distinct structures within late-stage spermatocytes as well as the pseudopods of spermatozoa within the spermatheca.

• Primary Identifier: Expr15541
• Subcellular Localization: In spermatozoa, both MFP2 and MSD-1 antibodies localize to the pseudopod, matching the known pattern in Ascaris spermatozoa. Within prophase and meiotically dividing spermatocytes, MFP2 and MSD-1 antibodies labelled discrete structures throughout the cytoplasm in a pattern strongly resembling FBs labelled by MSP antibodies. To verify that NSPH-2 and MSD-1 localized to FBs, we co-labelled male germline with anti-MSP and either anti-MSD-1 or anti-NSPH-2 antibodies. Within spermatocytes, the fully overlapping patterns revealed that NSPH-2 and MSD-1 are packaged along with MSP in the FBs. In spermatids, the MSP and MSD-1 were overlapping. In contrast, except in immature spermatids that still retained their FBs, the NSPH-2 and MSP patterns only partially overlapped. In spermatids, the NSPH-2 labelling was notably less robust which may suggest that a change in protein conformation or binding partners is blocking the antigenic site.

• Primary Identifier: Expr11975
• Pattern: Expression of INX-8, -9, -14, -21 and -22 overlapped closely throughout the gonad in wild-type adult hermaphrodites. In distal arms clusters of fine puncta were associated with each germ cell, as previously reported for INX-14 and INX-22 expression (Govindan et al. 2009). INX-14, -21 and -22 colocalized to puncta associated with long processes that extended from the DTC proximally, especially those running between germ cells and intercalating among them. At the distal limit of sheath cell coverage of germ cells, longer formations sometimes appeared associated with the apparent edge of the sheath cells. At the loop region of the gonad arm, individual puncta appeared in larger, higher-density aggregates. In the proximal arm puncta size was more variable, with many appearing to be considerably larger than the fine puncta seen in the distal arm. Although all innexins appeared to colocalize, INX-21 differed from the other innexins in that its expression level was relatively higher in the distal gonad compared to the proximal gonad. In contrast, the expression levels of INX-8, INX-9, INX-14, INX-22 appeared higher in the proximal gonad. Male gonads were examined and found to express INX-8, -9, -14, -21 and -22 as well. Presumptive gap junctions forming between both male distal tip cells and germ cells were detected with antibodies specific to INX-8/9, INX-14, INX-21, and INX-22. All 5 innexins are also expressed in the regions occupied by differentiated spermatocytes and sperm, with evidence of puncta formation. Examination of earlier developmental stages showed that all five innexins are expressed in the primordial gonad, consisting of somatic gonadal precursors Z1 and Z4 and the primordial germ cells Z2 and Z3. In a few serendipitous cases expression of INX-22 was detected prior to hatching, in pretzel-stage embryos. Early larval expression patterns differ from adults in that distinct, well-defined puncta potentially corresponding to gap junctions are less clearly discernible. In L2-L4 larval stages, germ cell innexins appear to be continually expressed, and somatic innexins are expressed predominantly in the DTC. In late L2 and early L3 stages, as the gonad arm lengthens, the migrating DTC seems to trail a process behind it that maintains contact with the germ cell compartment. The DTC appears to form gap junctions with germ cells at both long external processes and processes that intercalate between germ cells. At this time a second, more proximal focus of somatic innexin expression was sometimes detected. Though this second focus might represent extensions from the DTC, expression of inx-9::gfp suggested that other somatic cells besides the DTC might be involved. By the late L3 stage, the DTC no longer appears to be in contact with all of the germ cells. To better understand the contacts between germ cells and the DTC, we examined early L4-stage animals by TEM. We observed several processes trailing behind the DTC cell body on the outer edge of the germ line, as well as extensions that dig deeply between germ cells. The immunofluorescence results suggest that gap junctions form between the DTC and germ cells at both the outer and inner DTC arms.

• Primary Identifier: Expr10162
• Pattern: The expression pattern of FLAG::CDC-48.1 was observed in the early L4 stage, during which sperm are produced. FLAG::CDC- 48.1 was expressed from the bend to the proximal gonad, although it was predominantly expressed at the pachytene stage and in spermatocytes. FLAG::CDC-48.1 was not localized in sperm. It should also be noted that FLAG::CDC-48.1 was expressed in the distal tip cell in the mitotic region.

