Picture from Chatterjee I et al. (2005) Development "The Caenorhabditis elegans spe-38 gene encodes a novel four-pass integral ...."

Figure 9. The dynamic localization of SPE-38 requires MO fusion, but SPE-38 is not required for the dynamic localization of other sperm membrane proteins. (A) Confocal/multiphoton images of wild-type spermatids stained with DAPI (blue), anti-SPE-38 antibody (red) and the monoclonal antibody 1CB4 (Okamoto et al., 1985), which binds to an MO-associated antigen (green). Partial co-localization (yellow) of SPE-38 and 1CB4 staining is shown in the merged image. (B) Anti-SPE-38 immunolocalization in fer-1(hc17ts) mutant sperm, which fail to undergo MO fusion during spermiogenesis. Staining is detectable in fixed, permeabilized spermatids and spermatozoa, but not in live, non-permeabilized cells. (C) Localization of the 1CB4 antigen (green), SPE-9 (red) and DNA (blue) in wild-type and spe-38 sperm.

Picture from Chatterjee I et al. (2005) Development "The Caenorhabditis elegans spe-38 gene encodes a novel four-pass integral ...."

(A) Confocal/multiphoton images of wild-type spermatids stained with DAPI (blue), anti-SPE-38 antibody (red) and the monoclonal antibody 1CB4 (Okamoto et al., 1985), which binds to an MO-associated antigen (green). Partial co-localization (yellow) of SPE-38 and 1CB4 staining is shown in the merged image. (B) Anti-SPE-38 immunolocalization in fer-1(hc17ts) mutant sperm, which fail to undergo MO fusion during spermiogenesis. Staining is detectable in fixed, permeabilized spermatids and spermatozoa, but not in live, non-permeabilized cells. (C) Localization of the 1CB4 antigen (green), SPE-9 (red) and DNA (blue) in wild-type and spe-38 sperm.

Picture from Zhu G et al. (2003) Genetics "The Caenorhabditis elegans spe-39 gene is required for intracellular membrane ...."

Immunofluorescent localization of SPE-39 during spermatogenesis in males. From left to right a series of four corresponding images: a DIC image, DNA visualized by DAPI staining (blue), staining with the MO-specific monoclonal antibody 1CB4 (green), and staining with the anti- SPE-39 antibody N8 (red). (A-D) Wild-type spermatocyte in the process of spermatid budding. The arrows point to budding spermatids (s) and the anucleate residual body (rb). Bars, 5 um.

Picture from Zhu G et al. (2003) Genetics "The Caenorhabditis elegans spe-39 gene is required for intracellular membrane ...."

Figure 6. Immunofluorescent localization of SPE-39 in the male gonad. All worms were in a dpy-11 genetic background and the genotype with respect to spe-39 is indicated on A and E. From left to right, each row has a series of four corresponding images: a DIC image, DNA visualized by DAPI staining (blue), staining with the MO-specific monoclonal antibody 1CB4 (green), and staining with the anti-SPE-39 antibody N8 (red). d, distal end of gonad; p, proximal end of gonad. Bars, 50 um.

