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The transport of sperm in the cloaca and adjacent regions of the female red-spotted newt was examined. It was found that within 1 min after sperm were introduced into the vent, they progressed in a random pattern past the apertures of the spermatheca (the glandular, sperm storage organ that opens from the anterior roof of the cloaca) forward to the anterior end of the cloaca and on into the posterior regions of the hindgut and bladder. Sperm did not enter the dorsal recess of the cloaca into which the oviducts and ureters open. After 1 day, few sperm remained within the cloaca lumen. Sperm were not transported into the cloacae of artificially inseminated, anesthetized females without prior administration of norepinephrine to their cloacal mounds. Treatment of the cloacal mounds of naturally inseminated females with an antagonist of neuromuscular transmission (lidocaine) decreased the numbers of sperm in the anterior cloaca relative to those of saline-injected control specimens. Neither dead newt sperm nor live rabbit sperm entered the spermatheca. Rabbit sperm, however, entered the oviduct. It is argued that passive and active mechanisms of sperm transport work in concert. Contractions of smooth muscle, which may be initiated during courtship, probably serve to draw sperm passively into the cloaca and up to and beyond the apertures of spermathecal tubules, but sperm, once in the vicinity of those apertures, probably swim actively into them.

The motility of sperm was examined in vivo in the vas deferens, the spermatophore, and the spermatheca of the red-spotted newt and in in vitro preparations with variations in osmolality, hydrogen ion concentration, and concentrations of specific osmolytes. Sperm were motile within the spermatophore, but little or no evidence of motility was seen in the spermatheca or the vas deferens. Approximately 25% of sperm from the vas deferens became motile when dispersed in spermatic fluid plasma, the sperm-bearing liquid of the vas deferens, indicating crowding to be a possible motion-restraining factor. Fewer than 50% were motile in several saline media isosmotic with spermatic fluid plasma, whereas more than 90% became motile in distilled water or media at osmolalities near that of pond water. Motility in isosmotic solutions persisted beyond 12 hours, but at low osmolality ceased by 6 hours. When dispersed at higher osmolalities initial motility was low but increased to isosmotic levels by 12 hours. Responses to immersion in solutions of mannitol were similar to ones observed in saline solutions of equivalent osmolality. Dispersion in hydrogen ion concentrations between pH 4 and 9 did not affect the initial motility of sperm, but after 12 hours at pH 9, pH 4 or 5 movement was inhibited. In general, these data indicate a major role for osmolality in the enforced quiescence of sperm during storage and demonstrate that the low osmolality of pond water is primarily responsible for the activation of sperm in the spermatophore of the newt.

The objective of this study was to determine whether postnatal increases in rat Leydig cell number result from differentiation of precursor cells, division of existing Leydig cells, or both. Our approach was 1) to examine changes in the absolute number of Leydig cells and potential precursor cells (macrophages, pericytes, and mesenchymal, endothelial, and myoid cells) per testis on day 19 of gestation (day -2) and days 7, 14, 21, 28, and 56 postpartum; 2) to examine the frequency with which mesenchymal and Leydig cells divide during prenatal and postnatal development; and 3) to identify and examine the fate of the progeny of Leydig and mesenchymal cell divisions during prenatal and postnatal development. Stereological methods were used to show that mesenchymal cells comprised 44% of the total interstitial cell population and Leydig cells 16% on day -2, whereas by day 56 postpartum the relationship had reversed; mesenchymal cells comprised 3% and Leydig cells 49%. These results suggested a precursor-product relationship between mesenchymal and Leydig cells because no such reciprocal relationship was observed between Leydig cells and macrophages, pericytes, endothelial, or myoid cells. Autoradiographic analysis of [3H]thymidine incorporation into mesenchymal and Leydig cells was consistent with this interpretation. In a series of pulse-chase experiments, the percentage of labeled mesenchymal and Leydig cells was measured after a single injection of [3H]thymidine on days 2, 14, 28, and 56 postpartum, each followed by sampling at timed intervals (between 1 h and 14 days) thereafter. Starting on day 14, the percentage of labeled Leydig cells was approximately 1% immediately after injection of [3H]thymidine and increased significantly to approximately 6% by 6 days after injection. No such increase was observed when rats were similarly injected starting on days 2, 28, and 56 postpartum. The rise in Leydig cell labeling between days 14 and 28 postpartum did not result in a decline in the number of silver grains over labeled Leydig cell nuclei, indicating that the increase in the percentage of labeled cells was not caused by Leydig cell division. These observations led us to conclude that the increase in Leydig cell labeling from days 14 to 28 was the result of recruitment from a compartment of labeled mesenchymal cells. In contrast, our analysis indicated that from day 28 postpartum and thereafter until the mature number of Leydig cells is attained, Leydig cells are generated by division of morphologically recognizable Leydig cells.

Interleukin-3 (IL-3) is an activated T cell product that bridges innate and adaptive immunity and contributes to several immunopathologies. Here, we report the crystal structure of the IL-3 receptor α chain (IL3Rα) in complex with the anti-leukemia antibody CSL362 that reveals the N-terminal domain (NTD), a domain also present in the granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-5, and IL-13 receptors, adopting unique “open” and classical “closed” conformations. Although extensive mutational analyses of the NTD epitope of CSL362 show minor overlap with the IL-3 binding site, CSL362 only inhibits IL-3 binding to the closed conformation, indicating alternative mechanisms for blocking IL-3 signaling. Significantly, whereas “open-like” IL3Rα mutants can simultaneously bind IL-3 and CSL362, CSL362 still prevents the assembly of a higher-order IL-3 receptor-signaling complex. The discovery of open forms of cytokine receptors provides the framework for development of potent antibodies that can achieve a “double hit” cytokine receptor blockade. Refereed/Peer-reviewed

Golden hamster testes regress after short day exposure. The present study asks: 1) are Leydig cell numbers depleted during short days, and 2) if so, how are they replenished during recrudescence. Control hamsters were shown 14 h of light and 10 h of dark (LD 14:10) for 10 weeks (n = 12). Testicular regression was induced by LD 6:18 for 10 weeks (n = 4), and recrudescence by switching regressed hamsters to LD 14:10 for 3 and 5 weeks (n = 8 for each group). All hamsters were injected with [3H]thymidine [3 μCi/gm body wt., intraperitoneally (i.p.)] 1 h or 2 weeks before sacrifice. Leydig cell number per testis was determined by stereological analysis of sections of perfusion-fixed testes, and labeling indices were determined by autoradiography. Leydig cell numbers were reduced significantly from 18.2 × 106 in control to 9.0 × 106 in regressed testes (p < 0.05); then increased to 14.0 × 106 and 17.9 × 106 in 3- and 5-week recrudesced hamsters. The labeling index was nondetectable (n.d.) for regressed hamsters. In control and recrudescing hamsters the labeling index was measured at two times (t1 = 1 h vs. t2 = 2 weeks post-injection): in controls, t1 = 0.22 ± 0.15% (mean ± SEM) vs. t2 = 0.28 ± 0.22%; in 1 week recrudesced, n.d. vs. 1.92 ± 0.77% (p < 0.05); at 3 wk, n.d. vs. 4.58 ± 1.74% (p < 0.05); at 5 weeks, 1.92 ± 0.61% vs. 2.25 ± 0.59%. These results are indicative of the existence of interstitial precursor cells that divide, then differentiate, and thus replenish the Leydig cell population during testicular recrudescence.

Chapitre 13 ; International audience