• Primary Identifier: Expr9998
• Pattern: SPE-5 expression apparently occurs at a very low level or was somehow partly masked, so long exposures (~20 minutes) were required to visualize it in whole worm lysates, where a band appears in fem-3gf worms that contain sperm but is missing in fem-1lf worms that never contain sperm (data not shown). The SPE-5 protein is more prominent and easily detected (exposure times of less than 1 minute) in purified sperm (>90% spermatids) that were isolated from fem-3gf worms.
• Subcellular Localization: SPE-5 expression was coincident with spermatocytes, where it was distributed throughout the cytoplasm and excluded from the nucleus. A faint mottled pattern was observed in the cytoplasm, suggestive of punctate staining. During meiosis I, SPE-5 appears distributed throughout the cytoplasm, some of it in discrete dots. SPE-5 staining is observed in both spermatids and the residual body, and much of the staining in both regions is in discrete dots. Budded spermatids have distinct secretory membranous organelles (MO) just below their plasma membrane. Most of the SPE-5 signal is also localized near the plasma membrane, much of it as discrete dots that are adjacent to MOs. In certain favorable regions, it appears that there is one green dot per MO. These observations indicate that SPE-5 is located near the MO secretory vesicles of spermatids, consistent with mediating MO acidification.

• Primary Identifier: Expr9701
• Subcellular Localization: CED-4 appears to colocalize with mitochondria and CED-9 in secondary spermatocytes and spermatids. This CED-4 localization in spermatocytes and spermatids was observed with two independent CED-4 antibodies and by staining for CED-4::GFP.

• Primary Identifier: Expr1623
• Remark: No signal was detected using a fer-1 sense probe as a negative control.
• Pattern: The only tissue in which fer-1 message was detected was the testis, beginning in the loop and proceeding proximally. Based on DAPI-staining of nuclei to identify the stages of meiosis, expression begins when the cells enter meiosis and is confined to the primary spermatocytes. No signal was detected in the secondary spermatocytes, the mitotic spermatogonial cells, the spermatids or in any other tissue in the worm.

• Primary Identifier: Expr1163
• Pattern: All three gamete types of C. elegans (oocyte, male spermatocyte, and hermaphrodite spermatocyte) show meiosis-specific expression of HIM-3. No protein could be detected in somatic nuclei at any stage of development or in the distal tip nuclei that are undergoing mitotic divisions. A strong signal first appears in the transition zone where HIM-3 is associated with the chromatin of early meiotic prophase nuclei. Localization of HIM-3 becomes more discrete in pachytene and diplotene as the chromosomes synapse and desynapse. The protein remains associated with the chromosomes of oocytes at diakinesis; however, it could no longer be detected in postmeiotic sperm. The antibody identifies the first nuclei that are entering the pachytene stage of meiosis but does not stain the mitotic nuclei.
• Subcellular Localization: Nuclei. The him-3 protein localizes to the chromosome core and associates with both unsynapsed and synapsed chromosomes.

• Primary Identifier: Expr16180
• Pattern: comp-1 expression was visible in developing spermatocytes and spermatids in both males and hermaphrodites, and we observed no obvious differences in abundance between the two sexes.

• Primary Identifier: Expr1550
• Pattern: Expressed in testis, in both spermatogonial and spermatocytes, but not in spermatids. No staining was seen in somatic tissue.

• Primary Identifier: Expr1562
• Pattern: spe-27 message were detected in the testis. The transcript seems to be absent in spermatogonial cells undergoing mitosis, but is present in the spermatocytes. The message is also present in the residual bodies but absent in the spermatids. No hybridization to somatic tissues was observed.

• Primary Identifier: Expr15794
• Pattern: In males (and sperm-producing hermaphrodites), NHR-23::GFP was first observed in early pachytene spermatocytes, increased in intensity through late pachytene and became undetectable during late meiotic prophase with onset of chromatin condensation. NHR-23 was undetectable in meiotically dividing spermatocytes or mature spermatids.
• Subcellular Localization: NHR-23::GFP labeled most chromosomes along their entire length yet failed to label a region of DNA in each pachytene stage spermatocyte. Presumably NHR-23 is not labeling the X chromosome, which is transcriptionally silent during spermatocyte meiosis (Kelly et al., 2002).

• Primary Identifier: Expr13699
• Pattern: Crem-MSS-1::HA expression was first detected in large vesicles and on the plasma membrane of spermatocytes, with intensity increasing and localization restricted to secretory vesicles in mature spermatids. The secretory vesicles of nematode sperm, known as membranous organelles (MOs), fuse with the plasma membrane upon ejaculation and sperm activation.

• Primary Identifier: Expr10591
• Pattern: Wild-type male germ cells expressed CED-4 late in spermatogenesis during the transition from secondary spermatocyte to mature spermatids, consistent with observations in fly and mouse testes (Cagan, 2003).

• Primary Identifier: Expr15825
• Pattern: During spermatogenesis in L4 hermaphrodites and in males, both MIPs form puncta in spermatocytes, are eliminated into residual bodies, and are undetectable in mature sperm.

• Primary Identifier: Expr12948
• Pattern: msp-3 mRNAs are highly accumulated in the late pachytene and early spermatocytes, and their expression was dramatically reduced in spermatid and mature sperm.

0 Life Stages

1 Parents

Definition	Name	Synonym	Primary Identifier
a cell, of either sex, directly concerned in the production of a new organism.	germ cell		WBbt:0006796