Picture from Morrison, Kayleigh N. et al. (2021) MicroPubl Biol

Figure 1: (A) Overview of spermatogenesis and fibrous body-membranous organelle (FB-MO) development. (Top) Spermatocytes develop on a syncytial rachis then detach to undergo the meiotic divisions. Following anaphase II, components which are no longer needed partition to a central residual body (RB). (Bottom) Within meiotic prophase spermatocytes, fibrous bodies (FBs) develop on the cytosolic face of the Golgi-derived membranous organelle (MO). During the budding division, FB-MO complexes partition to the spermatids. MOs then dock with the plasma membrane as the FBs disassemble and release MSP (Major Sperm Protein) dimers into the cytosol. During sperm activation, MSP localizes to the pseudopod, and MOs fuse with the plasma membrane. (B) Alignment of C. elegans ZK265.3/NSPH-2 (Nematode Specific Peptide family, group H) and C04G2.9/NSPH-3.2 with Ascaris MFP2 (MSP Fiber Protein) performed using the EMBL-EBI Clustal Omega Multiple Sequence Alignment tool (Madeira et al., 2019). Peptide for the anti-NSPH-2 antibody is underlined. (C-D) DAPI and anti-NSPH-2 labelling in hermaphrodites with genetic knockouts of nsph-3.2 and nsph-2 respectively. (C) Left to right developmental sequence of developing spermatocytes and spermatids. (D) Hermaphrodite spermatozoa within spermatheca. No primary antibody controls were performed on nsph-3.2 knockout animals. (E-G) Sperm spreads and individually staged cells with (E) anti-MSP labelling (n=63, 9 experiments), (F) anti-NSPH-2 labelling (n=50, 7 experiments), and (G) labelling against the MFP1 homolog, MSD-1 (Major Sperm protein Domain containing) (n=46, 3 experiments). (H-I) Co-labelling of MSP with either (H) anti-NSPH-2 or (I) anti-MSD-1. In spermatozoa (enlarged), the patterns were either fully (top) or partially overlapping (bottom). For NSPH-2/MSP n=16; for MSD-1/MSP n=14. Scale bars = 10 mm for C, D and E-G left; 5 mm for H, I and E-G right. Abbreviations: Karyosome spermatocytes (K), Metaphase spermatocytes (M), Budding Figures (B), Spermatids (S), and Spermatozoa (Z).

Picture from Seidel HS et al. (2011) PLoS Biol "A novel sperm-delivered toxin causes late-stage embryo lethality and ...."

Figure 3. PEEL-1 localizes to fibrous body-membranous organelles. (A) Diagram of spermatogenesis and fibrous body-membranous organelle (FB-MO) development, adapted with permission from [16]. FB-MOs develop in pachytene spermatocytes as membrane-bound organelles having a head region separated by a collar-like constriction from a set of membrane folds (lower panel; red). As spermatogenesis proceeds, the membrane folds grow and extend into arm-like protrusions, enveloping bundles of polymerized Major Sperm Protein, referred to as fibrous bodies (hatched region). Coincident with budding of spermatids from the residual body, the membrane folds of FB-MOs retract, and the fibrous bodies depolymerize into the cytoplasm. The FB-free MOs then move to a position just inside the plasma membrane, and upon sperm activation, they fuse with the plasma membrane opposite the pseudopod. (B-F) Nomarski and fluorescence images of spermatocytes and sperm expressing PEEL-1::GFP. Panels in (B) show the proximal arm of a male gonad, oriented with pachytene spermatocytes towards the left (bracketed region). Arrow and arrowhead indicate primary and secondary spermatocytes, respectively. Panels in (C-F) show higher resolution images of the following stages: secondary spermatocyte (C), budding spermatids (D), unactivated spermatid (E), and activated spermatozoan (F). (G-K) Spermatids were dissected from peel-1(+) (G-I) or peel-1(D) (J-K) males and stained with anti-PEEL-1 (green) and the FB-MO marker, 1CB4 (red) [64]. Nuclei are stained with DAPI (blue).

Picture from Park BJ et al. (2001) Mol Biol Cell "Calreticulin, a calcium-binding molecular chaperone, is required for stress ...."

Figure 5. Immunolocalization of CRT-1 in sperm cells. (A-C) Wild-type sperm show normal staining with anti-CRT-1. (A) Wild-type spermatids budding from the residual body (rb) are stained with anti-CRT-1, whose nuclei are shown by DAPI staining. (B) Wild-type spermatid. (C) Male gonad containing spermatids and are stained with anti-CRT-1 antibodies. (D) crt-1(jh101) mutant spermatids that fail to be stained by anti-CRT-1 antibodies, indicating the absence of functional CRT-1 in the mutant worms. (E) mAb 1CB4 stains FB-MO complexes of the crt-1 sperm. Nomarski images of wild-type (F) and crt-1 (G) sperm, arrows point to the examples of nuclei that are off center in the cell.

