Retrograde ejaculation: can sperm retrieval be simpler and noninvasive?
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Adult↗
Biomedical subjects
J H Cleine
Publications and source records attributed to J H Cleine.
[Retrograde ejaculation: a treatable fertility problem].
Retrograde ejaculation was diagnosed in a 32-year-old man. This condition is an uncommon cause of male infertility. In most cases the only possible treatment is recovery of spermatozoa from the urine followed by artificial insemination; the couple underwent this treatment; the women received intrauterine insemination. In order to ensure the best possible survival of the sperm cells the pH (7.2-8.2) and the osmolality (300-380 mosmol/kg) of the urine were adjusted beforehand by administering bicarbonate and water to the man a few hours before ejaculation. A successful pregnancy ensued. When the couple desired another child the semen quality had deteriorated and in-vitro fertilisation with spermatozoa collected from urine was chosen as a treatment. A second successful pregnancy developed.
Adult↗
Replacement of posterior by anterior endoderm reduces sterility in embryos from inverted eggs of Xenopus laevis.
The genital ridges of Xenopus laevis tadpoles reared from eggs kept in an inverted position contain less than 40% of the number of primordial germ cells (PGCs) of controls (Cleine & Dixon, 1985). It has been suggested that this reduction is caused by the germ cells' ectopic position in the anterior endoderm of larvae from inverted eggs, from where they may be unable to migrate into the genital ridges (Cleine & Dixon, 1985). This hypothesis is tested here by interchanging anterior and posterior endodermal grafts between pairs of inverted embryos at the early tailbud stage. Replacement of anterior by posterior endoderm has no effect but replacement of posterior by anterior endoderm increases the number of PGCs in the genital ridges and significantly reduces the proportion of sterile embryos. In a control series, in which the same type of grafting was done with normal embryos, replacement of posterior by anterior endoderm reduced the number of germ cells to almost zero, but replacement of anterior by posterior endoderm nearly doubled it. These findings are explained in terms of the distribution of the germ cells in the endoderm at the time of grafting. The results firstly show that the position of the germ cells is crucial to successful migration and secondly they support the notion that germ plasm has a determinative role during early germ cell differentiation.
Animals↗
Normal fates and states of specification of different regions in the axolotl gastrula.
A fate map was constructed for four regions of the early gastrula of Ambystoma mexicanum using orthotopic grafts from donors labelled with FLDx (fluoresceinated-lysinated-dextran). The region around the animal pole gave rise to epidermis only and did not include prospective neural plate. The dorsal marginal zone contributed to cephalic endoderm and to the whole length of the axial mesoderm (notochord and somites), the lateral marginal zone to lateroventral and somitic mesoderm, and the ventral marginal zone to lateroventral mesoderm. It was found that the dorsal marginal zone contributed relatively more to the anterior regions of the mesodermal mantle and the ventral marginal zone more to its posterior parts. The same regions of the gastrula and also vegetal yolky tissue were cultured as explants and labelled with tritiated mannose. Their glycoprotein synthesis pattern was compared to those of the neurula tissues to which they contribute in vivo. Animal pole explants synthesized large amounts of the epidermis-specific marker epimucin. Dorsal marginal zone explants did not synthesize epimucin but did make amounts of S2 and S6 indicative of mesoderm, as well as the notochord-specific markers S2.2 and S3.2. Lateral marginal zone explants showed the same pattern as the dorsal marginal zone including the two notochord-specific markers, although they do not contribute to notochord in vivo. Ventral marginal zone explants were more variable in their behaviour. Yolky tissue from the vegetal hemisphere of the gastrula or the archenteron floor of the neurula synthesized mainly polydisperse material of high molecular weight rather than discrete glycoproteins. The results indicate that at the early gastrula stage states of specification exist which correspond to the three germ layers, ecto-, meso- and endoderm. The ectodermal specification of animal pole explants is quite robust and cannot easily be changed by variation of the culture conditions. However treatment with a concentrated pellet of vegetalizing factor does induce a change to mesodermal specification, which is clearly detectable in the pattern of glycoprotein synthesis. Similar inductive interactions between different regions of the early embryo are thought to occur during normal development.
Ambystoma mexicanum↗
The effect of egg rotation on the differentiation of primordial germ cells in Xenopus laevis.
Eggs of X. laevis were rotated (sperm entrance point downwards) either through 90 degrees (1 X 90 embryos) or 180 degrees in two 90 degrees steps (2 X 90 embryos) at approximately 25-30 min postfertilization after cooling to 13 degrees C. The embryos were kept in their off-axis orientation and cooled until the early gastrula stage. Rotation resulted in relocation of egg constituents with slight changes in the distribution of outer cortical and subcortical components and major changes in inner constituents where the heavy yolk and cytoplasm appeared to reorient as a single coherent unit to maintain their relative positions with respect to gravity. Development of rotated embryos was such that regions of the egg which normally give rise to posterior structures instead developed into anterior structures and vice versa. Germ plasm was displaced in the vegetal-dorsal-animal direction (the direction of rotation) and was segregated into dorsal micromeres and intermediate zone cells in 2 X 90 embryos and dorsal macromeres and intermediate zone cells in 1 X 90 embryos. In consequence, at the gastrula stage, cells containing germ plasm were situated closer to the dorsal lip of the blastopore after rotation--in 2 X 90 gastrulas around and generally above the dorsal lip. Hence, in rotated embryos, the cells containing germ plasm were invaginated earlier during gastrulation and therefore were carried further anteriorly in the endoderm to a mean position anterior to the midpoint of the endoderm. The number of cells containing germ plasm in rotated embryos was not significantly different from that in controls at all stages up to and including tail bud (stage 25). However at stages 46, 48 and 49 the number of primordial germ cells was reduced in 1 X 90 embryos in one experiment of three and in 2 X 90 embryos in all experiments. We tested the hypothesis that the decreased number of primordial germ cells in the genital ridges was due to the inability of cells to migrate to the genital ridges from their ectopic location in the endoderm. When anterior endoderm was grafted into posterior endodermal regions the number of primordial germ cells increased slightly or not at all suggesting that the anterior displacement of the cells containing germ plasm was not the only factor responsible for the decreased number of primordial germ cells in rotated embryos. Other possible explanations are discussed.
Animals↗
Regional specificity of glycoconjugates in Xenopus and axolotl embryos.
This paper reviews work on the presence, synthesis and developmental regulation of glycoconjugates (proteoglycans, glycoproteins and glycolipids) in the early amphibian embryo. In the axolotl there is little regional specificity of protein synthesis until the tailbud stage, but substantial regional specificity of glycoprotein synthesis from the end of gastrulation. Glycolipid synthesis is more uniform although a number of unique species are made in the epidermis. Isolated explants from axolotl early gastrulae show three types of behaviour in terms of glycoprotein synthesis, corresponding to the classical germ layers. Xenopus embryos at this stage show a higher degree of mosaicism. Changes of glycoprotein synthesis in response to mesodermal or neural induction follow the predicted course depending on the regional character of the induced tissue. The regional binding patterns of a number of lectins and monoclonal antibodies specific for particular carbohydrate determinants are presented and their significance discussed.
Ambystoma↗