Picture from Zhu G et al. (2003) Genetics "The Caenorhabditis elegans spe-39 gene is required for intracellular membrane ...."

Figure 7. Immunofluorescent localization of SPE-39 during spermatogenesis in males. All worms were in a dpy-11 genetic background and the genotype with respect to spe-39 is indicated on the six leftmost panels. From left to right, each row has a series of four corresponding images: a DIC image, DNA visualized by DAPI staining (blue), staining with the MO-specific monoclonal antibody 1CB4 (green), and staining with the anti- SPE-39 antibody N8 (red). (A-D) Wild-type spermatocyte in the process of spermatid budding. The arrows point to budding spermatids (s) and the anucleate residual body (rb). (E-L) spe-39 mutant spermatocytes. Occasionally, unsuccessful attempts to bud spermatids are observed (arrows in I). (M-P) A wild-type spermatid and (Q-X) spe-39 mutant spermatids. Nuclei are located off center in spe-39 spermatids (arrows in Q and U). Bars, 5 um.

Picture from Guo S et al. (1995) Cell "par-1, a gene required for establishing polarity in C. elegans embryos, encodes a ...."

Figure 5. Distribution of PAR-1 in the Gonad and Early Embryolmmunofluorescence image of gonads and embryos showing the distribution of the anti-PAR-l antibody (green) and nuclei (blue). (A) lmmunofluorescence image of a wild-type gonad showing strong fluorescence at the periphery of newly forming oocytes. Although we detect signal around the periphery of these young oocytes, the outer membrane signal is faint and is not apparent in this reproduction. Abbreviations: MN, meiotic nuclei; NFO, newly forming oocytes; MO, mature oocytes.(B) Double exposure of the same gonad shown in (A), showing the localization of anti-PAR-l relative to nuclei. (C) Double exposure showing absence of peripheral staining in par-1(it86) mutant gonad.(D-E) Wild-type 1- and 2-cell embryos, respectively, showing strong peripheral fluorescence in the posterior. (F-G) Mutant 1- and 2-cell par-l(lw39) embryos showing the absence of posterior peripheral staining and the reduction of cytoplasmic staining. The scale bar in (A)-(C) is 32 um; in (D)-(G), 8 um.

Picture from Arduengo PM et al. (1998) J Cell Sci "The presenilin protein family member SPE-4 localizes to an ER/Golgi derived ...."

Immunofluorescence localization of SPE-4 in sperm cells. Bar in A1, 5 um, applies to all panels. Each horizontal, lettered row contains a series of four or five corresponding images: Nomarski DIC to visualize cytology (#1 panel), DAPI staining to visualize nuclei (blue #2 panel), monoclonal antibody ICB4 staining to visualize the fibrous body-membranous organelle (FB- MO) complexes (RITC red #3 panel), affinity purified anti SPE-4 rabbit polyclonal antibody staining (FITC green #4 panel) and a photograph through a filter set that simultaneously passes the DAPI, RITC and FITC signals (yellow #5). The genotype of the cells depicted in each row is indicated at the bottom of the leftmost panel. All #4 panels were photographed and printed at the same exposure index except that E4 was printed at 50% exposure index relative to the other panels (see below). (A) Wild-type spermatids budding from the residual body (rb). Both 1CB4 staining (A3) and EU43 SPE-4 peptide antiserum staining (A4) occur in spermatids during their budding from the rb and these signals coincide to produce the yellow signal observed in A5. (B) A wild-type spermatid (lower right corner) and a spermatozoon with a single pseudopod (p at arrow) extending from its cell body. Staining with 1CB4 (B3) reveals that the punctate signal is mostly found in the cell body and a similar, overlapping signal (B4) is observed after staining with the 9910 SPE-4 antiserum.